MedChem 2015 - Новосибирский Институт Органической ...web.nioch.nsc.ru/medchem2015/images/MedChem_2015_-_book... · 2015. 7. 28. · 2nd Russian Conference

2nd Russian Conference on Medicinal Chemistry

2nd Youth School

6th Russian- Korean Conference “Current Issues of Biologically A ctive Compound Chemistry and Biotechnology”

MedChem 2015

http://web.nioch.nsc.ru/medchem2015/

Book of abstracts

Novosibirsk

Russian Conference on Medicinal Chemistry

Youth School -Conference on Medicinal Chemistry

Korean Conference “Current Issues of Biologically A ctive Compound Chemistry and Biotechnology”

MedChem 2015

/medchem2015/

Book of abstracts July 5-10, 2015

Novosibirsk , Russia

Russian Conference on Medicinal Chemistry

Conference on Medicinal Chemistry

Korean Conference “Current Issues of Biologically A ctive Compound

MedChem 2015

2nd Russian Conference on Medicinal Chemistry

6th Russian-Korean Conference "Current Issues of Biologically Active Compounds Chemistry and Biotechnology"

2nd Youth School-Conference on Medicinal Chemistry

Book of Abstracts

The most of abstracts are printed as presented, and all responsibilities should be addressed to authors.

Compiled by Dr. Denis A. Morozov

Revised by Prof. Konstantin P. Volcho, Dr. Olga I. Yarovaya, Dr. Oleg V. Ardashov, Dr. Artem D. Rogachev, Dr. Evgeny V. Suslov

© N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry

Organizing Committee

Prof. Nariman F. Salakhutdinov (Chairman) Corr. memb. Prof. Sergey O. Bachurin (Co-Chairman) Dr. Vladimir A. Nikonov (Co-Chairman) Prof. Andrey G. Pokrovsky (Co-Chairman) Dr. Byung Hun Um (Co-Chairman) Dr. Sang Hyun Lim (Co-Chairman) Dr. Mikhail P. Perelroyzen (Co-Chairman) Prof. Konstantin P. Volcho (Scientific Secretary) Prof. Alexander E. Prosenko Prof. Elvira E. Shults Prof. Alexey Ya. Tikhonov Dr. Sergey A. Popov Dr. Oleg V. Ardashov Dr. Tatyana M. Khomenko Dr. Denis A. Morozov Dr. Natalya A. Orlova Dr. Artem D. Rogachev Dr. Alexander V. Shpatov Dr. Evgeny V. Suslov Dr. Olga I. Yarovaya Irina L. Anisimova Konstantin Yu. Ponomarev Irina I. Popadyuk Anastasiya S. Sokolova

Advisory Board

Academician S. M. Aldoshin AcademicianI. P. Beletskaya Academician Yu. N. Bubnov Academician V. N. Charushin Academician O. N. Chupakhin Academician M. P. Egorov Academician A. I. Grigoriev Academician A. R. Khokhlov Academician A. I. Konovalov Academician V. I. Minkin Academician I. A. Novakov Academician V. N. Parmon Academician S. B. Seredenin Academician O. G. Sinyashin Academician V. A. Stonik Academician B. A. Trofimov Academician A. Yu. Tsivadze Academician M. S. Yunusov Academician N. S. Zefirov Corr.-Memb. S. O. Bachurin Corr.-Memb. A. G. Gabibov Corr.-Memb. U. M. Dzhemilev Corr.-Memb. A. V. Kutchin Corr.-Memb. N. E. Nifantiev Corr.-Memb. S. D. Varfolomeev Dr. B. H. Um Prof. J.-T. Lee

The organizers express their gratitude for the support of the Conference

Sponsors



MedChem-2015, July 5-10, 2015, Novosibirsk, Russia

6

TABLE OF CONTENTS

Searching for Natural Sesquiterpene Lactones – Sources of New Drug Substances

S.M. Adekenov .............................................................................................................................. 27

Mitochondrial Dependent Oxidative Stress Induced Cellular Hypoperfusion, Erythrocyte Metabolism, and Calcium Signaling in the Context of Neurodegeneration and Cancer: Recent Challenges

G. Aliev .......................................................................................................................................... 28

Contemporary Approaches for Pharmacological Intervention of Alzheimer’s Disease

S. Bachurin ..................................................................................................................................... 34

From Pharmaceutical Substances to Pharmaceutical Formulations. How Can Solid State Chemistry Help?

E.V. Boldyreva ............................................................................................................................... 35

Heterocyclic Motif in Medicinal Chemistry

V.N. Charushin and O.N. Chupakhin ............................................................................................ 36

Rational design of antibiotics and NSAIDs

M.A. Cooper .................................................................................................................................. 37

Mycobacterium tuberculosis “hybrid warfare”: virulence, pathogenicity, persistence and drug resistance. New counteraction concept.

V.N. Danilenko .............................................................................................................................. 38

The Fundamental Basis and Approaches for Dipeptide Drugs Development T.A. Gudasheva .............................................................................................................................. 39

Modern Developments in New Drugs and Their Delivery by Nanomaterials

J. R. Hwu, M. Kapoor and S.-C. Tsay ........................................................................................... 40

Ga-68 and Lu-177-labelled Peptide Conjugates in Cancer Theranostics

K. Kairemo ..................................................................................................................................... 41

Drug Development R&D Center “IPHAR”

V. Khazanov ................................................................................................................................... 42

New Frontiers in Brassinosteroids: from Convergent Synthesis and Highly Sensitive Analysis to Pharmacokinetics Studies

V. Khripach, V. Zhabinskii ............................................................................................................ 43

Antiviral Drugs Against Ebola Infection: Novel Targets and Perspective

O.I. Kiselev .................................................................................................................................... 44

Development of New Antivirals: Problems and Prospects

S. Kochetkov .................................................................................................................................. 45

Human Tyrosyl-DNA Phosphodiesterase 1: New Activities and Development of Enzyme Inhibitors as Anticancer Drugs

O. Lavrik ........................................................................................................................................ 46


7

The study on Effective compounds that fermented by Lactobacillus casei in chestnut (Castanea crenata) bur against skin aging caused by UVB

Jin-Tae Lee and Young-A Jang ...................................................................................................... 47

Metal-based drugs: State of the Art E. Milaeva ...................................................................................................................................... 48

Animal Models and Hightechnologal Phenotyping for Drug Discovery

M.P. Moshkin, S.E. Peltek, N.A. Kolchanov ................................................................................. 49

Seaweed fucoidans – a platform for new drugs discovery

N.E. Nifantiev, N.E. Ustyuzhanina, M.I. Bilan, A.I. Usov, A.S. Shashkov, A.G. Gerbst ............. 50

Industrial drug discovery – splendor, misery and revival V. Patchev ...................................................................................................................................... 51

Cell and molecular models for search and study antiviral and anticancer compounds

A.G. Pokrovsky .............................................................................................................................. 52

Way2Drug - Web-resource for Drug Discovery

D. Filimonov, D. Druzhilovsky, A. Rudik, A. Lagunin, A. Zakharov, A. Dmitriev, T. Gloriozova, P. Pogodin, and V. Poroikov ................................................................................. 53

Chemical, Biological and Disease space complementarity in Drug Discovery

Sham Nikam ................................................................................................................................... 54

Intracellular Targets and Therapeutic Efficacy of Antitumor Drugs: A Complex Liaison

А.А. Shtil ....................................................................................................................................... 55

Human Photosynthesis or the Unexpected Capacity of Melanin Molecule to Dissociate the Water Molecule: Implications as Alternate and Successful Treatment Strategies for the Neurodegenerative Disease

A. Solís Herrera, M. del Carmen Arias Esparza, P. E. Solís Arias and G. Aliev .......................... 56

Recent Studies on Bioactive Natural Products from Marine Organisms

V.A. Stonik .................................................................................................................................... 58

Search for socially important drugs in Favorsky Irkutsk institute of chemistry

B.A. Trofimov, A.V. Ivanov .......................................................................................................... 59

Therapeutic Nucleic Acids

V. Vlassov, M. Zenkova, D. Pyshnii, D. Stetsenko ....................................................................... 60

Molecular Design of Bivalent or Dual Action Drugs

N.S. Zefirov, V.A. Palyulin, O.N. Zefirova ................................................................................... 61

The Synthesis of Expected Epoxy-metabolites of (1R,2R,6S)-3-Methyl-6-(prop-1-en-2-yl)cyclohex-3-ene-1,2-diol

O. Ardashov, A. Pavlova, A. Genaev, G. Salnikov, D. Korchagina, I. Il’ina, K. Volcho, T. Tolstikova, N. Salakhutdinov .................................................................................................... 63

Astrocytes as Targets for Neuroprotection from Brain Injury

G.E. Barreto, G. Aliev .................................................................................................................... 64


8

New Derivatives of (-)-Cytisune - Selective Inhibitors of COX-2: in silico Screening with in vivo Verification

S.S. Borisevich, I.P. Tsypysheva, N.S. Makara, S.L. Khursan ...................................................... 66

Alkoxyamines for the in-situ Generation of Radicals as Theranostic Agents P. Brémond, G. Audran, S.R.A. Marque ....................................................................................... 67

New Effective and Selective Carboxylesterase Inhibitors Based on Polyfluoroalkylated 2-arylhydrazono-3-oxo Esters

Ya.V. Burgart, G.F. Makhaeva, N.P. Boltneva, N.V. Kovaleva, S.V. Lushchekina, E.V. Shchegol’kov, V.I. Saloutin, R.J. Richardson, O.N. Chupakhin ........................................... 68

Discovery of ZB40-0016 – a Novel GPR119 Agonist for Type 2 Diabetes Treatment M.A. Chafeev, R.N. Karapetian, I.N. Tyurenkov, E.V. Volotova ................................................. 69

Recyclizations of Pyrimidines as a Nontraditional Way of Pharmacophore and Biogenic Groups Introduction to the Molecule of a Biologically Active Compound

G. Danagulyan ............................................................................................................................... 70

Natural Products in Matrix Assisted Supramolecular Chirality Amplification

P.B. Drašar ..................................................................................................................................... 71

Mechanochemical Approaches to Creation of Drug Delivery Systems

A.V. Dushkin, T.G. Tolstikova, M.I. Khvostov, N.E. Polyakov, S.S. Khalikov, K.V. Gaidul ..... 72

"Fucolam" - the First in Russia Food Supplement Based on Fucoidan

S. Ermakova, T. Zaporozhets, M. Kusaykin, N. Besednova, T. Zvyagintseva .............................. 73

The Structural Peculiarities of Poly- and Oligosaccharides of Carrageenan from the Red Algae Tichocarpus crinitus and Their Ability to Induce in vivo Interleukin 10

A.A. Kalitnik, A.O. Kravchenko, A.O. Byankina, S.D. Anastyuk, I.M. Yermak ......................... 74

First Synthetic Macrocyclic Glycoterpenoids

V.E. Kataev .................................................................................................................................... 75

Mechanochemical Technology to Create of Targeted Delivery Drugs

S.S. Khalikov, Yu.S. Chistyachenko, A.V. Dushkin, I.A. Arkhipov, A.I. Varlamova, I.I. Glamazdin ..................................................................................................... 76

Arabinogalactan as a Drug Carrier

M.V. Khvostov, T.G. Tolstikova, S.A. Borisov, M.S. Biryukova, A.V. Dushkin, J.S. Chistyachenko, A.A. Chernonosov ......................................................................................... 77

New Chiral Catalyst for Henry Reaction

V. Konev, T. Khlebnikova, L. Malysheva and Z. Pai .................................................................... 78

In vitro Cytotoxicity of the Dehydroabietylamine Ammonium Salts

A. Kononova, K. Kovaleva, V. Korobeynikov, O. Yarovaya, S. Cheresiz, E. Semenova, A. Pokrovsky, N. Salakhutdinov .................................................................................................... 79

The Story of Amphipathic Antiviral Nucleosides

V.A. Korshun, A.A. Chistov, G.V. Proskurin, A.V. Ustinov ........................................................ 80


9

Development of Original Pluripotent Preparations for the Russian Pharmaceutical Market on the Subject of Import Substitution

Ya.A Kostyro, V.V. Kostyro, E.S. Gogol, V.V. Davaa, E.N. Petrova, A.O. Savinova, N.N. Trofimova, E.V. Stolpovskaya, L.A. Grischenko, L.A. Ostroukhova, S.A. Lepehova, K.V. Alekseev, V.A. Babkin, V.K. Stankevich, B.A. Trofimov ................................................... 81

Cytotoxicity, Intracellular Localization and Acute Intravenous Toxicity of Cluster Complexes Na4[{Re6Q8}(CN)6] (Q = S, Se or Te) – New Promissing Photoactive and X-ray Contrast Agents

A.A. Krasilnikova, A.O. Solovieva, K.E. Zubareva, T.N. Pozmogova, L.V. Shestopalova, K.A. Brylev, M.A. Shestopalov ..................................................................................................... 82

Targeting Hormone-Refractory Prostate Cancer with New Molecular Entities

K.V. Kudryavtsev .......................................................................................................................... 83

Sulfation of Polysaccharides and Triterpens by Sulfamic Acid

V.A. Levdansky, N.Yu. Vasil’eva, S.A. Kuznetsova, A.V. Levdansky, A.S. Kazachenko, B.N. Kuznetsov .............................................................................................................................. 84

Novel Biologically Active Compounds Based on (Indol-1-yl)maleimide Scaffold

S.A. Lakatosh ................................................................................................................................. 85

New Eudesmane-type Sesquiterpenes from the Marine-derived Fungus Penicillium thomii E.V. Leshchenko, Sh.Sh. Afiyatullov, O.I. Zhuravleva, V.A. Denisenko ..................................... 86

The Study of Drugs Action Mechanism via the Chemical Modeling of Drug – Enzyme and Drug - Receptor Interaction

T.V. Leshina, E.A. Khramtsova, M.A. Miranda ............................................................................ 87

Peucedanin in the Synthesis of Coumarin-chalcone Hybrids and Dicoumarins

A.V. Lipeeva, P.Z. Zanimkhanova, M.A. Pokrovsky, M.V. Khvostov, D.S. Baev, S.M. Adekenov, T.G. Tolstikova, A.G. Pokrovsky, E.E. Shults ................................................... 88

Soloxolone Methyl – a Novel Compound with Multiple Activities, Exhibited Anti-cancer, Anti-flu and Anti-inflammatory Activities in vitro and in vivo

E.B. Logashenko, A.V. Markov, O.V. Salomatina, N.F. Salakhutdinov, M.A. Zenkova ............. 89

Mechanochemically Assisted Extraction of Bio-active Compounds from Lignocellulosic Material I.O. Lomovskiy, O.I. Lomovsky, D.V. Orlov ................................................................................ 90

Evolution of Biological Activity from Usnic Asid to Its Derivatives

O.A. Luzina, D.N. Sokolov, M.E. Rakhmanova and N.F. Salakhutdinov ..................................... 91

Enhancement of Radical Producing Activity by Anticancer Quinone-chelator in the Presence of Metal Ions

I.D. Markova, O.Yu. Selutina, K.Yu. Fedotov, L.G. Fedenok, N.E. Polyakov, L.M. Weiner ...... 92

Identification of Target Genes Involved in Soloxolone Methyl-induced Carcinoma Cell Apoptosis with Microarray Data

A.V. Markov, E.B. Logashenko, A.E. Kel, O.V. Kel-Margoulis, O.V. Salomatina, N.F. Salakhutdinov, M.A. Zenkova ............................................................................................... 93


10

New Approaches to Cu (I) Catalyzed ATRA/ATRC Reactions for Stereoselective Synthesis of Functional Substituted Small Carbocycles: Convenient Building Blocks for New Agrochemicals and Drugs

A. Mikaelyan, A. Grigoryan and N. Asatryan ............................................................................... 94

The Synthesis of Chiral Fluorine Containing Compounds Based on Monoterpenoids

O. Mikhalchenko, D. Korchagina, K. Volcho and N. Salakhutdinov ............................................ 95

Design, Synthesis and Structure-anxiolytic Activity Relationship Study of New Pyrrolo[1,2-a]pyrazine TSPO Ligands

G.V. Mokrov, O.A. Deeva, A.S. Pantileev, V.V. Silman, T.A. Gudasheva, S.A. Yarkov, M.A. Yarkova and S.B. Seredenin ................................................................................................. 96

Protein Ruthenation and DNA Alkylation: New Ruthenium Complexes and Their Anticancer Activity

A.A. Nazarov, Y.N. Nosova and E.R. Milaeva ............................................................................. 97

Synthesis of Aromatic and Reduced Berberine derivatives and Their Hypolipidemic Activity

I.V. Nechepurenko, U.A. Boyarskikh, M.V. Khvostov, D.S. Baev, N.I. Komarova and N.F. Salakhutdinov ......................................................................................... 98

Discovery of Small Molecule Inhibitors of Tick-Borne Flaviviruses Reproduction

D.I. Osolodkin, L.I. Kozlovskaya, V.A. Palyulin, G.G. Karganova and N.S. Zefirov .................. 99

Computational Tools for Multi-target Drug Design

V.A. Palyulin, E.V. Radchenko, N.S. Zefirov ............................................................................. 100

Selective Functionalization of Eudesmane-type Methylenelactones by Cross-coupling Reaction with Xanthine Derivatives

S. Patrushev, E. Shults ................................................................................................................. 101

Approaches for Improvement of Drugs Solubility

G.L. Perlovich .............................................................................................................................. 102

Conjugation of N-Acetyl-L-Cysteine with Natural and Synthetic Naphthoquinones as a Short Route to Novel Bio-Active Products

S. Polonik, Yu. Sabutskii and E. Yurchenko ............................................................................... 103

Physicochemical Approaches to the Study of New Drug Delivery Systems Based on Naturally Occurring Polysaccharides and Oligosaccharides

N.E. Polyakov, O.Yu. Selyutina, A.V. Dushkin .......................................................................... 104

Modification of Deoxycholic Acid Ring A with S- and N-Containing Functional Groups

I.I. Popadyuk, O.V. Salomatina and N.F. Salakhutdinov ............................................................ 105

Synthesis, Modeling and Molecular Design of N,N-disubstituted 2-Aminothiazolines as a New Class of Butyrylcholinesterase and Carboxylesterase Inhibitors

E.V. Radchenko, G.F. Makhaeva, N.P. Boltneva, S.V. Lushchekina, I.V. Serkov, A.N. Proshin, V.A. Palyulin, and N.S. Zefirov ........................................................................... 106

Development of Thienopyridines as Anticancer Agents

J. Reynisson ................................................................................................................................. 107


11

Triazavirin – Antiviral Drug of a New Generation

V.L. Rusinov, V.N. Charushin, O.N. Chupakhin, O.I. Kiselev ................................................... 108

2-Cyano Substituted Derivatives of Glycyrrhetinic and Deoxycholic Acids: Synthesis and Biological Activities

O.V. Salomatina, I.I. Popadyuk, E.B. Logashenko, A.V. Markov, M.A. Zenkova and N.F. Salakhutdinov ........................................................................................ 109

Patenting Polymorphs at the European Patent Office

I. Seelmann ................................................................................................................................... 110

The Sea Urchin Embryo as a Promising Model in Screening for Antimitotic Compounds

M.N. Semenova ............................................................................................................................ 111

Synthesis of Natural Antimitotic Derivatives Using Parsley and Dill Seed Extracts

V.V. Semenov, D.V. Tsyganov, D.V. Demchuk, N.B. Chernysheva, M.N. Semenova .............. 112

From Viral Entry Mechanisms to the Entry-Arresting Antivirals Development against Human Enveloped Viruses (HIV, Flu, … Ebola)

A. Serbin, B. Bolshchikov, O. Alikhanova and V. Tsvetkov ...................................................... 113

New Approaches to Preparation of Composites of Betulin and its Diacyles with Improved Biological Activities

T.P. Shakhtshneider, M.A. Mikhailenko, S.A. Kuznetsova, Yu.N. Malyar, A.S. Zamay, V.V. Boldyrev ......................................................................................................... 114

Polyfluorinated Salicylates as Analogs of Known Drugs

E.V. Shchegol’kov, I.V. Shchur, Ya.V. Burgart, V.I. Saloutin, A.N. Trefilova, S.Yu. Solodnikov, O.P. Krasnykh, L.N. Markova, O.N. Chupakhin .......................................... 115

Development of Small Molecular Weight Mimetics of Glial Cell Line-derived Neurotrophic Factor for the Treatment of Parkinson’s Disease

Y.A. Sidorova, I. Suleymanova, J. Kopra, P. Piepponen, J.-M. Renko, M. Voutilainen, J. Sakki, R.K. Tuominen, M. Karelson and M. Saarma ............................................................... 116

3-D Microarray Based on Macroporous Polymer Monoliths for Detection of Human Genome Mutations Associated with Pregnancy Complications

E. Sinitsyna, A. Glotov, M. Danilova, E. Vlakh and T. Tennikova ............................................. 117

Synthesis of Heterocyclic Derivatives of Borneol as Potential Antiviral Agents

A. Sokolova, M. Semenova, O. Yarovaya, N. Salakhutdinov ..................................................... 118

Novel Pharmaceutical Compositions Enhances Denervation Properties of Botulinum Neurotoxin Type A

A.G. Strelnikov, V.V. Fomenko, D.S. Sergeevichev, E.A. Pokushalov, N.F. Salakhutdinov .... 119

Isomerization of α-Pinene Oxide to Campholenic Aldehyde Over Fe-and Al-containing Zeotype Materials

M.N. Timofeeva, V.N. Panchenko, A.A. Abel, Z. Hasan, K.P. Volcho, S.H. Jhung ................... 120


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Synthesis and Cytotoxicity Evaluation of Novel Benzene-fluorinated 2,2-Dimethyl-2,3-dihydro-1H-quinolin-4-ones

L. Politanskaya, E. Tretyakov, V. Shteingarts, L. Ovchinnikova, O. Zakharova and G. Nevinsky .................................................................................................... 121

Design and Synthesis of Polymeric Modules for the Antiviral Potentiation of Natural and Synthetic Small Molecule Resources

V. Tsvetkov, B. Bolshchikov, and A. Serbin ............................................................................... 122

Anti-mycobacterial Activity of Ag(0)-nanoparticles in Mouse Bone-Marrow Cells and Peritoneal Macrophages Infected with BCG Vaccine In Vitro

E.G. Ufimtseva, I.D. Ivanov and A.V. Popov .............................................................................. 123

Inhibition of Sonic Hedgehog Pathway in Breast Cancer Stem Cells Does Not Provide Extra Benefit in Cytotoxicity Resulted from Pd(II)-Saccharinate Complex of Terpyridine

E. Ulukaya, B. Cevatemre, D. Karakas, N. Aztopal, C. Icsel ...................................................... 124

Comparison of Anticonvulsant Effects of Lambertianic Acid Aamide and Memantine in Mouse Hippocampal Slices

S.O. Vechkapova .......................................................................................................................... 125

Poly(amino acid) Polymersomes as Potential Nanocontainers for Drug Delivery

E. Vlakh, N. Zashikhina, A. Hubina, T. Tennikova ..................................................................... 126

New Physiologically Active Compounds Synthesized From Verbenone

K. Volcho, I. Il’ina, O. Mikhalchenko, O. Ardashov, A. Pavlova, T. Tolstikova and N. Salakhutdinov ............................................................................................ 127

Development of Tetrazole-Containing Derivatives of Lupane Type Triterpenoids and Their Cytotoxicity

A.N. Volkova, N.I. Petrenko, M.A. Pokrovsky, E.E. Shults and A.G. Pokrovsky ...................... 128

Assessment of Triticain-α glutenase and Collagenase Activities for Use in Enzymatic Therapy Assays

A.A. Zamyatnin Jr., G. Aliev, N.V. Gorokhovets, V.A. Makarov, S.Yu. Morozov, L.V. Savvateeva, M.V. Serebryakova, A.G. Solovyev and E.Yu. Zernii ............................................ 129

Bactericidal Activity and Cytotoxicity of Substituted Cyclic Acetals

A.N. Kazakova, A.V. Tugarova, A.A. Kamnev, G.Z. Raskildina, S.S. Zlotskii ......................... 130

Phytochemical analysis, Hepatoprotective and Antioxidant Activities of Russian Nelumbo nucifera Leaves Against CCl4-Induced Hepatic Damage in Rats

M.S. Abdelhamid, E.I. Kondratenko, N.A. Lomteva .................................................................. 131

New Synthetic Potential in the Phenanthrene Series in the Treatment of Socially Significant Diseases

V. Abzianidze, K. Bolshakova, D. Krivorotov and V. Kuznetsov .............................................. 132

Synthesis of Natural Phaeosphaeride A derivatives

V. Abzianidze, K. Bolshakova, L. Chisty, A. Berestetskii and V. Kuznetsov ............................ 133


13

Macromolecular Characterization of Metal-polymer Nanobiocomposities by Aqueous Size-exclusion Chromatography with Multiple Detection

G. Aleksandrova, A. Boymirzaev, M. Lesnichaya, B. Sukhov and B. Trofimov ........................ 134

Water Soluble Bioactive Nanocomposites of Transition Metal Oxides with Optical and Magnetic Properties

G. Aleksandrova, A. Bogomyakov, E. Gasilova, S. Vityazeva, V. Dubrovina, B. Sukhov, V. Ovcharenko, B. Trofimov ....................................................................................................... 135

Pyrimidine Nucleoside Derivatives with Both Antitubercular and Antiviral Activity

I.L. Karpenko, L.A. Alexandrova ................................................................................................ 136

Phosphocholine Liposomes as a Tool for Delivery of Anti-Tuberculosis 2’-Deoxypyrimidine Nucleoside Derivatives into Mammalian Cells

L.A. Alexandrova, G.M. Sorokoumova, M.A Zaretskaya and I.L.Karpenko ............................. 137

Antioxidant Activity of Rare-Earth Carboxylates with 2,6-Di-tert-butylphenol Moiety

T.A. Antonenko, D.B. Shpakovsky, Yu.A. Gracheva, T.V. Balashova, M.N. Bochkarev, E.R. Milaeva .................................................................................................... 138

Interaction of Glycyrrhizic Acid with Lipid Membrane: Perspectives for Drug Delivery

I.E. Apanasenko, O.Yu. Selyutina, N.E. Polyakov, N.P. Isaev, A.V. Kim ................................. 139

Prospects of Plant Essential Oils Components Study for the Development of Original Drug Substances

G.А. Atazhanova .......................................................................................................................... 140

Study of Camphecin Influence on Mice Behavior in the Open Field Test A. Babina, V. Lavrinenko, O. Yarovaya, N. Salakhutdinov ........................................................ 141

Creation Medications on the Basis of Extractives from Wood of Siberian Larch Species

V.A. Babkin, L.A. Ostroukhova .................................................................................................. 142

Computer Simulation of 11-Phenylundeca-5Z,9Z-dienoic Acid Human Topoisomerase I Inhibitory Activity

D. Baev, V. D'yakonov, and T. Tolstikova .................................................................................. 143

The Influence of Organotin Compounds on Lipid Peroxidation in Liver of the Russian Sturgeon in vitro

N. Berberova, E. Mukhatova, V. Osipova, M. Kolyada, D. Shpakovsky, E. Milaeva ................ 144

Antiulcerative Activity of New Omeprazole Analogues

M. Biryukova, N. Zhukova, T. Tolstikova, O. Yarovaya, D. Sokolov and N. Salakhutdinov .... 145

One-pot ‘on-solvent’ Multicomponent Protocol for the Synthesis of Medicinally Relevant Pyrano[3,2-c]quinolone Scaffold

S.I. Bobrovsky, M.N. Elinson ...................................................................................................... 146

Degradation of B-domain Deleted Factor VIII in Serum Containing Medium

V. Bodyagin, D. Milekhin, and J. Krasotkina .............................................................................. 147

1-Phenyl-1,4-dihydrophosphinolines – New Type of Tertiary Phosphines

A. Bogachenkov, A. Dogadina and A. Vasilyev ......................................................................... 148


14

Design Salbutamol and Budesonide Solid Dispersion for Pulmonary Drug Delivery

E. Bogdanova, A. Ogienko, E. Boldyreva, S.A. Myz, A. Ogienko, B. Kolesov, V. Drebushchak, A. Manakov, Yu. Kovalenko, N. Trofimov, V. Boldyrev ............................... 149

The Comparison of Pharmacological Activity of Ibuprofen and Aspirin after Complexation with Arabinogalactans of Different Descent

S.A. Borisov, T.G. Tolstikova, M.V. Khvostov, A.V. Dushkin and J.V. Chistyachenko ........... 150

Alternative Desalting Approach for Semi Preparative Oligonucleotide Manufacturing on Industrial Scale

E. Bulgakova ................................................................................................................................ 151

Total Synthesis of Tiasporine B and C

A.S. Bunev, S.A. Sokov, Ya.I. Rudakova, and G.I. Ostapenko ................................................... 152

Synthesis of New Potential Inhibitors of Protein Kinases

A.S. Bunev, Ya.I. Rudakova, A.P. Pavlova and G.I. Ostapenko ................................................. 153

Synthesis of Novel Thiazole-containing Amino Acids

A.S. Bunev, Ya.I. Rudakova, S.A. Sokov and G.I. Ostapenko .................................................... 154

Tetracationic 1,4-Diazabicyclo[2.2.2]octane Derivatives as an Attractive Scaffold in Antimicrobial Drug Design

E. Burakova, I. Saranina, N. Tikunova, V. Silnikov .................................................................... 155

Mechanical Disordering of Cell Walls Supramolecular Structure

A.L. Bychkov, E.M. Podgorbunskikh, E.I. Ryabchikova, O.I. Lomovsky ................................. 156

Interaction of Tris(1-alkylindol-3-yl)methylium salts with Human Serum Albumin-the Docking Study

E.E. Bykov, N.A. Durandin, S.N. Lavrenov, M.N. Preobrazhenskaya ....................................... 157

New Approaches to Chemo- and Stereoselective Synthesis of Substituted Dihydropyrimidines or Dihydropyridines

V. Charushin, O. Fedorova, Yu. Titova, A. Vigorov, D. Gruzdev, V. Krasnov, I. Krivtsov, A. Murashkevich, G. Rusinov ...................................................................................................... 158

Arylethynyl Nucleosides as Potent Inhibitors of Tick-borne Encephalitis Virus Reproduction

A.A. Chistov, A.A. Orlov, A.V. Guz, P.P. Streshnev, S.V. Kutyakov, A.V. Ustinov, V.A. Korshun, D.I. Osoloskin, L.I. Kozlovskaya, G.G. Karganova ............................................ 159

Physicochemical Properties and Anti-opisthorchosis Effect of Mechanochemically Synthesized Supramolecular Complexes of Albendazole with Polysaccharide Arabinogalactan from Larch Wood

Yu.S. Chistyachenko, A.V. Dushkin, E.S. Meteleva, M.Y. Pakharukova, A.V. Katokhin, V.A. Mordvinov, N.E. Polyakov, M.V. Khvostov, T.G. Tolstikova ........................................... 160

Palladium(II) Complexes of S-methylcysteine and Methionine Derivatives: Synthesis and Biological Activity

S.G. Churusova, D.V. Aleksanyan, E.Yu. Rybalkina and V.A. Kozlov ..................................... 161

Synthesis of Pyrazolo[1,5-a]pyrimidines by Recyclization of the Pyrimidine Ring

G. Danagulyan and A. Tumanyan ................................................................................................ 162


15

Аdamantylation of 1,2,4-Triazolo[5,1-c][1,2,4]triazines as Approach for Synthesis of Antiviral Agents

S.L. Deev, T.S. Shestakova, I.A. Khalymbadzha, V.L. Rusinov, O.N. Chupakhin .................... 163

Design, Synthesis and Pharmacological Evaluation of Dipeptide Analogs of Alpidem, Potential Ligands of TSPO Receptor

O.A. Deeva, S.A. Yarkov, M.A. Yarkova, T.A. Gudasheva, S.B. Seredenin ............................. 164

One-pot Natural Monoterpene Alcohol Amination to Biologically Valuable Compounds Over Gold Catalysts

Yu.S. Demidova, I.L. Simakova, M. Estrada, S.A. Beloshapkin, E.V. Suslov, K.P. Volcho, N.F. Salakhutdinov, A.V. Simakov, D.Yu. Murzin ..................................................................... 165

MNPs Modified with APS and PMIDA for MRI in vivo

A. Demin, A. Pershina, V. Ivanov, O. Shevelev, K. Nevskaya, N. Shchegoleva, A. Minin, M. Uimin, A. Sazonov, V. Krasnov ............................................................................................. 166

Development of Drug Delivery System for Anticoagulant Warfarin on the Basis of Mesoporous Silica Particles: Adsorption as a Route of Preparation of the Drug Composite

E.S. Dolinina, E.V. Parfenyuk ..................................................................................................... 167

Structure-Function Relationships in Cobra Cardiotoxins: Antibacterial Activity

P.V. Dubovskii, A.A. Ignatova, Y.N. Utkin and A.V. Feofanov ................................................. 168

Improvement of Dissolution Properties of Valsartan by co-crystallization with Vitamin C via Mechanochemical Method

L. Du, A.V. Dushkin and W. Su .................................................................................................. 169

Natural and Synthetic nZ,(n+4)Z-Dienoic Fatty Acids: A New Method for the Synthesis and Inhibitory Action on Topoisomerase I and IIα

V.A. D'yakonov, L.U. Dzhemileva, A.A. Makarov, A.R. Mulyukova, U.M. Dzhemilev ........... 170

Stereoselective Synthesis of 11-Phenylundeca-5Z,9Z-dienoic Acid and Investigation of Its Human Topoisomerase I and IIα Inhibitory Activity

V.A. D'yakonov, L.U. Dzhemileva, A.A. Makarov, A.R. Mulyukova, U.M. Dzhemilev ........... 171

Cytotoxic Activity of Salicylic Acid-containing Ionic Liquids

K.S. Egorova, M.M. Seitkalieva, A.V. Posvyatenko and V.P. Ananikov ................................... 172

New Water-Soluble Antifungal Agents Based on 2-Amino-6-Methylpyrimidin-4(3H)-thione

A. Erkin, V. Krutikov and V. Tets ............................................................................................... 173

The Scallop Patinopecten yessoensis Enzyme Acting Hydrolysis of Fucoidans

M.S. Avtushenko, A.M. Zakharenko, S.P. Ermakova, T.N. Zvyagintseva ................................. 174

In vitro Anticancer Activity of Laminarans from Far Eastern Brown Seaweeds and Their Sulfated Derivatives

O. Malyarenko (Vishchuk), S. Ermakova, N. Shevchenko, T. Zvyagintseva ............................. 175

The Fucoidan, a Marine Sulfated Polysaccharide from Brown Alga Fucus evanescens, Prevents Colorectal Cancer Growth by Targeting PDZ-Binding Kinase/T-LAK Cell-Originated Protein Kinase

O. Vishchuk, D. Qiuhong, S. Ermakova, F. Zhu, T. Zvyagintseva .............................................. 176


16

Brown Algae Fucus evanescens as Sourse of Inhibitors of O-glycoside hydrolases

A.S. Silchenko, T.I. Imbs, M.S. Avtushenko, S.P. Ermakova, T.N. Zvyagintseva ..................... 177

Activity of Cytisine Derivatives Against Human Parainfluenza Virus Type 3 in vitro Experiments

V.A. Fedorova, I.P. Tsipisheva, A.V. Kovalskaya, A.N. Lobov, E.A. Nikolaeva, M.S. Yunusov, V.V. Zarubaev ..................................................................................................... 178

Methyl 3-benzoyl-4-oxo-1.4-dihydroquinoline-2-carboxylate: an in vitro Assessment of Biomembrane Permeability and Hydrolytic Stability

I. Fefilova, A. Trefilova, A. Boteva, A. Mayorov, O. Krasnykh ................................................ 179

Virtual and Experimental Screening of the Maillard Reactions Inhibitors in Class of Diphenyl Oxide Derivatives

V. Frantseva, P. Vassiliev, A. Spasov, V. Kuznetsova, O. Solovieva, Yu. Popov, V. Lobasenko, T. Korchagina and O. Efremova .......................................................................... 180

Cytotoxicity of Pomolic Acid Isoplated from Chamaenerion angustifolium Against Human Cancer Cells

T.S. Frolova,T.P. Kukina, N.V. Gubanova, O.I. Sinitsyna .......................................................... 181

Stereospecific 7α-alkylation and 7,7-bis (alkynation) 20 hydroxyecdysone. The first example spirocyclization of ecdysteroids

I.V. Galyautdinov, Z.R. Khairullina, V.P. Sametov, G.G. Gibadullina, V.N. Odinokov ............ 182

Antioxidative Activity of Silibum Marianum Cultivated in Mongolia

J. Ganbaatar, E. Lkhagvamaa, E.E. Shults ................................................................................... 183

Determining Oxidized Proteins in Human Serum by Micromethod

A.P. Godovalov and N.V. Vavilov .............................................................................................. 184

New Copper-catalyzed Synthesis and Acetylcholinesterase Inhibitory Activity of N-substituted 2-Aminoisonicotinic Acid

N.E. Golantsov, A.V. Varlamov, and L.G. Voskressensky ......................................................... 185

Novel Hydroxylaminoisoxazoles as Perspective Lipoxygenase Inhibitors

Yu. Gracheva, D. Vasilenko, E. Averina, M. Neganova, O. Redkozubova, E. Shevtsova, E. Milaeva ............................................................................................................. 186

New Chlorin E6 Based IR Photosensitizers for Photodynamic and Antimicrobial Therapy

O.I. Gushchina, E.A. Larkina, T.A. Nikolskaya, A.F. Mironov .................................................. 187

Synthesis and Analgesic Activity of Monoterpenoid Derived 2-Alkyl-4,7-dimethyl-3,4,4a,5,8,8a-hexahydro-2H-chromene-4,8-diols

I. Il’ina, A. Pavlova, D. Korchagina, T. Tolstikova, K. Volcho and N. Salakhutdinov .............. 188

Anticancer Activity of Some Plant Extracts on U87MG Human Glioblastoma Cell V.A. Isidorov, S. Bakier, J. Moskwa and M.H. Borawska .......................................................... 189

Study on Effects of Plant Extracts on Paenibacillus larvae, a Causative Agent of American Foulbrood: Preliminary Results

V.A. Isidorov, K. Buczek, M. Zambrzycka and I. Swiecicka ...................................................... 190


17

High Temperature High Resolution Gas Chromatography Coupled with Mass Spectrometry for Analysis of Biologically and Environmentally Important Organic Compounds

V.A. Isidorov ................................................................................................................................ 191

Mechanochemical Technology of Obtaining Amino Acid Containing Products

I.V. Ivanov, A.L. Bychkov, O.I. Lomovsky ................................................................................ 192

Anti-osteoporosis Activity of Artemisia capillaris Extract and its Major Compound Scoparone via Regulation of Osteoclastogenesis and Bone Resorption Activity

Sang-Hyun Lee, Yong-Beom Kwon, Dae-Kun Lee, Fang-Fang Wang, Myung-Hee Kang, Young-In Kwon, Young-Ho Kim, and Hae-Dong Jang .............................................................. 193

Turmeric Extract Fermented with Bifidobacterium Induces Apoptosis via Caspase-Dependent Pathway and Cell Cycle Arrest in HCT116 Human Colorectal Cancer Cells

Ji-Eun Park, Sang-Hyun Lee, Min-Ji Seo, P. Belousova, Myung-Hee Kang, Young-In Kwon and Hae-Dong Jang ........................................................................................... 194

Solvent-Free Hofmann Rearrangement under High-Vibration Ball-Milling Conditions

Zhijiang Jiang, Zhenhua Li, Weike Su ........................................................................................ 195

A New Thiazolidine-4-carboxylic Acids as a Potent Antitumor Substances. Syntheses and in vitro Investigations

D.S. Khachatryan, K.R. Matevosyan, S.Y. Frolova, A.V. Kolotaev, A.N. Balaev, M.A. Baryshnikova and V.N. Osipov .......................................................................................... 196

Innovative Technologies in the Creation of Plant Protection Products

S.S. Khalikov, N.G. Vlasenko, O.I. Teplyakova .......................................................................... 197

Complex Environmentally Friendly Products for the Protection of Potatoes

S.S. Khalikov, A.A. Malyuga, N.S. Chulikova ............................................................................ 198

Regioselective Au(Ш)-Catalyzed Cycloisomerization of Propargylaminomethyl- or Propargyloxymethyl 15,16-Disubstituted Methyllambertianate

Yu.V. Kharitonov, E.E. Shults ..................................................................................................... 199

Modified Fluorine-containing Indazolones: Synthesis and Anticancer Activity

T.S. Khlebnicova, Yu.A. Piven, F.A. Lakhvich, M.B. Golubeva, B.B. Kuzmitsky, M. Turks, V. Kumpins ................................................................................................................. 200

New Biological Active Selenium-Containing Antioxidants Based on Alkylated Phenols and Catechols

S. Kholshin, S. Yagunov, V. Cheblukova, N. Kandalintseva, and A. Prosenko ......................... 201

Antibacterial Property of 2-Aminoethylphosphonates

N. Khusainova and I. Galkina ...................................................................................................... 202

Synthesis and Inhibition of Mycobacterium tuberculosis by A-homoaminotriterpenoids

N.I. Medvedeva, E.F. Khusnutdinova, I.E. Smirnova, O.B. Kazakova ....................................... 203

Cycloaddition Reaction of 3-substituted Benzo[b]furans in Synthesis of Amaryllidaceae alkaloids and Their Derivatives

A.S. Kilmetiev, E.E. Shults .......................................................................................................... 204


18

Aziridinederivative of Grosheimin

A.S. Kishkentaeva, О.А. Nazarova, B.А. Abdigalymova, Ju.V. Gatilov, G.A. Atazhanova, S.M. Adekenov ............................................................................................... 205

Reliable Immunomodulatory Evaluation for Substances of Natural Origin

E. Kmoníčková, P. Drašar, J. Harmatha, M. Kverka, Z. Zídek ................................................... 206

A New Approach to the Automated Identification of Metabolites in Multi-vendor Datasets

D. Hardy, V. Lashin, R. Kubilius, J. Blanz, M. Trunzer and E. Kolovanov ............................... 207

ACD/Percepta – a Software Platform for ADME/TOX Property Prediction, Compound Profiling and Lead Optimization

P. Japertas, K. Lanevskij, A. Sazonovas, E. Kolovanov .............................................................. 208

Endosorb - New and Effective Tool for Purification of Proteins Contaminated with LPS

A. Morozov, M. Kopitsyna, A. Nuzhdina and I. Bessonov ......................................................... 209

Information-analytical system "IR-EXPERT"

T.A. Kornakova, K.S. Chmutina .................................................................................................. 210

Alkaloids of Papaver kuvajevii Schaulo et Sonnikova M.O. Korotkikh, N.A. Pankrushina, and O.I. Salnikova ............................................................. 212

Synthesis of Novel Coumarin-substituted Fluoroquinolones

S. Kotovskaya, N. Mochulskaya, Z. Baskakova, E. Nosova and V. Charushin .......................... 213

4-Quinolones: the Old Friends in a New Role

A. Boteva, I. Fefilova, A. Trefilova, S. Solodnikov, G. Ljushina, V. Maslova, O. Krasnykh .... 214

Complex Study of Phycobiliproteins and Sulfated Polysaccharides from Sterile and Reproductive Forms of Red Alga Ahnfeltiopsis flabelliformis

A. Kravchenko, S. Anastyuk, V. Glazunov, V. Isakov, W. Helbert, I. Yermak .......................... 215

Simulation of Bioavailability and Pharmacokinetics in Preclinical Studies

I. Yakovlev, V. Yuskovets, I. Narkevitch, A. Radilov and D. Krivorotov .................................. 216

Synthesis of Mimetics of Known GPCR Modulators with Peripheral Action

V. Kuznetsov, D. Krivorotov ....................................................................................................... 217

The Search for Novel Adaptogens Among Thiourea Amino Acid Derivatives

S. Ramsh, V. Kuznetsov, S. Petunov, D. Bobkov, A. Radilov and D. Krivorotov ..................... 218

Scots Pine Phloem Reaction to the Anthropogenic and Biotic Stress

T.P.Kukina, G.G. Polyakova, T.S. Frolova, O.I. Sal’nikova ....................................................... 219

Human Recombinant Factor VII For Replacement Therapy

P. Kustov, A. Orlov and J. Krasotkina ......................................................................................... 220

Synthesis and Studying of Biological Activity of 2-[2-(3-R-adamanthane-1-yl)-2-oxoethylidenhydrazino]-4-(het)aryl-4-oxobut-2-enoic Acids

A. Kuznetsov, N. Pulina, A. Krasnova ........................................................................................ 221


19

Synthesis and Biological Evaluation of 2,2,2-tris(1H-indol-3-yl)acetic acid Derivatives

S.N. Lavrenov, Ya.A. Solomatin ................................................................................................. 222

Synthesis and Anti-inflammatory Activity of Derivatives of 2-Hydroxy-4-oxo-4-thienyl-2-butenoic Acid

K. Lipatnikov, N. Pulina, A. Krasnova, T. Yushkova, and F. Sobin ........................................... 223

Novel Moronic Acid Semisynthetic Derivatives

I.Ya. Mainagashev, O.V. Salomatina, and N.F. Salakhutdinov ................................................... 224

Biofilm Probiotics

V. Melnikov, V. Chistyakov ........................................................................................................ 225

The Stereoselective Transformation of Eudesmanolide of (-)-α-Santonin

N. Merkhatuly, A.T. Omarova, A.K. Ibrayeva, D.D. Naushabekova, Zh.E. Suleimenova, L.T. Balmagambetova ................................................................................. 226

Liquid Phase Oxidation of Halo-vinyl Compounds on the Catalyst Ag/SiO2: Extremely Stable Water-Soluble AgxCly Cluster

N. Asatryan, A. Mikaelyan ........................................................................................................... 227

Water-soluble Metal Complexes of Pectin Polysaccharides with Antianemic Effect S. Minzanova, V. Mironov, A. Vyshtakalyuk, L. Mironova, V. Milyukov ................................. 228

Phlomisoic Acid in the Synthesis of Labdane Type Naphthoquinones and Biphenyl Substituted Spiroundecanes

М.Е. Mironov, М.A. Pоkrovsky, A.G. Pоkrovsky, E.E. Shults .................................................. 229

Chemical Modification of Cirsilineol Molecule

G.K. Mukusheva, G.M. Baisarov, S.M. Adekenov ..................................................................... 230

Synthesis of Novel Purine and 2-Aminopurine Conjugates with Heterocyclic Amines

V.V. Musiyak, D.A. Gruzdev, E.N. Chulakov, A.Yu. Vigorov, G.L. Levit, V.P. Krasnov ........ 231

Highly Stereoselective Synthesis of (2SR, 3aSR, 8aSR)-3a-methyloctahydrocyclohepta[b]pyrrol-4(1H)-one Derivatives

I.N. Myasnyanko, E.R. Lukyanenko, A.V. Kurkin ...................................................................... 232

Multicomponent Efficient Transformation of Salicylaldehydes, Dimedone and Coumarin into Substituted Tetrahydro-1H-xanthen-1-ones

R.F. Nasybullin, O.O. Sokolova and M.N. Elinson .................................................................... 233

5,5’-Modified 2’,5’-Dideoxyuridines: Synthesis and Biological Properties

S.D. Negria, I.L. Karpenko, S.N. Kochetkov, L.A. Alexandrova ............................................... 234

Menthyl Ester of Gamma-aminobutyric Acid as Potential Anticonvulsant Agent M. Nesterkina, I. Kravchenko ...................................................................................................... 235

Hemocoagulating Activity of Sulfur-Containing Monoterpenoids

S.V. Kiselev, L.E. Nikitina, V.A. Startseva, N.P. Artemova, A.V. Bodrov, А.А. Rakhmatullina, G.A. Yakubovskaya and V.V. Klochkov .................................................. 236


20

Synthesis of Compound of Ceftriaxone with L-α-Alanine

G.V. Novikova, N.A. Staloverova ............................................................................................... 237

Synthesis of 1-Methyl-9-phenyl-3,11-dioxatricyclo[5.3.1.02,4]undecane

A.M. Novoselov, E.R. Lukyanenko, D.S. Belov, A.V. Kurkin ................................................... 238

Synthesis of Novel Monoterpenoid and Methoxyphenyl Substituted Coumarins

T. Odarchenko, T. Khomenko, D. Korchagina, K. Volcho, N. Salakhutdinov ........................... 239

The Effect of Lithium, Cerium (III), Copper (I) Chlorides on the Synthesis of 1-(2-Chloro-3-quinolyl)-4-dimethylamino-2-(1-naphthyl)-1-phenyl-butan-2-ols

A. Omelkov, V. Fedorov .............................................................................................................. 240

Solubility of Powder Preparations from Quercetine Containing Plant Raw

D.V. Orlov, O.I. Lomovsky ......................................................................................................... 241

Antioxidative Activity of Novel Organogold(I) Complexes Based on 2,6-di-tert-butyl-4-mercaptophenol

V. Osipova, M. Polovinkina, N. Movchan, D. Shpakovsky, E. Milaeva ..................................... 242

Certain Short Peptide Analogues of Somatostatin

V.N. Osipov, A.N. Balaev, K.A. Okmanovich, A.V. Kolotaev and D.S. Khachatryan .............. 243

Mechanochemical, Microwave, Ultrasound assisted Isolation of Natural Alkaloids

N. Pankrushina, M. Korotkih, and A. Mikheev ........................................................................... 244

A New Method for Synthesis of Substituted Diazepines[1,4] Annelated with Indole Nuclei A.A Panov, S.A. Lakatosh ........................................................................................................... 245

Diversity of Solid Forms of Felodipine: Structure and Physicochemical Properties

A.O. Surov, G.L. Perlovich .......................................................................................................... 246

Using of Modern Preparative Chromatography Methods in Biotechnology

O.A. Pisarev ................................................................................................................................. 247

Mechanochemistry in Technology Bioconversion of Plant Raw Materials

E.M. Podgorbunskikh, A.L. Bychkov, O.I. Lomovsky ............................................................... 248

Synthesis and Cytotoxicity Evaluation of Benzene-fluorinated 2-Arylindoles

L. Politanskaya, V. Shteingarts,O. Zakharova and G. Nevinsky ................................................. 249

Molecularly Imprinted Polymers as New Generation of Sorbents with Properties of Artificial Receptors to Biologically Active Substances

I. Polyakova, N. Ezhova, A. Leshchinskaya, I. Garkushina, L. Borovikova, A. Osipenko, O. Pisarev ............................................................................................................... 250

Analgesic Activity of New Diazaadamantanes Containing Pinene Fragment K. Ponomarev, E. Suslov, A. Pavlova, O. Ardashov, D. Korchagina, A. Nefedov, T. Tolstikova, K. Volcho, N. Salakhutdinov ............................................................................... 251

Novel Synthetic Approaches to Organochalcogen Compounds with Potential Biological Activity

V.A. Potapov, M.V. Musalov, E.O. Kurkutov, R.S. Ishigeev, A.S. Filippov, M.V. Musalova, A.G. Khabibulina, and S.V. Amosova .............................................................. 252


21

Eudesmanolides of Inula helenium and Its Aminoderivatives with Cytotoxic Activity

S. Pukhov, L. Anikina, S. Afanasyeva and S. Klochkov ............................................................. 253

Anti-virus Activity Evaluation of Camphor Derivatives Using Pseudoviruses

Yu. Razumova, A. Sokolova, O. Yarovaya, A. Pokrovsky, N. Salakhutdinov ........................... 254

Virtual Screening for Novel Atg5-Atg16 Complex Inhibitors for Autophagy Modulation

J. Reynisson, A. Richardson, E. Robinson, E. Leung, A. Matuszek, N. Krogsgaard-Larsenn, D. Furkert, and M. Brimble .......................................................................................................... 255

The Nature of Known Drug Space

J. Reynisson ................................................................................................................................. 256

5′-(1,2,3-Triazolyl)substituted Lappaconitine Derivatives

V.E. Romanov, E.E. Shults .......................................................................................................... 257

Solvent-free Multicomponent Synthesis of Pyrano[2,3-dpyrimidines]

F. Ryzhkov, M. Elinson ............................................................................................................... 258

Synthesis and Investigation of 1,2,4-Triazolo[5,1-c]-1,2,4-triazines Antiviral Action

I.M. Sapozhnikova, N.R. Medvedeva, E.N. Ulomskii, and V.L. Rusinov ................................... 259

Thiadiazolo[1,5-a]pyrimidines as Perspective Precursors of Heterocycles with Potentially High Biological Activity

K. Savateev, E. Ulomsky, V. Rusinov ......................................................................................... 260

ω-Oxo-functionalized 20-Hydroxyecdysone Derivatives in the Synthesis of Aminoecdysteroids

R.G. Savchenko, S.A. Kostyleva, V.N. Odinokov ...................................................................... 261

Synthesis of 5-(2,4-Dimethylpyridin-4-yl)-trisnoroleanane

E.V. Salimova, E.V. Tretyakova, S.A. Kostyleva, R.M. Latypova, L.R. Miniyarova ................ 262

Synthesis and anti-TBEV Properties of 4-Aminopyrimidine N-oxides

K.N. Sedenkova, E.V. Dueva, D.I. Osolodkin, E.B. Averina, V.A. Palyulin, Yu.K. Grishin, T.S. Kuznetzova and N.S. Zefirov ............................................................................................... 263

Effect of Natural Polysaccharides and Oligosaccharides on Permeability of Cell Membrane – Implication in Drug Delivery

O.Yu. Selyutina, I.E. Apanasenko, N.E. Polyakov, A.G. Shilov ................................................. 264

Synthesis and Anti-acetylcholinesterase Potency of Onium Uracil Derivatives

V. Semenov, I. Galyametdinova, L. Saifina, I. Zueva, K. Petrov, V. Zobov, V. Reznik ............ 265

Synthesis and Biological Properties of Pyrrolylbenzoxadiazoles – Potential Nitric Oxide (II) Donors

Yu.P. Semenyuk, P.G. Morozov, E.V. Prazdnova, O.N. Burov .................................................. 266

Preparation of Solubilized Forms of Meloxicam

S.A. Myz, A.G. Ogienko, T.N. Drebushchak, T.P. Shakhtshneider, A.A. Ogienko, E.V. Boldyreva .................................................................................................... 267

The quantum-chemical DFT-calculating of the thermodynamic properties of arglabin molecule

S.T. Shamilova, L.K. Abulyaisova, G.A. Atazhanova, S.M. Adekenov ..................................... 268


22

Determination of the Enantiomeric Purity of Pemetrexed and Levalbuterol on the Macrocyclic Glycopeptide Bonded Phases

E.N. Shapovalova, I.A. Fedorova, A.A. Priporova, I.A. Ananieva, O.A. Shpigun ..................... 269

Ortho-dinitrobenzenes in the Synthesis of Biologically Active Benzimidazolones

A. Shchipalkin, D. Krivorotov and V. Kuznetsov ....................................................................... 270

Determination of Seven Flavonoids in Epimedium by Liquid Chromatography–Tandem Mass Spectrometry Method

O.A. Shevlyakova, K.J. Vasil’yev, A.A. Ihalaynen, A.M. Antokhin, V.F. Taranchenko, D.A. Mitrofanov, A.V. Aksenov, I.A. Shchetinina, I.A. Rodin, O.A. Shpigun .......................... 271

Phosphoramidates of Pyrimidine Nucleoside Analogues: Unexpected Products of Known Reaction

N.F. Zakirova, A.V. Shipitsyn ..................................................................................................... 272

Anti-influenza Activity of Usnic Acid Derivatives in vitro and in vivo

A.A. Shtro, V.V. Zarubaev, O.A. Luzina, D.N. Sokolov, N.F. Salakhutdinov ........................... 273

Nanoemulsions With Biologically Active Substances

E.G. Shubenkova .......................................................................................................................... 274

Modification of Cyprofloxacine with Indol-1-ylmaleimide Derivatives. Synthesis and Study of the Antibacterial Activity

A.Yu. Simonov, S.A. Lakatosh, E.P. Mirchink, E.B. Isakova ..................................................... 275

Search of the Bioactivity of Extracts of Thymus serpyllum L. Growing in Yakutia

S.V. Sivtceva, Yun Jeong Choe, Kyu Lee, Ho Young Kang, Su In Cho and Zh.M. Okhlopkova ............................................................................................... 276

Antioxidants as a criterion of EXTRA VIRGIN olive oil quality

N.V. Sizova .................................................................................................................................. 277

Triphenylantimony(V) Catechlolates Derived from Alkyl Gallates: Synthesis and Antioxidant Activity

I.V. Smolyaninov, A.I. Poddel’sky and N.T. Berberova ............................................................. 278

Synthesis and Biological Activity of Conjugates of Substituted Indoles with Phenothiazine-3,7-Diamine

V.B. Sokolov, A.Yu. Aksinenko, T.V. Goreva, T.A. Epishina, V.V. Grigoriev, A.V. Gabrel’yan, D.V. Vinogradova, M.E. Neganova, P.N. Shevtsov, E.F. Shevtsova and S.O. Bachurin ........... 279

The Biological Properties of the New Agent for Photodynamic Therapy And Luminescent Bioimaging Based on Molybdenum Complexes

A.O. Solovieva, K.E. Zubareva, A.A. Krasilnikova, L.V. Shestopalova, E.V. Vorontsova, K.A. Brylev, A.F. Poveshchenko, M.A. Shestopalov .................................................................. 280

New Acyclic Nucleoside Phosphonates Bearing Unsaturated Fragments in the Chain. Synthesis and Antiviral Properties

P.N. Solyev, M.V. Jasko and M.K. Kukhanova .......................................................................... 281

Pentacyclic Triterpenoid Bioregulators – a New Class of Hepatoprotective Agents

I. Sorokina, N. Zhukova, and T. Tolstikova ................................................................................. 282


23

Bioflavonoid’s and Carotenoid’s Based Compositions of Enhanced Solubility and Antioxidant Activity

L.P. Suntsova, A.V. Dushkin, N.E. Polyakov, I.E. Apanasenko, Е.S. Меtеlеvа, V.I. Evseenko ...................................................................................................... 283

Chimeric Structures Based on 1-(oxiran-2-ylmethyl)-1H-indole-3-carbaldehyde as New Antidiabetic Agents

K.F. Suzdalev, A.A. Spasov, S.A. Tsirulnik, N.I. Chepljaeva and V.A. Kuznetsova ................. 284

In Silico And In Vitro Search For Inhibitors Of Dipeptidyl Peptidase-4 Among Adamantane Derivatives

A. Taran, P. Vassiliev, A. Spasov, N. Cheplyaeva, Yu. Popov, V. Lobasenko, E. Shishkin and V. Mokhov ......................................................................................................... 285

Polyphosphate Nucleotide Derivatives Functionalized at Terminal Phosphate Group

Yu. Tarasenko, P. Chalova and T. Abramova .............................................................................. 286

Chemical Composition of the Essential Oils of the Plant Family Apiaceae from Buryatian and Mongolian Flora

V. Taraskin, B. Urbagarova, G. Tykheev and L. Radnaeva ........................................................ 287

Novel Dosage Forms on the Basis of 1,2,4-Thiadiazole Derivative and β-Cyclodextrins

O. Silyukov, M. Chislov, A. Proshin, E. Chibunova, T. Volkova and I. Terekhova ................... 288

Effect of Acid Modification of Montmorillonite Clays on Synthesis of Octahydro-2H-chromen-4-ol from Vanillin and Isopulegol

M.N. Timofeeva, K.P. Volcho, V.N. Panchenko, O.S. Mikhalchenko, N.F. Salakhutdinov, K.V. Kochegarova, A. Gil, M.A. Vicente .................................................................................... 289

Efficient Route to 18-[(5-Bromomethyl)oxazol-2-yl]-18-norisopimaradiene and its Derivatives

M.A. Timoshenko, Yu.V. Kharitonov, E.E. Shults ..................................................................... 290

Inhibition of Copper and Iron Induced Hydroxyl Radical Formation by Metal Chelator Deferiprone

V.A. Timoshnikov, T.V. Kobzeva, N.E. Polyakov, G.J. Kontoghiorghes ................................. 291

The Development of a New Innovative Anticoagulant Drug

D. Tovbin, D. Tarasov ................................................................................................................. 292

Antiviral Activity of Preparation from Rice Husk and Green Tea

E.G. Trofimova, O.I. Lomovsky, E.I. Kazachinskai, T.V. Teplyakova ...................................... 293

A Computation Modeling for Rational Design and Synthesis of Oligomer Compounds Capable of Arresting the Viral Fusion Machinery

V. Tsvetkov, B. Bolshchikov and A. Serbin ................................................................................ 294

Molecular Modeling of New G-quadruplex Inhibitors of HIV Infection

V. Tsvetkov, A. Varizhuk, V. Oslovsky and G. Pozmogova ....................................................... 295

Parameters for Describing Conformational Rearrangements in G-quadruplexes

V. Tsvetkov, A. Varizhuk, G. Pozmogova .................................................................................. 296


24

5-Aminotetrazole in the Synthesis of 6-Ar(Het)-3,5-diamino-1,2,4-triazines. Lamotriginum and Its Analogues

E.N. Ulomsky, V.L. Rusinov, E.K. Voinkov, V.V. Fedotov, K.V. Savateev .............................. 297

Mechanochemical Activation of Brown Coal and modification of Its humic acids

T.S. Urazova, A.L. Bychkov, O.I. Lomovsky ............................................................................. 298

Electrophilic Fluorination of 1-Ethyl-7-(4-R-piperazin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic Acids with NF-Reagents

A.N. Usoltsev, P.A. Zaikin, G.I. Borodkin, V.G. Shubin ............................................................ 299

Synthesis of bi(hetero)arenes Potential Physiologically Active Compounds

I. Utepova, O. Chupakhin, M. Trestsova, D. Kucheryavaya, A. Musikhina ............................... 300

Synthesis of Nopinane-annelated Pyridines

E. Vasilyev, A. Agafontsev and A. Tkachev ............................................................................... 301

Calculation of the Membership Function in the Consensus Prediction of the Pharmacological Activity of Chemical Compounds

P. Vassiliev ................................................................................................................................... 302

Regulating the NMDA-receptors Activity by Lambertianic Acid Amide

S.O. Vechkapova, T.A. Zapara, T.G. Tolstikova, A.S. Ratushnyak ............................................ 303

A New Look at the Synthesis of 5-Halogen Derivatives of Morpholino Uridine

I. Vohtancev, Yu. Tarasenko, V. Silnikov and T. Abramova ...................................................... 304

Ethylnitrocyanacetic Ester as the Latent Form of the Nitroacetonitrile

E.K. Voinkov, K.V. Savateev, V.V. Fedotov, E.N. Ulomskiy, V.L. Rusinov ............................ 305

Computer-aided Prediction of Bioactivity of Glucose-modified Benzocaine

T.G. Volkova, I.O. Talanova, A.M. Ryzhakov, A.I. Tsarkova .................................................... 306

Influence of Pectins Isolated from Different Raw Materials on Blood Parameters of Rats

A. Vyshtakalyuk, S. Minzanova, L. Mironova, V. Zobov, V. Mironov ...................................... 307

Mechanochemically-Assisted Extraction of Water Solutable Pectin from Gardenia Jasminoides

W. Xu, A.V. Dushkin and W. Su ................................................................................................. 308

New Vitamin E Analogues: Sulfides and Selenides Based on 5-Hydroxy-2,3-Dihydrobenzofurans

S. Yagunov, S. Kholshin, N. Kandalintseva and A. Prosenko ..................................................... 309

Silver Nanoparticles Stabilized in a Poly(ethylene oxide) Matrix – a Prospective Antimicrobial Agent for Hydrocolloid Wound Dressings

O.V. Yakutkina, A.V. Kharenko, S.V. Antonov, Yu.M. Korolev ............................................... 310

Consensus screening of new inhibitors of glycogen phosphorylase in class of adamantane derivatives

L. Yanalieva, P. Vassiliev, A. Spasov, N. Cheplyaeva, E. Vorobiev, Yu. Popov, V. Lobasenko, E. Shishkin and V. Mokhov ................................................................................. 311


25

Pyrophosphate Analogs Suppress Phosphorolytic Activity of Wild-type and AZT-resistant HIV-1 Reverse Transcriptase

M.K. Kukhanova, A.N. Korovina, N.N. Usanov, S.N. Kochetkov, D.V. Yanvarev ................... 312

Synthesis of Novel Non-natural Conformationally Restricted Cyclopropane Amino Acids of Spiro[2.3]hexane series

N. Yashin, E. Averina, T. Kuznetsova, N. Zefirov ...................................................................... 313

The trolox ionic conjugates with amines - a new type of functionalized bioactive compounds

Yu.V. Yushkova, E.I. Chernyak, S.V. Morozov, I.A. Grigorèv ................................................. 314

Synthesis and Biological Evaluation of Novel Tyrosyl-DNA Phosphodiesterase 1 Inhibitors with a Benzopentathiepine Moiety

A.L. Zakharenko, T.M. Khomenko, S.V. Zhukova, O.A. Koval, O.D. Zakharova, R.O. Anarbaev, N.A. Lebedeva, D.V. Korchagina, N.I. Komarova, V.N. Vasiliev, J. Reynisson, K.P. Volcho, N.F. Salakhutdinov and O.I. Lavrik ................................................. 315

Novel Camphor- based Compound of Broad Range of Anti-influenza Activity

V.V. Zarubaev, A.V. Garshinina, S.A. Boldyrev, V.A. Fedorova, Y.R. Orshanskaya, A.A. Shtro, A.S. Sokolova, O.I. Yarovaya, N.F. Salakhutdinov ................................................. 316

Mechanochemical-assisted extraction of polysaccharides and alkaloids from Sophora Flavescens Ait

Q. Zhang, J. Yu, W. Su ................................................................................................................ 317

Synthesis and Cytotoxicity of New Functionalized Fluorinated 1,4-Naphtoquinones

S. Zhivetyeva, L. Goryunov, V. Shteingarts, O. Zakharova and G. Nevinsky ............................ 318

Synthesis and pharmacological activity of trifluoromethylphenyl substituted imidazo[1,2-a ]benzimidazoles

V.A. Anisimova, O.N. Zhukovskaya ........................................................................................... 319

Synthesis and Antimicrobial Activity of N-(2-alkoxy-2-oxoethyl) Pyridinium Chloride and Tetrafluoroborates

O.E. Zhuravlev, A.A. Belyaeva, and L.I. Voronchihina .............................................................. 320

AUTHOR INDEX............................................................................................................................ 321


26


27

Searching for Natural Sesquiterpene Lactones – Sources of

New Drug Substances

S.M. Adekenov

JSC “International Research and Production Holding “Phytochemistry” 100009, Republic of Kazakhstan, Karaganda,

E-mail: [email protected]

Among terpenoid family of compounds the most interesting, in terms of sources of new drug

substances, are sesquiterpene γ-lactones.

Over the last years we have studied 37 plant species of Kazakhstan flora in regard of the

containing of sesquiterpene lactones. First a chemical study of 13 plant species was conducted;

from which 42 compounds were extracted; among them 22 belong to sesquiterpene lactones. 17

sesquiterpene lactones are identified for the first time, mainly, in studied endemic plant species.

Thus, from Artemisia halophila Krasch two new compounds were isolated and studied. They are

1β-acetoxy-7α,6,11β(Н)-eudesm-4(5)-en-6,12-olide (1) and 1β-acetoxy-3β-hidroxy-6,11β,7α(Н)-

eudesm-4(5)-en-6,12-olide (2) and artesin was identified for the first time. From Tanacetopsis

pjataevae (Kovalevsk.) Karmysch new sesquiterpene lactone 3-oxo-10β-hidroxy-5,7α,4,6,11β(Н)-

guai-1(2),11(13)-dien-12,6-olide was discribed (3) and first identified β-isoepoxiestafiatin was

found. For the first time ever from Artemisia porrecta Krasch. ex Poljak and Artemisia karatavica

Krasch. et Abol ex Poljak sesquiterpene lactones herbolide А, 1β,9β-dihydroxy-5,7α,6,11β(Н)-

eudesm-4(15)-en-12,6-olide and 1α(10α)-epoxy-9β-hidroxy-6β,7α(Н)-germacr-4(5)-en-12,6-olide

were isolated and identified, and from Hieracium bectauatensis Kupr. - jacquinelin. For the fist

time sesquiterpene lactone cnicin was discovered in Centaurea iberica Trev. ex Spreng, and

costunolide, dihydroсostunolide, hanphyllin and artesin - in Artemisia semiarida (Krasch. et Lavr.)

Filat. First isolated argracin from Artemisia aralensis Krasch was found.

OO

O

O

HO

O

O

O

O

O

O

HO

H

O

(1) (2) (3)

On the basis of sesquiterpene lactones of arglabin, tourneforine, argolide, grosheimin,

alantolactone, isoalantolactone more than 30 new polyfunctional, heteroatom containing derivatives

were synthesized. Herewith, biological screening of sesquiterpene lactones and its derivatives was

conducted in regards of antibacterial, anti-inflammatory, fungicide, phagocytosis stimulating,

membrane-protective, antiviral, cytotoxic activity. It was established, that studied compounds have

certain biological activity


28

Mitochondrial Dependent Oxidative Stress Induced Cellular Hypoperfusion,

Erythrocyte Metabolism, and Calcium Signaling in the Context of

Neurodegeneration and Cancer: Recent Challenges

G. Aliev 1,2

1GALLY” International Biomedical Research Institute Inc., San Antonio, TX, USA 2Department of Health Science and Healthcare Administration,

University of Atlanta, Atlanta, GA USA

E-mail:[email protected], [email protected], [email protected]

Neurodegeneration [Stroke and Alzheimer disease (AD)] and cancer are fastly becoming one of

the leading causes of age-associated disability, dementia, and death. In addition, the Centers for

Disease Control and Prevention (CDC) and the National Center for Health Statistics recently

reported that AD has surpassed diabetes as a leading cause of death and is now considered the sixth-

leading cause of death in the United States.

Oxidative stress induced mitochondrial DNA over-proliferation and/or deletion of the organ

and/or tissues, especially the mitochondrial energy demands, have been implicated in the

pathogenesis of several diseases, including AD, tumor growth, and metastasis. Decline in

mitochondrial function during the development and maturation of the neurodegeneration, tumor

growth, and metastases characterizing the tissue oxygen deficiency may lead to cellular energy

defects, which will compensate vital cellular components and regulators. The hyper-expression of

the enzymes such as NOS induce the production of unwanted large amounts of free radicals that

cause the oxidative stress, cellular change, and particularly the concomitant mitochondrial lesions

and decline in normal organ function. The present study has determined if an intimate, i.e. causal,

relationship between oxidative stress and mitochondrial damage and/or vascular lesions occurs

before the development of human AD, in animal models that mimic human neuro-degeneration and

human colorectal carcinoid cancer or malignant brain cancer.

Our in situ hybridization and ultrastructural analysis by using electron microscopy techniques of

the mitochondria (mitochondria with electron dense matrix, mitochondrial-derived lysosomes)

showed that mitochondria with the destructions and lysosomal associated lipofuscin granules

appear to be main key features of hippocampal damaged neurons, vascular endothelial cells,

astrocytes and macro- and microphages’ in human AD, aged Tg (+) mice, 2 vessel occlusion

model of the brain hypoperfusion, and malignant primary and metastatic brain cancer as well as

colorectal cancer metastasis to the liver. The abnormally damaged mitochondria appear to be a

permanent hallmark and primary feature in all of the cellular components; in situ hybridization

analysis with mouse and human mtDNA probes found a large amount of deleted mtDNA in human

AD and in all models that mimic human AD (mice, rats etc.) hippocampus, brain cortex and cancer


29

tissues compared to aged controls. The majority of these mtDNA deletions containing colloidal gold

particles were found in mitochondrial-derived lysosomes in regions closely associated with

lipofuscin and/or tumor growth regions, and suggests that proliferation, deletion, and duplication of

mtDNA occurs in mitochondria, many of which have been fused with lysosomes in human AD,

Tg(+) mice, and malignant tumors. In addition, the biopsy and perfused brain tissue samples from

AD and the animals’ models that mimic human AD as well as biopsy that obtained from the cancer

patients were dominated by abnormal mitochondria as compared to a control group. In situ

hybridization with a chimeric cDNA probe for the 5kb common deletion indicated that the 5kb

mtDNA is increased at least 3 and 4 fold respectively in AD and malignant tumor cases as

compared to controls. In quantitative analysis of the mtDNA deletion and 8OHG in the same cases,

we found a strong significant positive correlation (r=0.934). Only hippocampal and cortical

vulnerable neurons as well as malignant cancer tissues showed immunopositive staining for RNA

oxidation markers visualized by using 8-OHG-staining, nitric oxide syntheses (NOSs: nNOS, iNOS

and eNOS), and all oxidative stress markers. The mitochondrial DNA over-proliferation and

deletion detected by using electron microscopic cytological techniques suggests that unsuccessful

irregularities of the cell cycle is also the hallmark of neoplasm; early mitochondrial dependent cell-

cycle pathophysiology in AD may recruit oncogenic signal transduction mechanisms and hence, can

be viewed as an abortive neoplastic transformation. This observation indicates that the oxidative

stress markers seen in the AD brain and malignant cancer selectively affects the population of

vulnerable neurons, vascular EC, and perivascular cells, suggesting that oxidative stress induced

mitochondrial DNA overproliferation and/or deletion plays a key role in the pathogenesis of AD

and cancer. The common features on the mitochondrial abnormality were seen on the brain during

tumorigenesis and AD indicating that mitochondrial DNA overproliferation and/or deletion are the

key initiating factors for development, maturation, and progression of neurodegeneration as well as

tumor growth and/or metastases. In addition, our later studies demonstrated that the mitochondrial

dependent brain cellular damage and therefore development of the neurodegeneration has NCS1

(neuronal calcium sensor-1) dependent pathway. NCS1 is a Ca2+-myristoyl switch protein of the

NCS protein family involved in synaptic plasticity and neurotransmission via Ca2+-dependent

regulation of dopamine D2 receptor and associated G-protein coupled receptor kinase (GRK)-2.

Hyper expression of NCS1 in synaptic terminals results in accumulation of membrane-bound

protein and its redundant regulatory activity associated with neurological disorders that manifest as

Alzheimer’s, Parkinson’s, multiple sclerosis (MS), cerebral palsy (CP) and possible other diseases.

By using functional and electron microscopic techniques we have found that the mitochondrial

lesions appeared to be the primary hallmark of the human glioblastoma. Vessel endothelium from

tumor tissues shows the damage of mitochondrion cristae. The mitochondria derived lysosomes

appeared to be permanent feature of the glial cells or derived tumor cells. The lipid laden tumor


30

cells and surrounding cells often show a different degree of mitochondria abnormality (such as

mitochondria with broken cristae, presence of edema in their matrix, disruption of inner and

external mitochondrial membrane). Moreover, giant mitochondria also appeared to be permanent

features of tumor growth and metastases indicating involvement all of compensatory pathway for

the survival. Comparative characteristics of marginal and central portion of tumor tissues obtained

from patients undergoing surgery with diagnosis of the primary glioblastoma showed that the

distance area of this tumor tissue is characterized by heterogeneous distribution of damage in the

very structure of the mitochondria. Central regions of tumor tissues in almost all of area shows

astrocytes with clusters of mitochondria derived lysosomes. The same patterns of cellular and

subcellular damage were seen in spinal cord tumor.

One of big challenge for the diagnosis of the dementia and/or neuro-degeneration appeared to be

the actual absence of the peripheral markers. Our extended study showed that age-related defects in

erythrocyte 2, 3-diphosphoglycerate metabolism/antioxidant status as well as the energy state of

erythrocytes in Alzheimer dementia can be used as a successful peripheral markers not only for the

diagnosis but also probing for markers regarding the “effectiveness” of the treatment. We have

found that erythrocytes (red blood cells: RBC) are considered as passive "reporter cells" for the

oxidative status of the whole organism, not active participants in mechanisms of AD pathogenesis

and are not well studied in AD. The aim of this work is to assess whether the antioxidant status and

energy state of RBC from elderly individuals change in AD. We measured levels of key products

and enzymes of oxidative metabolism in RBC from AD (n = 12) and non-Alzheimer dementia (NA,

n = 13) patients, as well as in cells from age-matched controls (AC, n = 14) and younger adult

controls (YC, n = 14). Parameters of the adenylate system served to evaluate the energy state of

RBC. In both aging and dementia, oxidative stress in RBC increased and exhibited elevated

concentrations of H₂O₂and organic hydroperoxides, decreased the GSH/GSSG ratio and

glutathione-S-transferase activity. Reductions in the ATP levels, adenine nucleotide pool size (AN)

and adenylate energy charge accompanied these oxidative disturbances. The patterns of changes in

these indices between groups strongly correlated with each other, Spearman rank correlation

coefficients being r(s) = 1.0 or -1.0 (p < 0.01). Alterations of the RBC parameters of oxidative stress

and adenylate metabolism were nonspecific and interpreted as age-related abnormalities. Decreased

glutathione peroxidase activity in RBC may be considered as a new peripheral marker for AD.

The main reason failing treatment of neurodegenerative diseases and cancer appeared to be

difficulty delivering drugs into the injured and affected tissues. Our future studies are aimed to

show that injection of silver nanoparticles in the brain lead to leaking on the inter endothelial

contact and luminal plasma membrane, and therefore elucidate the possibility of penetrating into the

cerebrovascular, neuronal, and glial cell which are especially damaged in AD and/or brain cancer.


31

Our clinical study showed the preservation and improvement of cognitive tasks in depressed and

demented patients after 24, 36 and 60 month follow up of combined pharmacological (especially

the combination of the diseases and mitochondrion specific compounds) and non- pharmacological

treatment. The study group consisted of 156 medically ill and physically disabled patients with mild

to moderate dementia and depression. Patients were treated with antidepressants, cholinesterase

inhibitors, and NMDA antagonists, along with their regular medication regimen. Non-

pharmacological intervention was centered on a home-based program of physical and cognitive

exercises as well as with vitamins and supplements (multivitamins, vitamin E, L-methylfolate,

alpha-lipoic acid, acetyl-L-carnitine, omega-3, and coenzyme Q-10 and QIAPI-1) and diet

modification. Cognitive assessments were performed yearly. After 60 months of treatment,

performance of all tasks remained at or above baseline. The MMSE, Cognistat–Attention,

Cognistat–Judgment, and RFFT - Total Unique Designs demonstrated significant improvement. Our

results also demonstrate that the arrest in cognitive decline in demented or depressed patients with

multiple medical co-morbidities for 60 months.

Our study, for the first time, demonstrated the pattern of oxidative stress induced mitochondrial

DNA over-proliferation and/or deletion as well as mitochondrial enzyme activities, oxidative stress

markers in erythrocytes during the development of human AD, and animals that mimic human AD,

colorectal cancer in liver metastasis, and malignant brain cancers. We conclude that mitochondrial

lesions, especially mitochondrial DNA abnormalities, erythrocytes enzymes and antioxidants

patterns are responsible for cell viability which can be used as new diagnostic tools and/or criteria

for the earlier detection of diseases and future considerations for this approach will enable us to

open new pathways, not only for the better understanding of BBB homeostasis which most likely

plays a key role in the development of AD, but also for the development of new and more specific

treatment strategies that will be more powerful and effective in bringing a cure for this devastating

disease. Thus, our research involving the conjugation of the silver nanoparticles with

mitochondrially-specific drugs would help to diminish the lesions that occur in AD and/or tumor

tissues. Future investigations addressing the application of a combined, integrative treatment

models as well as peripheral markers in clinical practices are warranted.

Acknowledgements: This study was supported by “GALLY” International Biomedical Research

Consulting LLC, San Antonio, TX, USA.


32

Selected Key References:

Tikhonova L. A., Kaminsky Y. G., Li Y., Reddy V.P., Solomadin I. N., Kosenko E. A. and

Aliev G. Impacts of Аβ25–35 on membrane stability, energy metabolic and antioxidant enzymes in

erythrocytes of different ages. Am J Alzheimers Dis Other Demen. 2014, Vol. 29(8): 685-695

[Epub ahead of print, 2014 May 16. pii: 1533317514534757. ].

Shevtsova E.F., Vinogradova D.V., Kireeva E.G., V. Prakash Reddy, Aliev G. and Bachurin

S.O. Dimebon Attenuates the Rat-Brain Mitochondrial Permeabilization. Curr Alzheimer Res.

2014;11(5):422-429. [Epub ahead of print May 4].

Aliev G., Ashraf G.Md., Kaminsky Y.G., Sheikh I.A., Sudakov S. K., Yakhno N.N., Benberin

V.V. and Bachurin S.O. Implication of the Nutritional and Non-Nutritional Factor in the Context of

the Preservation of The Cognitive Performance in Demented/Depressed and Alzheimer Patients.

Am J Alzheimers Dis Other Demen. 2013, 28(7): 660-670. first published on October 1, 2013 as

doi:10.1177/1533317513504614

Kosenko E.A., Solomadin I.N., Tikhonova L.A., P. Reddy, Aliev G. and Kaminsky Y.G.

Pathogenesis of Alzheimer Disease: Role of Oxidative Stress, Amyloid-β Peptides, Systemic

Ammonia and Erythrocyte Energy Metabolism. CNS & Neurological Disorders – Drug Targets,

2014, 13(1), 112-119. [2013 Sep 16, Epub ahead of print].

Aliev G., K. Shahida, C.K. Hua Gan S.K. Firoz, A. Khan, A.M. Abuzenadah, W. Kamal, M.A.

Kamal, Tan Y., Qu X and M. Reale. Alzheimer disease and type 2 diabetes mellitus: the link to

tyrosine hydroxylase and probable nutritional strategies. CNS & Neurological Disorders-Drug

Targets. 2014, 13 (3), 467-477 [Epub ahead of print, 2013 Sep 18].

Aliev, G., Obrenovich M.E., Tabrez S., Jabir N.R., Reddy V.P., Li Y., Burnstock G., Cacabelos

R. and Kamal M.A. Link between Cancer and Alzheimer Disease via Oxidative Stress Induced by

Nitric Oxide-Dependent Mitochondrial DNA Overproliferation and Deletion. Oxidative Medicine

and Cellular Longevity. vol. 2013, Article ID 962984, 19 pages, 2013. doi:10.1155/2013/962984.

Aliev, G. Palacios H.H., Cacabelos P., Cacabelos R., Burzynski G., and S.R. Burzynski.

Mitochondria Specific Antioxidants and their Derivatives in the Context of the Drug Development

for Neurodegeneration and Cancer. Drug Des 2012, 2: 103, doi: 10.4172/2169-0138.1000103

Aliev, G. “The Role of Oxidative Stress, Mitochondria Failure, and Cellular Hypoperfusion in

the Context of Alzheimer Disease: Past, Present and Future”. Monograph Book: Nova Science

Publishers, Inc., New York, 2013, ISBN: 978-1-61942-878-2. PP. 1-426.

https://www.novapublishers.com/catalog/product_info.php?products_id=31801

Kotelnikova R. A., Grigorev V. V., Smolina A. V., Faingold I. I., Mishchenko D. V., Vankin G.

I., Zamoiskii V. L., Poletaeva D. A., Markova N. A., Romanova V. S. , Kotelnikov A. I., Aliev G.

and S. O. Bachurin. Design of a hybrid nanostructure based on fullerene C60 and biologically active


33

substance as one of the ways for modeling physiological properties of compounds. Russ. Chem.

Bull., Int. Ed., 09/2014, No. 10:2375-2382.

Iomdina E.N., Khoroshilova-Maslova I.P., Robustova O.V., Averina O.A., Kovaleva N.A., Aliev

G., Reddy V.P,. Zamyatnin Jr. A.A., Skulachev M.V., Senin I.I. and V.P. Skulachev.

Mitochondria-targeted antioxidant SkQ1 reverses glaucomatous lesions in rabbits. Front Biosci

(Landmark Ed), 2015, Jan 1, V. 20: 892-901. DOI No:10.2741/2817.

Solís Herrera A., Arias Esparza M. del. C., Ashraf G.M., Zamyatnin Jr. A.A. and G. Aliev.

Beyond mitochondria as the energy source of the cell? Central Nervous System Agents in

Medicinal Chemistry (CNSA-MC), Central Nervous System Agents in Medicinal Chemistry, 2015,

15, 32-41 [Feb 2., Epub ahead of print].

Baksheeva V.E., Nazipova A.A., Zinchenko D.V., Serebryakova M.V., Senin I.I., Permyakov

S.E., Philippov P.P., Li Y., , Zamyatnin Jr. A.A., Zernii E.Yu. and Aliev G. Ca2+-myristoyl switch

in neuronal calcium sensor-1: A role of c-terminal segment. CNS Neurological Disorders and Drug

Targets, CNS Neurol Disord Drug Targets. 2015 Feb 25. [Epub ahead of print].

Torrente D., Cabezas R., Avila M., Sanchez Y., Morales L., Ashraf G.M., Barreto G.E., J.

Gonzalez and Aliev G. Mechanisms of PDGFRα promiscuity and PDGFRβ specificity in

association with PDGFB. Frontiers in Bioscience (Elite edition) 01/2015; 7: in press.

Cabezas R., Avila M.F., Vega-Vela N.E., Echeverria V., González J., Hidalgo O.A., Santos

A.B., Aliev G. and Barreto G.E. Growth factors and astrocytic metabolism: Possible roles for

PDGF. Medicinal Chemistry, 2015 (in press: BSP-MC-2014-HT4-3).


34

Contemporary Approaches for Pharmacological Intervention of

Alzheimer’s Disease

S. Bachurin

Institute of Physiologically Active Compounds, Severny pr.1, Chernogolovka, 142432, Russia

Alzheimer’s disease (AD) is the most prevalent form of age-related dementia, which affects about 5% of individuals over the age of 65 and more than 16% over the age of 80. The number of people living with dementia worldwide in 2013 is estimated at 44.35 million, reaching 75.62 million in 2030 and 135.46 million in 2050. The global economic impact of Alzheimer’s disease is shown by the worldwide cost of $ 604 billion, which exceeds 1% of gross world product. It can be seen as the most significant health crisis in the 21st century [1]. At the present time only few medicines have been approved for AD treatment, though the market for AD is currently estimated at $ 5-8 billion annually with projections that show the market potential will surpass $ 20 billion by 2020 [2].

The analysis of contemporary approaches for discovery of novel efficient agents for AD treatment permits to outline the following main trends:

The development of therapeutic agents acting on the main stages in pathogenesis of AD. These agents are called “disease modifying drugs (DMD)”. Such agents should slow the progression of structural damages and produce the improvement of cognition functions in AD patients that persists even after abolishment of the agent [3]. The main strategies in the development of DMD focused on blockade of pathogenic beta-amyloid oligomers formation, in particular, beta- and gamma- secretases inhibition, alpha-secretase activation, and beta-amyloid deposition. The alternative is the prevention of neurofibrillary tangles formation related to abnormal phosphorilation of tau-proteins.

The multifactorial nature of AD determines strong interest for the developing novel multi-targeted drugs for AD. Among them special attention is focused on compounds acting simultaneously on acetylcholinesterase and monoaminoaxidase, different subtypes of glutamate and serotonin receptors, targets of beta-amyloid formation and degradation pathway, pharmacophores which possess additionally NO-generating and antioxidant activities, and many others [4].

Search and study of novel potential biotargets related to the pathogeneses of AD. The analysis of date presented in “Thomson Reuters Integrity” database in fall 2014 permitted to outline about 140 potential biotargets, which are used currently for AD drug discovery [4]. Recent years number of new promising mechanisms for treating of AD pathology has been proposed. In particular, stabilization of mitochondrial functioning, prevention of pathological protein aggregation in brain (proteinopathy), activation of endogenous mechanism of nervous cell protection, stimulation of neurogenesis and autophagy. Some original agents, which utilize such mechanisms as novel efficient neuroprotectors and pro-neurogenic compounds have been reported.

During the last 10 years more than 20 compounds passed phase 2 clinical trails but none of them passed phase 3. As a result of very low outcome from clinical trails of innovative drugs and correspondingly high risk for investments the already known drug repositioning for AD treatment is discussed now as a very attractive area for fast development of AD-therapeutics [5]

The examples of the most promising current investigational agents are presented and discussed from the point of structure-activity relationships. [1]. The Global Impact of Dementia 2013-2050 [2]. http://www.acimmune.com/content/?p=22 [3]. Salomone S.et al., Br. J. Clin. Pharmacol., 2012, l73: 504-517. [4]. Kukharsky M., et al., in ”Neurodegenerative disease” M. 2014, Part 2: 137-162 (russ). [5]. Corbet at al., Nature Rev.Drug Disc., 2012., 11: 833-846.


35

From Pharmaceutical Substances to Pharmaceutical Formulations. How Can

Solid State Chemistry Help?

E.V. Boldyreva1,2

1 Novosibirsk State University, 630090, Russian Federation, ul. Pirogova, 2,

2 Institute of Solid State Chemistry and Mechanochemistry SB RAS, 630128, Russian Federation, ul. Kutateladze, 18


A way from drug substance to drug form is long and often difficult. One needs not only to

synthesize an active pharmaceutical compound, but to obtain this compound in the form which

would make it possible to test its bioavailability, toxicity, and to develop the form(s) optimized for

administration, manufacturing, storage. At all the stages of drug research and development the

approaches and techniques of solid state chemistry may be helpful. This will be illustrated by the

present contribution. Examples will be given of the applications of solid state chemistry and

mechanochemistry for “dry synthesis” and “dry extraction” of pharmaceutical compounds from

natural resources or wastes. A special emphasis will be made on obtaining new drug forms without

modifying the molecular formula of the active pharmaceutical ingredient (control of polymorphism

of pharmaceutical substances; obtaining pharmaceutical compounds as amorphous solids; control of

particle size and shape distribution; cryotechnologies for obtaining micro-balls of nanoparticles, e.g.

for per nasal delivery). Different approaches to solubilization of drugs will be considered, including

the manufacturing of salts, co-crystals, composites with excipients. Examples of using non-ambient

temperature and pressure conditions in order to improve drug forms will be given.

References [1]. Е.В. Болдырева Физическая фармация и ее возможности, Наука в России, 2014, 3, 26-35; [2]. Е.G. Zewak, А.G. Ogienko, Е.V. Boldyreva et al. Application of methods of physical pharmacy for the

improvement of solid drug forms, Theoretical Foundations of Chemical Engineering, 2013, 14(8),.480-492; [3]. Е.Г. Зевак, А.Г. Огиенко, Е.В. Болдырева, и др. Дизайн нанокомпозитов сальбутамол/глицин для

ингаляционной терапии: лекарственная форма нового поколения без использования хладонов, Нанотехнологии и охрана здоровья, 2013, 5, №3 (16), 30-43;

[4]. E.G. Zevak, A.G. Ogienko, E.V. Boldyreva et al. Salbutamol-Glycine Composite Microballs for Pulmonary Drug Delivery, RDD Europe 2013, 2, 329-334;

[5]. А.Г. Огиенко, Е.Г.Богданова, Е.В. Болдырева, и др. Высокоэффективные лекарственные формы нового поколения: фармацевтические композиции сальбутамола и будесонида для использования в порошковых ингаляторах, Разработка и регистрация лекарственных средств, 2015, 1 (10), февраль, 40-47;

[6]. A. Markel, A. Achkasov, T. Alekhina, и др. Effects of the alpha- and gamma-polymorphs of glycine on the behavior of catalepsy prone rats, Pharmacology, Biochemistry and Behavior, 2011, 98, 234–240;

[7]. Т.П. Шахтшнейдер, С.А. Мызь, М.А. Михайленко, и др. Композиция бетулина с биосовместимыми носителями с улучшенной динамикой растворения. Инновации в медицине: основные проблемы и пути их решения. Регенеративная медицина и новые биосовместимые материалы: материалы III Международного форума // Под общей редакцией М.А. Садового и Е.В.Мамоновой. Новосибирск: АНФПО Новосибирский академический центр человека, 2014. С. 263-267.


36

Heterocyclic Motif in Medicinal Chemistry

V.N. Charushin1,2 and O.N. Chupakhin1,2

1Postovsky Institute of Organic Synthesis, Ural Branch of the Russian Academy of Sciences,

Ekaterinburg, Russia. 2Ural Federal University, Ekaterinburg, Russia,


Development of new drugs, effective against socially important diseases (antivirals,

antibacterials, including antitiberculosis, cardio-vascular agents and others), appears to be the actual

task of medicinal chemistry. As far as the drugs of synthetic origin are concerned, the role of

heterocyclic compounds can hardly be overestimated, as illustrated by numerous examples.

Effective methods for the synthesis of promising antivirals in the series of heterocyclic

compounds, both nucleosides and non-nucleosidederivatives, have been developed. They involve

the synthesis of antiviral compounds of the azoloazine family, such as anti-influenza drug

Triazavirin (I) [1], launched in Russia in 2014, and its numerous derivatives, which can be regarded

as analogues of heterocyclic bases of DNA. Also chemo-enzymatic methods, leading to ribo- and

2'-deoxyribo nucleosides of the benzimidazole family, have been advanced. A number of

compounds of this series (II) proved to inhibit progression of herpes virus at subtoxical

concentrations [2].

NN

NN

N

O

RSNO2

Cat-

+

-

I

N

NF

O

OH R"'

OH

R'

R"

¸

-

IIR'''= H, OH

N

N SR

-

III

F

R'

R"

N

O

z

COOH

¸

-

IV

A number of pyrimidines (III) [3], and condensed azolo[1,2,4,5]tetrazines [4] have been

established to possess a high antituberculosis activity.

Recent advances in the synthesis of highly effective antibacterials of the fluoroquinolone family

IV, including their enantiomerically pure forms (levofloxacin, moxyfloxacin and others) will also

be discussed [5]. [1]. I. Karpenko, S. Deev, O. Kiselev, V. Charushin, V. Rusinov, E. Ulomsky, E. Deeva, D. Yanvarev, A. Ivanov,

O. Smirnova, S. Kochetkov, O. Chupakhin, M. Kukhanova. Antimicrobial Agents and Chemotherapy, 2010, vol. 54, No. 5, p. 2017-2022.

[2]. I.D. Konstantinova, O.M. Selezneva, I.V. Fateev, et al. Synthesis, 2013, vol. 45, p. 272-280. [3]. M.A. Kravchenko, E.V. Verbitsky, I.D. Medvinsky, et al. Bioorganic and Medicinal Chemistry Letters, 2014,

vol.24, p. 3118-3120. [4]. O.B. Bekker, V.N. Danilenko, R.I. Eshmetova et.al. Russian Patent 2545458 (2015). [5]. V.N. Charushin, G.N. Lipunova, E.V. Nosova, O.N. Chupakhin. Fluoroquinolones: Synthesis and Application. In:

Fluorine in Heterocyclic Chemistry, V. Nenajdenko (Ed.), vol. 2, Springer, 2014, p. 111-180.


37

Rational design of antibiotics and NSAIDs

M.A. Cooper

Institute for Molecular Bioscience, The University of Queensland, St Lucia, Australia


The lack of effective treatments for multi-drug resistant bacteria is an urgent health issue, with

the world approaching a ‘post-antibiotic’ era. There are over 1.2 million annual infections by Gram-

positive methicillin resistant S. aureus (MRSA) and drug resistant S. pneumoniae in the US alone,

leading to 18,000 deaths. We postulated that membrane-targeting motifs could be appended to

drugs acting on membrane-anchored targets, enhancing membrane binding and concomitantly

increasing drug concentration at the target site. We selected vancomycin to test this approach, as

this prototypical glycopeptide inhibits peptidoglycan formation by binding lipid II.

Over 300 analogues have been prepared in an extensive medicinal chemistry campaign. The best

compounds are 10- to 100-fold more active than vancomycin, with excellent plasma and

microsomal stability, minimal inhibition of CYP450 enzymes, negative in Ames mutagenicity

testing, and non-haemolytic. Mouse pharmacokinetic profiles are consistent with once daily dosing

in man, and the compounds show equivalent or superior efficacy to the reference antibiotics

vancomycin and daptomycin in multiple mouse infection models. For example, a single sub-

cutaneous 10 mg/kg dose in an MRSA thigh infection model was equivalent to 200 mg/kg

vancomycin, with greater than 100,000-fold reduction in bacterial load (cfu/thigh) at 24h.

Optimisation to select a preclinical candidate is in progress.

In addition we have explored the interface between infection, innate immunity and sterile

inflammation, with a focus on the role of short chain fatty acids and synthetic ligands in ischemia

reperfusion injury, and on development and validation of ligands to block inflammasome activation.

Proof-of-concept molecules in various inflammatory disease indications will be discussed.

References [1]. “A small molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases,”

R.C. Coll, et al. Nature Medicine, 2015, DOI: 10.1038/NM.3806 [2]. Surface stress sensors for rapid and ultrasensitive detection of active free drugs in human serum, R. M.

Counago, et al., Nature Nanotechnology., 2014, doi: 10.1038/nnano.2014.33 [3]. Imperfect coordination chemistry facilities metal ion release in the Psa permease, R. M. Counago, et al.,

Nature Chem. Biol., 2013, doi:10.1038/nchembio.1382 [4]. Identification of anti-tubercular benzothiazinone compounds by ligand-based design, T. Karoli et al., J

Med Chem, 2012, 55, 7940-4. [5]. Fix the antibiotics pipeline M.A. Cooper & D.A. Shlaes, Nature, 2011 472, 32. [6]. Nanomechanical detection of antibiotic-mucopeptide binding in a model for superbug resistance, J.W.

Ndieyira, et al., Nature Nanotechnology, 2008, 3, 691-696.


38

Mycobacterium tuberculosis “hybrid warfare”: virulence, pathogenicity,

persistence and drug resistance. New counteraction concept.

V.N. Danilenko

Vavilov Institute of General Genetics, Gubkin str. 3, 119991, Moscow, GSP-1, Russia

E-mail: [email protected].

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, is responsible for considerable worldwide human morbidity and mortality. Nearly a third of the world’s population is infected with persistent or dormant Mtb, referred to as latent tuberculosis. Reactivation of latent infection is the major source of active tuberculosis in adults. The main obstacles in the global control of tuberculosis are linked to the emergence of multi-drug resistant strains and the therapeutic failure of persistent infection treatment using conventional anti-tuberculosis drugs. But these are only the tip of the iceberg.

The mechanisms of Mtb adaptation to external factors in human organism, including antibiotic treatment, is more complicate and often unpredictable to our modern knowledge. It is obvious today that the phenomena of persistence, antibiotic tolerance, multidrug resistance, virulence and pathogenicity are connected to each other. The flexible regulation of Mtb metabolism in response to environmental changes (hypoxia, shift from aerobic to anaerobic conditions etc.) plays a significant contribution to the virulence of Mtb.

The advancement of molecular genotyping tools and whole genome sequencing technology allows us to identify more subgroups among the classic Mtb genotype lineages. A correlation between these subgroups and virulence and pathogenicity properties is observed. It has been suggested that some lineages and subgroups are more prone to develop new adaptive forms with enhanced virulence and multidrug resistance.

To address these problems it is crucial to: develop new molecular diagnostic tools based on functional biomarkers (those taking part in determining the described Mtb properties, such as toxins-antitoxins systems); create new databases of SNPs in the mentioned group of genes and bioinformatics tools for their analysis; conduct whole-genome sequencing of reference strains, representing all the known genotypic lineages and subgroups; develop next generation drugs targeting key regulators of the complex process of forming the phenotypes of persistence, tolerance, virulence and pathogenicity. References [1].Danilenko V.N. et al. Bacterial eukaryotic type serine-threonine protein kinases: from structural biology to targeted

anti-infective drug design. // Curr Top Med Chem. 2011. Vol. 11, № 11. Pp. 1352–136918 pp. [2].Prozorov A.A., Zaichikova M.V., Danilenko V.N. Mycobacterium tuberculosis mutants with multidrug resistance:

History of origin, genetic and molecular mechanisms of resistance, and emerging challenges // Russ J Genet. SP MAIK Nauka/Interperiodica, 2012. Vol. 48, № 1. Pp. 1–1414 pp.

[3].Prozorov A.A., Zaichikova M.V., Danilenko V.N. Systems of genes and proteins affecting mycobacteria virulence and their homologs participation in conjugation of Mycobacterium smegmatis // Russ J Genet. SP MAIK Nauka/Interperiodica, 2013. Vol. 49, № 1. Pp. 110–12516 pp.

[4].Prozorov A.A. et al. The virulence factors of Mycobacterium tuberculosis: Genetic control, new conceptions // Russ J Genet. Pleiades Publishing, 2014. Vol. 50, № 8. Pp. 775–79723 pp.

[5].Bekker O.B. et al. Synthesis and activity of (+)-usnic acid and (−)-usnic acid derivatives containing 1,3-thiazole cycle against Mycobacterium tuberculosis // Med Chem Res. Springer US, 2015. Pp. 1–1313 pp.

[6].Maslov D.A. et al. Resistance to pyrazinamide in Russian Mycobacterium tuberculosis isolates: pncA sequencing versus Bactec MGIT 960. // Tuberculosis 2015 (In Press).

[7].Patent RU 2547598 C1 (application 2013155216/10, 12.12.2013), “Method of genotyping strains Mycobacterium tuberculosis” by Alekseeva M.G., Danilenko V.N., Zaichikova M.V. and Zakharevich N.V.


39

The Fundamental Basis and Approaches for Dipeptide Drugs Development

T.A. Gudasheva

Zakusov Research Institute of Pharmacology, Russia, Moscow, Baltiyskaya str., 8

E-mail [email protected]

Peptidergic drugs have a special place among other medicines. This is because regulatory

peptides form a global hierarchic system of regulation that cover immune, nerve and endocrine systems of organism. The peptides have the wide diverse of physico-chemical properties of the side chains and, as a result, high information density that allow them to interact with a large number of pharmacological receptors. The unity of ribosomal synthesis allows peptides to be a universal information transmitter in living systems. Many medicines are developed on the basis of peptides. Peptide medicines usually have a high activity, low toxicity due to metabolism into natural amino acids, they rarely initiate tolerance and dependence.

Dipeptide drugs have advantage over oligopeptide in stability in biological liquids, in ability to penetrate through biological barriers and, as a result, in the possibility of oral administration. Dipeptides have a low multyfinctionality and low cost of synthesis.

Dipeptide direction in pharmacology based on the hypothesis that active site of polypeptide chain that interacts with receptor is usually a beta-turn. The size of active site is a compromise between recognition accuracy and dissociation rate. The position of active site on the peptide chain dictated by geometry of peptide-receptor complex. The most important role in the peptide recognition plays the central dipeptide fragment of beta-turn that is the most deeply penetrate into receptor (according to geometric reasons). This explains why non-peptide medicines with peptidergic mechanisms of action have molecular weight similar to dipeptides.

On this basis we developed approach named drug-based peptide design that is the movement from non-peptide drugs structure to their topological dipeptide analogs. For instance, starting from classical nootropic drug Piracetam structure we developed dipeptide medicine Noopept (N-phenylacetyl-L-prolyl-glycine ethyl ester) which come to Russian pharmaceutical market in 2006. Noopept laid a bridge between racetames and memory peptide Vasopressine, and led to discovery of a new neuropeptide cyclo-L-prolylglycine. Starting from benzamide neuroleptic Sulpiride structure we developed dipeptide neuroleptic Dilept (N-Caproyl-L-prolyl-L-tyrosine methyl ester) that successful passed stage 2 of clinical trials and connected the group of benzamide neuroleptics and endogenous neuroleptic Neurotensin.

Another approach for dipeptide drugs development consists of design of dipeptide mimetics of regulatory peptides and proteins active sites. These sites usually represent beta-turns of peptide chain. For example, starting from neuropeptide Cholecystokinine-4 and using Shemiakin-Ovchinnikov-Ivanov topochemical principle we developed potential selective anxyolytic Anxyopept that is on the stage of receiving of permission for clinical trials. On the base of beta-turns of the loops of endogenous neuroprotective proteins neurotrophines in the present time we are developing their dipeptide mimetics.

Thus our dipeptide direction resulted in a development of a line of dipeptide medicines that are high active, non-toxic and available orally.

This work was partically supported by the Russian Scientific Foundation (project 14-15-00596) and Russian Foundation for Basic Research (project 15-04-04485).


40

Modern Developments in New Drugs and Their Delivery by Nanomaterials

J. R. Hwu1,2, M. Kapoor1 and S.-C. Tsay2

1Department of Chemistry, National Tsing Hua University, 300, Hsinchu, Taiwan 2Department of Chemistry, National Central University, 320, Jhongli City, Taiwan


Paclitaxel-conjugated nanoparticles (NPs) of three different types have been synthesized in our

laboratory by use of Fe3O4 and Au as the cores. Possessing the polyethylene glycol (PEG)-SH

spacer and the phosphate joining unit, the new paclitaxel-P(=O)(OH)-PEG-S-Fe-NP nanomaterials

functioned as a prodrug of paclitaxel, which was liberated in the presence of phosphodiesterase. A

new class of targeted anti-cancer drugs is thus established by exploitation of these conjugated

nanomaterials [1].

Moreover, single-stranded DNA of mixed deoxy-adenine (dA), -thymine (dT), -guanine, (dG),

and -cytosine (dC) was used for the first time to wrap around functionalized single-walled carbon

nanotubes. Their external surface was covalently attached with multiple triazole–(ethylene glycol)–

dA ligands. This approach of hybridization involved the formation of hydrogen bonds between dT

of single-stranded DNA and dA of functionalized carbon nanotubes. The degree of functional

ligands was one dA-containing ligand per 54 carbon atoms on nude carbon nanotubes [2].

Advancements of nanoparticles and nanotubes as vehicles provide two new platforms for

delivery of targeted drugs and biologically active compounds. These two strategies have great

potential in biomedical applications and drug development.

References [1] J. R. Hwu, Y. S. Lin, T. Josephrajan, M.-H. Hsu, F.-Y. Cheng, C.-S. Yeh, W.-C. Su, and D.-B. Shieh, J. Am. Chem. Soc. 2009, 131, 66−68. [2] J. R. Hwu, M. Kapoor, R.-Y. Li, Y.-C. Lin, J.-C. Horng, and S.-C. Tsay, Chem. Asian J. 2014, 9, 3408−3412.


41

Ga-68 and Lu-177-labelled Peptide Conjugates in Cancer Theranostics

K. Kairemo

Department of Nuclear Medicine & Molecular Radiotherapy, Docrates Cancer Center,

Helsinki, Finland

Neuroendocrine neoplasms (NENs) constitute a heterogeneous group of tumours that frequently express cell membrane-specific peptide receptors, such as somatostatin receptors (sst), and of which gastroenteropancreatic (GEP), carcinoid and pancreatic islet cell tumours exhibit the highest expression of sst receptors. Radiolabelled receptor-binding somatostatin analogues (octreotide and lanreotide) act as vehicles to guide radioactivity to tissues expressing sst receptors, and can thus be used for their diagnosis and treatment. Five somatostatin receptor (sst) subtype genes, sst1, sst2, sst3, sst4 and sst5, have been cloned and characterised. The five sst subtypes all bind natural somatostatins-14 and -28 with high affinity. Endocrine pancreatic and endocrine digestive tract tumours also express multiple sst subtypes, but sst2 predominance is generally found. However, there is considerable variation in sst subtype expression between the different tumour types and among tumours of the same type. The predominant expression of sst2 receptors on pancreatic endocrine or carcinoid tumours is essential for the control of hormonal hypersecretion by the octapeptide somatostatin analogues such as octreotide and lanreotide. Somatostatin and its octapeptide analogues are also able to inhibit proliferation of normal and tumour cells. The high density of sst2 or sst5 on pancreatic endocrine or carcinoid tumours further allows the use of radiolabelled somatostatin analogues for in vivo visualisation. The predominant expression of sst2 receptors in these tumours and the efficiency of sst2 receptors to undergo agonist-induced internalisation is also essential for the application of radiolabelled octapeptide somatostatin analogues.

A number of radioisotopes with different physical properties have been used for their diagnosis and treatment. Currently, [111In-DTPA0]octreotide, [90Y-DOTA0,Tyr3]octreotide, [177Lu-DOTA0Tyr3]octreotate, [111In-DOTA0]lanreotide and [90Y-DOTA0]lanreotide can be used for this purpose. The administration of high doses of the Auger electron and gamma-emitter [111In-DTPA0] octreotide in patients with metastatic tumours has been associated with considerable symptomatic improvement but relatively few and short-lived objective tumour responses. The use of another radiolabelled somatostatin analogue coupled with 90Y, a pure b-emitter, [90Y-DOTA0,Tyr3]octreotide(90Y-DOTATOC, OctreoTher), was associated with 10–30% objective tumour response rates, and appears to be particularly effective in larger tumours.

However, treatment with [177Lu-DOTA0Tyr3]octreotate (177Lu-DOTATATE), which has a higher affinity for the sst2, resulted in approximately 1/3 complete or partial tumour responses; this radiopharmaceutical is particularly effective in smaller tumours. Furthermore, treatment using both 90Y-DOTATOC and 177Lu-DOTATATE seems promising, as the combination of these radiopharmaceuticals could be effective in tumours bearing both small and large lesions. 177Lu-DOTATATE and 90Y-DOTATOC are already included as a part of Nordic Guidelines of the management of NETs. In this work I describe our patient material where we applied standard protocols both for dose planning and dose calculations based on serial imaging.

The prostate-specific membrane antigen (PSMA) is a type II membrane glycoprotein that is present primarily in the prostate and is overexpressed during all stages of the androgen-insensitive or the metastatic cancer of this organ. Because of its high expression during the development and progression of a malignancy, PSMA is considered to be a good target for the imaging and treatment of prostate cancer. Several investigators have reported the use of radio-halogenated (e.g., 18F, 125I, etc.) and 99mTc-labeled small molecule inhibitors for the imaging of PSMA. In this work I describe our patient material where we applied standard protocols for prostate cancer diagnostics using Ga-68-PSMA-positron emission tomography (PET)/CT.


42

Drug Development R&D Center “IPHAR”

V. Khazanov

OOO “IPHAR”, 643021, Russia, Tomsk, str. Elizarovykh, 79/4


“IPHAR” company is an R&D center of a private pharmaceutical group, which includes a

licensed pharmaceutical manufacturing facility and project companies. Promising candidate

molecules, chosen by the Expert Board of IPHAR, undergo full-scale development that, if

successful, ends with state registration and technology transfer to industrial production. Usually,

proprietary or third-party laboratories, engaged in fundamental research, act as sources of new

molecules. For the development of every new drug a new spin-off company is created, which owns

intellectual property for the drug and later commercializes it. Drug development is performed in

accordance with industrial quality standards GLP, GCP and GMP. “IPHAR” company is the only

Russian full-scale drug development center that is currently undergoing national GLP accreditation.

The company has all the necessary departments, including vivaria for specific-pathogen-free

animals, pharmaceutical department (including synthesis laboratory, analytical chemistry

laboratory, formulation and manufacturing technology laboratories), preclinical trials departments

(including toxicology laboratory, specific activity laboratory, morphology laboratory,

pharmacokinetics laboratory, cell technology laboratory, etc.), clinical trials department (including

data management group, biostatistics and medical writing groups), pharmaceutical department

(including synthesis laboratory, formulation laboratory, analytical laboratory), pilot manufacturing

department (for substances and dosage forms), quality management department, archive (for

documents and laboratory samples), management department (including legal services and project

management units).

IPHAR is an active participant in the government program of pharmaceutical industry

development (Pharma 2020), performing 8 drug development projects. In the last 4 years IPHAR

has completed about 40 projects of drug development for domestic and foreign customers. In 2013

IPHAR was the first Russian company to be accredited by government foundation “Skolkovo” as a

drug development center.


43

New Frontiers in Brassinosteroids: from Convergent Synthesis and Highly

Sensitive Analysis to Pharmacokinetics Studies

V. Khripach, V. Zhabinskii

Laboratory of Steroids, Institute of Bioorganic Chemistry NAS Belarus, 220141, Belarus, Minsk,

Kuprevich, 5/2


Brassinosteroids (BS), the first group of plant hormones having a steroid structure, are a subject

of extensive studies as potential leads for new generation agrochemicals and medications [1,2].

Chemical synthesis is the only way to prepare BS in sufficient amount for practice and despite the

progress made in this area, preparation of many of them (and especially the most active BS of

campestane series) is still rather complicated task because of the necessity to construct de novo the

side chain bearing four chiral centers.

Here we report a new efficient convergent synthetic procedure based on commercially available

BS (24-epibrassinolide and 24-epicastasterone) as starting materials and new selective methods for

the side chain construction. Using these approaches, a number of natural-type BS, their analogs,

isotope-, enzyme- and fluorescent-labeled derivatives have been synthesized and used for

physiological studies and as tracers in analytical measurements of BS.

As a part of our work of the latter direction, a new ELISA for quantification of 6-keto-

brassinosteroids has been developed. Its combination with two other immunoenzymatic assays for

brassinosteroids was used to determine individual compounds of this series. In addition, a direct

method of brassinosteroid analysis was proposed. It has the advantage of requiring no sample

pretreatment steps such as extraction with organic solvents and chromatography.

Another ELISA test-system has been developed for measuring of 24-epibrassinolide (EBl) in

physiological liquids and tissues of animals and applied for studies of pharmacokinetics of our new

anticholesterolemic agent Decrechol having EBl as an active ingredient. Being more sensitive than

the HPLC-MS and due to very low active doses of Decrechol, it was found to be an easy and

convenient tool to follow the changes of the drug concentration in animals in the course of

preclinical trials.

References [1] V. A. Khripach, V. N. Zhabinskii, N. B. Khripach, in Brassinosteroids (Eds.: S.Hayat, A.Ahmad), Kluwer

Academic Publishers, Dordrecht, 2003, pp. 189-230. [2] V. N. Zhabinskii, N. B. Khripach, V. A. Khripach, Steroids 2015, 97, 87-97.


44

Antiviral Drugs Against Ebola Infection: Novel Targets and Perspective

O.I. Kiselev

Federal State Institute of Influenza, MPH Russian Federation, Saint Petersburg

Antiviral drugs are fast growing field of research and development stimulated by appearance of

new infectious threats. Recent revision of potential in a choice of drugs for EVD treatment showed dramatically narrow set of medicines to cure this fatal disease. Also it is obvious that clinical trials of drugs against highly pathogenic virus is a difficult problem from various points: random outbreak, very short time for decision to use any kind of new drug, strong responsibility to work with fatal infection, very narrow therapeutic window etc. Even trials on monkey can’t be the reason of underpinning for recommendations and approval. As an example of fast and responsible enactment for consideration concerning new drug application for treatment of Ebola is compelling by the WHO to approve the use of experimental treatments in infected patients using humanized monoclonal antibodies cocktail ZMab. Several compounds have been trialed in the recent past on animal models in non-human primates and appear promising for human use in future protocols for EVD treatment.

It is obviously, that there is a great demand for drugs as small molecular inhibitors of viral and cellular functions important for viral propagation. Ebola virus has an unexpected high level of viral load (up to 7 and more Ig) and high viral proteins saturation in blood stream as secretory product from infected cells. Drugs developed against Ebola should be addressed to those paradoxes to be effective at any stage of the disease.

There are a real line of other promising new drugs recently trailed and submitted to approval: Small interfering RNA molecules (siRNA) targeting Ebola virus VP24, VP35 and polymerase

(L) genes. To improve pharmaceutical properties of siRNA unique encapsulation in stable nucleic acid lipid particles (SNALPs) had been used.

TKM-Ebola had recently got the approval by US Food and Drug Administration for prevention and treatment of Ebola infection. PMOPlus (Positively charged phosphorodiamidatemorpholino oligomers) successfully trialed on animals including monkey with high protective activity. AVI-7537 targeting the VP24 gene of Ebola virus and AVI-7288 targeting NP.

Novel adenosine analogue BCX 4430 an adenosine analogue had been proofed as a drug, which can be used intravenously in a patients with hard conditions.

Very attractive drug with extremely high activity in vitro was identified as a blocker of Ebola virus interaction with cellular receptor NPC1 cholesterol transporter.

T-705, (favipiravir) a pyrazinecarboxamide derivative, known as an anti-influenza drug was trialed on animal models/ this drug is active as an inhibitor od viral RNA-polymerase capable to decrease virus load in peripheral blood. Drug is a strong inhibitor of Marburg virus replication.

Triazavirine (and related triazolopyrimidines/triazolopurines) developed in National Influenza Center (Saint Petersburg, Russia) was recently registered and approved for hard influenza cases treatment. The drug was testing in NIH (USA) against WNHFV and proofed as a drug with wide spectrum of antiviral activity. This is a strong inhibitor of Marburg virus replication. Now it is under the control animal trails for anti-Ebola virus activity.

There are several new unexpected observations in testing of drugs by high throughput screening procedure testing known drugs on anti-Ebola virus activity. Among these discovery is a drug Clomifen and derivatives.

Clinical development and pathogenesis of EVD is too complicated and require step by step accurate application of antiviral drugs in combination with others for prevention of the most dangerous complications such as sepsis and vascular leakage syndrome and mortality as well as. There is a great demand for the line of drugs for choice according to requirements of highly targeted treatment according to the stage of disease and syndromes development.


45

Development of New Antivirals: Problems and Prospects

S. Kochetkov

Engelhardt Institute of Molecular Biology Russian Academy of Sciences, 119991 Russia, Moscow, Vavilov st., 32


Infectious diseases are the second leading cause of mortality in the world, and among them, the viral diseases are prevalent. The intracellular localization of the virus, peculiar properties of viral entry and replication and rapid development of resistance determine the complexity of the antiviral therapy. Among the more than 2,000 species of viruses of particular concern are etiological agents of the so-called socially significant infections – respiratory viral infections, AIDS and viral hepatitis B and C. The present report deals with the development of potential antiviral agents against human immunodeficiency (HIV) and hepatitis C (HCV) viruses.

HIV studies Despite the numerous drawbacks, nucleoside inhibitors of reverse transcriptase (NRTIs),

particularly 3’-azido-3’-deoxythymidine (AZT, Zidovudine) remains one of the key drugs used in the treatment and prevention of HIV infection. A strategy in searching for new effective and safe AZT agents among latent (depot) forms of AZT has yielded its positive results. Synthesis and preclinical studies of such prodrug, AZT 5’-aminocarbonylphosphonate, have demonstrated that it releases AZT when taken orally. Pharmacokinetic studies have shown a prolonged action potential of AZT aminocarbonyl phosphonate. It has been recommended for clinical trials; preliminary results of the first stage are promising. Phosphonate chimera of two well-known NRTIs, namely AZT and 3TC, can be regarded as a depot form of both Zidovudine and Lamivudine. This newly synthesized compound suppressed HIV-1 infection both in vitro (in cell cultures infected by HIV-1) and ex vivo (human tonsil tissues infected by HIV-1) more efficiently than did other phosphonates of NRTIs. Moreover, the new compound was not toxic for tissue cells, thus making the chimeric phosphonate strategy a valid approach for the development of anti HIV-1 compound heterodimers.

HCV studies A diverse collection of more than 40 derivatives of benzohydroxamic (BHAs) and pyridine

hydroxamic acids (PHAs) of various structural groups were synthesized and tested against HCV in full-genome replicon assay. Some of these compounds demonstrated an exceptional activity, suppressing viral replication at sub-micromolar concentrations. The compounds were inactive against key viral enzymes in vitro assays, suggesting host cell inhibition target(s). Both classes of hydroxamic acids decreased viral RNA concentration in hepatocyte cell line and simultaneously caused hyperacetylation of α-tubulin that pointed to inhibition of histone deacetylase 6 (HDAC6) as a component of their antiviral activity. The correlation between these two processes points to HDAC6 as perspective cellular target for hepatitis C therapy.

The work was supported by Russian Science Foundation, project No 14-50-00060


46

Human Tyrosyl-DNA Phosphodiesterase 1: New Activities and Development of

Enzyme Inhibitors as Anticancer Drugs

O. Lavrik1,2

1Institute of Chemical Biology and Fundamental Medicine, 630090, Russia, Novosibirsk, Lavrentiev Av., 8

2Department of Natural Sciences, Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogov St., 2


Tyrosyl-DNA phosphodiesterase 1 (TDP1) is responsible to process topoisomerase 1 (TOP1) –

DNA adducts as well as to hydrolyze a variety of other DNA 3′-substituents. We have shown

recently that TDP1 can initiate repair of apurinic/apyrimidinic (AP) sites located in the internal

positions of DNA generating breaks with the 3΄- and 5΄-phosphate termini [1-3]. This activity was

not observed for TDP1 SCAN mutant responsible for neurodegeneration. Polynucleotide kinase

phosphatase, Polβ and DNA ligase coordinated with XRCC1 are completing repair of the AP sites.

The AP site cleaving activity of TDP1 is shown to be stimulated by the key modulator of base

excision repair (BER) – poly(ADP-ribose)polymerase 1 (PARP1). The data suggest a role of TDP1

in the new APE1-independent BER pathway in mammals. This activity can contribute to repair of

AP sites particularly in ssDNA structures or in the context of cluster-type lesions. The specific real-

time fluorescent detection of the AP-site cleaving activity of TDP1 was developed, which is not

sensitive to AP site cleavage by APE1 and useful to evaluate a biological significance of new TDP1

function to initiate cleavage of AP sites. TDP1 is a promising target for antitumor therapy based on

TOP1 poison-mediated DNA damage. The row of novel benzopentathiepins and other compounds

were synthesized and tested as TDP1 inhibitors using an original oligonucleotide-based

fluorescence assay [4]. The benzopentathiepines showed IC50 values in the range of 0.2–6.0 µM.

The specificity of these inhibitors was investigated by using other target DNA repair proteins such

as PARP1, PARP2, AP-endonuclease 1 and DNA polymerase beta. The study of cytotoxicity of

these compounds revealed that their action leads to the apoptotic cell death of cancer cells.

Therefore the new class of very effective inhibitors of TDP1 was elaborated, which are very

prominent to improve cancer therapy based on TOP1 poison–mediated DNA damage.

Financial Support: RSCF (14-24-00038), RFBR (14-04-00268); MCB Program of RAS; Ministry of Education and Science of Russian Federation, State Agreement №14.604.21.0018, ID RFMEFI60414X0018.

References [1]. Lebedeva N.A., Rechkunova N.I., Lavrik O.I. FEBS Lett., 2011, 585, 683-686. [2]. Lebedeva N.A., Rechkunova N.I., El-Khamisy S.F., Lavrik O.I. Biochimie, 2012, 94, 1749-1753. [3]. Lebedeva N.A., Rechkunova N.I., Ishchenko A.A, Saparbaev M., Lavrik O.I. DNA Repair, 2013, 12, 1037– 1042. [4]. Zakharenko A., Khomenko T., Zhukova S., Koval O., Zakharova O., Anarbaev R., Lebedeva N., Korchagina D.,

Komarova N., Vasiliev V., Reynisson J., Volcho K., Salakhutdinov N., Lavrik O. Bioorg. Med. Chem,. 2015, 23, 2044-2052.


47

The study on Effective compounds that fermented by Lactobacillus casei in

chestnut (Castanea crenata) bur against skin aging caused by UVB

Jin-Tae Lee1 and Young-A Jang2

1Department of Cosmeceutical Science, Daegu Haany University,

Gyongbuk Province, Gyongsan City, Republic of Korea

E-mail: [email protected] or [email protected]

People have long been in contact with cosmetic culture and cosmetics is changing with the times. Cosmetics is settled as a necessity the body clean and glorifying a simple function in young and healthy skin product. In now days, people is getting interested in the variety functional product because demand of cosmetics industry expanded and become high-quality in response to the development of raw materials, the development of cosmetics from industrial development technologies. Functional cosmetic industrial is, which is defined as a small quantity batch, environment friendly, high value-added, and a knowledge-based technology, depending on the raw material as a materials for medicine or functional foods. In recent years, cosmetic industry and academia lead research trends to search useful components obtained from the fermentation of various natural products or to apply the studied physiological activity for cosmetic material. Particularly, anti-wrinkle or known immune function components have been reporting that they involve the control of human's physiology or maintaining of homeostasis, so removing active oxygen (oxygen free radical) showed to play an important role in health maintenance such as anti-aging, anti-inflammatory activities.

Skin is composed of the epidermis and dermis. Epidermis is the most outer layer and composed of the skin epithelial layer which works for skin protection. Dermis, which is consist in a support layer of connective tissue cells, are at the bottom of epidermal cell. Extracellular matrix (ECM) in dermal cell is composed of collagen type I for major parts which provides the tension and elasticity to the skin, and some collagen type Ⅱ, elastin, glycoprotein, and fibronectin.

Autocrine and paracrine are cytokines produced by UV stimulus and may cause internal aging or various skin cancer by the complex inflammation or immune response through serious damages in dermal connective tissue. NF-κB activation induces the gene expression of matrix-degrading enzymes such as MMP-1 (matrix metalloproteinase-1), MMP-3 (stromelysin-1), and MMP-9 (92KDa gelatinase), eventually leads these matrix metalloproteinases (MMPs) accumulation. References [1]. Uitto J.: Collagen. In : Fitzpatrick TB, Eisen AZ, Wolff K, Ferrdberg IM, Austen KF, eds. Dermatology in genneral

medicine. 4th ed. Vol. 1. New York: McGraw-Hill (1933) pp.299-314 [2]. Record I. R. and Dreosti I. E, Konstantinopoulos M and Buckley R. A.:1991, The influence of topical and systemic

vitamin E on ultraviolet ligth-induced skin damage in hairless mice., Nutr. Cancer, 16:219-226. [3]. Kawaguchi Y, Tanaka H, Okada T, Konishi M, Ito M and Asai J.:1996, The effects of ultraviolet A and reactive

oxygen species on the mRNA expression of 72kDa type Ⅳ Collagenase and its tissue inhibitor in cultured human dermal fibroblasts., Arch. Dermatol. Res., 288:39-44.

[4]. Vincenti M. P., White L. A., Schrone D. J., Benbow U., Brinckerhoff C. E.:1996, Regulating expression of the gene for matrix metalloproteinase-1(Collagenase): mechnisms that control enzyme activity, transcription, and mRNA stability., Crit. Rev. Eukaryot. Gene Expr., 6:391-411.

[5]. Westermarck J., Kahari V. M.:1999, Regulation of matrix metalloproteinase expression in tumor invasion., FASEB J., 13:781-792.

[6]. White L. A., Brinckerhoff C. E.:1995, Two activator Protein-1 elements in matrix metalloproteinase-1 promoter have different effects on transciption and bind Jun D, c-Fos and Fra-2., Matrix Biol., 14:715-725.


48

Metal-based drugs: State of the Art

E. Milaeva

Moscow State Lomonosov University, Lenin Hill, 1-3, Moscow, 119991, RUSSIA


The specific chemical properties of metal-based drugs impart innovative pharmacological

profiles to this type of therapeutic agents, most likely in relation to novel biomolecular mechanisms.

The development of novel metallodrugs requires clearer understanding of their physiological

processing and boimolecular mode of actions. The presentation will focus on a novel approach to

construction of organometallic/coordination compounds with physiological activity based on

computer-aided design, new synthetic approaches and extensive biological screenings. The presence

of metal atom allow extensive modification including coordination to the targeted specific groups to

control and tune toxicity-activity profiles.

The protein target-focused syhthesis of metal complexes RML (M = Fe, Mn, Co, Cu, Zn, Ni, Pt,

Ru, Sn, Au; R = pharmacophore organic group; L = end-functional group) and their physico-

chemical properties and biological activity have been studied. New synthetic design includes two

major approaches: a) Incorporation of the Ru, Au, Sn into the molecule of the well-known organic

anticancer drug. Such combination will increase affinity to cancer cells targets and as result the

compound will perform anticancer specific mode of action; (b) Modification of the known

anticancer active organometallic/coordination compound by a cytoprotective functional organic

group in order to reduce the non-specific toxicity towards the normal cells.

The multifactor antiprolifirative and antioxidative activities assay of compounds has been

performed by using DPPH, CUPRAC-tests, liposome system and enzymatic methods

(lipoxygenase, gluthathione reductase, thioredoxine reductase, tubulin); the model reactions of fatty

acids peroxidation; ex vivo lipid peroxidation in mitochondria isolated from Wistar rat brain and

liver, in vitro lipid peroxidation in rat liver homogenates. The in vivo study was performed for the

hit compounds.

The financial support of RFBR (13-03-12460-ofi_m2, 14-03-01101, 14-03-00611, 15-03-03057), RSF (14-13-00483), Program “Medicinal Chemistry” of Russian Academy of Sciences is gratefully acknowledged.

References A.A. Nazarov, S.M. Meier, O. Zava, Y.N. Nosova, E.R. Milaeva, C.G. Hartinger, P.J. Dyson. Dalton Trans., 2015, 44, 3614. E.R. Milaeva, D.B. Shpakovsky, Y.A. Gracheva, T.A. Antonenko, D.I. Osolodkin, V.A. Palyulin, P.N. Shevtsov, M.E. Neganova, D.V. Vinogradov2, E.F. Shevtsova. J. Organomet. Chem., 2015. 782, 96. D.B. Shpakovsky, C.N. Banti, E.M. Mukhatova, Yu.A. Gracheva, V.P. Osipova, N.T. Berberova, D.V. Albov, T.A. Antonenko, L.A. Aslanov, E.R. Milaeva, S.K. Hadjikakou. Dalton Trans., 2014, 43, 6880. E.R. Milaeva, D.B. Shpakovsky, Y.A. Gracheva, S.I. Orlova, V.V. Maduar, B.N. Tarasevich, N.N. Meleshonkova, L.G. Dubova, E.F. Shevtsova, Dalton Trans., 2013, 42, 19, 6817. I. Ozturk, A. Metsios, S. Filimonova-Orlova, N. Kourkoumelis, S. Hadjikakou, M. Manos, A. Tasiopoulos, S. Karkabounas, E. Milaeva, N. Hadjiliadis, Med. Chem. Res., 2012, 21, 11, 3523. D. Shpakovsky, C. Banti, G. Beaulieu-Houle, N. Kourkoumelis, M. Manoli, M. Manos, A. Tasiopoulos, S. Hadjikakou, E. Milaeva, K. Charalabopoulos, T. Bakas, I. Butler, N. Hadjiliadis, Dalton Trans., 2012, 41, 48, 14568. E. Milaeva, Curr. Topics Med. Chem., 2011, 11, 21, 2703.


49

Animal Models and Hightechnologal Phenotyping for Drug Discovery

M.P. Moshkin, S.E. Peltek, N.A. Kolchanov

The Federal Research Center “Institute Cytology and Genetics SB RAS”, Novosibirsk

Availability of laboratory animal genetic lines, modelling human diseases, as well as possibility

for suitable diagnostics of a pathological condition during the process of experimental therapy are

of key importance for effective work in the field of drug discovery. Already today, the collection of

Genetic Resources Center (GRC) for laboratory animals in the Institute of Cytology and Genetics of

Siberian branch of the Russian academy of sciences allows for development of experimental models

of almost all socially important human diseases and answers the modern requirements for research

and pre-clinical trial of novel drugs. The variety of animal models are accompanied by a wide range

of high-technology methods for phenotyping that among others includes automatic monitor of

various animal behaviors, in vivo study of morphology and physiology function by magnetic

resonance imaging, metabolome analysis on the base of non-invasive nuclear magnetic resonance

spectroscopy, luminescent bioimaging, flow cytometry, clinical biochemistry and histology

analysis. The use of GRC animal model collection and methods allowed to uncover previously

unknown preventive effects of known drugs (for example ”Reishi”), and also to begin research for

therapeutic potential of novel chemical compounds and nano-objects. As a public facility, the GRC

is open for collaboration with all scientific institutions in the field of research and testing of novel

preventive medicine and drugs for treatment of diseases.


50

Seaweed fucoidans – a platform for new drugs discovery

N.E. Nifantiev,1 N.E. Ustyuzhanina,1 M.I. Bilan,2 A.I. Usov,2 A.S. Shashkov,3 A.G. Gerbst1

Laboratories of Glycoconjugate Chemistry,1 Plant Polysaccharides2 and NMR Spectroscopy,3 N.D.

Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, E-mail: [email protected]

Natural polysaccharides fucoidans were found in many species of brown seaweeds and in some

echinoderms. Fucoidans represent an intriguing class of fucose-enriched sulfated polysaccharides,

they have been tested in different experimental models showing anti-coagulant, anti-tumor,

immunomodulatory, anti-inflammatory, and anti-complement properties. Chemical structures of

fucoidans depend primarily on used source of polysaccharides. However, a sulfated polysaccharide

fraction isolated from a given species of algae may be a mixture of structurally different polymers.

Thus the structure-activity relationship of fucoidans is still an unresolved issue.

Fucoidans often consist from selectively O-sulfated α-L-fucopyranose residues linked through

(1→2)-, (1→3)- and (1→4)-linkages with the presence of branched points that makes difficult their

detail structural characterization. Recently we have investigated the anti-inflammatory, anti-

coagulant, anti-angiogenic, and anti-adhesive activities of nine different fucoidans isolated from

Laminaria saccharina (renamed as Saccharina latissima) L. digitata, Cladosiphon okamuranus,

Fucus evanescens, F. vesiculosus, F. serratus, F. distichus, F. spiralis, and Ascophyllum nodosum

as pool samples. We found that the different profiles of their biological activities depend on

variations of structural features. Among the most active compounds studied, those extracted from L.

saccharina have been characterized by high anti-angiogenic and anti-coagulant activities in vitro as

well as their ability to block selectin-mediated inflammation in vivo.

It should be noted that the structure of fucoidans differ remarkably from human ligands involved

in the development of above listed diseases and processes, thus the pharmacophore fucoidan

fragments are the efficient mimetics of these human ligands and thus can be considered as potential

drugs. To assess structures of pharmacophore fucoidan fragments we conduct chemical

modification of fucoidans as well as systematic synthesis of corresponding short and long chain

linear and branched selectively and per-O-sulfated oligofucosides and study their conformational

characteristics and biological properties. Obtained results will be summarized in this lecture.

This work is supported by Russian Scientific Foundation (grant 14-50-00126). Selected publications: Part 1: J. Carbohydr. Chem., 19 (2000) 1151-1173; Synthesis, (2006) 4017-4031; Focus Article: Mendeleev Commun. 17 (2007) 57-62; Glycobiology, 17 (2007) 541-552; Tetrahedron Lett. 49 (2008) 5877–5879; Chapter “Synthesis of fucoidan fragments” In: "Progress in the synthesis of complex carbohydrate chains of plant and microbial polysaccharides", N.E. Nifantiev Ed., ISBN 978-81-7895-424-0; 2009, 131-154; Part 11: Carbohydr. Res. 346 (2011) 540-550; PLoS ONE 6(2): e17283. doi:10.1371/journal.pone.0017283; Mar. Drugs, 11 (2013) 2444-2458; Glycobiology 24 (2014) 1265-1274.


51

Industrial drug discovery – splendor, misery and revival

V. Patchev

Center for Clinical Studies, University Hospital Jena, Germany

07747 Jena, Salvador-Allende-Pl. 27


Evolving knowledge on the human genome generated great expectations in target-based drug

discovery. Scrutiny of the productivity and attrition rates of the pharmaceutical industry during the

past 10 years, however, has cooled down the exuberant enthusiasm and ignited a discussion on the

appropriate relationship between investment and commercial success. Indeed, several analyses have

shown that not only the projected return on investment, but also the predictions concerning the

strategic line of approach and promising indications have been missed.

Inability to transfer data obtained in-vitro and in animal models of disease into working

paradigms of clinical development has been recognized as another major flaw of industrial drug

discovery. Unwarranted reliance on mechanistic concepts of human disease and exaggeration of the

importance of individual molecular mechanisms of pathogenesis delivered heavy blows to

premature optimism that could not be overcome by the effort of translational medicine. Thus,

target-based discovery is consistently losing its aura of a fertile universal approach to the solution of

complex medical issues.

The present contribution attempts an analysis of the causes of previous failures and outlines

emerging alternative approaches (such as phenotypic discovery, drug re-positioning and open

innovation) that apparently may close the gap between medical need and drug discovery.


52

Cell and molecular models for search and study

antiviral and anticancer compounds

A.G. Pokrovsky

Department of medicine, Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova 2


Viral diseases and cancer take third position among all reason of mortality in the world. There

are more than one hundred drugs for treatment these diseases are in market; nevertheless many viral

diseases and types of cancer don’t have till effective drugs. We use different approaches for search

and study of compounds with antiviral and anticancer activity.

Traditional model for searching compound with antiviral activity include cell lines permissive

for each type of virus. We have developed a pseudovirus-cell based system for high-throughput

screening of antiretroviral compounds. Briefly, lentiviral particles are produced by a multi-plasmid

transient transfection in 293T cells and are pseudotyped with vesicular stomatitis virus G protein

mediating fusion to a wide range of target cells. Different classes of HIV and influenza virus

inhibitors can be tested using this model, either at the stage of pseudovirus production (in producer

cells) or at the infection stage (in target cells).. The system has already been tested for the classic

nucleoside reverse transcriptase inhibitors and the interfering RNA inhibitors of HIV-1 expression.

A GFP-based lentiviral vector was used for quantitative characterization of inhibition activity of

tested compounds using flow cytometry approach, however, the former can be easily replaced by a

luciferase-based vector, and the quantification stage can be performed in plate reader format. The

described system, with minor modifications, can work as a platform for study of virus entry

inhibitors for a wide range of medically important viruses. Lentiviral cores can be easily

pseudotyped, for example, with influenza virus hemagglutinin protein (HA-HIV pseudoviruses) or

with fliloviruses (Marburg and Ebola) as well as a number of other heterologic viral fusion proteins.

These pseudovirus preps can be used for the infection of permissive cell lines or primary cells

expressing the specific receptors/coreceptors for the envelope proteins used for pseudotyping, and

the compounds potentially able to inhibit the efficient virus entry (fusion inhibiting peptides,

soluble receptor.coreceptor analogs for concurrent inhibition, etc) can be tested in this model, again,

using GFP- or luciferase-based approach.

Compounds with anticancer activity were studied using different types of cancer cell lines and

subsequent determination of cytotoxicity and apoptosis induction. Molecular mechanism of

anticancer activity examine using real-time PCR, that allow easily and reliably analyze expression

of a focused panel of genes related to apoptosis.


53

Way2Drug - Web-resource for Drug Discovery

D. Filimonov,1 D. Druzhilovsky,1 A. Rudik,1 A. Lagunin,1,2 A. Zakharov,1,3

A. Dmitriev,1 T. Gloriozova, P. Pogodin,1,2 and V. Poroikov1,2

1Department for Bioinformatics, Institute of Biomedical Chemistry, 119121, Russia, Moscow,

Pogodinskaya Street, 10/8 2Department of Biochemistry, Medico-Biologic Faculty, Pirogov Russian National Research

Medical University, 117997, Russia, Moscow, Ostrovityanova Street, 1 3Chemical Biology Laboratory, NCI-Frederick, National Institutes of Health, 21702, USA,

Frederick, MD, Boyles Street, 376


Chemo- and bioinformatics today deal with a "data deluge" derived from the availability of high-

throughput screening results obtained with postgenomic technologies. Plethora of (Q)SAR and

molecular modeling methods designed to assist researcher in effective retrieval of the available data

sometimes leads to a difficult challenge: which method(s) to choose? The quality of (Q)SAR

models depends on (1) representativity of the training set; and (1) accuracy and predictivity of the

methods used for analysis of (quantitative) structure-activity relationships. Since the reasonable

quality of our computational tools PASS and GUSAR has been already demonstrated in many

internal and independent studies, we decided to create web-resource Way2Drug

(www.way2drug.com), which provide several tools for bioactivity prediction jointly with the

interface for input of novel information by the user. This additional information may be used to

improve the quality of the (Q)SAR models by re-training the predictive tools.

Currently a few predictive tools are presented at Way2Drug portal: PASSOnline predicts over

4000 kinds of biological activity; GUSAROnline predicts acute rat toxicity and interaction with

antitargets; CLC-Pred predicts cytotoxicity for tumor and normal cell-lines; DIGEP-Pred predicts

drug-induced changes of gene expression profiles; SOMP predicts sites of metabolism by five

major drug-metabolizing isoforms of P450; SMP predicts substrate/metabolite specificity; B.B.B

prediction - to calculate the passive permeability of compounds through the blood-brain barrier. We

also presented the results of bioactivity predictions for 2100 phytoconstituents from 50 Ayurvedic

medicinal plants at our resource, to provide numerous researchers with a key regarding the most

promising directions of further research of these natural compounds.

The study is partially supported by Russian Scientific Foundation grant No. 14-15-00449.


54

Chemical, Biological and Disease space complementarity in Drug Discovery

Sham Nikam

Head of Extra Value Generation Drug Discovery Unit, Pharmaceutical Research Division, Takeda Pharmaceutical Company Ltd., Fujisawa, Kanagawa 251-8555, Japan

Drug discovery process has changed significantly over last several years to address significant

attrition at different stages of Discovery and Development. Failures in Phase II, i.e., Human POC

are expensive and lead to significant losses. Major efforts are being made choose the right target

and chemical matter. Advances in informatics are helping us redefine some parameters that include

better design of chemical matter and create new repositioning hypothesis for old chemical matter.

How incremental changes over the years in chemical space have impacted biological target space

that ultimately led to new therapeutic options will be presented. Key aspects of repositioning

current or terminated chemical assets will also be mentioned.


55

Intracellular Targets and Therapeutic Efficacy of Antitumor

Drugs: A Complex Liaison

А.А. Shtil

Blokhin Cancer Center, Moscow, Russian Federation

The paradigm of the intracellular target, a critical aspect in modern strategies of antitumor drug

design, is discussed. The concept of targeted therapy postulates that the interaction of the drug with

a biomacromolecule (target) in the cell is a key event in triggering signaling cascades that lead to

cell death. Apparently, the molecules important for cell proliferation and/or survival are considered

antitumor drug targets. Accordingly, the pharmacological blockade of certain targets such as growth

factor receptors with small molecular weight inhibitors of protein kinases has proved the principle.

When the target is indeed critical for the biology and clinical manifestations of the individual tumor

type, the new generation drugs showed their efficiency (Gleevec as a remarkable example). But is

the problem ultimately solved? Should we expect that inactivation of one particular target is

necessary and sufficient for killing tumor cells that survived the selection in the course of natural

history of the disease plus multiple rounds of chemotherapy? Must we identify the targets for

efficient conventional drugs that had not been conceived to antagonize a specific target?

Based on the analysis of drug-DNA complex formation I highlight key problems such as the

uncertainty of target identification (due to binding of drugs to a variety of biomacromolecules), the

multiplicity of targets and the complexity of death signaling pathways. Nevertheless, the idea of

preferred (that is, the most affine) targeting by the drug (specifically, the structure of the drug-target

complexes) is considered a major valuable tool for proceeding from an empirical screening of

chemical libraries to rational drug design. Thus, the concept of the drug target is indispensable for

elucidating molecular mechanisms of therapeutic efficacy of drug candidates.


56

Human Photosynthesis or the Unexpected Capacity of Melanin Molecule to

Dissociate the Water Molecule: Implications as Alternate and Successful

Treatment Strategies for the Neurodegenerative Disease

A. Solís Herrera,1 M. del Carmen Arias Esparza,1

P. E. Solís Arias1 and G. Aliev2,3

1Human Photosynthesis Study Center, R & D & I Department, López Velarde 108, Centro, Aguascalientes, Aguascalientes, C.P. 20000, México

2GALLY” International Biomedical Research Institute Inc., San Antonio, TX, USA 3Department of Health Science and Healthcare Administration, University of Atlanta, Atlanta, GA

USA


Cellular bioenergetic pathways have altered in complex ways thereby making it difficult to be

able to generate and use energy from the oxidation of glucose. This is the exact spot where

mitochondria play an important role through oxidative phosphorylation [1.3]. Currently, cell

biology is based upon glucose being used as a main source of energy. During a descriptive study

about the three leading causes of blindness in the world, the ability of melanin to transform light

energy into chemical energy through the dissociation of water molecule was unraveled. Initially,

during 2 or 3 years; we tried to link together our findings with the widely accepted metabolic

pathways already described in metabolic pathway databases, which have been developed to collect

and organize the current knowledge on metabolism scattered across a multitude of scientific

articles. However, the literature on cellular metabolism is extensive but rarely conclusive evidence

is available, and also it is important to state that one would expect these databases to contain largely

the same information, but the contrary is true. Furthermore, in a single well-known metabolic

process as the tricarboxylic acid (TCA) or Krebs Cycle; which is found in nearly every biology and

chemistry curriculum, described in 1937; even for this apparently well-studied process there is

considerable disagreement between at least five databases. In brief, of the nearly 7000 reactions the

databases jointly contain only 199 are described in the same way to all five databases. Thereby try

to integrate chemical energy from melanin with the supposedly well-known bioenergetic pathways

is easier said than done; and the lack of consensus about metabolic network constitutes an

insurmountable barrier. Finally, after years of unsuccessful results, we realized that the chemical

energy that is released through the dissociation of water molecule by melanin represents just over

90% of cell energy requirements, because the cell utilizes this energy in many various ways; in as

much so that both glucose and ATP have biological functions related only to biomass and not so

much with energy. Our finding about the unexpected intrinsic property of melanin to transform


57

photon energy into chemical energy through the dissociation of water molecule, a role performed

supposedly only by chlorophyll in plants, questions seriously the sacrosanct role of glucose and

thereby mitochondria as an energy and power source of the cell [1,2].

The extraordinary ability of the eukaryotic cell to capture light energy and transform it into

chemical energy through the dissociation of the water molecule, as it occurs in plants, was

identified incidentally in the human retina during an investigation that began in 1990 about of the

three leading causes of blindness in the world: Glaucoma, Diabetes and Macular Degeneration

related to age [1.3].

The reason for the research was that are the same causes of blindness since the middle of last

century, and since the incidence and prevalence is the same, it is conceivable that the treatments are

not working.

The ability to perform photosynthesis begins to decline at age 26 at the rate of 10% every decade

going into free fall after the 50s. But it also decreases with the cold, as in plants with pesticides,

herbicides, contaminated water, fertilizers, drugs with a high apparent volume of distribution,

trauma, anesthetic agents, metals, etc [1-3].

Macular degeneration related to age (AMD) is considered from the 80s as the equivalent of

Alzheimer disease (AD) in the eye and booth illnesses can be explained in terms of energy, thereby

the pharmacological modulation of human photosynthesis by using Melanin Precursor (“QIAPI-1”)

opened a completely new and unexpected powerful way that has allowed us to significantly

improve therapeutics outcome during the course of different neurodegenerative diseases such as

AD, Parkinson's, Huntington's, Macular degeneration, and cerebral palsy. Therefore we are

confident that the future investigations addressing the application of Melanin Precursors based

treatment protocols in a large scale clinical practices are warranted.

This study was supported by Human Photosynthesis Study Center, Aguascalientes, México and “GALLY” International Biomed. Res. Consulting LLC, San Antonio, TX, USA.

References: [1]. Solís Herrera A., Arias Esparza M. del. C., Ashraf G.M., Zamyatnin Jr. A.A. and G. Aliev. Beyond mitochondria as

the energy source of the cell? Central Nervous System Agents in Medicinal Chemistry (CNSA-MC), Central Nervous System Agents in Medicinal Chemistry, 2015, 15, 32-41 [Feb 2., Epub ahead of print].

[2]. Aliev G., Ashraf G.Md., Kaminsky Y.G., Sheikh I.A., Sudakov S. K., Yakhno N.N., Benberin V.V. and Bachurin S.O. Implication of the Nutritional and Non-Nutritional Factor in the Context of the Preservation of The Cognitive Performance in Demented/Depressed and Alzheimer Patients. Am J Alzheimers Dis Other Demen. 2013, 28(7): 660-670. first published on October 1, 2013 as doi:10.1177/1533317513504614

[3]. Aliev G., Solis-Herrera A., Li Y Kaminsky Y.G., Yakhno N.N., Nikolenko V.N., Zamyatnin Jr. A.A., Benberin V.V., and S.O. Bachurin. Human Photosynthesis, the Ultimate Answer to the Long Term Mystery of Kleiber’s Law or E=M ¾: Implication in the Context of Gerontology and Neurodegenerative Diseases. Open Journal of Psychiatry, 2013, 3, 408-421. http://dx.doi.org/10.4236/ojpsych.2013.34045.


58

Recent Studies on Bioactive Natural Products from Marine Organisms

V.A. Stonik

Pacific Institute of Bioorganic Chemistry of Far-Eastern Branch of the Russian Academy of Sciences, 690022, Vladivostok-22, Russia


During several recent years, our studies on marine organisms from the Pacific Ocean waters and

their natural products were happily developing in the both directions: i) search for producers of new

natural products followed by the isolation of the corresponding compounds and ii ) investigation of

bioactivities of the isolated substances.

Marine organisms, especially sponges, echinoderms and other invertebrates are characterized by

alkaloids, steroids, different lipids with unique structural features that are products of unusual

biosyntheses. As result of our recent studies on marine organisms collected from the North-west

Pacific areas and from Vietnamese waters about 100 new polyhydroxylated sterols, steroid and

triterpene glycosides, different alkaloids, cerebrosides, terpenoids were isolated and structurally

elucidated. New marine natural compounds were proved to be potent immunomodulators, inductors

of apoptosis and autophagy of tumor cells, anti-inflammatory agents, and activators of

mitochondrial functions in cardiomyocytes. Molecular mechanisms of their actions were studied

jointly with scientists from the Republic of Korea and Germany.

Some recent publications: [1]. Dyshlovoy S. A., Hauschild J., Amann K., Tabakmakher K. M., Venz S., Walther R., Guzii A. G., Makarieva T.

N., Shubina L. K., Fedorov S. N., Stonik V. A., Bokemeyer C., Balabanov S., Honecker F., Amsberg G. Marine alkaloid Monanchocidin А overcomes drug resistance by induction of autophagy and lysosomal membrane permeabilization. Oncotarget 2015, 13, 1–14.

[2]. Dyshlovoy S. A., Venz S., Shubina L. K., Fedorov S. N., Walther R., Jacobsen C., Stonik V. A., Bokemeyer C., Balabanov S., Honecker F. Activity of aaptamine and two derivatives, demethyloxyaaptamine and isoaaptamine, in cisplatin-resistant germ cell cancer. Journal of Proteomics, 2014, 96, 223−239.

[3]. Jeong S. H., Kim H. K., Song I.-S., Lee S. J., Ko K. S., Rhee B. D., Kim N., Mishchenko N. P., Fedoryev S. A., Stonik V. A., Han J. Echinochrome A protects mitochondrial function in cardiomyocytes against cardiotoxic drugs. Marine Drugs, 2014, 12, 2922−2936.

[4]. Lyakhova E. G., Kolesnikova S. A., Kalinovsky A. I., Dmitrenok P. S., Nam N. H., Stonik V. A. Further study on Penares sp. from Vietnamese waters: minor lanostane and nor-lanostane triterpenes . Steroids 2015, 96, 37–43.

[5]. Popov R. S., Ivanchina N. V., Kicha A. A., Malyarenko T. V., Dmitrenok P. S., Stonik V. A. Metabolite profiling of polar steroid constituents in the Far Eastern starfish Aphelasterias japonica using LC−ESI MS/MS. Metabolomics, 2014, 10, 1152−1168.

[6]. Santalova E. A., Denisenko V. A., Dmitrenok P. S., Drozdov A. L., Stonik V. A. Cerebrosides from a Far Eastern glass sponge Aulosaccus sp. Lipids 2015, 50, 57–69.

[7]. Stonik V. A., Fedorov S. N. Marine low molecular weight natural products as potential cancer preventive compounds. Marine Drugs, 2014, 12, 636−671.


59

Search for socially important drugs in Favorsky Irkutsk institute of chemistry

B.A. Trofimov, A.V. Ivanov

A.E. Favorsky Irkutsk Institute of chemistry, 664036, Russia, Irkutsk, Favorsky Str., 1


Search for socially important drugs was and remains a cardinal point of basic and applied

investigations of A.E. Favorsky Irkutsk institute of chemistry. Among the innovative drugs

launched by the institute are:

1. Antituberculosis medicine of new generation, trade name Perchlozone (jointly developed with

“Pharmasyntez” company and St.-Petersburg research institute of phthisiopulmonology), registered

by 09.11.2012, № ЛП-001899, commercially produced from 2013.

2. The efficient antidote of carbon monoxide, trade name Acyzole (jointly developed with

“Aсyzole Pharma”, Moscow) registered by 23.12.2005, № ЛС-001061, commercially produced

from 2005.

Both drugs are included in State List of Vital and Essential Medicines.

Close to the drug shelf is original antiatherosclerosis agent Agsular that possesses high

anticoagulant and hypolipodemic activity, comparable with import drugs like Vessel Due F®,

Zokor® and Liprimar®.

Under clinical trials is the active heamopoiesis stimulator Cobazole.

The super antiseptics Anavidine and Anavidine M, the institute invention, are now industrially

manufactured.

An efficient natural antioxidant Dihydroquercetin (from Siberian larch), first isolated in the

institute, is now produced on bench scale.

A widespread application as drug delivery and membranotropic agent is now getting

polysaccharide Arabinogalactan, also bench-scale produced, patented by the institute.

The institute is also leading in the field of medicinal nanobiocomposites which are intended

to be used as cytostatics, nonnarcotic analgetics, antianemia drugs, magnet-controlled and delivery

medicines.

The long-term projects for search of anti-HIV and antituberculosis drugs based on original

synthetic methodologies are being implemented in the institute (jointly with “Pharmasyntez”,

“Vector” and other companies and institutions).


60

Therapeutic Nucleic Acids

V. Vlassov, M. Zenkova, D. Pyshnii, D. Stetsenko

Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of

Sciences, 8, Lavrentiev Avenue, Novosibirsk 630090, Russia E-mail: [email protected]

Identification of genetic programs and proteins responsible for development of diseases,

discovery of mechanisms of gene expression regulation by RNAs and elaboration of methods of

nucleic acid synthesis opened possibilities for development of new generation of nucleic acid

therapeutics, “smart drugs” capable of specific silencing of unwanted genes expression, correction

of mutated genetic programs and modulation of functions of specific proteins.

Antisense oligonucleotides and siRNAs can arrest expression of target genes in mammalian cells

and several oligonucleotide therapeutics have been approved for use in clinics. Design of improved

versions of modified oligonucleotide conjugates capable of entering into bacterial cells and into

specific mammalian cells will open possibilities for development of a broad range of therapeutics

for treatment of different diseases caused by inappropriate functions of cellular genetic programs,

such as cancer, and diseases caused by infectious agents, having their own unique genetic programs.

Aptamers, capable of binding to specific proteins, proved to be potential therapeutics, capable of

interfering with functions of the target proteins in vivo. These compounds are easy to design and

manufacture, and further development of modified versions of aptamers will provide compounds

competitive with antibodies in different biomedical applications.

Recent development of the CRISPR – Cas9 system for manipulating DNA opened possibilities

for correction of genetic programs in cells in culture and in vivo. This system and stem cell

technologies provide a broad range of potential applications in therapy, such as correction of

genetic defects, introduction of new programs in the cell genome and inactivation of viral genomes

inserted in cellular genetic programs.

Success in the field of design of nucleic acid based therapeutics will be determined by

development of chemistry of nucleic acids, needed for synthesis of appropriate oligonucleotide

analogs and by development of techniques providing possibilities for efficient targeting specific

cells and deliver nucleic acid therapeutics into cells, to their targets.


61

Molecular Design of Bivalent or Dual Action Drugs

N.S. Zefirov, V.A. Palyulin, O.N. Zefirova

Department of Chemistry, M. V. Lomonosov Moscow State University, Moscow 119991, Russia

Institute of Physiologically Active Compounds RAS, Chernogolovka, Moscow Region, 142432, Russia

E-mail: [email protected];

Design of new drugs is a sophisticated process involving the application of molecular modeling

and QSAR techniques, molecular docking and virtual screening. The joint application of these

approaches provides deeper understanding of both ligand-receptor interactions and structure-

activity relationships as well as facilitates lead finding and further optimization.

For the above discussed design the molecular models of all closed and open forms of ionotropic

(NMDA and AMPA) and metabotropic (mGluR1-8) glutamate receptors, adenosine, melatonin,

GABAA and GABAC receptors and tubulin have been either built or refined. The docking of known

agonists, antagonists, modulators, and channel blockers into the models was used to reveal binding

modes of ligands and to explain known structure-activity relationships.

Dual action of a drug can be sub-classified into its action on two different biotargets and action

on two different sites of the same biotarget. The dual drug approach will be illustrated using (a) dual

action conjugate of colchicine with a “simplified” taxol analogue interacting with tubulin and (b)

the bivalent positive modulator of AMPA receptor. Indeed, this approach was successfully used for

synthesis of dual action compounds active against human lung carcinoma А549 cell line as well as

in the design of new neuroprotectors with cognition enhancing properties (extraordinary high

potency of some designed compounds starting from picomolar concentration had been revealed –

absolute record among all currently known positive AMPA receptor modulators).

References [1] N.S.Zefirov, V.A. Palyulin. Molecular Modelling of Central Nervous System Receptors. Mendeleev Commun. 2010, 20, 243-248. [2] O.N. Zefirova, E.V. Nurieva, H. Lemcke, A.A. Ivanov, D.V. Shishov, D.G. Weiss, S.A. Kuznetsov, N.S. Zefirov. Design, synthesis and bioactivity of putative tubulin ligands with adamantane core. Bioorg. Med. Chem. Lett. 2008, 18, 5091–5094.


62


63

The Synthesis of Expected Epoxy-metabolites of

(1R,2R,6S)-3-Methyl-6-(prop-1-en-2-yl)cyclohex-3-ene-1,2-diol

O. Ardashov, A. Pavlova, A. Genaev, G. Salnikov, D. Korchagina, I. Il’ina, K. Volcho,

T. Tolstikova, N. Salakhutdinov

N.N. Vorozhtsov Novosibirsk institute of organic chemistry, 630090, Russia, Novosibirsk,

Lavrentiev av., 9


Recently [1] we found that (1R,2R,6S)-3-methyl-6-(prop-1-en-2-yl)cyclohex-3-ene-1,2-diol 1

displayed a potent antiparkinsonian activity and possessed low acute toxicity.

In order to study compound 1 pharmacokinetics it is important to synthesize its metabolites. In

this work approaches to synthesis of four expected metabolites – all possible epoxides 2 - 5 were

developed. The structure of epoxide 4 was determined by using isomerisation of its diacetate

derivative to ortoether 6 with rigid framework, NOESY-NMR technique and DFT-calculation.

OH

OH

1

OH

OH

2

OH

OH

3

OH

OH

4

OH

OH

5

O

OAc

6

O O

O O

H H

O

OH

The investigation of antiparkinsonian activity of all the epoxides was carried out and data

obtained will be presented.

References [1] O.V. Ardashov, A.V. Pavlova, I.V. Il’ina, E.A. Morozova, D.V. Korchagina, E.V. Karpova, K.P. Volcho, T.G. Tolstikova, N.F. Salakhutdinov, J. Med. Chem. 2011, 54, 3866–3874.


64

Astrocytes as Targets for Neuroprotection from Brain Injury

G.E. Barreto,1 G. Aliev2,3

1Departamento de Nutrición y Bioquímica, Facultad de Ciencias, Pontificia Universidad Javeriana,

Bogotá D.C., Colombia. Email. [email protected] 2 GALLY Biomedical Research Consulting LLC., 7733 Louis Pasteur Drive, #330, San Antonio, TX,

78229, USA. 3School of Health Science and Healthcare Administration, University of Atlanta, E. Johns Crossing,

#175, Johns Creek, GA, USA 30097. Email: [email protected]

Astrocytes are the most numerous cell types in the brain and also perform many duties that are

essential for continuous neurological functioning. They also act as providers for stabilized activity

of neurons within the brain. There is strong experimental evidence that astrocytes protect neurons

against traumatic brain-induced injury. While astrocytes are generally considered more resistant to

damage than neurons are to such disturbances, few studies offer insight into how changes of

astrocytic support functions may lead to loss of protection. Mitochondria are one of the primary

targets of injury in astrocytes. We hypothesize that brain-induced mitochondrial damage in

astrocytes can impair astrocyte function and contribute to neuronal injury. In this regard, better

protecting astrocytic functions may represent a potential strategy against brain neuro-degeneration.

Results from our lab show that astrocytes treated with both mitogens (PDGF) and conditioned

medium isolated from adipose tissue-derived mesenchymal stem cells protect in an in vitro model

of Parkinson’s Disease and scratch assay injury. Furthermore, these effects were accompanied by a

reduction in reactive oxygen species production and important changes in cell morphology.

Improving astrocytes function by hyper expression of chaperones and/or dampening their reactive

state might be considered such a strategy as well. In conclusion, our findings indicate that focusing

particularly on the two-sided role of reactive astrocytes is an important experimental paradigm

helpful in eradicating damage from protective mechanisms after brain injury.

Keywords: Astrocytes, mitochondria, brain injury, mesenchymal stem cells, growth factors, neuroprotection. References [1]. Cabezas R, Avila-Rodriguez M, Vega-Vela NE, Echeverria V, Gonzalez J, Hidalgo OA, Santos AB, Aliev G,

Barreto GE. Growth Factors and Astrocytic Metabolism: Possible Roles for Platelet Derived Growth Factor. Medicinal Chemistry, 2015, In Press.

[2]. Avila-Rodriguez M, Garcia-Segura LM, Cabezas R, Torrente D, Capani F, Gonzalez J, Barreto GE. Tibolone protects T98G cells from glucose deprivation. J Steroid Biochem Mol Biol., 2015, In Press.

[3]. Cabezas R, Avila MF, Gonzalez J, El-Bachá RS, Barreto GE. PDGF-BB Protects Mitochondria from Rotenone in T98G cells. Neurotoxicity Research, 2015, In Press.

[4]. Torrente D, Avila MF, Cabezas R, Morales L, Gonzalez J, Barreto GE. Paracrine factors of human Mesenchymal Stem cells increase wound closure and reduce reactive oxygen species production in a traumatic brain injury in vitro model. Hum Exp Toxicol., 2014.


65

[5]. Barreto GE, White RE, Xu L, Palm CJ, Giffard RG. Effects of heat shock protein 72 (Hsp72) on evolution of astrocyte activation following stroke in the mouse. Exp Neurol. 238(2):284-296, 2012.

[6]. Adelson JD, Barreto GE , Xu L, Ouyang I, Naserke T, Djurisic M, Xiong X, Shatz CJ, Giffard RG. Neuroprotection from stroke in the absence of MHCI or PirB. Neuron, 73(6):1100-7, 2012.


66

New Derivatives of (-)-Cytisune - Selective Inhibitors of COX-2: in silico

Screening with in vivo Verification

S.S. Borisevich, I.P. Tsypysheva, N.S. Makara, S.L. Khursan

Ufa Institute of Chemistry of Russian Academy of Sciences, 71, Pr. Octyabrya, Ufa, 450054,

Russian Federation


Some derivatives of the quinolizidine alkaloid (−)-cytisine known for the high

neuropharmacological activity, possess the antiinflammatory properties [1]. This data is the basis

for the focused search of perspective antiinflammatory agents among its new derivatives. So, in

silico screening of inhibition of COX-1 and COX-2 by derivatives of 12-N-methylcytisine (А2),

presented in a figure 1 is carried out Schrodinger Suite 2014-4.

Figure 1 – The library of the derivatives of 12-N-methylcytisine (А2)

Five structures for carrying out in vivo screening are chosen on the basis of the docking results.

In vivo screening of antiinflammatory activity

(the model of the carrageenan-induced raw edema

in the rats [2]) has shown that all choosen

compounds А2, А168, А169, А179 and А182,

are effective at the second phase of an

inflammation induced COX-2. And activity of

these compounds is comparable or surpasses the

Voltaren activity. Thus, on the basis of the in

silico screening and in vivo verification we can draw a conclusion, that the leading compound А179

is the selective inhibitor of COX-2 which locating in the active site is presented in the figure 2.

This study was supported by the Russian Foundation for Basic Research (project No. 14-04-97035-povolz’e-a)

References [1] E. G. Pérez, C. Méndez-Gálvez, B. K. Cassels, Nat. Prod. Rep., 2012, 29, 555. [2] R. Vinegar, W. Schreiber, R. Hugo, J. Pharm. Exper. Ther., 1969, 168, 96.

Figure 2 – A179 in the active site of COX-2 активном сайте ЦОГ-2


67

Alkoxyamines for the in-situ Generation of Radicals as Theranostic Agents

P. Brémond, G. Audran, S.R.A. Marque

Institute of Radical Chemistry (UMR CNRS 7273), Aix-Marseille Université

13397 Marseille Cedex 20, France


Alkoxyamines R1R2NO––R3 are able to undergo homolysis upon chemical activation to release a

stable nitroxide R1R2NO•, which can be used for Overhauser-enhanced MRI, and a transient alkyl

radical R3•, which can be used for killing tumor cells. By combining diagnostic and therapeutic

activities into a single low-molecular

weight molecule, alkoxyamines are

new and promising theranostic tools.

We have developed this concept

recently, and we also proved that

they are reliable and controllable

sources of radicals generated in-situ

and able to exhibit interesting

biological and imaging properties.

[1-3]

References [1]. "Alkoxyamines: First Steps Towards a New Family of Theranostic Agents Against Cancer" Moncelet, D.; Voisin,

P.; Bouchaud, V.; Massot, P.; Parzy, E.; Audran, G.; Franconi, J.-M.; Marque, S. R. A.; Brémond, P.; Mellet, P. Mol. Pharmaceutics 2014, 11, 2412-2419.

[2]. "Alkoxyamines: Past, Present and Future" Audran, G.; Brémond, P.; Marque, S. R. A. Chem. Commun. 2014, 50, 7921-7928.

[3]. "Alkoxyamines: a New Family of Pro-Drugs against Cancer. Concept for Theranostics." Audran, G.; Brémond, P.; Franconi, J.-M.; Marque, S. R. A.; Massot, P.; Mellet, P.; Parzy, E.; Thiaudière, T., Org. Biomol. Chem. 2014, 12, 719-723.

enzymaticactivation

R2

R3

R1 O N

homolysis

NOR1

R3

R2

+

highly labilealkoxyamine

nitroxidealkyl

radical

DIAGNOSTICTHERAPEUTIC

THERANOSTIC

R2

R3

R1 O N

highly stable alkoxyamine

peptide


68

New Effective and Selective Carboxylesterase Inhibitors Based on

Polyfluoroalkylated 2-arylhydrazono-3-oxo Esters

Ya.V. Burgart,1 G.F. Makhaeva,2 N.P. Boltneva,2 N.V. Kovaleva,2 S.V. Lushchekina,2,3

E.V. Shchegol’kov,1 V.I. Saloutin,1 R.J. Richardson,4 O.N. Chupakhin1

1 I.Ya. Postovsky Institute of Organic Synthesis UB RAS, 620990, Russia, Yekaterinburg,

S. Kovalevskoy Str., 22. 2Institute of Physiologically Active Compounds RAS, 142432, Russia, Chernogolvka, Severny, 1 3N.M. Emanuel Institute of Biochemical Physics RAS, 119334, Russia, Moscow, Kosygina Str., 4

4Toxicology Program, University of Michigan, 48109, USA, Michigan, Ann Arbor E-mail: [email protected]

Carboxylesterases (EC 3.1.1.1, CaE) are key enzymes of hydrolytic metabolism for major therapeutic agents that contain ester, carbamate or amide groups. These are structurally diverse drugs from different pharmacological groups: e.g., antiplatelet agents; angiotensin-converting enzyme inhibitors; statins; central nervous system agents; antiviral agents; and anticancer drugs. The use of CaE inhibitors as co-drugs can considerably improve the therapeutic efficacy of known drugs and reduce their side effects. The aim of our investigation is to search for new effective and selective CaE inhibitors among the polyfluorinated alkyl 2-arylhydrazono-3-oxo esters. We have synthesized a series of compounds 1-6 and their non-fluorinated analog 7:

O

R1

O

ON

NHR2

R3

1 (R1 = CF3, R2 = Et, R3 = Н), 5 (R1 = CF2H, R2 = Me, R3 = 4-Me),

2 (R1 = CF3, R2 = Et, R3 = 4-OMe), 6 (R1 = C3F7, R

2 = Me, R3 = 4-OMe), 3 (R1 = CF3, R

2 = Et, R3 = 4-Me), 7 (R1 = Me, R2 = Et, R3 = Me). 4 (R1 = CF3, R

2 = Et, R3 = 2-NO2),

Inhibitory activity of compounds 1-7 against porcine liver CaE and structurally related enzymes,

e.g., human erythrocyte acetylcholinesterase (EC 3.1.1.7, AChE,) and horse serum butyrylcholinesterase (EC 3.1.1.8, BChE) was studied. Esterase profile analysis of compounds 1-6 showed that the CF3-containing ethyl esters 1-4 had the maximum activity against CaE in the nanomolar range (IC50 = 5–13 nM). Their inhibitory activity against BChE and AChE was in the micro- and millimolar range, i.e., compounds 1-4 were highly selective CaE inhibitors. Their low anti-AChE activity suggests low acute cholinergic toxicity. The increase in the length of polyfluoroalkylated substituent (6) and the replacement of trifluoromethyl by difluoromethyl radical (5) led to a significant reduction in anti-CaE activity. The non-fluorinated analog 7 had no inhibitory activity against all esterases. Molecular docking using AutoDock 4.2.6 showed effective binding of these compounds to the CaE active site. Thus, highly effective and selective inhibitors of CaE have been found in the series of polyfluorinated alkyl 2-arylhydrazono-3-oxo esters suggesting their use for modulating the biotransformation of ester-containing drugs.

This work was financially supported by Program 9 of DCMS RAS “Medicinal Chemistry” and Program of UB RAS (Project 15-21-3-5). The work of Lushchekina S.V. was supported by scholarship of the Foundation "Dynasty".


69

Discovery of ZB40-0016 – a Novel GPR119 Agonist for

Type 2 Diabetes Treatment

M.A. Chafeev, R.N. Karapetian, I.N. Tyurenkov, E.V. Volotova

Chemical Diversity Research Institute, Khimki, Russian Federation

In recent years the G protein-coupled receptor 119 (GPR119) has received significant attention

from the pharmaceutical industry. The activation of GPR119 increases the intracellular

accumulation of cAMP, leading to enhanced glucose-dependent insulin secretion from pancreatic β-

cells and increased release of the gut peptides GLP-1 (glucagonlike peptide 1), GIP (glucose-

dependent insulinotropic peptide) and PYY (polypeptide YY). Due to its mechanism of action

GPR119 agonists are promising new therapeutic approaches for diabetes and related diseases.

Several companies have GPR119 agonists in preclinical and clinical studies for type 2 diabetes.

In the presented work synthesis and biology activity of a novel GPR119 agonist ZB40-0016 are

described. Initial HTS resulted in a promising series of small molecules with a structurally novel

cyclic sulfone fragment with IC50 for several compounds in 0.4-2.0 uM range. After the lead

optimization process a new orally-active GPR119 agonist ZB40-0016 was discovered. Significant

structural changes were made in order to enhance its potency from several hundred nanomol to

single digit nanomol range and to improve microsomal stability.

Cl

NH

O

SO O

NO

S

N

N

Cl N

O

F O

O

O

С530-0315 - initial HTS hit ZB40-0016 IC50 405 nM IC50 7 nM

The synthetic pathway for ZB40-0016 includes parallel preparation of two building blocks with

consequent coupling. OHOH TfO O

S

OO

Nboc

OH

Nboc

O

F

BO

O

S

N

N

Cl N

O

F O

O

O

S

N

O

F O

O

O

boc

ZB40-0016 ZB40-0016 demonstrates activity in-vivo in several models of diabetes and has low toxicity.

Currently ZB40-0016 is considered as a clinical candidate for Phase one study.


70

Recyclizations of Pyrimidines as a Nontraditional Way of Pharmacophore and

Biogenic Groups Introduction to the Molecule of a Biologically

Active Compound

G. Danagulyan 1,2

1Russian-Armenian (Slavonic) State University, 0051, Armenia, Yerevan, H. Emin St., 123

2Scientific-Technological Center of Organic and Pharmaceutical Chemistry of the National Academy of Sciences of the Republic of Armenia, 0014, Yerevan, Azatutyan Ave, 26


It is known that the reactions used in medical chemistry, as a rule, are based on well-proven and

simple transformations, such as the reactions of substitution, condensation, cyclization.

The suggested presentation focuses on rearrangements of pyrimidinium iodide under the

influence of different nitrogen-containing nucleophilic reagents proceeding through heterocycle

recyclization. The described transformations result in derivatives of pyridine, 1,2,4-triazole and

fused systems containing biogenic and pharmacophoe fragments of the initial amine reagent.

The presented results prove the possibility of using nontraditional transformations, including the

reactions proceeding due to recyclizations, in medical chemistry. At the same time knowledge of

pyrimidine ring recyclizations is of great practical significance, in particular, for prediction of

schemes of possible directions of metabolism of the used medications of pyrimidine series in the

body.

The work has been accomplished within the framework of the joint Russian-Armenian

grant number 13RF-087 of the Committee on Science of the Ministry of Education and Science of

the Republic of Armenia and Russian Foundation for Basic Research (RFBR) as well as with

financial support from the Committee on Science of Armenia (theme 13-1D334).

The author thanks academician RАS, Prof. Oleg N. Chupakhin for support and collaboration.


71

Natural Products in Matrix Assisted Supramolecular Chirality Amplification

P.B. Drašar

Department of Chemistry of Natural Products, University of Chemistry and Technology, Prague,

CZ 166 28, Czech Republic, Praha 6 - Dejvice, Technická, 5


The amplification of supramolecular chirality in different solvent mixtures (the “Sergeants and

Soldiers” Principle [1]) caused by dynamic non-covalent bonding assemblies formation [2] has been

studied on supramolecular synthons of achiral porphyrin core with chiral substituents (sugars,

steroids) in meso positions.

Initial observation, that some porphyrin derivatives with chiral meso substituents exhibit very

high and time dependent specific optical rotation [3], resulted in synthesis of a series of compounds

of the above mentioned type and deeper study of the phenomenon.

In some cases just a different chirality on one carbon in the sugar substituent (e.g. D-gluco → D-

galacto) totally reversed the CD Cotton effect of substituted porphyrin, and CD spectra were

moreover strongly dependent on the polarity and ionic strength of the solvent matrix [4]. The chiral

moieties were recently extrapolated to multisubstituted, rather complicated brassinolides and

ecdysones [5]. The study of formation of chiral superassemblies was supported by computer

modeling.

We believe that the above mentioned synthetic constructs form the information-rich

superassemblies that have the potential in helping to understand biological processes and drug

delivery and bioavailability. There are being studied semisynthetic supramolecular synthons that

contain several types of chiral natural products, in general.

The work was done in collaboration with the Institute of Bioorganic Chemistry in Minsk and Universities Tor Vergata and La Sapienza in Rome.

References [1] M. M. Green, M. P. Rediy, R. J. Johnson, G. Darling, D. J. O’Leary, and G. Willson, J. Am. Chem. Soc. 1989, 111,

6452–6454. [2] L. J. Prins, P. Timmerman, and D. N. Reinhoudt, J. Am. Chem. Soc. 2001, 123, 10153–10163. [3] P. Štěpánek, M. Dukh, D. Šaman, J. Moravcová, L. Kniežo, D. Monti, M. Venanzi, G. Mancini, and P Drašar, Org.

Biomol. Chem. 2007, 5, 960–970. [4] D. Monti, M. Venanzi, E. Gatto, G. Mancini, A. Sorrenti, P. Štěpánek, and P. Drašar, New. J. Chem. 2008, 32,

2127–2133. [5] H. A. Zhylitskaya, V. N. Zhabinskii, R. P. Litvinovskaya, R. Lettieri, D. Monti, M. Venanzi, V. A. Khripach, and P.

Drašar, Steroids 2012, 77, 1169–1175.


72

Mechanochemical Approaches to Creation of Drug Delivery Systems

A.V. Dushkin,1 T.G. Tolstikova,2 M.I. Khvostov,2 N.E. Polyakov,3 S.S. Khalikov,4 K.V. Gaidul5

1 Institute of Solid State Chemistry SB RAS, 630128, Russian Federation, Novosibirsk, Kutateladze, 18

2N.N.Vorozhtsov Novosibirsk Institute of Organic Chemistry SB RAS, 630090, Russian Federation, Novosibirsk, Lavrentieva pr., 10

3 Institute of Chemical Kinetics and Combustion SB RAS, 630090, Russian Federation, Novosibirsk, Institutskaya, 3

4 A.N.Nesmeyanov Institute of Organoelement Compounds of Russian Academy of Sciences, 119991, Russian Federation, Moscow, Vavilova str., 28

5 Institute of Clinical Immunology SB RAMS, 630099, Russian Federation, Novosibirsk, Yadrintsevskaya str., 14


Pharmacological and physicochemical properties of mechanochemically synthesized

supramolecular systems were investigated in the Institutes of RAS and RAMS. Actually such type

of systems appears as the "guest-host" complexes, where the "guest" is a drug molecule and the

"host" is a carrier particle – usually macromolecule of polysaccharides, micelle of glycosides, silica

particles, etc. The increasing of pharmacological effect by using of such structures might be

achieved through improved delivery of drug molecules to the active centers of the cell receptors. It

was demonstrated that plant metabolites - glycyrrhizic acid and polysaccharide arabinogalactan, as

well as nano-structured colloidal silica are very effective “carriers”.

To obtain such kind of solid dispersions of drugs and auxiliary substances the original solid state

mechanochemical technology was developed. The supramolecular complexes are formed by solid-

phase synthesis or by dissolving the obtained solid dispersion in aqueous media. The studies of

several tens of widely used medicinal agents of different pharmacological classes show that the

inclusion of drug molecules in these supramolecular systems can significantly improve the

efficiency and safety of their actions.

References

[1]. A.V. Dushkin, T.G. Tolstikova, M.V. Khvostov, G.A. Tolstikov, Complexes of polysaccharides and glycyrrhizic

acid with drug molecules. Mechanochemical synthesis and pharmacological activity // in book «The Complex World of

Polysaccharides», ed. by Dr. D.N. Karunaratne. - Publisher: InTech. 2012, p. 573-602.


73

"Fucolam" - the First in Russia Food Supplement Based on Fucoidan

S. Ermakova,1 T. Zaporozhets,2 M. Kusaykin,1 N. Besednova,2 T. Zvyagintseva1

1 G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian

Academy of Sciences, Vladivostok, Russia 2 G.P. Somov Research Institute of Epidemiology and Microbiology, Siberian Branch of Russian

Academy of Medical Sciences


Marine algae are ancient photosynthetic organisms that constitute the largest group in the plant kingdom. They are used for functional food, cosmetic additives, supplements productions, and in traditional medicine due to taste, prophylactic and therapeutic effects. Algae contain microelements and iodine-containing organic compounds, as well as vitamins, mannitol more than terrestrial plants. Polysaccharides of algae are especially valuable substances. Some of them (for example agarose, carrageenans, and alginates) have found widespread application. At the moment polysaccharides synthesized by brown algae (laminarans and especially fucoidans) are of greatest interest. The general term "fucoidan" is used to integrate the molecules differenced in composition, structure, and in degree of sulfation, acetylation, etc. Analogs of these polysaccharides have not been found on the land till now. Fucoidans long since attracted attention due to diverse biological activity, low toxicity, and plant origin [1, 2]. It is important because of contamination and side effects of the preparation produced from animals (for example, heparin). A large number of publications devoted to the study of antitumor, anticoagulant, antimutagenic activities, and immunostimulatory, antiinfective and antioxidant properties of these polysaccharides. However, despite the obvious prospects for exploitation in medicine, none of fucoidan is declared yet as a drug. The reason is that the structural diversity of fucoidans is extremely large. Structural investigation of fucoidans is of great difficulties because of variety of monosaccharide composition, different types of glycosidic linkages, presence of large numbers of non-carbohydrate substituents. There are only a small number of fucoidans with established basic elements of the chemical structure. Unfortunately, uncharacterized crude preparations are often used even in scientific research. In G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences in 2006 the suplement "Fucolam" (No 77.99.23.3.У.739.1.06, Russia) was registered. The "Fucolam" was created based on structurally characterized fucoidan from the brown alga Fucus evanescens, synthesizing from 12 to 15% of the polysaccharide, The biological effects of the "Fucolam" are studied in detail. It was established that the "Fucolam" has probiotic, hepatoprotective, glucose and cholesterol lowering effects. The results about this will be presented. This work was supported by RFBR Grant #14-04-93003.

References: [1] M. Kusaykin, I. Bakunina, V. Sova, S. Ermakova, T. Kuznetsova, N. Besednova, T. Zaporozhets, T. Zvyagintseva,

Biotechnol J. 2008, 3, 7, 904-915 [2] O. Berteau, B. Mulloy, Glycobiology. 2003, 13, 6, 29R-40R


74

The Structural Peculiarities of Poly- and Oligosaccharides of Carrageenan from

the Red Algae Tichocarpus crinitus and Their Ability to Induce in vivo

Interleukin 10

A.A. Kalitnik, A.O. Kravchenko, A.O. Byankina, S.D. Anastyuk, I.M. Yermak

Pacific Institute of Bioorganic Chemistry, Far-East Branch of Russian Academy of Sciences, Vladivostok, Russia, fax: 7(4232) 31-40-50


Сarrageenans are a family of natural water-soluble sulfated galactans extracted from numerous

species of red alga and have a wide application in food and medical industry. The basic structural

of these polysaccharides units are disaccharide, carrabiose, consisting of alternating β-1,3- and α-

1,4-linked galactose residues. The terms κ- and ι-carrageenans are used for polysaccharides which

contains of κ- (G4S-DA), λ- (G2S-D2S6S) and ι-carrabiose moieties (G4S-DA2S), respectively as

main components. However natural carrageenans are often hybrid of more than one of these units

and made of several carrabiose moieties. The hybrid nature of carrageenans at the molecular level

and also their molecular weight are responsible for changes in biological and physico-chemical

properties of carrageenans compared with those of their homopolymeric ideal types.

The purpose of the current study was to investigate the structural peculiarities of carrageenan

from Tichocarpus crinitus based on the analysis their oligosaccharides obtained by mild acid

hydrolysis and study the ability of studied samples to induce in vivo synthesis of anti-inflammatory

cytokine IL-10.

We have shown that the gelling polysaccharides isolated from the vegetative forms of T. crinitus

belong to κ/β-carrageenans type (DA-G4S/ G-DA) with predomination (80%) of the carrabiose

units of κ-type. Oligosaccharides of κ/β-carrageenans were obtained by mild acid hydrolysis,

purified and fractionated by gel-permeation chromatography. The structural peculiarities of

oligosaccharide mixture of κ/β- carrageenan from the red algae T. crinitus were investigated by

negative-ion MALDI-TOFMS. According to results of analysis in structure of oligosaccharides are

presented β- (DA-G), ι- (G4S-DA2S) and γ-(G-D6S) carrageenan (the precursor of β-carrageenan)

blocks. It is known that carrageenans excited a biosynthesis of different mediators of immune

system such as interleukins, TNF and others. IL-10 has great potential for use in the treatment of

inflammatory and immune illnesses. The effect of source polysaccharide with molecular weight

(MW 400 kDa) and oligosaccharides (1.7 kDa) on the in vivo production of anti-inflammatory

cytokine IL-10 was examined. It was shown that both studied samples of carrageenan previously

administrated orally into mice for 7 days significantly stimulate induction of IL-10 and increase IL-

10 levels in mice sera in comparison with control group of animals.


75

First Synthetic Macrocyclic Glycoterpenoids

V.E. Kataev

A.E.Arbuzov Institute of Organic and Physical Chemistry Kazan Scientific Center, Russian Academy of Sciences

420088, Russian Federation, Kazan, Arbuzov str., 8


First macrocyclic glycoterpenoids were synthesized on the basis of Stevia rebaudiana glycoside

steviolbioside 1, its isomeric aglicon diterpenoid isosteviol 2, trehalose, and glucuronic acid.

The search for synthetic approaches for obtainig glycoterpenoids pictured above was not as

simple as it may seem to someone, so it is described in detail in the report.

The study was supported by Russian Scientific Foundation (grant 14-05-00015).


76

Mechanochemical Technology to Create of Targeted Delivery Drugs

S.S. Khalikov,1 Yu.S. Chistyachenko,2 A.V. Dushkin,2 I.A. Arkhipov,3

A.I. Varlamova,3 I.I. Glamazdin3

1Federal State Institution of Science Nesmeyanov Institute of Organoelement Compounds Russian

Academy of Sciences- 28, Vavilov str., Moscow, Russia 2Federal State Institution of Science Institute of Solid State Chemistry and Mechanochemistry

Siberian Branch of the Russian Academy of Sciences - 18, Kutateladze str., Novosibirsk, Russia 3All-Russian Research Institute of Fundamental and Applied Parasitology animals and plants named after Scriabin Russian Academy of Sciences -28, Bolshaya Cheremushkinskaya str.,

Moscow, Russia


Helminthiases are especially dangerous parasitic diseases of humans, animals and plants caused by parasitic worms. Known methods of treatment of helminthiasis of animals based on a wide range of anthelmintic drugs, many of which, because of their poor solubility, often do not provide the necessary efficiency and to achieve it we have to use their excessive dose [1]. Promising innovative directions in addressing the solubility is to create effective drugs based on known anthelmintic by preparing solid dispersions of these substances with water-soluble polymers [2,3]. Such dispersions in water and physiologically active media form supramolecular complexes, under consideration as a Drug Delivery System, the proportion of publications which becomes dominant [4].

In this paper we will present research data on mechano-chemical modification of certain substances anthelmintic benzimidazole derivatives (Carbendazim, Albendazole, Fenbendazole) with water-soluble polymers. The formation of supramolecular complexes is confirmed on the basis of physical and chemical methods of analysis, namely, the formation of complexes of the type "guest-host" is due to hydrogen power, van der Waals interactions between the functional groups of components, forces, hydrophobic interaction, and so on.

Anthelmintic activity of the synthesized complexes were carried out on laboratory models in white mice experimentally infested Trichinella spiralis, Hymenolepis nana and Fasciola hepatica, respectively. It was shown that the supramolecular complex Albendazole: Arabinogalaktan = 1: 10 showed 100% effectiveness against all species studied helminths reduction in dosage is 9-10 times.

Will discuss the results of the study of the properties of anthelmintic benzimidazole innovative drugs in laboratory models and production experience. References: [1]. Arkhipov I.A. Anthelmintics: pharmacology and application. - Publishing House of Agricultural Sciences. - 2009.

- 109 p. [2]. Khalikov S.S., Meteleva E.S., Dushkin A.V. et al. Mechanochemical modification of the properties of

anthelmintic drugs // Chemistry for sustainable development. -2011. - T. 19. - № 6. C. 699-703. [3]. Dushkin A.V., Suntsova L.P,, Khalikov S.S. Mechanochemical technology to increase the solubility of drugs //

Basic Research. - 2013. - № 1 (part 2). - S. 448-457. [4]. De Jong W.I., Born P.I.A. Drug delivery and nanoparticles: Applications and hazards // Inter. J. Nanomedicine. -

2008. - V. 3.- N 2. - P. 133-149.


77

Arabinogalactan as a Drug Carrier

M.V. Khvostov,1 T.G. Tolstikova,1 S.A. Borisov,1 M.S. Biryukova,1 A.V. Dushkin,2

J.S. Chistyachenko,2 A.A. Chernonosov3

1Laboratory of Pharmacological Research, N.N. Vorozhtsov Institute of Organic Chemistry SB RAS, 630090, Russia, Novosibirsk, Lavrent’eva str., 9

2Institute of Solid State Chemistry and Mechanochemistry SB RAS, 630128, Russia, Novosibirsk, Kutateladze str., 18

3Institute of Chemical Biology and Fundamental Medicine, SB RAS, 630090, Russia, Novosibirsk, Lavrent’eva str., 8


Worldwide use of natural polysaccharides for complexation with drugs as a way for their

bioavailability and stability improvement is firmly became a part of global pharmaceutics. Here we

suggest expand the list of well-known compounds by a novel one that is Arabinogalactan (AG)

from Larix Sibirica or Gmelini. It has branched structure and is able to form inclusion complexes

with drugs that has low aqueous solubility. Its molecular mass is 15-20 kDa and LD50 >5000 mg/kg.

AG possesses good water solubility without any structure modifications. By a solid phase synthesis

we have obtained and investigated pharmacological properties of AG complexes (mass ratio

10/20:1) with drugs possesses different effects: anticoagulant Warfarin (WF), NSAID Aspirin

(ASA) and hypotensive drug Nifedipine (NF). All experiments were carried out in vivo on Wistar

rats and albino mice with standard pharmacological tests application. In the experiment involving

WF:AG complex the significant influence of complexation on WF’s pharmacokinetics after single

or repeated intragastric dosing was found. Effective ASA per os single analgesic and antiplatelet

dose in the AG complex was reduced in a half (25 mg/kg). 30 days introduction of the ASA

complex at doses (in terms of ASA) of 25 and 50 mg/kg reveals no ulcerogenic effect on rat’s

stomach mucosa whereas ASA at its effective dose of 50 mg/kg provoke mucosa damage.

Hypotensive NF intravenous dose in its AG complex was reduced to 10 times and oral dose to 2

times. Obtained pharmacokinetic parameters revealed that NF’s concentration in blood plasma was

two times higher in first 30 min after oral introduction to rats in the case of NF:AG complex

compared to free NF in equal dose. In all cases aqueous solubility of drugs in AG complexes was

increased (WF 5.3 times; ASA 1.1 times; NF 8.5 times). Thus, presented approach is a simple and

effective way increasing drug’s bioavailability and safety.


78

New Chiral Catalyst for Henry Reaction

V. Konev, T. Khlebnikova, L. Malysheva and Z. Pai

Boreskov Institute of Catalysis Siberian Branch of RAS,

630090, Russia, Novosibirsk, Akad. Lavrentiev Pr. 5


β-Nitro alcohols are the convenient precursors of chiral β-amino alcohols widely used in

producing and development of the contemporary pharmaceutical agents. Efficient method of chiral

β-nitro alcohols synthesis is the asymmetric version of aldehydes and lower nitroalkanes

condensation catalyzed by the copper(II) complex salts with chiral ligands of various structural

types. A number of the efficient catalysts of asymmetric reactions were prepared using the optically

active natural alcohols, amino acids, and terpenes derivatives as chiral constituents of metal

complexes [1]. However, there are few examples of the natural diterpenes use for the synthesis of

chiral inductors. Earlier, we had developed the synthesis of the enantiomerically pure phosphorus-

and nitrogen-containing compounds from natural diterpene - levopimaric acid [2].

As a result of our further investigations in the area of the resin acids transformations, we have

developed the synthetic method for the producing of diamine 1 [3], which was used for the series of

aminophenols preparation by a two-step synthesis. It was found that in the reaction of 4-

nitrobenzaldehyde with nitromethane the Cu(II) complex with aminophenol 2 promotes the

formation of 2-nitro-1-(4-nitrophenyl) ethanol in 95% yield and the 75% ee. Using the same

catalyst, the condensation of nitromethane with benzaldehyde derivatives 3 having donor or

acceptor substituents results in formation of β-nitro alcohol 4 with 40-95% yields and 45-82% ee.

Thus, the optically pure tetradentate ligand 2 have been synthesized from resin acids mixture.

The Cu(II) complex with ligand 2 catalyzes the condensation of aromatic aldehyde with

nitromethane to form nitro alcohol with moderate to high chemical and optical yields.

Acknowledgements The work is supported by RAS Program of basic researches, project N 5.7.3. References [1] H. Blaser, Chem. Rev. 1992, 92, 935-952 [2] T. Khlebnikova, N. Karpyshev, O. Tolstikova, A. Tolstikov, Chirality. 2004, 16, 40-50 [3] V. Konev, T. Khlebnikova, Z. Pai, Chem. for Sustainable Development. 2011, 19, 165-168


79

In vitro Cytotoxicity of the Dehydroabietylamine Ammonium Salts

A. Kononova,1 K. Kovaleva,1,2 V. Korobeynikov,1 O. Yarovaya,1,2 S. Cheresiz,1

E. Semenova,1 A. Pokrovsky,1 N. Salakhutdinov1,2

1Novosibirsk State University, 630090, Russia, Novosibirsk Pirogova St. 2.

2N.N. Vorozhtsov Novosibirsk Institute of Organic chemistry SB RAS, 630090, Lavrentiev av., 9, Novosibirsk, Russia


Chemotherapy remains the therapeutic modality of choice in the treatment of a number of types

of cancer. The search for the novel compounds with potential anti-tumor activity is essential. Due to

the interest in the synthesis of ammonium salts, different members of this class of compounds were

shown to posess a high biological activity. We, thus, attempted to synthetize ammonium salts of

dehydroabietylamine with various organic acids and study their cytotoxic activity in the cell lines of

different origin. For obtaining ammonium salts, we selected several aliphatic, aromatic,

heteroaromatic, and natural acids, as well as cinnamic acid derivatives. Formation of ammonium

salts was confirmed by IR spectroscopy.

Cytotoxicity of all synthetized compounds was studied in several transformed (cancer) cell lines

including. MCF7 and MDA-MB-231 human breast cancer cells and U87MG human malignant

glioma cells. Cell cultures were incubated with different concentrations of compounds for 48 h,

then, the cell viability was assessed by colorimetric MTT assay. Out of several dozens of

compounds screened, several compounds possessed high cytotoxic activity in the cancer cells lines

(as demonstrated by their CD50 values in the range of concentrations 8-25 mcg/mL).


80

The Story of Amphipathic Antiviral Nucleosides

V.A. Korshun, A.A. Chistov, G.V. Proskurin, A.V. Ustinov

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997, Russia, Moscow, Miklukho-

Maklaya st., 16/10


Alkynylated nucleosides find diverse applications in bioorganic chemistry [1]. E.g., alkynyl

linker appeared useful for fluorescent dye attachment in epoch-making DNA sequencing techniques

[2, 3]. Two decades ago we prepared 5-(1-pyreneethynyl)-2′-deoxy-uridine (1), the first nucleoside

containing fluorescent polyaromatic hydrocarbon conjugated to nucleobase through a triple bond

[4]. Suddenly, these nucleosides (e.g., compounds 1–3) were recognized as a new class of antiviral

compounds with non-nucleoside mode of action [5, 6]. More careful studies revealed that perylene

residue in this type of compounds (e.g., nucleosides 3, 4) appears to be crucial prerequisite for

potent antiviral activity against broad spectrum of enveloped viruses (herpes simplex virus types 1

and 2, hepatitis C virus, various strains of influenza virus A etc.) [7, 8]. The suggested mechanism

of action for these nucleosides includes inhibition of virus-cell fusion by stabilizing the positive

curvature of cellular and viral membranes, and the

compounds were called RAFIs (rigid amphipathic fusion

inhibitors) [7]. However, an alternative photosensitizing

mechanism of antiviral action for 5-(3-peryleneethynyl)-2′-

deoxy-uridine (3) was suggested [9] and recently supported

with some experimental data [10]. At present, we are

synthesizing related compounds by varying of

polyaromatics, rigid linker, nucleobase, and a carbohydrate

part of molecule. These are used for structure–activity

relationship studies and for further elucidation of a

mechanism of antiviral action. The research was supported by Russian Science Foundation (project no. 15-15-00053).

References [1] V.A. Korshun, E.V. Manasova, and Y.A. Berlin, Russ. J. Bioorg. Chem. 1997, 23, 300–362. [2] J.M. Prober et al., Science 1987, 238, 336–341. [3] D.R. Bentley et al., Nature 2008, 456, 53–59. [4] V.A. Korshun et al., Russ. J. Bioorg. Chem. 1996, 22, 807–809. [5] V.L. Andronova et al., Russ. J. Bioorg. Chem. 2003, 29, 262–266. [6] M.V. Skorobogatyi et al., Org. Biomol. Chem. 2006, 4, 1091–1096. [7] M.R. St.Vincent, C.C. Colpitts, A.V. Ustinov et al., Proc. Natl. Acad. Sci. USA 2010, 107, 17339–17344. [8] C.C. Colpitts, A.V. Ustinov et al., J. Virol. 2013, 87, 3640–3654. [9] F. Vigant, A. Hollmann, and B. Lee, Chem. Biol., 2010, 17, 1049–1051. [10] F. Vigant et al., J. Virol., 2014, 88, 1849–1854.


81

Development of Original Pluripotent Preparations for the Russian

Pharmaceutical Market on the Subject of Import Substitution

Ya.A Kostyro,1 V.V. Kostyro,1 E.S. Gogol,1 V.V. Davaa,2 E.N. Petrova,1 A.O. Savinova,3

N.N. Trofimova,1 E.V. Stolpovskaya,1 L.A. Grischenko,1 L.A. Ostroukhova,1 S.A. Lepehova,4

K.V. Alekseev,5 V.A. Babkin,1 V.K. Stankevich,1 B.A. Trofimov1

1A.E. Favorsky Institute of Chemistry, Siberian Branch of Russian Academy of Science, Irkutsk

2Republican Medical College, Kyzyl, Tuva Republic; 3LLC "Specialized Industrial Company IrIOCh", Irkutsk;

4Irkutsky Scientific Center of Surgery and Traumatology, Irkutsk; 5V.V. Zakusov Research Institute of pharmacology, Moscow, Russia

E-mai; [email protected]

Considering the current political and economic situation in the Russian Federation the Order of

RF Ministry of Industry and Trade № 656 "Adoption of the medical sector plan to implement

"import substitution" in the pharmaceutical industry of the Russian Federation" was issued on

31.03.2015, according to the Order the quota of import pharmaceuticals has to be reduced in 10-

50% for the treatment of socially dangerous diseases.

The original pharmaceutical active substances, which have antimicrobial [1], lipid-lowering and

anticoagulating [2-5], anti-inflammatory and wound healing activities [6, 7], are elaborated in the

Irkutsk Institute of Chemistry SB RAS. On their bases the variety of pharmaceutical formulations

are developed for the treatment of such socially dangerous diseases as atherosclerosis, acute and

chronic venous insufficiency, diabetes mellitus, infected wounds and burn injuries.

Preclinical studies have revealed their performance targets to be applied in practical medicine.

All the original pharmaceutical substances are non-toxic, pluripotent and effective as modern

medicines as the most of import drugs for the treatment of the above diseases.

At present, great efforts are undertaken to enter the original domestic preparations into RF

pharmaceutical market that is completely in line with the tasks of "import substitution" in the

pharmaceutics.

This work was supported by the Program of the Presidium of Russian Academy of Sciences "Fundamental sciences for medicine" (2009-2014 years) References [1]. RU Patent 2144024 Stankevich V.K. et al., 2000. [2]. RU Patent 2319707 Kostyro Ya.A. et al., 2008. [3]. RU Patent 2468789 Kostyro Ya.A et al., 2012. [4]. RU Patent 2532915 Kostyro Ya.A et al., 2014. [5]. RU Patent 2541808 Kostyro Ya.A et al., 2015. [6]. RU Patent 2513186 Kostyro Ya.A et al., 2014. [7]. RU Patent application 2014142414. Kostyro Ya.A. et al., 2014.


82

Cytotoxicity, Intracellular Localization and Acute Intravenous Toxicity of

Cluster Complexes Na4[{Re6Q8}(CN)6] (Q = S, Se or Te) – New Promissing

Photoactive and X-ray Contrast Agents

A.A. Krasilnikova,1,3 A.O. Solovieva,3 K.E. Zubareva,3 T.N. Pozmogova,1,3 L.V. Shestopalova,1

K.A. Brylev,1,2 M.A. Shestopalov1,2,3

1Novosibirsk State University, 2 Pirogova Str., 630090 Novosibirsk, Russia. 2Nikolaev Institute of Inorganic Chemistry SB RAS, Novosibirsk, Russia.

3Federal State Budgetary Scientific Institution «Scientific Institute of Clinical and Experimental Lymрhology», 2 Timakova Str., 630060 Novosibirsk, Russia.


Octahedral rhenium cluster complexes may have considerable potential as therapeutic and diagnostic drugs due to their luminescent and X-ray contrast properties. However, their potential biological effects and toxicity in vitro and in vivo are rather far from being understood. Thus, the aim of our research was to study cytotoxicity, intracellular localization in vitro and acute intravenous toxicity in vivo of several chalcocyanide octahedral rhenium cluster complexes of the general formula Na4[{Re6Q8}(CN)6] (Q = S, Se or Te) to evaluate, which cluster complex is more promissing for biomedical application.

It has been established that IC50 for Na4[{Re6S8}(CN)6] and Na4[{Re6Se8}(CN)6] were 208.5±10.4 µМ and 212.5±10.62 µМ, respectively. While IC50 for Na4 [{Re6Te8}(CN)6] was 720.7±36 µМ, so the telluride cluster was the safest agent for cell culture. However it is likely that concentration of rhenium clusters higher than 100µM is too high to be used in practical medical applications.

All three investigated cluster complexes penetrated into cells in vitro, presumably by diffusion pathway. Confocal microscopic study showed that complexes localized in nucleus and cytoplasm of the cells. Moreover, TEM microphotographs of the cells with Na4[{Re6Se8}(CN)6] and Na4[{Re6Te8}(CN)6] were very contrasting, that confirm the possible application of these cluster complexes as the alternative contrast agents for TEM.

The study of acute intravenous toxicity in vivo also confirmed that Na4[{Re6Te8}(CN)6] was the safest hexarhenium cluster complex for biomedical application. Mice showed a good tolerance of the intravenous injection of this complex at a dose up to 300 mg(Re)/kg and LD50 for this complex was equal to 1081.84±82.57 mg(Na4[{Re6Te8}(CN)6])/kg (or 506.54±38.66 mg(Re)/kg). This dose is 3 times higher than maximal presumable clinical dose for vascular contrast agents and approximately 500 times higher than presumable dose for contrast additive in biodegradable pins. Morphological analysis has shown that liver is target organ for all investigated cluster complexes.

Therefore, we may conclude that octahedral rhenium cluster complexes Na4[{Re6Q8}(CN)6] (Q = Se, S or Te), are potential agents for photodynamic therapy of superficial tumors, as biomarkers in biology and as alternative contrast agents for TEM, since they penetrate cells and have low toxic effects in vitro and in vivo. Na4[{Re6Te8}(CN)6] is the most promissing cluster complex for biomedical applications. Acknowledgements: This work was supported by the Russian Science Foundation (Grant №14-14-00192).


83

Targeting Hormone-Refractory Prostate Cancer with New Molecular Entities

K.V. Kudryavtsev

Department of Chemistry, M.V. Lomonosov Moscow State University, 119991, Russian Federation,

Moscow, Leninskie Gory, 1, stroenie 3


Chemotherapeutical treatment of hormone-refractory prostate cancer (HRPC) was of little value

until recent years but novel emerging targets and pathways crucial for tumor growth tend to change

the situation. In the course of our research we synthesized well-defined β-proline oligomers A and

small-molecule imidazolylthiazoles B and studied their influence on proliferation of HRPC cell

lines and cancers cell cycle progression [1-3].

Most potent poly-β-prolines A induced caspase-dependent apoptosis of PC-3 cells through the

cell cycle arresting at G1 phase and mitochondrial damage at low micromolar and submicromolar

concentrations [2]. 2-(1H-Imidazol-1-yl)-4-(3-(trifluoromethyl)phenyl)thiazole (B, R = m-CF3)

inhibited tubulin polymerization associated with an anti-Aurora A activity in PC-3 and DU-145 cell

lines that induced an arrest of the cell cycle at G2/M phase in a time-dependent fashion and a

subsequent increase of hypodiploid sub-G1 phase (apoptosis) [3]. Structure-activity relationships

(SARs) for both compounds sets as well as mechanistic study details will be considered in the

report.

References [1] K.V. Kudryavtsev, P.M. Ivantcova, A.V. Churakov, S. Wiedmann, B. Luy, C. Muhle-Goll, N.S. Zefirov, and S. Bräse, Angew. Chem. Int. Ed. 2013, 52, 12736-12740. [2] K.V. Kudryavtsev, C. C. Yu, P.M. Ivantcova, V.I. Polshakov, A.V. Churakov, S. Bräse, N.S. Zefirov, and J.H. Guh, Chem. Asian J. 2015, 10, 383-389. [3] C.C. Yu, S.P. Liu, J.L. Hsu, J.T.A. Hsu, K.V. Kudryavtsev, and J.H. Guh, J. Biomed. Sci. 2015, 22, 2.


84

Sulfation of Polysaccharides and Triterpens by Sulfamic Acid

V.A. Levdansky,1,2 N.Yu. Vasil’eva,2 S.A. Kuznetsova,1,2 A.V. Levdansky,1

A.S. Kazachenko,1 B.N. Kuznetsov1,2

1 Institute of Chemistry and Chemical Technology SB RAS, 660036, Russia, Krasnoyarsk,

Akademgorodok, 50-24 2 Siberian Federal University, 660041, Russia, Krasnoyarsk, Svobodniy, 79


Sulfated polysaccharides and triterpenes have good prospects of application in industry and

medicine. Conventional sulfation methods are mainly based on the use of quite aggressive sulfation

agents (sulfuric acid, sulfur trioxide, chlorosulfonic acid) and toxic solvents (pyridine etc.).

New green methods of microcrystalline cellulose (MCC), arabinogalactan (AG), betulin,

3-acetate betulin and betulinic acid sulfates synthesis are described. They are based on the

application of low-toxic and low-corrosive sulfation agent – complex of sulfamic acid with urea.

At the first time it was suggested to carry out the sulfation of MCC and AG with sulfamic acid in

the presence of base catalyst-urea in such solvents as dioxane, diglyme, DMFA. In contrast to

conventional sulfation agents (H2SO4, SO3 etc.) the sulfamic acid is a stable non-hydroscopic and

non toxic crystalline solid which acidity is comparable with sulfuric acid. Sulfamic acid can form

with urea a donor-acceptor complex, which has a high sulfatizing reactivity.

Sulfated MCC and AG were isolated in the form of ammonium, sodium and potassium salts. The

degree of substitution of sulfated MCC reaches to 1.40–1.54 in diglyme and DMFA solvents. The

most effective sulfation of MCC takes place in diglyme. The yield of MCC sulfate prepared in

diglyme was by 2.5 times higher than the yield of MCC sulfate obtained in DMFA.

AG sulfates with a high content of sulfur (11.5–11.8 % wt.) were obtained in dioxane solvent at

85–95 oC, at ratio AG/SA (g/mmol) – 1:14 and time 2.5–3.0 h.

At the first time it was suggested to carry out the sulfation of triterpenes (betulin, betulinic acid

and 3-acetate betuline) by sulfamic acid in the presence of urea. Sulfated triterpenes are obtained in

acidic form and as ammonium, sodium and potassium salts.

Obtained MCC and AG sulfates were characterized by elemental analysis, 13C NMR, FTIR,

SEM, AFS and XPS methods.

Sulfates of betulin, 3-acetate betulin and betulinic acid were characterized by elemental analysis, 13С NMR and FTIR.

The anticoagulant activity of obtained MCC sulfates is comparable with the activity of the

commercial the drug heparin.


85

Novel Biologically Active Compounds Based on (Indol-1-yl)maleimide Scaffold

S.A. Lakatosh

Gause Institute of New Antibiotics

ul. Bol'shaya Pirogovskaya 11, Moscow, 119021, Russia,


Indolylmaleimide derivatives are an interesting group of natural and synthetic compounds

posessing the valuable biological properties. The first examples were the alkaloid strurosporine (1),

a very potent yet unselective inhibitor of various proteinkinases and an attitumor antibiotic

rebeccamicyn (2), sharing the indolocarbazole core. The further development was targeted mostly at

the proteinkinases, the enzymes, regulating the intracellular sygnals transduction, and led to the

different derivatives with improved potency and selectivity against a specific target (e.g. 3, 4). In

our approach we started from a close but a different core structure namely 3,4-(indol-1-

yl)maleimide or 3-(indole-1-yl)-4-(indol-3-yl)maleimide or the related polyannelated structures.

Several series of the potenetially biologically active derivatives were synthesized. The screenining

revealed the interesting types of activities. The gramines 5 and (indol-1-yl) maleimide derivatives 6

were the inhibiting proteinkinase C α, preventing the tumor cells from MDR development. Several

compoundsof the type 7a,b were active against Leushmania parasites. The series of staurosporine

analogues 8 were very potent and selective PIM-1 protein kinase inhibitors and had also

pronounced antitumor activity against a set of the leukemia cell lines.


86

New Eudesmane-type Sesquiterpenes from the Marine-derived

Fungus Penicillium thomii

E.V. Leshchenko,1 Sh.Sh. Afiyatullov,2 O.I. Zhuravleva,1,2 V.A. Denisenko2

1Far Eastern Federal University, Suhanova 8, Vladivostok 690950, Russian Federation 2G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian

Academy of Sciences, Prospect 100-let Vladivostoku 159, Vladivostok 690022, Russian Federation


Marine-derived fungi are sources of secondary metabolites with an original chemical structure

and promising biological and pharmacological properties [1]. The total number of new natural

products from marine-derived fungi currently exceeds 1000. An algae and sea grass are the most

prevalent source of marine fungi for chemical studies. Recently the main producers of new

compounds have been the fungi of the genus Penicillium and Aspergillus [2]. So, in our search for

fungal secondary metabolites with novel chemical structures and a biological activity we have

investigated marine-derived fungus Penicillium thomii associated with sea grass Zostera marina

(Sea of Japan).

The fungus P. thomii was cultured for 21 days on specially modified rice medium. The EtOAc

extract of the mycelia was purified by a combination of Si gel column chromatography and

reversedphase-HPLC to yield four new eudesmane-type sesquiterpenes (1-4) together with one

known eudesmane derivative (5). Their structures were established by 2D NMR spectroscopy and

HRMS.

The known compound 5 was isolated from the whole plant of Saussurea deltoidea mainly

distributed in Asia and Europe [3]. Many eudesmane-type sesquiterpenes have been isolated from

ground plants and show inhibitory effects on NO production in lipopolysaccharide-activated

macrophages [4-5].

The study was supported by the Far Eastern Federal University project No. 14-08-01-34_u, by the program grant from the Russian Foundation for Basic Research (RFBR № 14-04-00910-a.

References: 1. Blunt J.W., Copp B.R., Keyzers R.A., Munro M.H., Prinsep M.R. Nat. Prod. Rep. 2015. V. 32, No. 2. P. 116-211. 2. Rateb M.E., Ebel R. Nat. Prod. Rep. 2011. V. 28, No. 2. P. 290-344. 3. Xu J.-J., Huang H.-Q., Zeng G.-Z., Tan N.-H. Fitoterapia. 2012. V. 83, No. 6. P. 1125-1130. 4. Liu Y., Ma J., Wang Y., Donkor P.O., Li Q., Gao S., Hou Y., Xu Y., Cui J., Ding L., Zhao F., Kang N., Chen L.,

Qiu F. Eur. J. Org. Chem. 2014. V. 2014, No. 25. P. 5540-5548. 5. Xu J., Ji C., Zhang Y., Su J., Li Y., Tan N. Bioorg. Med. Chem. Lett. 2012. V. 22, No. 4. P. 1660-1663.


87

The Study of Drugs Action Mechanism via the Chemical Modeling of Drug – Enzyme and Drug - Receptor Interaction

T.V. Leshina,1 E.A. Khramtsova,1 M.A. Miranda2

1Institute of Chemical Kinetics and Combustion, 630090, Russia, Novosibirsk, Institutskaya 3 2Instituto de Tecnologґıa Quґımica, UPV-CSIC, Universidad Politґecnica de Valencia, Spain.


Interest in the cellular and molecular mechanisms of drugs action is great and it is increasing constantly. The main reasons for this interest are related to the fact that knowledge of the mechanisms allows one to apply remedies more conscious and targetedly. Moreover, it makes possible to avoid time-consuming screening, due to, for example, the establishment of structure-property correlation. According to the modern concepts, drug action occurs through the interaction of a drug with a specific receptor protein or an enzyme. These processes include the steps of a so-called “weak (non-covalent) interactions” (hydrogen bonds, dipole-dipole and van der Waals force, charge-transfer complexes). The chemical transformations of drugs are also possible. We have proposed an approach to model the separate stages of these processes by means of the reaction in solution or organized media. The key goal of such simulation is to observe the drug’s structure influence on the elementary stages of the drug action processes. This approach allows us to identify the principal factors that determine the effectiveness of a drug by comparing the effect on the individual stages and the overall effect. Our approach is based on two major assumptions. Firstly, we suggest the reactivity of active intermediates would not depend on their generation pathway. Secondly, we used photo irradiation for the activation of model processes. If the activation barrier in the drug-receptor or drug–enzyme complexes decreases through the charge–transfer complex formation, it shows up in model reaction as a result of the increasing of drug’s oxidation potential in its excited state. Whereas simulative processes involve the reaction of electron or “electron – proton” transfer, we can apply both spin chemistry methods to study short-lived active paramagnetic intermediates and photochemistry methods to study the appearance of “weak interactions”. In particular, this approach has been used to study the nature of the well –known phenomenon – the impact of chiral centers on the drug’s reactivity. In spite of very intensive investigation of chirality phenomenon, many questions remain without answers. So, there is no understanding about how chiral compounds appeared in nature and about the origin of the difference in chemical reactivity of isomers. We have attempted to study the nature of the chiral centers influence by the example of widely-used non-steroidal anti-inflammatory drug (NSAID) – (S)-naproxen and its less active (R)-isomer. The literature proved reliable difference in the impact of naproxen optical isomers and some other NSAIDs on the cyclooxygenase activity (enzyme which is responsible for the synthesis of molecules producing inflammations). It was shown that cyclooxygenase demonstrates two kinds of activity: carries out the cyclization of substrate and electron transfer. Trying to trace the influence of the naproxen’s optical configuration on its binding with amino acid residues in the active site of the enzyme, we used a model process: the photoinduced electron transfer in the linked systems: the dyads containing acceptor – (S)- or (R)- naproxen and donor (S)- N–methyl-2-pyrrolidine connected by various bridges. The analysis of the CIDNP effects and the quantum yields and kinetics of dyad’s fluorescence quenching has shown that the rates of complete and partial (exciplex) charge transfer, which simulate the stage of binding between the donor and acceptor, are really different for (S)- and (R)-naproxen. We can assume that the smaller efficiency of quenching by electron transfer which is typical for the (S)-isomer, is also the cause of its weaker reversible binding to the active site of the receptor. The latter is the result of the biochemical studies of the binding processes for the (S)- and (R)-optical isomers.

The work was supported by the RFBR (grant 14-03-00192).


88

Peucedanin in the Synthesis of Coumarin-chalcone Hybrids and Dicoumarins

A.V. Lipeeva,1 P.Z. Zanimkhanova,2 M.A. Pokrovsky,3 M.V. Khvostov,1 D.S. Baev,1

S.M. Adekenov,2 T.G. Tolstikova,1 A.G. Pokrovsky,3 E.E. Shults1

1 N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, Siberian Branch, Russian Academy of Sciences, Lavrentjev Avenue 9, 630090 Novosibirsk, Russia

2 International Research and Production Holding “Phytochemistry”, 100009,Karaganda, Kazakztan

3 Novosibirsk State University, Pirogova St. 2, Novosibirsk 630090, Russia


Natural and synthetic coumarins represent an important group of biological activity compounds

and its therapeutic effects depend on the presence and position of substituent. Earlier we supplied

method of synthesis of natural coumarin peurutenicin 1 [1] and 7-hydroxy-6-cyanocoumarin 2 from

accessible plant furocoumarin peucedanin 3 [2]. Different coumarin derivatives were obtained by

Suzuki and Pd-catalyzed amination reactions of triflates 4,5 [3-4].

Herein we report about the synthesis of 7-alkynilcoumarins 6-15 by cross-coupling reaction of

triflates 4,5 with terminal alkynes. Conditions for obtaining of 7-ethynylcoumarins 16,17 were

found. By the CuAAC reaction of alkynes 16,17 with 2-(azidoalkoxy)chalcones 18,19 coumarin-

chalcone hybrids 20-21 were obtained. 1,2,3-Triazole linked chromen-2,3-dihydrofurocoumarins

22,23 were synthesized by CuAAC reaction of alkynes 16,17 with 2-azidooreoselone 24.

OHO

R1

OOO

MeO

O OR2O

R1

O

3 1:R1=C(O)OMe; 2: R1= CN

4: R1= C(O)OMe, R2= Tf;5: R1= CN, R2 = Tf;

a или b

66-78%

a: Tf2O, Pyb: TsCl, NEt3, THFc: Pd(PPh3)2Cl2,CuI, Et3N, ДМФА, 135-140оС, 10 ч.

O

R1

O

R

c

6-17 (44-72%)

R= Ph (6,7), 4-MeOC6H4 (8,9), 3-NHAc-4-O2MeC6H3 (10,11),2,3,4-(MeO)3C6H2 (12,13), Si(CH3)3(14,15), H (16,17).

O

R1

O

R1 = C(O)OMe, CN;

O

R1

O

H 16,17

OO

O

ON3

OO

O

ONN

NO

HOO

H3CO

X NN N

O O

R1

O

OH O

H3CO X N3

aa22, 23(63-75%)20, 21 (52-58%)

18, 19 24

a: CuSO4, sodium ascorbate, CH2Cl2-H2O, 20 40oC

The biological activity of synthesized compounds would be discussed.

This work was supported by RFBR (grant 15-03-06546)

References 1. Shults, E.E., Petrova, T.N., Shakirov, M.M. et.al., Khimiya v interesakh ustoichivogo razvitiya

(Chem. Sustainable Develop.), 2003, vol. 11, p. 683. 2. Osadchii, S.A., Shults, E.E., Shakirov, M.M., and Tolstikov, G.A., Izv. Akad. Nauk, Ser. Khim., 2006, p. 362. 3. Makhneva E. A., Lipeeva A. V., Shults E. E., Shakirov M. M., Tolstikov G. A.. Russ. J. Org. Chem., 2012, Vol. 48,

pp 1094-1102. 4. Makhneva E. A., Lipeeva A. V., Shults E. E., Russ. J. Org. Chem., 2014, Vol. 50, pp 662-669.


89

Soloxolone Methyl – a Novel Compound with Multiple Activities, Exhibited

Anti-cancer, Anti-flu and Anti-inflammatory Activities in vitro and in vivo

E.B. Logashenko,1 A.V. Markov,1 O.V. Salomatina,2 N.F. Salakhutdinov,2 M.A. Zenkova1

1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch Russian Academy of Sciences, 8, Lavrentjeva ave., Novosibirsk, 630090, Russian Federation

2N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, Siberian Branch Russian Academy of Sciences, 9, Lavrentjeva ave., Novosibirsk, 630090, Russian Federation

Some of the triterpenoids like oleanolic and glycyrrhetinic acids are known to possess a wide spectrum of biological activities. However, the activities of these naturally occurring molecules in respect to the particular molecular targets are weakly exhibited. Recently, soloxolone methyl (SM) (2-cyano-3,12-dioxo-11-deoxo-18βH-glycyrrhet-1(2),9(11)-dienoate) by combined modification of A, C and E rings of glycyrrhetinic acid was synthesized.

Analysis of biological action of SM in vitro showed that this compound induces the death of cancer cells by triggering of mitochondrial pathway of apoptosis.

Here, we studied spectrum of SM activities using various in vivo models of disease. Intraperitoneal injections of SM caused two fold decrease of the number of cancer cells in ascite of mice with Ehrlich’s ascite tumor; increase to 24% the life span of mice with limphosarcoma as compared to the control. Intraperitoneal injections of SM emulsions in 10% Tween-80 and Cremophore–EL in mice with ascite tumor Krebs-2 suppress tumor growth to 76 and 57%, respectively.

Many serious human diseases are associated with dysregulation of inflammatory processes which is accompanied with overproduction of reactive oxygen species or reactive nitrogen species in the affected tissues. We showed that SM caused significant and dose-dependent decrease of NO production in LPS activated J-774 murine macrophages. Incubation of the cells in the presence of SM resulted in the decrease of NO production to the level in the control cells. In vivo, using murine model of acute inflammation, we showed that SM significant decreases the value of the inflammation index.

It was showed the antiviral activity of SM. The mechanism by which SM protects cells from infection with influenza A virus (IAV) was investigated: SM treatment leads to a reduction in the number infected MDCK and human lung cells, causing 10-fold decrease of virus titer relative to control. Analysis of different SM treatment protocols suggested that the antiviral effect of SM was limited to an early step in the virus replication cycle.

Obtained data indicated that novel derivative of glycyrrhetinic acid SM exhibits spectrum of biological activities: anti-cancer, anti-inflammatory and antiviral in vitro and in vivo and may consider as multitarget therapeutic.


90

Mechanochemically Assisted Extraction of Bio-active Compounds from

Lignocellulosic Material

I.O. Lomovskiy, O.I. Lomovsky, D.V. Orlov

Institute of solid state chemistry and mechanochemistry, 630090, Russia,

Novosibirsk, Kutateladze St.,18 E-mail: [email protected]

Mechanochemical treatment includes different types of mechanical loading is widely spread for

controlling physicochemical properties of solid compounds. As result the product of treatment has

high surface area, increased surface energy, decreased diffusion difficulties, high number of

contacts of reagents. The properties of product (so-called mechanocomposite) are altered from usual

mixture of fine powdered components.

Usage of mechanochemical approaches allow to realize complex waste-free processing of plant

based on increasing of extraction yield of low-molecular bioactive substances [1]. One of the main

parameter of low-molecular compounds is the thermal and mechanical stability of target

components. It was shown, that there are conditions that allow production of mechanocomposites

without degradation of active components.

The mechanocomposites posses the increased yields of active components. The yield can be

increased 2 times for serotonin [2], 1.5 times for green tea catechins [3].

The properties of product of mechanochemical treatment strongly depends on intensity and type

of mechanochemical loading and reagents. In case plastic deformation of cellulose matrix of

material there are solid-state reactions between substances that occur during mechanical loading.

This approaches are used to produce other products: chelate complexes of silica [4], salt forms of

triterpene acids, hypericin’s salt [5], ecdysteroids glycosides.

References [1]. Lomovsky O., Lomovsky I. Mechanochemically Assisted Extraction // in Enhancing Extraction Processes in the

Food Industry, ed. by N. Lebovka, E. Vorobiev, F. Chemat, NY - London: CRC Press, 2011, [2]. Lomovskiy O.I., Moshkin M.P. Serotonin containing powder from plant raw // 13th annual conference YUCOMAT

2011, Herceg Novi, p.156. [3]. I.O. Lomovsky Stability of Green Tea Catechins in the Solid Phase and Aqueous Solutions // Chemistry for Sustainable Development, V. 20 (2012), p 215 - 220

[4]. Shapolova E.G., Lomovskiy O.I. Mechanochemical solubilization of silica dioxide by polyphenolic compounds from plant material // Chem. of Plant Raw Mat., 2011, N. 4, p. 145-152. [5]. Lomovskii, A.A. Politov Mechanochemical neutralization of hypericin in Hypericum perforatum studied by spectroluminescence // Chemistry of Natural Compounds, Vol. 50, No. 2, May, 2014, p. 247-250.


91

Evolution of Biological Activity from Usnic Asid to Its Derivatives

O.A. Luzina,1 D.N. Sokolov,1 M.E. Rakhmanova1,2 and N.F. Salakhutdinov1,2

1Novosibirsk Institute of Organic Chemistry, 630090, Russia, Novosibirsk, Lavrentèv ave., 9

2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova st., 2


Chemical modification of natural compounds to obtain new promising bioactive compounds is

one of the most popular tendencies in medicinal chemistry. Usnic acid is an abundant secondary

metabolite of lichens, which shows a variety of biological activities, including antibacterial,

antiviral and cytotoxic. Its usage in the pharmacopoeia is restricted by moderate level of activities

and high hepatotoxicity. We developed some methods for modification of usnic acid structure in

order to enhance several types of biological effects.

HO

OH

OO

OO

OH

O

HO

OO

O O

OH

O

OH

HO

OH

O

OO

OH

HO

O

OO

HO

N

N

R

O

OH

O

O

OO

OH

R

HO

OH

OO

O

O

NHR

SN

NH2

HO

OH

O

O

OO

OH

HO

OH

O

OO

OH

SN

NH N

R

SR

The study was supported by the Russian Foundation for Basic Research (grants № 13-03-00810

and № 15-03-10051)


92

Enhancement of Radical Producing Activity by Anticancer Quinone-chelator in

the Presence of Metal Ions

I.D. Markova,1 O.Yu. Selutina,1 K.Yu. Fedotov,1 L.G. Fedenok,1

N.E. Polyakov,1 L.M. Weiner2

1Institute of Chemical Kinetics and Combustion, Institutskaya St. 3, Novosibirsk, 630090, Russia;

2Weizmann Institute of Science, Rehovot, 76100, Israel.


Anthraquinones, known as anthracycline antibiotics, are widely employed in cancer therapy.

Currently, elucidation of the mechanisms of drug activity at the molecular level and finding ways to

increase their efficiency is an important scientific problem. At the present work the quinone which

can form complexes with various metal ions: (2-phenyl-4-

(butylamino)naphtho[2,3-h] quinoline-7,12-dione) has been

synthesized to study the effect of chelation of metal ions on the

generation of free radicals in photoinduced processes. The purpose of

the present work is to figure out how the binding with the metal ion

affects the ability of the quinone to generate free radicals. We tried to investigate this question by

using the NMR and CIDNP (chemically induced dynamic nuclear polarization) techniques.

The chelate complex formation of this quinone with the metal ions (Fe2+, Ca2+ and Zn2+) was

proved using the optical absorption and NMR methods. It was demonstrated that binding with metal

ions results in significant changes in absorption spectrum of quinone. The stoichiometry of chelate

complexes with these metal ions is 2:1. The stability constants and extinction coefficients of all

complexes were calculated.

To study the influence of metal binding on the efficiency of the generation of free radicals by

NMR and CIDNP techniques, the photoinduced oxidation of NADH and its synthetic analog, 2,6-

dimethyl-3,5-dicarbomethoxy-1,4-dihydropyridine (DHP) by quinone was investigated in the

presence of Zn and Ca ions. The significant increase of polarization intensity and the yield of

reaction products after photolysis was observed in the presence of these metal ions. Increase in the

CIDNP intensity and the yield of reaction product allows one to suggest that the reactivity of the

quinone increased in the presence of metal ions. Taking into account that the free radicals

production is the key step of anticancer activity of quinones, the present results indicate the

perspectives of its practical application.

O

O

N

Ph

N

H


93

Identification of Target Genes Involved in Soloxolone Methyl-induced

Carcinoma Cell Apoptosis with Microarray Data

A.V. Markov,1 E.B. Logashenko,1 A.E. Kel,3 O.V. Kel-Margoulis,3 O.V. Salomatina,2

N.F. Salakhutdinov,2 M.A. Zenkova1

1 Institute of Chemical Biology and Fundamental Medicine, Siberian Branch Russian Academy of Sciences, 8, Lavrent’ev ave., Novosibirsk, 630090, Russia

2 N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, Siberian Branch Russian Academy of Sciences, 9, Lavrent’ev ave., Novosibirsk, 630090, Russia

3 GeneXplain GmbH, 10B, Am Exer, Wolfenbuettel, 38302, Germany E-mail: [email protected]

Chemical modification of natural bioactive compounds and metabolites is important area in drug development. Triterpenoids are considered as promising building blocks for synthesis of new highly active drugs due to their wide spectrum of biological activities and low toxicity. New glycyrrhetinic acid derivative methyl 2-cyano-3,12-dioxo-11-deoxo-18βH-glycyrrhet-1(2),9(11)-dieneoate (Soloxolone methyl), was obtained by the direct modification of A- and C-rings of glycyrrhetinic acid. We showed that Soloxolone methyl (SM) displays high antiproliferative activity with respect to cancer cells, inducing cell death by caspase-dependent intrinsic apoptosis pathway [1].

To find the molecular targets of SM we investigated its effect on human carcinoma cells KB-3-1 by gene expression profiling using HumanHT-12 v4 BeadChip (Illunmina, USA). The analysis of obtained raw data using geneXplain platform 3.0 (GeneXplain GmbH, Germany) showed that cell incubation with SM resulted in significant time-dependent changes of expression levels of 311 genes (fold change >2, p<10-3). The functional analysis of differentially expressed genes in GeneOntology database detected significantly over-represented GO terms (p<10-10) included response to endoplasmic reticulum stress, cellular response to stress, response to oxidative stress, regulation of apoptotic process, etc. Analysis of promoter [-500 bp, +100 bp] regions of detected up- and down-regulated genes in TRANSFAC 2012.3 database revealed a set of transcription factors (for instance, SRF, USF1, USF2, ATF3, JUN), which probably respond to SM action. Using TRANSPATH database, we identified as well a set of master regulators (as example, p22phox, p70S6K2, pkmyt1, cxcr4, LAMA5) which seems like activated by SM.

The obtained data confirm the ability of SM to induce of apoptosis of cancer cells as shown previously [1]. Incubation of carcinoma cells in the presence of SM leads to EPR stress, overexpression of p53-regulated proapoptotic genes, activation of Egr1/ATF3/NAG1 and KLF4/p21 signaling pathways. Intracellular targets of SM were identified.

This work was supported by RAS programs “Molecular and Cellular Biology” and “Basic Sciences to Medicine” (No. 10), RFBI No. 12-04-31254, RFBI No. 14-03-3008, Scholarship of the President of the Russian Federation SP-408.2012.4, Grant in Support of Scientific Schools SS-1350.2014.4

References: 1. E.B. Logashenko et al. (2011) Synthesis and pro-apoptotic activity of novel glycyrrhetinic acid derivatives, ChemBioChem, 12(5): 784-94.


94

New Approaches to Cu (I) Catalyzed ATRA/ATRC Reactions for Stereoselective

Synthesis of Functional Substituted Small Carbocycles: Convenient Building

Blocks for New Agrochemicals and Drugs

A. Mikaelyan,1 A. Grigoryan,2 and N. Asatryan1

1Department of Chemical Technologies and Environmental Engineering, National Polytechnic

University of Armenia, 0009(Zip code), Republic of Armenia, Yerevan, Teryan St., 105 2Department of Medical Chemistry, Yerevan State Medical University After Mkhitar Heratsi,

0025(Zip code), Republic of Armenia, Yerevan, Koryun St., 2


Transition metal catalyzed atom transfer radical (ATR) reactions, were proven to be one of the

most powerful methods for the synthesis of carbocycles. Active catalysts derived from copper

complexes are very attractive for industrial application. In the present work we suggest catalytic

diastereoselective preparation of a wide variety of derivatives of small carbocycles from readily

available starting materials or reaction intermediates. Two step procedures for the creating

cyclobutane subunits are elaborated: sequential addition-cyclization (ATRA/ATRC) reactions for

1,3-alkadienes; or ATRA to unsaturated substrates and next reductive cyclopropanation of 1,3-

dihalides via metals [1]. The crucial role of solvent in ATR reactions as a compatible co-ligand,

homogenising and stabilizing media for catalyst was reported [2]. The most known catalytic

systems showed the several drawbacks: prolonged reaction time and relative high temperatures for

complete conversion. More polar solvent DMSO, are controlled the activity of copper salts and

stabilized reaction intermediates, they even can change the structure of catalyst by coordination

with metal centre. Application of DMSO as solvent-coligand in observed ATRA/ATRC reactions

brings to extremely increase the activity of catalytic complex. Further transformations of obtained

products bring to a number of new agrochemicals and drugs with improved properties.

X

Cl

Cl

O

O R

Cl

O

O RCl3C

Cl

Acknowledgments: This work was supported by SCS MES RA, research project № SCS

13Ap_2e021.

References [1] A.R. Mikaelyan, Russian Journal of Organic Chemistry. 2010, 46(11), 1662-1666 [2] F. Bellesia, A.J. Clark, F. Felluga, A. Gennaro, A.A. Isse, F. Roncaglia, and F. Ghelfi, Adv. Synth. Catal. 2013, 355, 1649-1660


95

The Synthesis of Chiral Fluorine Containing Compounds

Based on Monoterpenoids

O. Mikhalchenko,1 D. Korchagina,1 K. Volcho1,2 and N. Salakhutdinov1,2

1Novosibirsk Institute of Organic Chemistry, 630090, Russia, Novosibirsk, Lavrentjev ave., 9

2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova St., 2


Previously we have studied reactions of monoterpenoid 1 with various aromatic aldehydes in the

presence of K10 montmorillonite clay. It was found [1] that hexahydrochromenes 2 are formed as

the main products in these reactions. It was shown that some of the obtained products exhibit high

analgesic activity in vivo in combination with low toxicity [2].

In this work, we have developed a simple and efficient method for synthesizing analogs of

hexahydrochromenes 2 containing fluorine instead of the hydroxy group. We carried out the

reactions of monoterpenoid 1 and aldehydes in the presence of boron trifluoride etherate and water

to get fluorine-containing compounds 3. Boron trifluoride etherate acts both as a catalyst and as a

source of fluorine in these conditions. Various aliphatic and aromatic (including those containing

phenolic group) aldehydes can be successfully used in the reaction with diol 1.

Isopulegol 4 and citronellal 5 in the same conditions can also be used as starting monoterpenoids

in the synthesis of fluorinated products leading to the octahydrochromenes 6.

OHCHO

4 5

O R

F

H

6

RCHO RCHO

Introduction of the fluorine atom in the biologically active compounds can lead to increased or

changed their physiological activity. For example, fluorine containing compounds 3 in contrast to

hydroxy analogs 2 showed anti-influenza activity.

This work was supported by the Russian Science Foundation (project № 15-13-00017).

References [1] Il'ina, Irina V.; Volcho, Konstantin P.; Mikhalchenko, Oksana S.; Korchagina, Dina V.; Salakhutdinov, Nariman F. // Helvetica Chimica Acta, 2011 , vol. 94, № 3 p. 502-513; [2] Mikhalchenko O., Il’ina I., Pavlova A., Morozova E., Korchagina D., Tolstikova T., Pokushalov E., Volcho K., Salakhutdinov N. // Medicinal Chemistry Research, 2013, vol. 22, № 6, p.3026–3034.


96

Design, Synthesis and Structure-anxiolytic Activity Relationship Study of New

Pyrrolo[1,2-a]pyrazine TSPO Ligands

G.V. Mokrov, O.A. Deeva, A.S. Pantileev, V.V. Silman, T.A. Gudasheva, S.A. Yarkov,

M.A. Yarkova and S.B. Seredenin

Federal State Budgetary Institution “Research Zakusov Institute of Pharmacology”,

125315, Russia, Moscow, Baltiyskaya St., 8


Translocator protein 18 kDa (TSPO) is a new target for creating effective anxiolytics free from

side effects of benzodiazepines [1]. This receptor activates cholesterol transport from the outer to

the inner mitochondrial membrane that is the rate-limiting step of neurosteroids biosynthesis.

Neurosteroids are potent positive allosteric modulators of GABAA receptor, which plays an

important role in the pathophysiology of anxiety disorders; they have their own binding sites on

GABAA, which are different from the benzodiazepine [2].

In this work we designed a new type of TSPO ligands relating to 1-arylpyrrolo[1,2-a]pyrazine-3-

carboxamides 1. Using molecular docking program AutoDock 4 was shown the high theoretical

affinity to TSPO for new ligands (Ki 10-8-10-6 M). Convenient and efficient synthetic method was

proposed to obtain target compounds 1. The key step of this method is [3+3]-cycloaddition of

arylpyrrolketones 2 to 2-azidoacrylamides 3.

Radioligand in vitro binding studies were performed to investigate the affinity of two

synthesized compounds against TSPO. Both compound demonstrated high TSPO affinities

comparable with that of specific TSPO ligand PK11195 (Ki 5.2*10-8 and 5.3*10-7 M).

Anxiolytic activities of synthesized compounds were investigated in Balb/c mice using the open-

field test and in CD-1 mice using elevated plus-maze test. Some of compounds displayed high level

of anxiolytic efficacy comparable with that of diazepam. The involvement of TSPO receptor in the

mechanism of anxiolytic activity new compounds was proved by antagonism of the most active

compound with TSPO selective inhibitor PK11195.

References [1] R. Rupprecht, V. Papadopoulos, G. Rammes, T.C. Baghai, J. Fan, N. Akula, G. Groyer, D. Adams, M. Schumacher,

Nat. Rev. Drug Discovery 2010, 9, 971−988. [2] R. Rupprecht, Psychoneuroendocrinol. 2003, 28, 139–168.


97

Protein Ruthenation and DNA Alkylation: New Ruthenium Complexes and

Their Anticancer Activity

A.A. Nazarov, Y.N. Nosova and E.R. Milaeva

Department of Medicinal Chemistry and Fine Organic Syntheses, M. V. Lomonosov Moscow State

University, 119991, Russia, Moscow, Leninskie gory 1/3


Alkylating agents have dominated cancer chemotherapy for many years and many examples of

alkylating agents have been approved for cancer treatment. Platinum complexes are often

considered also as alkylating agents as they form coordination bonds to DNA nucleobases. The

potency of platinum complexes in the chemotherapy is unprecedented and they are used to

treat many types of tumors. However, their disadvantages, in particular resistance

development and side effects, have driven the development of metal complexes comprising

metal centers other than platinum. Ruthenium compounds take a prominent position in this

development process as two compounds have entered clinical trials. Organometallic

ruthenium complexes are among the most promising compounds in preclinical development

and although some of them are essentially non-cytotoxic in cancer cell lines, they have

demonstrated anticancer activity in vivo. The ruthenium scaffold can be equipped with

functionalities that provide specific properties. In his work, we have designed a new compound

that features the ruthenium organometallic framework conjugated with organic alkylating ligand.

Such compounds should be able to alkylate DNA and at the same time coordinate to proteins

through the ruthenium center. The anticancer activity of the complexes is widely driven by the

cytotoxicity of the alkylating moiety. In a mass-spectrometry experiment, cross-linking of DNA and

protein fragments by a new ruthenium complexes was observed.[1]

The financial support of Russian Science Foundation (14-13-00483) is gratefully

acknowledged.

References [1] A.A. Nazarov, S.M.Meier, O. Zava,; Y.N. Nosova, E.R. Milaeva, C.G. Hartinger,; P.J. Dyson, , Dalton Trans., 2015, 44, (8), 3614-3623.


98

Synthesis of Aromatic and Reduced Berberine derivatives and Their

Hypolipidemic Activity

I.V. Nechepurenko,1 U.A. Boyarskikh,2 M.V. Khvostov,1 D.S. Baev,1

N.I. Komarova1 and N.F. Salakhutdinov1

1N.N. Vorozhtzov Novosibirsk Institute of Organic Chemistry of the Siberian Branch of Russian

Academy of Sciences, 630090, Russia, Novosibirsk, ave. Lavrentjeva, 9.

2Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of Russian Academy of Sciences, 630090, Russia, Novosibirsk, ave. Lavrentjeva, 8.


The decrease of total cholesterol and triglycerides levels in the blood is considered the main

criterion in the prevention of cardiovascular diseases. The statins currently used for this purpose

inhibit HMG-CoA reductase, leading to significant side effects: elevated liver function tests,

myopathy, myalgia and in some cases rhabdomyolysis. Therefore, the search continues for novel

cholesterol-lowering compounds with different to statins mechanism of action. Berberine 1, a

widespread plant isoquinoline alkaloid, has been reported to possess cholesterol- and triglyceride-

lowering effects. It is thought that berberine lowers cholesterol by up-regulation of LDLR

expression in liver cells.

For the study of structure - hypolipidemic activity relationships we have synthesized a number of

aromatic and reduced berberine derivatives with various substituents in C-9 and C-12 positions. It

was tested the influence of berberine derivatives on the LDLR mRNA expression in human

hepatoma cells HEPG2. For active compounds, their ability to lower cholesterol and triglycerides

levels were studied in vivo on a model of Triton WR1339-induced hyperlipidemia.

This work was supported by the Russian Foundation for the Basic Research, project No 15-03-

10051


99

Discovery of Small Molecule Inhibitors of Tick-Borne Flaviviruses

Reproduction

D.I. Osolodkin,1,2 L.I. Kozlovskaya,1 V.A. Palyulin,2 G.G. Karganova1 and N.S. Zefirov2

1Chumakov Institute of Poliomyelitis and Viral Encephalitides, 142782, Russia, Moscow

2Department of Chemistry, Lomonosov Moscow State University, 119991, Russia, Moscow, Leninskie Gory, 1/3


Tick-borne viral diseases represent a significant health concern due to severity of clinical

manifestations and limited options for prophylaxis and treatment. The most important among them

is tick-borne encephalitis, widely distributed in Europe, Russia and China; several different viruses

belonging to the same Flavivirus genus are circulating in local natural foci. Efficient small molecule

antiviral drugs against these diseases are required to provide a therapy complimentary to

vaccination and immunoglobulin treatment.

Inhibition of viral fusion is a prominent mechanism of action of antiviral drugs. Putative fusion

inhibitors for flaviviruses can be identified with the help of virtual screening. In this report we

present an overview of our recent efforts on identification and structure-based design of small

molecule inhibitors of tick-borne encephalitis virus (TBEV) reproduction, acting on the fusion stage

[1,2], and analysis of their mode of inhibition based on molecular dynamics simulations [3].

Different classes of heterocyclic compounds, including 4-aminotetrahydroquinazolines [1], 1,4-

dihydropyridines [2], pyrido[2,1-b][1,3,5]thiadiazines [3], arylethynyl nucleosides, are shown to

possess the antiviral activity against TBEV, Omsk haemorrhagic fever virus, and Powassan virus at

submicromolar concentrations while being low toxic in the cell cultures. A putative binding mode

of an active compound in the pocket of envelope protein E was thoroughly studied by molecular

dynamics simulations, and structural basis for fusion inhibition was suggested. The findings of our

studies prepare the ground for further design of more advanced inhibitors of flavivirus reproduction.

The studies presented are partially supported by RFBR grants no. 14-03-31566, 14-03-

00469.

References [1] K. N. Sedenkova, E. V. Dueva, et al., Organic & Biomolecular Chemistry. 2015, 13, 3406-3415 [2] D. I. Osolodkin, L. I. Kozlovskaya, et al., ACS Medicinal Chemistry Letters. 2013, 4, 869-874 [3] E. V. Dueva, D. I. Osolodkin, et al., Molecular Informatics. 2014, 33, 695-708


100

Computational Tools for Multi-target Drug Design

V.A. Palyulin, E.V. Radchenko, N.S. Zefirov

Department of Chemistry, Lomonosov Moscow State University, Leninskie Gory, 1/3,

Moscow, 119991, Russia,

Drug design is a sophisticated process involving in the first steps the application of various

computational techniques including molecular modelling and QSAR, molecular docking and virtual

screening. Until recently the main criterion in the design of drug molecules was based on their

selective interaction with one binding site of one particular biotarget, while interactions with other

biotargets were considered as side effects. However in some cases simultaneous interaction of drug

molecules with two (or more) different biotargets is necessary for the proper therapeutic effect. The

Molecular Field Topology Analysis (MFTA) developed by the authors can be used as a QSAR tool

to optimize the activity and selectivity profiles using individual endpoint models and/or specially

constructed multi-target selectivity parameters. Recent advances of MFTA allow one to apply this

tool to the design of new antioxidants, enzyme inhibitors, receptor agonists, antagonists and

modulators, virus entry inhibitors, etc.

In MFTA the local features are compared for different molecules in the framework of a

supergraph approach which is based on the topological alignment of all training set structures

having similar scaffolds. MFTA models reveal the local molecular properties/features (partial

atomic charges, steric parameters, H-bond donor and acceptor ability, local lipophilicity, etc.) which

are necessary for the interaction of ligands with each biotarget. Alignment of these features can be

instrumental in the design of molecules binding to several desired biotargets while not binding to a

set of forbidden ones.

In combination with molecular modelling and molecular dynamics simulation MFTA

demonstrates a good performance in the design of either selective or multi-target compounds with

respect to a wide range of target macromolecules. Real world examples are given which confirm the

applicability of considered approaches for multi-target drug design.


101

Selective Functionalization of Eudesmane-type Methylenelactones by Cross-

coupling Reaction with Xanthine Derivatives

S. Patrushev,1,2 E. Shults1,2

1Laboratory of Medicinal Chemistry, N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry,

SB RAS, 630090, Russian Federation, Novosibirsk, Lavrentuev Ave, 9 2Department of Organic Chemistry, Novosibirsk State University, 630090, Russian Federation,

Novosibirsk, Pirogova St., 2


Xanthine derivatives exhibit a wide range of useful biological properties, which include affinity

to adenosine receptors, phosphodiesterase inhibition, antiproliferative and analgesic activities [1-3].

In recent years these compounds are considered as acetylcholinesterase inhibitors and have potential

in the treatment of vascular dementia or Alzheimer's disease [4-5].

Herein, we present a synthesis of novel hybrid compounds containing fragments of xanthine and

eudesmanolides by Pd-catalyzed cross-coupling reaction. We obtained the data on the effect of

conditions (the influence of catalytic system, base, additive, solvent and temperature), the structure

of lactones 1-3 and 8-bromo derivatives of xanthine 4a-e on the ratio of products 5a-e, 6a-e, 7a-e,

8a-e or 9a, 10a. Conditions for the selective formation of compounds 6a,c,d,e (yield > 90%) and

10a (yield 76%) were found.

Inhibiton data of acetylcholinesterase by novel hybrid compounds will be presented.

O

Y

O

H

HO

Y

O

H

H

H

1-3 N

N

N

N R2

OR1

O

6a-e 48-92%8a-c 53%10a 76%

OO

YH

H

H

N

NN

N

R2

OO

R1

5a-e 44-52%7a-c 46%9a 52%

i) 4a-e (1.2 eq), Pd(OAc)2 (4 mol %), o-Tol3P (16 mol %), NEt3, DMF, 120oC, 20h;

ii) 4a-e (1.5 eq), Pd(OAc)2 (4 mol %), o-Tol3P (16 mol %), TBAB (1 eq), NEt3, DMF, 140oC, 40h.

XY=CH2 1; XY=OCH2 2; XY=CCl2CH2 3; R1=R2=Me 4a;R1=Н, R2=Me 4b; R1=Me, R2=Н 4c; R1=Bu, R2=Me 4d;R1=Me, R2=Bu 4e; XY=CH2, R1=R2=Me 5a,6a;XY=CH2, R1=Н, R2=Me 5b,6b; XY=CH2, R1=Me, R2=Н 5c,6c;XY=CH2, R1=Bu, R2=Me 5d,6d; XY=CH2, R1=Me, R2=Bu 5e,6e;XY=OCH2, R1=R2=Me 7a,8a; XY=OCH2, R1=Н, R2=Me 7b,8b;XY=OCH2, R1=Me, R2=Н 7c,8c; XY=CCl2CH2, R1=R2=Me 9a,10a

X

N

N N

N

O

O

R1

Br

R2

ii)

4a-e

X X

i)

This work was supported by the Russian Science Foundation (project No 14-13-00822).

References [1] A.J. Szentmiklosi, A. Cseppento, R. Gesztelyi, et al., Curr. Med. Chem. 2011 18 (24) 3695; [2] R.B. Derek, R.S.A. Jonathan, J.C. Brendan, et al., J. Med. Chem. 1994 37 (4) 476. [3] K.S.S. Praveena, S. Durgadas, N.S. Babu, et al., Bioorg. Chem. 2014 53 8. [4] G.M. Cunha, P.A. Farias, G.S. Viana, Behav. Pharmacol. 2002 13 (2) 149. [5] T. Mohamed, W. Osman, G. Tin, P.P.N. Rao, Bioorg. Med. Chem. Lett. 2013 23 (15) 4336.


102

Approaches for Improvement of Drugs Solubility

G.L. Perlovich

Division of Physical Chemistry of Drugs, Krestov’s Institute of solution chemistry of the Russian

Academy of Sciences, Akademichesky str., 1, Ivanovo, 153045, Russia


One of the key goals in pharmaceutics is solving the problem of poor drug solubility. Solubility

is a very important property of drug substances which determines both the optimal therapeutic doses

and probable side effects. It is worth mentioning that the analysis of up-to-date data bases of drug

compounds shows that biological activity properties and solubility values are inversely

proportional. In other words, the substances highly affinitive to the receptors are very poorly soluble

in aqueous solutions. Therefore, it is necessary to create well soluble forms in order to strengthen

the position of the drug compounds on the market. There are a number of approaches to producing

such forms. However, one of the most promising techniques is obtaining solvatomorphic

modifications, cocrystals and crystallosolvates/crystallohydrates in particular.

The last decade saw the development of an approach of producing soluble pharmaceutical

systems based on cocrystal design. One of the advantages of cocrystals is their high thermodynamic

stability and essential improvement of solubility in comparison with individual active

pharmaceutical ingredient (API). Additional valuable advantages of cocrystal formation for the

pharmaceutical industry include the possibility of extending the life cycles of old APIs and the

opportunity of intellectual property protection. Moreover, selection of different components at

cocrystal design gives opportunity to carry out “fine adjustment” of cocrystal properties and, as

consequence, to create a wide spectrum of products on the market.

In the presentation it will be considered the questions connected with design, screening,

crystal structures and optimization of various properties of pharmaceutical cocrystals.

This work was supported by the Russian Scientific Foundation (№14-13-00640).


103

Conjugation of N-Acetyl-L-Cysteine with Natural and Synthetic

Naphthoquinones as a Short Route to Novel Bio-Active Products

S. Polonik, Yu. Sabutskii and E. Yurchenko

G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, 690022, Russia, Vladivostok, Prospect 100-let Vladivostoku, 159


1,4-Naphthoquinones are widely distributed in Nature as metabolites of various living organisms1,2. These substances have wide spectrum of bioactivity: anticancer, antibacterial, antiprotozoal, antimalarial and others1,2. Some of them are using in medicine. Quinones exert their actions through radical reactions: as prooxidants, reducing oxygen to reactive oxygen species; as antioxidants and as electrophiles, forming covalent bonds with tissue nucleophiles. In most cases naphthoquinones have low solubility that prevent their wide application and biotesting.

In order to improve the quinone water solubility we conjugated various 5,8-dihydroxy-1,4-naphthoquinones (naphthazarines) with sulfur amino acid N-acetyl-L-cysteine (HSCH2CHNHAcCO2H, NAC-SH)1–3. N-Acetyl-L-cysteine is a non-toxic water soluble pharmaceutical drug widely used as mucolitic agent, detoxifier for detoxification of paracetamole overdosing and as a precursor of gluthatione. Therefore, conjugation of substituted hydroxy-1,4-naphthoquinones with a N-acetyl-L-cysteine led to new useful S-amino acid derivatives. The two type conjugates were created. The first type conjugate 1 has NAC-SH attached to quinone core through S-bond3,4, and in the second type conjugate 2 NAC-SH connects with quinone nucleus by methylene chain5. These conjugates have a better water solubility and structurally related with bio-active echinochrome 3 – the active principle of Russian cardioprotective drug HistochromeTM 6.

All the compounds were tested on cytotoxic activity against mouse Ehrlich carcinoma cells and

were nontoxic in vitro in concentrations up to 100 µM. The antioxidant activities of the obtained NAC-SH conjugates were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay in 30% and 96% water ethanole and compared with echinochrome, ascorbic and gallic acids. Structure-antioxidant activity relationships are discussed. References [1]. R.H. Thomson, Naturally occurring quinones, 3rd ed.; Chapman & Hall: London, 1987. [2]. R.H. Thomson, Naturally occurring quinones, 4rd ed Blackie Academic and Professional: London – NY, 1997. [3]. Y. Sabutskii, N. Polonik, V. Denisenko, P. Dmitrenok, S. Polonik, Rus. J. Org. Chem. 2013, 49, 1157-1164. [4]. Y. Sabutskii, S. Polonik, Rus. J. Org. Chem. 2013, 50, 755-756. [5]. Y. Sabutskii, S. Polonik, V. Denisenko, P. Dmitrenok, Synthesis. 2014, 46, 2763-2770. [6]. N. Mishchenko, S. Fedoreev, V. Bagirova, Pharm. Chem. J. 2003, 37, 48-53.


104

Physicochemical Approaches to the Study of New Drug Delivery Systems Based

on Naturally Occurring Polysaccharides and Oligosaccharides

N.E. Polyakov,1 O.Yu. Selyutina,2 A.V. Dushkin3

1Institute of Chemical Kinetics and Combustion, 630090, Russia, Novosibirsk, Institutskaya St., 3 2Novosibirsk state University, 630090, Russia, Novosibirsk, Pirogova St.,2

3Institute of Solid State Chemistry and Mechanochemistry, 630128, Russia, Novosibirsk, Kutateladze St., 18


Use of natural oligosaccharides and polysaccharides as complexants for delivery of drug

compounds significantly increases their therapeutic activity and storage stability, and also reduces

the side effects. Application of modern physical methods allows to obtain unique information about

the structure, dynamics, and physicochemical properties of supramolecular inclusion complexes of

the drugs. The report will presents some examples illustrating the possibilities of physical methods

to study various drug delivery systems (DDS) in the solid phase and in solution, as well as examples

of application of these techniques for elucidation of the mechanisms of increasing the

bioavailability of drug compounds in the complexes. Interesting and practically important results

were obtained using the natural polysaccharide arabinogalactan and an oligosaccharide -

glycyrrhizic acid, as the DDS, with a wide range of drug compounds. The formation of inclusion

complexes, their structure and properties have been studied in solid state by X-ray diffraction and

thermal analysis, and in solutions by gel permeation chromatography, HPLC, SPR and NMR. In

particular, it was demonstrated that:

- Complexation increases by tens or hundreds times the solubility of poorly soluble drugs and

their bioavailability [1].

- Complexation significantly increases the stability of “guest” compounds including oxidative

stability and photostability [2].

- DDS has an impact on the permeability of cell membranes [3]. These results allowed to

propose the mechanisms of enhancement of drugs activity in supramolecular complexes.

References [1]. Y. Chistyachenko, A. Dushkin, N. Polyakov, M. Khvostov, T. Tolstikova, G. Tolstikov, N. Lyakhov, Drug

Delivery. 2014 (doi: 10.3109/10717544.2014.884655). [2]. I. Apanasenko, O. Selyutina, N. Polyakov, L. Suntsova, E. Meteleva, A. Dushkin, P. Vachali, P. Bernstein,

Archives of Biochemistry and Biophysics. 2015, 572, 58-65 (DOI: 10.1016/j.abb.2014.12.010). [3]. O. Selyutina, N. Polyakov, D. Korneev, B. Zaitsev, Drug Delivery, 2014 (doi:10.3109/10717544.2014.919544).


105

Modification of Deoxycholic Acid Ring A with S- and N-Containing

Functional Groups

I.I. Popadyuk,1 O.V. Salomatina,1 and N.F. Salakhutdinov1,2

1 N. N. Vorozhtsov Novosibirsk institute of organic chemistry SB RAS, 630090, Russian Federation,

Novosibirsk, acad. Lavrent’ev ave., 9 2 Novosibirsk State University, 630090, Russian Federation, Novosibirsk, Pirogova Str., 2


Design of new drug by chemical transformation of natural metabolites is an actual medicinal

chemistry problem [1]. Modification of the steroid skeleton by different heteroatom containing

functional groups leads to increasing of the native and/or occurrence of a new biological activity,

which is primarily due to the heteroatom ability to form additional hydrogen bonds and

consequently to increase the hydrophilicity of the original molecule and enhance receptor binding

[2].

In this work, new deoxycholic acid derivatives were synthesized by modification of the steroid

skeleton ring A. Key intermediates in the synthesis were methyl C-3β-epoxy-12-oxocholan-24-oate

2 and methyl 2-hydroxymethylene-3,12-dioxocholan-24-oate 3 obtained by interaction between C-3

carbonyl group of compound 1 and dimethylsulfoxonium methylide and a condensation reaction of

compound 1 and with methyl formate respectively. Target compounds were obtained with high

yields via epoxide ring opening reaction between 3-epoxyderivative 2 and S- and N-nucleophiles,

and condensation of 2-hydroxymethylene derivative 3 with primary and secondary amines.

Structures of all new compounds were confirmed by 1H and 13C NMR and high resolution mass

spectrometry.

This work was supported by RFBR № 14-03-31408 References [1] D. J. Newman and G. M. Cragg, J. Nat. Prod. 2012, 75, 311−335 [2] M. Ibrahim-Ouali, L. Rocheblave, Steroids. 2010, 75, 701-709


106

Synthesis, Modeling and Molecular Design of N,N-disubstituted

2-Aminothiazolines as a New Class of Butyrylcholinesterase and

Carboxylesterase Inhibitors

E.V. Radchenko,1 G.F. Makhaeva,2 N.P. Boltneva,2 S.V. Lushchekina,2,3

I.V. Serkov,2 A.N. Proshin,2 V.A. Palyulin,1,2 and N.S. Zefirov 1,2

1 Department of Chemistry, Lomonosov Moscow State University,

119991, Russia, Moscow, Leninskie gory, 1/3 2 Institute of Physiologically Active Compounds Russian Academy of Sciences,

142432, Russia, Chernogolovka, Severny proezd, 1 3 Emanuel Institute of Biochemical Physics Russian Academy of Sciences,

119334, Russia, Moscow, Kosygina str., 4


Serine esterase enzymes present promising pharmacological targets in a variety of conditions. In

particular, selective butyrylcholinesterase (BChE) inhibitors can be used for the cognition-

enhancement therapy of Alzheimer’s disease, while selective carboxylesterase (CaE) inhibitors are

employed as modulators of hydrolytic metabolism of ester- or amide-containing drugs. The esterase

inhibitors based on the 5-halomethyl-2-aminothiazoline scaffold were found to open wide

possibilities for the control of their esterase profile. The compounds did not inhibit

acetylcholinesterase (AChE) while the most potent compounds against BChE and CaE had IC50

values of 0.22 to 2.3 µM. Docking simulations provided insight into differences in their binding and

activity. Using the Molecular Field Topology Analysis (MFTA) method, the models for the

relationships between their structure and inhibitor activity toward AChE, BChE and CaE were built.

Molecular design was performed and the focused libraries of potentially active and selective

inhibitors of butyrylcholinesterase and carboxylesterase were proposed.

This work was supported by the Russian Foundation for Basic Research (grants no. 14-03-

01063 and 15-03-09084), the Medicinal Chemistry program of the Russian Academy of Sciences,

and the Dynasty Foundation grant to S.V.L.

References [1] G.F. Makhaeva, N.P. Boltneva, S.V. Lushchekina, O.G. Serebryakova, T.S. Stupina, A.A. Terentiev, I.V. Serkov,

A.N. Proshin, S.O. Bachurin, and R.J. Richardson, Eur. J. Med. Chem. 2015, in press. [2] E.V. Radchenko, G.F. Makhaeva, N.P. Boltneva, O.G. Serebryakova, I.V. Serkov, A.N. Proshin, V.A. Palyulin, and

N.S. Zefirov, Russ. Chem. Bull. 2015, in press.


107

Development of Thienopyridines as Anticancer Agents

J. Reynisson

School of Chemical Sciences, University of Auckland, New Zealand

The phosphoinositide specific-phospholipase C – γ enzymes are plausible anticancer targets

implicated in cell motility important to invasion and dissemination of tumour cells [1]. Virtual high

throughput screening was performed against this target and the hit compounds tested in a

biochemical and cell based assays [2]. The most active compounds were in the single digit micro-

molar range, which translated into ~15 µM for the most active compound in functional assays in

cells [2]. These compounds were tested against the National Cancer Institute’s human tumour cell

line panel (NCI60) panel of human tumour cell lines [3]. A class of thieno[2,3-b]pyridines showed

excellent growth arrest with derivative 1 (see figures) giving GI50 = 58 nM for the melanoma MDA-

MB-435 cell line [4]. The biological effect was further investigated finding that 1 inhibits DNA

synthesis in the nanomolar range, arrests the cell cycle in the G2/M phases and has severe

morphological changes on tumour cells [5]. Furthermore, structure activity relations studies (SAR)

revealed even more cytotoxic derivatives with GI50 values in the teens [6, 7].

NS

O

HN

Cl

ONH2

Derivative 1, GI50 = 58 nM (melanoma MDA-MB-435) Currently we are conducting further mechanistic experiments as well as preliminary

pharmacokinetic and toxicological investigations to elucidate the anticancer properties of the

thieno[2,3-b]pyridines.

[1] V. Kölsch, P.G. Charest, R.A. Firtel, J. Cell Sci., 121 (2008) 551-559. [2] J. Reynisson, W. Court, C. O’Neill, J. Day, L. Patterson, E. McDonald, P. Workman, M. Katan, S.A. Eccles, Bioorg.

Med. Chem., 17 (2009) 3169-3176. [3] R.H. Shoemaker, Nat. Rev. Drug Dis., 6 (2006) 813-823. [4] L. Feng, I. Reynisdóttir, J. Reynisson, Eur. J. Med. Chem., 54 (2012) 463-469. [5] E. Leung, J.M. Hung, D. Barker, J. Reynisson, Med. Chem. Comm., 5 (2014) 99-106. [6] H.J. Arabshahi, E. Leung, D. Barker, J. Reynisson, Med. Chem. Comm., 5 (2014) 186-191. [7] J.M. Hung, H.J. Arabshahi, E. Leung, J. Reynisson, D. Barker, Eur. J. Med. Chem., 86 (2014) 420-437.


108

Triazavirin – Antiviral Drug of a New Generation

V.L. Rusinov,1 V.N. Charushin,1,2 O.N. Chupakhin,1,2 O.I. Kiselev3

1Ural Federal University, Ekaterinburg, Russia,

2Organic Synthesis Institute, Ural Branch of RAS, Ekaterinburg, Russia 3Influenza Institute, Saint-Petersburg, Russia


Development of new effective drugs for prevention, treatment and cure of viral diseases is an actual task of medicinal chemistry due to a high danger of viral infections, their variability and appearance of highly pathogenic agents. As a result of joint research studies, carried out in Ural Federal University, Organic Synthesis Institute of the Ural Branch of the Russian Academy of Sciences, and Influenza Institute of the Ministry of Health Care, a new original class of non-nucleoside antiviral compounds, which are effective against influenza viruses, herpes, and a number of hemorrhagic fever diseases, has been advanced. The data obtained provide a good basis for the development of new effective antiviral drugs.

The first drug of this family is known as Triazavirin (sodium salt of 2-methylthio-6-nitro-1,2,4-triazolo[5,1-с]-1,2,4-triazin-7-one, dehydrate). It has passed successfully the full course of clinical trials as the drug against influenza virus, and has been registered on 28.08.2014 and involved in the Register of Drugs of the Russian Federation.*

Use of Triazavirin in antiviral therapy results in a lower duration of the main symptoms of influenza, facilitates normalization of the body temperature and decreases considerably the level of viruses isolated from patients.

In order to establish a plausible mechanism for antiviral action of Triazavirin, the target and its metabolism in organism, a computer modeling has been used to determine 3D-structure of potential sites for binding of the drug with hemagglutinin of influenza virus A/California/04/2009(H1N1). The data on reactivity of 6-nitroazolo-1,2,4-triazines proved to be a basis to suggest plausible conversions of Triazavirin and to carry out a number of model experiments, such as reduction of the nitro group (A) and oxidation of the alkylthio fragment (B). Also interactions with N- and S-nucleophiles, such as lysine, arginine, cysteine, and glutathione, which are capable to displace alkylthio or the nitro group (C), аs well as N-alkylation or N-glycosilation reactions (D) have been considered.

NN

NN

N

O

CH3SNO2

Na+

-Nu-

(A)

(B)

(C)

(С) [H][O]

Nu- 1

2

3 4

5

67

(D)

_________________________

*The drug has been launched into production by “Medsynthez” Enterprise (Novouralsk, Sverdlovsk Region) jointly with the Ural Center of Biopharmaceutical Technologies in the frames of cooperation with the «Skolkovo» Innovation Foundation.


109

2-Cyano Substituted Derivatives of Glycyrrhetinic and Deoxycholic Acids:

Synthesis and Biological Activities

O.V. Salomatina,1 I.I. Popadyuk,1 E.B. Logashenko,2 A.V. Markov,2 M.A. Zenkova,2

and N.F. Salakhutdinov1,3


Novosibirsk, acad. Lavrent’ev ave., 9 2Institute of Chemical Biology and Fundamental Medicine, SB RAS, 630090, Russian Federation,



One of the most important and perspective direction of medicinal chemistry, allows to get new

effective drugs, is a synthetic modification of natural metabolites. The study of the compounds that

have reliable data of biological activity is the most acceptable [1]. 18βH-Glycyrrhetinic acid (GA)

(plant metabolite) and deoxycholic acid (DCA) (animal metabolite) were chosen as a starting

material for further investigation, because they are widespread in nature, possesses a high

enantiomeric purity and a broad spectrum of native biological activities (anti-inflammatory,

antiviral, anticancer, immunostimulatory). Also, GA and DCA are biogenetic relatives, as both are

products of squalene bio-cyclisation, therefore a series based on GA and DCA of compounds was

synthesized using the following strategies: building up a 2-cyano-3-oxo-1-ene moiety in the A ring

and creating different carbonyl group and/or double bond in ring C

These derivatives and intermediates were then screened in vitro for their ability to inhibit NO

production and for their cytotoxic activities. Structures of all new compounds were confirmed by 1H

and 13C NMR and high resolution mass spectrometry.

This work was supported by program RFBR № 14-03-31408 References [1] D. J. Newman and G. M. Cragg, J. Nat. Prod. 2012, 75, 311−335[2] M. Ibrahim-Ouali, L. Rocheblave, Steroids. 2010, 75, 701-709


110

Patenting Polymorphs at the European Patent Office

I. Seelmann

European Patent Office, Germany, 80335 Munich, Bayerstrasse 34


For a Patent to be granted the subject-matter of the application must meet a number of criteria.

For the European Patent these criteria are laid down in the European Patent Convention (EPC). The

most important substantial requirements to be met are:

Clarity (Article 84 EPC)

Disclosure of the invention (Article 83 EPC)

Novelty (Article 54 EPC)

Inventive step (Article 56 EPC)

Unity of invention (Article 82, Rule 44 EPC)

For applications relating to polymorphic forms of pharmaceutical compounds a number of

frequently occurring issues has been identified. Some of them are specific for the “polymorph”

while others apply more generally to compounds as such.

Particular deficiencies in the patent application regularly occur in the following situations:

A new crystalline form is merely defined as “new form X”.

A polymorph is defined by unusual or unreliable parameters.

No clear description of the measurement method is given.

Seed crystals are used to obtain a new crystalline form.

Additional objections are usually raised in view of the prior art (usually pre-published

documents):

A (crystalline) form of the same compound is already disclosed in the literature, but can not be

compared with or distinguished from the “new form”.

Only a different form of the same compound is already known, but the new polymorph is merely

an alternative to the known (crystal) form.

A different form of the same compound is disclosed in the literature, but more than one “new”

form is claimed in the same patent application.


111

The Sea Urchin Embryo as a Promising Model in

Screening for Antimitotic Compounds

M.N. Semenova

Koltsov Institute of Developmental Biology RAS, 119334, Russian Federation,

Moscow, Vavilov St., 26


The sea urchin embryo is considered as a promising simple organism model for studying the

effects of antiproliferative agents. Due to the abundance and accessibility of gravid animals,

straightforward artificial spawning, fertilization and rearing, rapid synchronous development,

embryo optical transparency and high penetrability to different chemicals sea urchin embryos are

extensively used in a wide range of biological and ecological tests of natural products and synthetic

compounds. The well understood sea urchin embryogenesis makes it possible to suggest the

mechanism of compound effect from the phenotype of treated embryos. Based on these benefits sea

urchin embryos were included in the list of model organisms for medicinal chemistry.

We have devised a phenotypic in vivo sea urchin embryo assay for the rapid evaluation of

potential tubulin modulators [1-2]. The assay is highly reproducible yielding rapid information on

antiproliferative, antimitotic, cytotoxic, and microtubule destabilizing activities of the molecules

along with their solubility and permeability potential. The sea urchin embryo tests do not require

complicated laboratory tools and costly chemical reagents. The experiments fulfill the requirements

of biological ethics: post-spawned adult sea urchins and intact embryos are returned to their natural

habitat. Importantly, the data generated by the sea urchin embryo assay correlate well with the

results obtained by conventional cell-based and in vitro tubulin polymerization assays.

Biological effects of more than 1000 synthetic molecules and natural products have been

analyzed by the assay during the past 10 years. More than 300 antimitotics with microtubule

destabilizing mode of action have been identified, including ~70 compounds with effective

concentration values of ≤5 nM. Their potency was further confirmed by cytotoxicity assessment on

60 human cancer cell lines at National Cancer Institute (USA). 14 compounds were included in

tubulin list as tubulin-targeting agents, and 4 molecules were selected for further in vivo tests in

mice. In addition, we have revealed a series of small molecules with tubulin independent

antiproliferative activity as well as with unique effects on the sea urchin embryo development and

motility. [1]. M. N. Semenova, A. Kiselyov., and V. V. Semenov, BioTechnique,. 2006, 40, 765-774 [2]. V.V. Semenov, M.N. Semenova, Russ.Chem.Rev., 2015, 84, 2, 134-158


112

Synthesis of Natural Antimitotic Derivatives Using

Parsley and Dill Seed Extracts

V.V. Semenov, D.V. Tsyganov, D.V. Demchuk, N.B. Chernysheva, M.N. Semenova

N. D. Zelinsky Institute of Organic Chemistry RAS, 119991, Russian Federation,

Moscow, Leninsky Prospect, 47 E-mail: [email protected]

Analogs of antimitotic natural products combretastatin A-4 (CA4), podophyllotoxin (PT) and

flavanoids were synthesized using allylpolyalkoxybenzenes from dill and parsley seed oil. The

targeted molecules were evaluated in vivo in a phenotypic sea urchin embryo assay for antimitotic

and microtubule destabilizing activity. Structure–activity relationship studies identified mostly

active molecules with 3,4,5-trimethoxyphenyl and 3,4-methylenedioxy-5-methoxyphenyl rings as

potent antiproliferative agents. The effective threshold concentrations (EC) resulting in mitotic

abnormalities in the sea urchin embryos were 0.5–1 nM. These molecules displayed high

cytotoxicity against a panel of 60 human cancer cell lines including multi-drug resistant cells.

Cytotoxic effect of tested compounds was attributed to microtubule destabilization resulted in cell

cycle arrest followed by apoptotic cell death.

Considering these encouraging data from phenotypic and mechanistic studies, some compounds

may prove to be lead candidates for further in vivo studies to assess its potential as an anti-tumor

agents.


113

From Viral Entry Mechanisms to the Entry-Arresting Antivirals Development

against Human Enveloped Viruses (HIV, Flu, … Ebola)

A. Serbin,1,2 B. Bolshchikov,2 O. Alikhanova1 and V. Tsvetkov1,2

1Biomodulators RC, Health RDF, 117042, Russia, Moscow, Blvd. Adm. Ushakova, 14-209

2A.V. Topchiev Institute of Petrochemical Synthesis RAS, 119991, Russia, Moscow, Leninsky Pr., 29


An analysis of modern knowledge of viral entry mechanisms reveals evidently that many

viral particles (virions) penetrate into various permissive cell, using similar molecular

intermediates and routes independently of the viruses belonging to different viral families.

1. No less than 80% of human viruses of various families use the cholesterol-enriched “raft”-

microdomains on plasma membranes as general portals to entry. Thus a “raft”-tropic approaches to

antivirals design can be very promising [1].

2. As the “cytotargeting” parasites, viruses usually accumulate an excess of positive charge on

envelope glycoprotein spikes to be electrostatic-driven toward cells surface that have an counter

(negative charge) potential. At the earliest steps of attachment – adsorption many viruses use the

heparansulfate or other negative charge-enriched receptors. Therefore, the electrostatic factors can

be targets for an anti-entry neutralization (for instance, through anionic polymer antivirals) [2].

3. After adsorption (or after endocytosis) many enveloped viruses initiate a fusion of virion’s

envelope with cell (subcell) membranes, using nano-machineries of similar molecular architecture.

Particularly, the genetically nonequivalent but uniformly- constructed Class I fusion systems are

applied by retro- (HIV…), orthomixo- (Influenza…), paramixo-, filo- (Ebola…) and other viruses.

All these entry-mediated mechanisms were taken into consideration in our research [1-3]. Now

the several generations of HIV, Influensa A/B, and Herpes entry inhibitors are designed on a nano-

competent level, synthesized and successfully evaluated in vitro. The key mechanisms are studied

via docking and molecular dynamics, and first series of computation-predicted substances for anti-

Ebola evaluations are prepared as well. [1] Egorov Y., Serbin A., Alikhanova O., et al. Antiviral Research. 2007. 74(3):A49 [2] Patents: USA 5880154A, RUS 2281297, 2270690, 2315617; Appl Biochem Microbiol. 2012. (9):723-739 [3] Tsvetkov V.B., Serbin A.V. et al. J Comp-Aid Mol Des. 2012. 26(12):1369-1388; 2014. 28(6):647-673


114

New Approaches to Preparation of Composites of Betulin and its Diacyles with

Improved Biological Activities

T.P. Shakhtshneider,1,2 M.A. Mikhailenko,1 S.A. Kuznetsova,3,4 Yu.N. Malyar,3

A.S. Zamay,3 V.V. Boldyrev1,2

1 Institute of Solid State Chemistry and Mechanochemistry SB RAS, Novosibirsk, Russia 2 Novosibirsk State University, Novosibirsk, Russia

3 Institute of Chemistry and Chemical Technologies SB RAS, Krasnoyarsk, Russia 4 Siberian Federal University, Krasnoyarsk, Russia

Betulin and the related compounds found in the outer bark of the birch tree have attracted an increasing interest in the pharmaceutical formulation, due to their versatile biological activity. The poor solubility of betulin and its derivatives in water has limited their applications. Therefore new approaches are required to increase rate of dissolution and solubility of betulin and its derivatives. The purpose of this work was to obtain the composites of betulin and its esters, betulin diacetate (BDA) and betulin dipropionate (BDP), with improved dissolution properties and study their pharmacological activities.

Betulin and its esters were obtained directly from birch bark according to the original methods [1, 2]. Arabinogalactan was isolated from larch wood [3].

The composites of betulin, BDA, and BDP with water-soluble polymers, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG) and arabinogalactan (AG), were prepared by ball-milling the mixtures of the components in a SPEX 8000 mixer mill (CertiPrep Corp., USA). It was found that in the case of the mechanocomposites, the solubility of betulin and its derivatives increased as compared to the initial substances. Possible reasons of increasing the solubility are disordering of crystal structures of the drugs and formation of molecular complexes with the polymers. The obtained composites were non-toxic and exhibited antitumor properties against the ascites adenocarcinoma Ehrlich cells. In vivo tests showed that mechanocomposites of betulin with PVP and PEG exhibited improved gastroprotective properties. Moreover, betulin-PEG mechanocomposite demonstrated an excellent effect on acute renal failure in rats compared to raw betulin.

The composites of betulin esters with AG were prepared also by dissolution the mixtures of the components in water and subsequent solvent evaporation. The products obtained were amorphous thin films readily soluble in water. In vitro studies revealed that the composites prepared as the water-soluble films exhibited the highest antitumor activity against lung adenocarcinoma in comparison with the initial substances [4]. It was shown that the composites of BDA and BDP with AG, especially in film form, showed targeted antitumor effect, causing apoptosis in cancer cells without affecting healthy cells. The work was partly supported by the RFBR (grant No. 14-03-31900) and the RF Ministry of Education and Sciences (grant No. RFMEFI60714X0031).

References [1]. Kuznetsova S.A., Kuznetsov B.N., Red’kina E.S., Sokolenko V.A., Skvortsova G.P. R.F. Patent 2324700, May

20, 2008. [2]. Kuznetsova S.A., Skvortsova G.P., Malyar Yu.N., Vasil’eva N.Yu., Kuznetsov B.N. R.F. Patent 2469043,

December 10, 2012. [3]. Kuznetsova S.A., Kuznetsov B.N., Mikhailov A.G., Skvortsova G.P. R.F. Patent 2273646, April 6, 2006. [4]. Kuznetsova S.A., Shakhtshneider T.P., Mikhailenko M.A., Malyar Yu.N., Boldyrev V.V. R.F. Patent 2541153,

December 04, 2013.


115

Polyfluorinated Salicylates as Analogs of Known Drugs

E.V. Shchegol’kov,1 I.V. Shchur,1 Ya.V. Burgart,1 V.I. Saloutin,1 A.N. Trefilova,2

S.Yu. Solodnikov,2 O.P. Krasnykh,2 L.N. Markova,2 O.N. Chupakhin1

1 I.Ya. Postovky Institute of Organic Synthesis UB RAS., S. Kovalevskoy str. 22,

Yekaterinburg, 620137, Russia 2 Perm National Research Polytechnic University., Komsomolsky Av. 29, Perm, Russia, 614990


Acetylsalicylic acid (ASA) and its analogues present the most popular modern drugs with an unique combination of beneficial pharmacological effects, which include anti-inflammatory, analgesic and antipyretic properties, the ability to reduce platelet aggregation, the efficiency in the prevention of cardiovascular diseases, diabetes and cancer. With the goal to explore the diversity of potent ASA derivatives characterized by an optimized pharmacological profile, we have developed the effective original method for polyfluorosalicylic acids synthesis based on the available polyfluorobenzoic acids. The polyfluorinated analogs of known drugs (methyl-salicylate, sodium

salicylate, aspirin, salicylamide) were obtained using the conversions of hydroxyl and carboxyl groups in polyfluorosalicylic acids. The synthesized compounds were evaluated for lipophilicity (computationally, ChemDraw). The ability to pass through the biological membranes was estimated for a selected set of samples in Parallel Artificial Membrane Assay (PAMPA). The calculated values logP and clogP for fluorinated derivatives are remarkably higher than values for the corresponding non-fluorinated analogs. However the logP/clogP do not correlate with permeability values obtained from PAMPA for tri- and tetrafluoro-ASA since these derivatives are very susceptible to hydrolysis in the conditions of the PAMPA experiment. The permeability values for tri- and tetrafluorosalicylic acids at pH5 are about two-times higher than those for ASA which implies that the fluorinated derivatives will be absorbed in the corresponding environment of GIT more efficiently than ASA. The anti-inflammatory activity of the polyfluorinated salicylates was evaluated in the model of acute inflammation (Carrageenan induced paw edema, Wistar rats). Tetrafluorosalicylic acid, its acetyl derivative and some other compounds in series were found be effective at 50 and 25 mg/kg on the level of ASA or higher. Even more inspiring data was obtained in the “Hot Plate” test for tri- and tetrafluorosalicylic acids (25 mg/kg): they increased the nociceptive response time on the level of Diclofenac. Several derivatives were evaluated for their ability to inhibit the cyclooxygenase-1 and it was found that this enzyme is likely not a primary target of the synthesized compounds. An approximate acute toxicity values for several derivatives were assessed (i.p., CD-1 mice) and found to be close to that of ASA. Thus, the compounds with a superior pharmacological effect while not succeeding toxicity of acetylsalicylic acid were found. This work was financially supported by RFBR (№ 14-03-31541), UB RAS (№ 15-21-3-5), the Program «The PNRPU - EPFL collaboration on Metabolism and Diabetes» supported by «Neva» foundation.


116

Development of Small Molecular Weight Mimetics of Glial Cell Line-derived

Neurotrophic Factor for the Treatment of Parkinson’s Disease

Y.A. Sidorova,1 I. Suleymanova,1 J. Kopra,2 P. Piepponen,2 J.-M. Renko,2 M. Voutilainen,1

J. Sakki,2 R.K. Tuominen,2 M. Karelson3 and M. Saarma1 1 Institute of Biotechnology, Viikki Biocenter, University of Helsinki, Helsinki, Finland

2Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, Finland

3Tartu, University, Tartu, Estonia & GeneCode Ltd, Tallinn, Estonia

Parkinson’s disease (PD) is caused by degeneration and progressive loss of dopaminergic (DA) neurons in the specific region of brain called substantia nigra pars compacta. It affects up to 6 million people world-wide impairing their quality of live and imposing significant economic burden to the society. Currently no cure for this disease is available. Existing therapeutic strategies based on dopamine replacement alleviate PD symptoms, but do not influence its cause or in other words do not prevent or slow down degeneration of DA neurons. Glial cell line-derived neurotrophic factor (GDNF) is one of the few molecules able to protect and repair DA neurons in vitro and in vivo in animal models of PD. GDNF protein and the related factor neurturin were tested in six clinical trials, but the results have been controversial, that is (at least partly) attributed to poor pharmacological properties of these proteins. In particular, they do not penetrate blood-brain-barrier and diffuse poorly from the site of injection because of high affinity to extracellular matrix, thus their delivery to damaged DA neurons is problematic. To overcome these difficulties we decided to develop small molecules acting similarly to GDNF with improved pharmacological characteristics (GDNF mimetics).

Using a combination of rational drug design methods and a panel of biological tests ranging from high-throughput luciferase reporter-based assay to low-throughput DA neurons survival assay we identified several candidate GDNF mimetics. These molecules selectively activated GDNF receptors and downstream intracellular signalling in immortalized cells. One of them supported survival of cultured mouse embryonic DA neurons and stimulated release of dopamine after injection into the mouse brain.

In pilot neuroprotection experiment in unilateral 6-hydroxydopamine (6-OHDA) model of Parkinson’s disease this GDNF mimetic delivered directly to the brain dose-dependently alleviated motor disbalance caused by the selective death of DA neurons in one side of substantia nigra pars compacta with similar efficacy to GDNF protein. Currently we analyse the number of DA neurons and the density of their fibers in the brains of animals treated with GDNF mimetic, vehicle and GDNF. Since pharmacokinetics studies demonstrated that this compound penetrates blood brain barrier with high efficiency we plan to access the activity of the systemically delivered compound in both neuroprotection and neurorestoration paradigms in 6-OHDA model of PD. In addition, we will test other GDNF mimetics in vivo and subsequently optimize active molecules using medicinal chemistry approaches to improve their efficacy and safety.

To summarize we identified a small molecule acting similarly to neurotrophic factor GDNF in vitro and in vivo. This compound might pave way to the development of novel disease-modifying treatment against PD. Supported by FP7-PEOPLE-2013-IAPP GA N 612275, Parkinson's UK Innovation grant - K-1408.


117

3-D Microarray Based on Macroporous Polymer Monoliths for Detection of

Human Genome Mutations Associated with Pregnancy Complications

E. Sinitsyna,1,2 A. Glotov,3 M. Danilova,3 E. Vlakh1,2 and T. Tennikova1,2

1 Institute of Chemistry, Saint-Petersburg State University, 198504, Russia, St. Petersburg, Universitetsky pr,. 26

2 Institute of Macromolecular Compound, Russian Academy of Sciences, 199004, Russia, St. Petersburg, Bolshoy pr., 31

3 D.O. Ott Research Institute of Obstetrics and Gynecology, Russian Academy of Medical Sciences, 199034, Russia, St. Petersburg, Mendeleyevskaya line, 3


The application of microarray technology makes possible the analysis of various DNA changes:

translocation, duplications, deletions, microdeletion and single nucleotide substitutions. The

development of low-density biochips, especially for routine diagnostics in evidence-based

medicine, is still important. The most significant, after genetic diseases, are the health problems

related to a predisposition to cardiovascular affections. Among those, preeclampsia as pregnancy

complication caused by thrombophilia occupies a particular place.

The aim of present study was to confirm the analytical potential of new 3-D platforms based on

macroporous polymer layers for development of DNA microarray. The results of experiments

revealed that the new type of microarray might be successfully used to detect the most frequent

variants of nucleotide substitutions in human genome. Particularly, the correct determination of

single nucleotide replacements in F5 (Leiden G/A, rs6025), MTHFR (C/T, rs1801133) and ITGB3

(T/C, rs5918), involved in coagulation, and COMT (C/G, rs4818), TPH2 (T/A, rs11178997), PON1

(T/A rs854560), AGTR2 (C/A, rs11091046) and SERPINE1 (5G/4G, rs1799889) genes, associated

with pregnancy complications, was observed. The effect of such parameters as amount and type of

oligonucleotide probe and amount of PCR product on signal-to-noise ratio, as well as mismatch

discrimination was analyzed. Sensitivity and specificity of mutation detections using developed

microarray were determined. The comparison of two independent methods, namely, Next

Generation Sequencing (NGS) and microarray technology based on the developed macroporous

polymer layers, allowed the conclusion that the biochip demonstrated higher accuracy and

specificity in the case of ins/del variation.

Acknowledgements

This work was financially supported by grant of St. Petersburg State University (project 0.37.682.2013) and by Fellowship of President of Russian Federation (SP-2763.2015.4) for Dr. E. Sinitsyna.


118

Synthesis of Heterocyclic Derivatives of Borneol as Potential Antiviral Agents

A. Sokolova,1,2 M. Semenova,1,3 O. Yarovaya,1,2 N. Salakhutdinov1,2

1N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, 630090 Russia,

Novosibirsk, Lavrentjev Ave., 9 2Novosibirsk State University, 630090, Russia, Novosibirsk Pirogova St., 2

3 Novosibirsk State Pedagogical University, 630126, Russia, Novosibirsk, Viluiskaya St., 28


We previously reported the synthesis and antiviral activity of compounds based on camphor1,2.

Borneol, a terpene and natural bicyclic organic compound that has shown analgesic, anti-

inflammatory, antibacterial activities3. The aim of this work, syntheses of a set of borneol

derivatives and investigation of antiviral activity.

Scheme 1. Reagents and conditions: (i) CH2Cl2 (dry), (COCl)2, 3-chloropropionic acid, r.t., Ar; (ii) borneol (1), 3-chloropropionic acid chloride , pyridine, r.t., atm Ar; (iii) CH3CN, DBU, refluxe; (iv) MeOH, Et3N, corresponding heterocycle, r.t.

Scheme 2. Reagents and conditions: (i) CH2Cl2 (dry), chloroacetyl chloride, r.t., Ar; (ii) MeOH, dimethylaminel, Py,

r.t.; (iii) MeOH, CH3I, Et3N, r.t; (iv) MeOH, Et3N, corresponding heterocycle, r.t.

The antiviral activity of these compounds was studied against influenza virus A/California/7/09

(H1N1)pdm09 in MDCK cells. The highest efficacy in virus inhibiting was shown

for compound 9.

This work was supported by the RFBR (N 15-03-00193 А, 15-03-10051) and by Russian Science Foundation (N 15-13-17 Creation of new anti-influenza drugs for resistant strains by chemical transformations of terpenes) References [1]. Sokolova A.S. et al. 2013, Bioorg. Med. Chem. vol. 21, pp. 6690–6698 [2]. Sokolova A.S. et al 2014 Bioorg. Med. Chem. Vol. 22, pp. 2141-2148. [3]. Liu R. et al 2011 Neuroscience Vol. 176, pp 408-419


119

Novel Pharmaceutical Compositions Enhances Denervation Properties of

Botulinum Neurotoxin Type A

A.G. Strelnikov,1 V.V. Fomenko,2 D.S. Sergeevichev,3 E.A. Pokushalov,1 N.F. Salakhutdinov2

1Arrythmology Department, Academician E.N. Meshalkin State Research Institute of Circulation

Pathology, 630055, Russia, Novosibirsk, Rechkunovskaya str., 15 2Laboratory of Physiologically Active Substances N.N. Vorozhtsov Novosibirsk Institute of Organic

Chemistry SB RAS, 630090, Russia, Novosibirsk, Lavrentiev ave., 9 3 Laboratory of Experimental Surgery and Morphology, Academician E.N. Meshalkin State

Research Institute of Circulation Pathology, 630055, Russia, Novosibirsk, Rechkunovskaya str., 15


Pharmaceutical compositions of botulinum neurotoxin and chitozan useful for treating cardiac

arrhythmia, in particular having a high therapeutic effect, an increased lasting effect and reduced

side effects. There was discovered that chitozan composition achieves increased pharmacological

activity of the botulinum toxin type A, a desired therapeutic effect already achieved by a single

dose, a prolongation of the botulinum toxin effect, while a reduction of botulinum toxin side effects.

Further, compositions allow preparing mixtures with the desired properties for personalized

medicine directly into the clinic and present a prolonged activity when maintained in the solution

ready to the introduction.


120

Isomerization of α-Pinene Oxide to Campholenic Aldehyde Over

Fe-and Al-containing Zeotype Materials

M.N. Timofeeva,1 V.N. Panchenko,1 A.A. Abel,1 Z. Hasan,2 K.P. Volcho,3 S.H. Jhung2

1Boreskov Institute of Catalysis SB RAS, 630090, Russia, Novosibirsk, Pr. Ak. Lavrentieva 5

2Kyungpook National University, Daegu 702-701, Republic of Korea, Sankyuck-Dong, Buk-Ku 3Novosibirsk Institute of Organic Chemistry SB RAS, 630090, Russia, Novosibirsk,

ave. Ak. Lavrentjeva, 9


Nowadays, considerable attention is focused on the development of new solid Lewis catalysts,

which can be applied at industrial level. The main goal of the design of solid Lewis acids is the

substitution of the traditional homogeneous acid catalysts, such as ZnCl2, AlCl3, FeCl3 etc. Zeolites

and zeotype materials containing transition metal ions are the promising systems as Lewis acids.

These materials can be used as catalysts for processes of isomerization of terpenes, for example,

isomerization of terpene oxides, which is one of the important reactions for synthesis of

intermediates for production of drugs, vitamins and fragrances.

α-Pinene oxide (PO) is one of the important product in industry, which isomerizes rapidly in the

presence of acids, thereby forming many products.

The aim of this study is to investigate the dependence of activity and selectivity of the

rearrangement of PO to campholenic aldehyde (CA) on the textural and acid-base properties of new

Fe-containing materials, such as Fe-containing mesoporous mesophase silica materials (Fe-MMM-

2), microporous Fe-containing nickel phosphate molecular sieves (Fe-VSB-5) [1], and Al-

containing porous metal-benzenetricarboxylates (Al-BTCs), such as MIL-X(Al) (X = 96, 100 and

110) and isostructural family MIL-100(М) (М – Al, Fe and Cr) [2]. The catalytic performance of

these materials was investigated with a combination of physicochemical and catalytic approaches.

The investigation of Lewis acidity was done by EPR and IR spectroscopy using 2,2',6,6'-

tetramethyl-1-piperidinyoxyl radical (TEMPO) and benzonitrile/pyridine as the probe molecules,

respectively.


121

Synthesis and Cytotoxicity Evaluation of Novel Benzene-fluorinated

2,2-Dimethyl-2,3-dihydro-1H-quinolin-4-ones

L. Politanskaya,1 E. Tretyakov,1 V. Shteingarts,1∗ L. Ovchinnikova,2

O. Zakharova3 and G. Nevinsky3

1N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SB RAS, 630090, Russia,

Novosibirsk, Lavrentjeva av., 9 2 Institute of Cytology and Genetics, SB RAS, 630090, Russia, Novosibirsk, Lavrentjeva av., 10

3 Institute of Chemical Biology and Fundamental Medicine, SB RAS, 630090, Russia, Novosibirsk, Lavrentjeva av., 8


Quinolone derivatives, namely 2,3-dihydro-4(1H)-quinolinones, are known to possess a broad

scope of pharmacological properties including anti-ulcer, anti-inflammatory, anticancer activity [1,

2] and pain-blocking properties. We have developed the two-step procedure for preparation of

fluorinated 2,2-dimethyl-2,3-dihydro-1H-quinolin-4-ones including the Sonogashira cross-coupling

of fluorinated ortho-iodanilines with 2-methylbut-3-yn-2-ol and the following PTSA-catalyzed

hydration-cyclization of prepared ortho-alkynylanilines.

The cytotoxicity evaluation of compounds 1(a─i) against human myeloma, human mammary

adenocarcinoma, human hepatocellular carcinoma HepG2 epithelial tumor cells, normal mouse

fibroblasts and Chinese hamster Ag 17 cells was performed. It has been revealed that the

polyfluorinated compounds 1(f,g) inhibit the growth of three tumor mammalian cell lines, the

average IC50 value towards tumor cell lines being approximately 2-fold lower than that towards

normal cells. Mutagenic and antioxidant properties of the compounds using Salmonella tester strain

were studied. It was shown, that the compounds are well antioxidants. In addition all fluorinated

compounds efficiently suppress spontaneous and H2O2-induced mutagenesis of bacterial cells.

The work is supported by the RFBR (grant № 14-03-00108) and by the interdisciplinary grant from SB RAS № 98. References [1] B. Nammalwar, R. Bunce, Molecules. 2014, 19, 204-232. [2] N. Gill, A. Kaur, R. Arora, V. Dhawan, M. Bali, Curr. Res. Chem. 2012, 4, 88–98.

∗ Deceased


122

Design and Synthesis of Polymeric Modules for the Antiviral Potentiation of

Natural and Synthetic Small Molecule Resources

V. Tsvetkov,1,2 B. Bolshchikov,2 and A. Serbin1,2

1 A.V. Topchiev Inst. of Petrochemical Synthesis RAS, 119991, Russia, Moscow, Leninsky Pr., 29

2 Biomodulators RC, Health RDF, 117042, Russia, Moscow, Blvd. Adm. Ushakova, 14-209


The “traditional” antiviral drugs are based mainly on small molecules (SM) from natural or

(semi-)synthetic origins. Although such molecules can be very relevant anti-metabolites or local

suppressors of small-size active centers of enzymes/receptors, they are incapable of efficient

interference with larger-scale biopolymeric targets [1]. At the same time, just the macromolecular,

the protein / nucleic acid (NA) structures and their nano-complexes play key roles in viral life

cycles and pathogenesis. Therefore, in addition to the SM-drugs, the next generation strategies for a

“size-adequate” (poly-/oligomer)-based drug design are required [1]. In this scope the synthetic

polymeric modules (PM) were developed on the principles of: (1) mimicry to polymeric backbone

of NA / polysaccharides; (2) a protein-like but over-genetic (artificial) programming the PM toward

required structure-functionality due to (2.1) intra-chain monomer units regulation, (2.2) chain length

controlling, and (2.3) forming the side-chain variable sequences, that involved some optimal

combinations of SM-derived species [2].

For instance, these principles were realized on basis of RAFT-controlled free-radical alternating

cyclocopolymerization with sequential step-by-step inserting the SM-species in side-positions.

Quantum chemistry modeling the chain propagation routes cleared the mechanisms and conditions

for switching the PM’s isomery from “Pyran” (polysaccharide-like) to “Furan” (NA-like) mimicry,

where the last is preferable for an antiviral interferon-inducing activity [2]. The PM(multi-SMi)-

coupled derivatives were synthesized, modeled via docking and molecular dynamics [3], and

optimized. In contrast with moderately active (SI<10) or ineffective SMs (tested among: carboxylic

and sulfonic acids, camphor-family terpenoids-, adamantan-, cholesten- related synthetic

derivatives, CCR5 and CXCR4 peptidomimetics, etc.), the PM(multi-SMi)s possess a strongly

amplified, and unique-broaden antiviral potency both as antiviral immunoadjuvants and direct entry

inhibitors of retro- (HIV), orthomixo- (influenza-A/B), paramixo-, herpes- and other human viruses

(with SI up to 30000). Therefore, it can be very promising prospect to biomedical advancements of

natural and synthetic SM-resources on the PM platform.

References [1] Serbin A.V., Karaseva E.N., Tsvetkov V.B., et al. Macromol Symp. 2010, 296(1):466-477; [2] Serbin A.V., Veselovsky A.V., Tsvetkov V.B. Appl Biochem Microbiol. 2012. (9):723-739 [3] Tsvetkov V.B., Serbin A.V. J Comp-Aid Mol Des. 2012. 26(12):1369-1388; 2014. 28(6):647-673


123

Anti-mycobacterial Activity of Ag(0)-nanoparticles in Mouse Bone-Marrow

Cells and Peritoneal Macrophages Infected with BCG Vaccine In Vitro

E.G. Ufimtseva,1 I.D. Ivanov2 and A.V. Popov2

1Department of Medical Biotechnology, The Research Institute of Biochemistry, 630117, Russia, Novosibirsk, Timakova St., 2

2Department of Drug Metabolism and Pharmakinetic, The Institute of Molecular Biology and Biophysics, 630117, Russia, Novosibirsk, Timakova St., 2/12


Tuberculosis, with the intracellular Gram-positive parasite Mycobacterium tuberculosis as the

causative agent, accounts for about 2 million deaths annually. Currently, the development of new

anti-tuberculous drugs intended for prevention and treatment of tuberculosis are required because of

the emergence and spread of high-virulence strains of mycobacteria that possess multidrug and

extensive drug resistance. It is well known antimicrobial action of silver nanoparticles (NPs) on

Gram-negative bacteria [1]. We have investigated anti-mycobacterial activity of Ag(0)-NPs with

different size (2 nm, 40 nm, and 10-15 nm (Verity Technology, and “Argonika”, Vektor-Pro,

Novosibirsk, Russia, respectively)) in mouse BALB/c bone-marrow cells and peritoneal

macrophages infected with the Bacillus Calmette-Guérin vaccine (BCG, a strain of attenuated live

M. bovis, the Institute of Microbiology and Epidemiology, Moscow, Russia) in vitro as in the work

[2], because good concordance was found between the results on the sensitivity of M. bovis BCG

and M. tuberculosis to diverse compounds [3]. The Ag(0)-NPs were used at the 0.05, 0.5, and 5.0

µg/ml concentrations in the growth medium. After 4, 24, and 48 h of incubation with the Ag(0)-NPs

or isoniazid at the 5 and 10 µM concentrations acid-fast BCG-mycobacteria were stained by Ziehl-

Neelsen method in mouse cells. We have found that the 10-15 nm and 40 nm Ag(0)-NPs at the 5.0

µg/ml concentration for 48 h of incubation have demonstrated considerable bactericidal effect on

BCG-mycobacteria in host cells. On confocal fluorescent images of these host cells we have

observed colocalization of lipoarabinomannan-detected BCG-mycobacteria and host cell lysosomes

stained by the LysoTracker Red DND-99 dye, and lack of its colocalization with the clursters of the

Ag(0)-NPs. Immunofluorescence assay didn’t detected production of the proinflammatory

cytokines IFNγ and IL-1α both in these host macrophages and in mouse control cells. Therefore,

bactericidal action of the Ag(0)-NPs was not regulated by these cytokines. Our findings of anti-

mycobacterial activity of the Ag(0)-NPs attract research and clinical interest due to the antibiotic

resistance of modern pathogenic mycobacteria. References

[1]. J. Morones, J. Elechiguerra, A. Camacho, et al., Nanotechnology. 2005, 16: 2346-2353. [2]. E. Ufimtseva, Russian Journal of Infection and Immunity. 2014, 4: 213-220. [3]. S. Franzblau, M. DeGroote, S. Cho, et al., Tuberculosis. 2012, 92: 453-488.


124

Inhibition of Sonic Hedgehog Pathway in Breast Cancer Stem Cells Does Not

Provide Extra Benefit in Cytotoxicity Resulted from

Pd(II)-Saccharinate Complex of Terpyridine

E. Ulukaya,1 B. Cevatemre,2 D. Karakas,2 N. Aztopal,2 C. Icsel3

1Uludag University, Faculty of Medicine, Department of Clinical Biochemistry,

16059, Bursa, Turkey 2Uludag University, Faculty of Arts and Sciences, Department of Biology, 16059, Bursa, Turkey

3Uludag University, Faculty of Arts and Sciences, Department of Chemistry, 16059, Bursa, Turkey

Cancer stem cells are a minör fraction of a tumor tissue and is thought to be responsible for the

recurrence of the disease some months or years after the therapy. These cells are resistant to

classical chemotherapeutics due to aberrant expressions of some pathways (Sonic Hedgehog, Wnt,

Notch) that involve in the survival/self-renewal and proliferation of these cells. The inhibition of

these pathways are expected to increase the patient survivals.

We propagated MCF-7 cell line to increase the stem cell fraction and then formed

mammospheres that are enriched in cancer stem cells. The mammospheres were not full of cancer

stem cells but they provided quite a reasonable resource to work on. Characteristic cell surface

markers (CD44+/CD24-) were determined by flow cytometry. The sphere formation assay (SFA)

was performed to assess the effect of treatments on the CSC self-renewal capacity. Ten micromolar

cyclopamine was used to inhibit the Sonic Hedgehog pathway. This dose was also used in

combination with various doses (3.12 – 100 micromolar) of the Pd (II) complex for 48 h.

The addition of sonic hedgehog inhibitor, cyclopamine, did not change the mammosphere

forming ability, compared that of Pd (II) complex treatment alone. Interestingly, the result was the

same for both parental MCF-7 cells and cancer stem cells-enriched mammospheres.

This results imply that the inhibition of sonic hedgehog pathway does not provide an extra

cytotoxic effect to a Pd(II) complex that kills the cells by damaging their DNA. The possible reason

for this may be relatively inadequate expression of sonic hedgehog pathway in these cancer model.

In fact, in our cancer stem cell model, Wnt pathway is much more active than this pathway, thereby

the inhibition of Wnt pathway results in enhanced cytotoxic activity of the same Pd(II) complex.

This study is supported by TÜBİTAK (The Scientific and Technological Research Council of Turkey) with the project numbered 212T147


125

Comparison of Anticonvulsant Effects of Lambertianic Acid Aamide and

Memantine in Mouse Hippocampal Slices

S.O. Vechkapova

Design Technological Institute of Digital Techniques, Siberian Branch of the Russian Academy of

Sciences, Russia, Novosibirsk, Rzhanova st., 6


The most common neurodegenerative disorders, such as Alzheimer’s, Parkinson’s, multiple

sclerosis, epilepsy may share a common pathway: excessive NMDA receptor activation causing

excitotoxicity [1]. Currently, drugs with glutamatergic mechanisms of action are intensely being

developed for the treatment of cognitive disorders and neurodegenerative processes. One approach

to solving this problem is chemical transformation of biologically active plant metabolites, which

are renewable resources. The lambertianic acid is produced by the Siberian cedar (Pinus sibirica R.

Mayr), could easily be extracted from its galipot and needles and has proved to be a promising

compound that could be used in the synthesis of valuable pharmaceutical reagents [2].

The aim of this work was to compare the effects of lambertianic acid amide (AmLA) and

memantine on the induced epileptiform activity in CA1 hippocampal area. Memantine is a low-

affinity voltage-dependent uncompetitive antagonist at NMDA-receptors, which is used for the

treatment of Alzheimer's disease [3]. The experiments were performed on the hippocampal slices of

ICR male mice with the help of standard electrophysiological techniques. Stimulation of Schaffer

collaterals and registration of induced population spikes of pyramidal neurons in the CA1 field were

conducted with the use of glass microelectrodes, filled with saline.

Both AmLA in concentration of 170 µМ and memantine in concentration of 100 µМ normalize

the epileptiform activity caused by magnesium-free medium. Also AmLA did not affect the

initiation of NMDA-dependent synaptic potentiation. The similarity of the effects of AmLA with

memantine allows to suggest that it acts in conformable manner, i.e. by blocking the NMDA

receptor ion channel much like low affinity voltage-dependent uncompetitive antagonists.

The work was supported by the basic project of fundamental research of RAS IV 35.1.5 and the program of Presidium of RAS FSM-46 "Fundamental Sciences for medicine". References: [1] Lau A., Tymianski M. Glutamate receptors, neurotoxicity and neurodegeneration. Pflugers Arch, 2010, V.60(2),

P.525-542. [2] Tolstikova T.G., Sorokina I.V., Voevoda T.V., Shults E.E., Tolstikov G.A. Nootropic activity of lambertianic acid

derivatives. Doklady Biological Sciences, 2001, V. 376(2), P. 8-9. [3] Sonkusare S.K., Kaul C.L., Ramarao P. Dementia of Alzheimer's disease and other neurodegenerative disorders –

memantine, a new hope. Pharmacol Res, 2005, V. 1(1), P. 1-17.


126

Poly(amino acid) Polymersomes as Potential Nanocontainers for Drug Delivery

E. Vlakh,1 N. Zashikhina,2 A. Hubina,1 T. Tennikova1

1Institute of Chemistry, Saint-Petersburg State University,

198504, Russia, St. Petersburg, Universitetskiy pr. 26 2 Institute of Macromolecular Compounds, Russian Academy of Sciences,

199004, Russia, St. Petersburg, Bolshoy pr. 31 E-mail: [email protected]

It is known that the conventional application of drugs is associated with limited effectiveness,

poor biodistribution, and lack of selectivity. These drawbacks can be overcome by the development

of polymer-based drug delivery system with controlled drug release. One of perspective kinds of

polymer nanoparticles is polymersomes. Such nanobeads are formed from amphiphilic block

copolymers as a result of their self-assembly. Comparatively to liposomes, polymersomes represent

hollow spheres, which contain an aqueous medium in the core surrounded by a bilayer hydrophobic

membrane. The advantage of polymersomes is the possibility to encapsulate both hydrophilic and

hydrophobic drugs [1]. However, contrary to liposomes, polymersomes have higher membrane

stability and lower membrane permeability that can be controlled and modulated by varying block

lengths.

Poly(amino acids) is related to the class of biocompatible and biodegradable polymers that are

very attractive for the different fields of bioapplication. The traditional approach for synthesis of

poly(amino acids) is the polymerization of α-amino acid-N-carboxyanhydrides (NCA) with

applications of amines as initiators [2].

In the present work the synthesis of different amphiphilic block-copolymers was carried out.

Poly(glutamic acid) and polylysine were applied as hydrophilic blocks, whereas polyphenylalanine

and polyleucine were chosen as hydrophobic components. The block copolymer obtained was

characterized with gel-permeation chromatography (GPC) and NMR. The self-assembly of

amphiphilic block copolymer was studied depending on pH and polymer concentration. The

morphology of self-assembled nanoparticles was evaluated using transmission electron microscopy

(TEM). The mean size and size distribution of prepared polymersomes were analyzed using

dynamic light scattering (DLS). The formation of nanparticles from 60 to 350 nm was observed.

The encapsulation of model compounds was studied under variation of pH, time and concentration

of target molecules and the encapsulation efficiency was calculated. Finally, the positive example of

encapsulation of several water-soluble anticancer drugs was demonstrated.

This work was supported by grant of Russian Scientific Foundation (№ 14-50-00069). References

[1]. Lee J.S., Feijen J., Journal of Controlled Release, 2012, 161, 473–483. [2]. Cheng J., Deming T.J, Topics in Current Chemistry, 2012, 310, 1–26.


127

New Physiologically Active Compounds Synthesized From Verbenone

K. Volcho,1,2 I. Il’ina,1,2 O. Mikhalchenko,1 O. Ardashov,1,2 A. Pavlova,1

T. Tolstikova1 and N. Salakhutdinov1,2

1Novosibirsk Institute of Organic Chemistry, 630090, Russia, Novosibirsk, Lavrentjeva ave., 9

2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova St., 2


Verbenone is pinane monoterpenoid showing pheromone activity in bark beetle. It can be

synthesized by allylic oxidation of α-pinene, the main component of pine turpentine.

We found that epoxidation of verbenone with following transformations makes it possible to

obtain numerous monocyclic and polycyclic compounds.

Some of obtained products demonstrated highly promising anti-Parkinsonian, anticonvulsant or

analgesic activity [1-3].

References [1] O. V. Ardashov, A. V. Pavlova, I. V. Il’ina, E. A. Morozova, D. V. Korchagina, E. V. Karpova, K. P. Volcho, T. G.

Tolstikova, N. F. Salakhutdinov, J. Med. Chem. 2011, 54, 3866–3874. [2] O. Mikhalchenko, I. Il’ina, A. Pavlova, E. Morozova, D. Korchagina, T. Tolstikova, E. Pokushalov, K. Volcho, N.

Salakhutdinov, Med. Chem. Res. 2013, 22, 3026–3034. [3] I. Il’ina, O. Mikhalchenko, A. Pavlova, D. Korchagina, T. Tolstikova, K. Volcho, N. Salakhutdinov, E. Pokushalov.

Med. Chem. Res. 2014, 23, 5063-5073.


128

Development of Tetrazole-Containing Derivatives of Lupane Type Triterpenoids

and Their Cytotoxicity

A.N. Volkova,1 N.I. Petrenko,1 M.A. Pokrovsky,2 E.E. Shults1 and A.G. Pokrovsky2

1Novosibirsk Institute of Organic Chemistry, SB RAS,

630090 Novosibirsk, Lavrentjev Ave. 9, Russia 2Novosibirsk State University,

630090 Novosibirsk, Pirogova St., 2 E-mail: [email protected]

Synthetic transformations of the available lupane-type triterpenoids are of interest for the

preparation of pharmacologically valuable compounds [1]. Earlier we obtained various betulin

nitriles the transformation of which opens a convenient way to the nitrogen heterocycles of the

lupan series [2]. Throughout researches on the chemical modification of betulin derivatives, we

synthesized tetrazole-containing triterpenoids.

The transformations of terpenoid nitriles 1,3 into tetrazole compounds 2,4 was carried out by the

1,3-dipolar cycloaddition reaction of compounds 1,3 with dialkylammonium azide, obtained in situ

by the reaction between triethylamine hydrochloride and sodium azide. The yields of compounds

2,4 are composed to 64-76%. The structure and molecular formula of compounds 2,4 are confirmed

by the spectral data.

The cytotoxicity of the new type of tetrazole-containing triterpenoids derivatives will be

discussed.

This work was supported by the Russian Science Foundation (project № 14-13-00822). References [1] R.-Y. Kuo, K. Qian, S. L. Morris-Natschke, and K.-H. Lee, Nat. Prod. Rep., 2009, 26, 1321–1344. [2] A. N. Antimonova, N. V. Uzenkova, N. I. Petrenko, M. M. Shakirov, E. E. Shul’ts, G. A. Tolstikov, Russ. J. Org.

Chem. 2011, 47, 4,589–601.


129

Assessment of Triticain-α glutenase and Collagenase Activities for Use in

Enzymatic Therapy Assays

A.A. Zamyatnin Jr.,1,2 G. Aliev,3,4 N.V. Gorokhovets,1 V.A. Makarov,1 S.Yu. Morozov,2

L.V. Savvateeva,1 M.V. Serebryakova,2 A.G. Solovyev2 and E.Yu. Zernii2

1Sechenov First Moscow State Medical University, Institute of Molecular Medicine,

119991, Moscow, Russia 2Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology,

119991, Moscow, Russia 3”GALLY” International Biomedical Research Consulting LLC, San Antonio, TX, 78229, USA

4School of Health Science and Healthcare Administration, The University of Atlanta, Johns Creek, GA, 30097, USA


Insufficient and/or improper protein degradation results in the development of various human

pathologies. Proteolytic enzymes aimed to improve insufficient proteolytic activity were suggested

for the treatment of protease deficiency-induced disorders. Since in some cases human degradome

is incapable to support degradation of the entire target protein(s), other organisms can be used as a

source of proteases exhibiting additional activities distinct from human enzymes, and plants are

perspective candidates for this source. Present study revealed that recombinant cysteine protease

Triticain-α from wheat (Triticum aestivum L) refold in vitro into an autocatalytically activated

protease possessing glutenase and collagenase activities at acidic or close to neutral pH levels at the

temperature of human body. Mass-specrometry analysis of the products of Triticain-α-catalyzed

gluten hydrolysis revealed multiple cleavage sites within the sequences of gliadin toxic peptides, in

particular, in the major toxic 33-mer α-gliadin-derived peptide initiating inflammatory responses to

gluten in Celiac Disease (CD) patients. Importantly that Triticain-α was found to be relatively stable

in the conditions simulating stomach environment. Thus, we are confident that Triticain-α can be

used as a basic compound for development of (i) pharmaceuticals for oral administration aimed at

release of the active enzyme into the gastric lumen for CD treatment, and (ii) topically active

pharmaceuticals for wound debridement applications.


130

Bactericidal Activity and Cytotoxicity of Substituted Cyclic Acetals

A.N. Kazakova,1 A.V. Tugarova,2 A.A. Kamnev,2 G.Z. Raskildina,1 S.S. Zlotskii1

1 Ufa State Petroleum Technological University,

Kosmonavtov Str.,1, Ufa 450062, Russian Federation 2 Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of

Sciences, 410049, Russian Federation, Saratov, Prospect Entuziastov, 13


Cyclic acetal and benzocyclic acetal fragments are present in a number of molecules with high

biological activity, which are used in medical chemistry.

We have synthesized the model cyclic acetals containing oxygen and nitrogen atoms in the side

chain and have analyzed their bactericidal activity and toxicity.

Primary screening showed the ability of the 1,2,3 compounds to influence free radical oxidation

processes.

1 2

О

О СН3

СН3

3

ОО

Сl

4

OH

O OO

5

OH

O OO

OH

O OO

6O O

ClO

O O

OCl

Cl

7 8

O O

NH

9O O

N N

NN

Cl

Cl

The prospectivity of creation of the antioxidants based on these compounds was proved,

suppressing the accumulation of oxidation products in the blood.

For estimate of biotoxicity of a number of substituted 1,3-dioxo cycloalkanes the bacterial strains

were used. In 0.1% concentration 2-isopropyl-1,3-dioxolan 4 and 4-chloromethyl-1,3-dioxan 5

inhibit bacterial growth by ~50%.

The cytotoxicity of the derivatives of 5-oxy-1,3-dioxolane 6,7 and 4-oxy methyl-1,3-dioxolane

1-3,8,9 in relation to Du145 human prostate carcinoma cells proved to be lower than the

cytotoxicity of tubercidin (standard).

We can assume that the cyclic acetal fragment without any self-dependent toxicity promotes

molecule movement through cell membranes. The functional groups connected with 1,3-dioxo

cyclanic nucleus (phenols, aryl amines and others) remain active in these molecules.

This work was supported by the President of Russian Federation (the scholarship of the President of RF to the young scientists and PhD students to 2015-2017, the order from March 10, 2015, # 184)


131

Phytochemical analysis, Hepatoprotective and Antioxidant Activities of Russian

Nelumbo nucifera Leaves Against CCl4-Induced Hepatic Damage in Rats

M.S. Abdelhamid, E.I. Kondratenko, N.A. Lomteva

Department of Molecular Biology, Genetics and Biochemistry, Faculty of Biology, Astrakhan State

University, 414000, Russia, Astrakhan, Shaumyan Square, 1


The aim of present study was to investigate the hepatoprotective and antioxidant activities of the

ethanol extract of Russian Nelumbo nucifera leaves (NNLE) against CCl4-induced hepatic injury in

female albino rats. Phytochemical screening of NNLE was performed by GC-MS analysis. Rats

were treated intragastrically with NNLE at doses of 50 and 100 mg/kg body weight (BW) after

administration of CCl4 for 49 consecutive days. Silymarin, a known hepatoprotective drug, was

used as standard at 50 mg/kg BW. Alanine aminotransaminase (ALT), aspartate aminotransaminase

(AST), alkaline phosphatase (ALP), albumin (ALB), total protein (TP), Total bilirubin (TB), urea

and total cholesterol (TC) were estimated in serum of rats. The antioxidant activity of NNLE was

evaluated by the estimation of malondialdehyde (MDA) level and the activity of catalase (CAT) in

liver tissues. GC-MS chromatogram of NNLE showed thirty-eight peaks which indicating the

presence of thirty-eight phytocomponents. The major constituents are 1-hexadecanol(10.47%),13-

docosenoic acid-methyl- ester,(Z) (7.88%), 2-naphthol-1-(p-chlorophenylazo) (5.38%), quercetin-

7,3',4'-trimethoxy (4.61%), docosane (4.43%), anodendroside-A (4.31%), colchifoleine (3.73%), 6-

octadecenoic acid -methyl-ester (3.39%) and (-)-loliolide (3.00%). Administration of NNLE at 50

and 100 mg/kg BW after CCl4 toxicity caused a significant decrease in serum ALT, AST, ALP, TB,

urea and TC ,while a significant increase in serum ALB and TP compared to CCl4 treated control.

Furthermore, administration of NNLE at 50 and 100 mg/kg BW produced a significant decrease in

the level of MDA, while a significant increase in the activity of CAT in liver tissues compared to

control. Our results suggested that NNLE has hepatoprotective and antioxidant activities against

CCl4-induced hepatic toxicity in rats due to its contents of bioactive compounds.

Key words: Nelumbo nucifera leaves, ethanol extract, hepatoprotective activity, antioxidant

activity, carbon tetrachloride, GC-MS analysis.


132

New Synthetic Potential in the Phenanthrene Series in the

Treatment of Socially Significant Diseases

V. Abzianidze,1 K. Bolshakova,2 D. Krivorotov,1 and V. Kuznetsov1

1Chemical modeling laboratory, Research Institute of Hygiene, Occupational Pathology and

Human Ecology, Federal Medical Biological Agency, p/o Kuz’molovsky, Saint Petersburg, 188663, Russian Federation

2All-Russian Institute of Plant Protection, Russian Academy of Agricultural Sciences, Pushkin, Saint Petersburg, 196608, Russian Federation


The development of the chemistry of multiple-function natural and medicinal compounds

requires studying the characteristics of reactions behavior, e.g. for optimization of pharmacological

characteristics. As objects of research interest are some semi-synthetic drugs of the phenanthrene

series influencing CNS. Synthesis of their prodrugs is important due to a possible modification of

receptor activity, ability to penetrate through the protective barriers of the body and prolonged

effect.

3-Monophosphate and 3-benzoates (1-3) of 17-(cyclopropylmethyl)-4,5α-epoxy-3,14-

dihydroxymorphinan-6-one A and its 6-methylene analog B have been synthesized for the first

time.

OOH

OH

H

N

R

(CH3O)2P(O)Clp-ClC6H4C(O)Cl

OO

OH

H

N

R

P

O

O O1 - R = O

2 - R = CH2

3 - R = CH2

R = O [1]

TEA TEA

OO

OH

H

N

R

O

Cl

A - R = O

B - R = CH2

Although the metabolic profile of substances A and B is hard to predict, simulation of some

pharmacokinetic parameters and ability to cross the BBB has been provided using standard

ACD/Percepta software. The differences in the results obtained determining pharmacological

properties of the compounds discussed are presented. Compound R Type Simulation results of ACD/Percepta software, oral administration of A, B, 1-4

bioavailability, F, % Тмах BBB penetration А O phenol 99%- calcd. / 22% - exp. 95 min – calcd. / 60 min – exp. CNS active В CH2 phenol 99% - calcd. / 41% - exp. 95 min – calcd. / 90 min – exp. CNS active 1 O benzoate 98% - calcd. 153 min CNS active 2 CH2 benzoate 63% - calcd. 321 min CNS active - O alkylphosphate 99% - calcd. 71 min CNS active 3 CH2 alkylphosphate 99% - calcd. 77 min CNS active

Thus, some of the compounds synthesized may have prolonged central effects and may serve as

a basis for drug development in the treatment of socially significant deseases.

References [1] S. Lazar, S. Jabbouri, C. Moisand, S. Noel-Hocquet, J. Meunier, C. Ropars, G. Guillaumet, Eur.J.Med.Chem. 1994, 29, 45-53


133

Synthesis of Natural Phaeosphaeride A derivatives

V. Abzianidze,1 K. Bolshakova,2 L. Chisty,1 A. Berestetskii,2 and V. Kuznetsov1



2All-Russian Institute of Plant Protection, Russian Academy of Agricultural Sciences, Pushkin, Saint Petersburg, 196608, Russian Federation


Many drugs for treating human diseases have been discovered from natural sources. Captivated

by the unique molecular structure and promising antitumor activity of natural phaeosphaeride A [1],

we have launched its chemical studies.

Thus, we have conducted alkoxylation of phaeosphaeride A in an alkaline medium. The products

formed were slightly unexpected.

ON

OOH

OHO

ONH

OOH

OH

OROR

+ROX

H2O

LiOH

1a2a3a

2b3b

1bR = CH3 (1a,1b), C2H5 (2a,2b), CD3 (3a,3b)

X = H (1a,1b,2a,2b), D (3a,3b)

ONH

OOH

OH

OR

OR

The absolute structure of the compounds obtained was

determined by X-ray analysis of a methoxy derivative. The

presumable mechanism of the reaction is shown in the

following scheme.

ON

OOH

OHO

RO-

H2O

O C-

N

OOH

OHO

OR

ON

OOH

OH

OR

RO-

ONH

OOH

OH

OR

O

R

LiOH

H+

-CH3O-

δ+

δ+

δ−

δ−

The structure of the compounds synthesized has been proved using modern spectral methods.

References [1] Maloney, K.N., Hao, W., Xu, J., Gibbons, J., Hucul, J., Roll, D., Brady, S.F., Schroeder, F.C. & Clardy J., Org. Lett. 2006, 8, 4067-4070.


134

Macromolecular Characterization of Metal-polymer Nanobiocomposities by

Aqueous Size-exclusion Chromatography with Multiple Detection

G. Aleksandrova,1 A. Boymirzaev,2 M. Lesnichaya,1 B. Sukhov,1 and B. Trofimov1

1Favorskii Irkutsk Institute of Chemistry, Siberian Branch, Russian Academy of Sciences, 664033

Russia, Irkutsk, Favorsky str, 1 2Department of Chemical-Technology, Namangan Institute of Engineering and Technology, 160115

Uzbekistan, Namangan, Kasansai str, 7


Size-exclusion chromatography (SEC) is one of the most powerful methods for investigation and determination of average molar mass (MM) and molar mass distribution (MMD) of polymers [1].Biopolymers, defined broadly as biosynthesized materials, play a central role in medicine and pharmacy with applications in tissue engineering, regenerative medicine, and drug-carrier systems. Such materials are advantageous in biomedical and pharmaceutical applications because of their inherent properties of biocompatibility and biodegradation. This work aimed to investigate the influence of the noble metals nanoparticles formed within the polysaccharide matrix on the change of molar mass parameters of the galactose-containing polysaccharides: arabinogalactans (AG), galactomannans (GM), and carrageenans (CG). MM parameters of the prepared metalbiopolymer composites were studied by means of SEC taking advantage of three hyphenated detection systems.

The SEC system was composed of an Agilent 1100/1260 Series chromatograph with a quaternary pump with degasser (G1311B), an autosampler (G1329A), two Ultrahydrogel Linear columns (300 x8 mm) from Waters (USA) connected in series, a differential refractometer (RID10A, Shimadzu), and a MALLS detector (Mini DAWN TriStar, Wyatt Technology Corporation) with a semiconductor laser diode at 690 nm. The SEC conditions were as follows: degassed 0.1 M NaNO3 was used as eluent, the sample concentration was 1-4 mg/mL, injection volume varied from 50 to 200 ml, flow rate was maintained at 0.5 mL/min, and the column and detector temperature were kept at 40°C Pullulan and PEO and dextran standards were used for calibration of the column system. Data acquisition and processing were carried out by use of the Astra software, version 5.3.4.20.

Formation of the disperse nanoparticles was accompanied by appearance of new links between the in situ formed metal particles and the polysaccharide stabilizers. The polysaccharides acted as reducing agents, being oxidized in the course of zerovalence metal formation [2]. The SEC analysis taking advantage of hyphenated three-detector technique was extremely informative mean to study the changes of molar mass parameters in the course of formation of metal nanocomposites with polysaccharides. It was shown that the composites formation was accompanied by the decrease of molar mass of the polymers and change of their polydispersity index, due to a combination of redox reactions involving the noble metals precursors and the alkali-induced degradation.

The interaction of metal nanoparticles with the macromolecules of the stabilizer was directly confirmed. The nature and the content of the metal both influenced the observed changes of the polymer molecular parameters. In detail, increase of the metal nanoparticles amount enhanced the fraction of aggregated macromolecules in the sample. The degradation processes were majorly reflected in the molar mass decrease in the cases of GM and CG. Self-assembly of the nanoparticles and the macromolecule to form a nanocomposite is a result of a set of factors including the dimensional effect of the nanoparticles.

The work has been carried out at partial support of RFBR (grant 14-43-04127 r_sibir_a). References [1]. A. Boymirzaev, Khim. Rast. Syr’ya, 2009, no. 2, p. 19. [2]. L. Grishchenko, S. Medvedeva, G. Aleksandrova, L. Feoktistova, A. Sapozhnikov, B. Sukhov, and B. Trofimov,

Russ. J. Gen. Chem., 2006, vol. 76, no. 7, p. 1111.


135

Water Soluble Bioactive Nanocomposites of Transition Metal Oxides with Optical and Magnetic Properties

G. Aleksandrova,1 A. Bogomyakov,2 E. Gasilova,3 S. Vityazeva,4 V. Dubrovina,4 B. Sukhov,1

V. Ovcharenko,2 B. Trofimov1 1 A.E. Favorsky Irkutsk Institute of Chemistry SB RAS, Favorsky st. 1, 664033, Irkutsk, Russia

2Institute International Tomography Centre SB RAS, 630090, Institutskaya 3, Novosibirsk, Russia 3 Institute of Macromolecular Compounds, RAS, Bolshoy pr., 31, 199004, St._Petersburg, Russia 4Anti-Plague Research Institute of Siberia and Far East, Trilissera 78, 664047, Irkutsk, Russia


Metal-polymer nanocomposites are a very special group of hybrid composite materials whose polymer nanostabilizing matrix is a key part of their aggregate structure. Studies on regularity formations of nanoparticles, their specific structural and optical properties sequent of their nanosizes, present multiple fundamental information about the nanosized state of the substances [1]. Applications of nanotechnologies to biological systems involve primarily the development of new biocompatible nanosized materials and integrated study of their uncommon biologically active properties. We developed a method for nanosized oxides synthesis on the basis of unique natural nanostable polymer matrix of arabinogalactan. The objective of the present work is the design of multifunctional nanobiocomposites with oxides of transition metal cores stabilized by arabinogalactan and the analysis of their optical and magnetic characteristics properties.

A new general approach to its synthesis has been developed and experimental conditions for the synthesis of nanosized biocomposites with variable sizes and grades of dispersiveness have been optimized. Arabinogalactan, polysaccharide of Siberian larch, was used as stabilizing basis and iron and transition metals (Co, Ni, Mn, Cu, Bi) oxides were taken as nanosized cores. Nanobiocomposites have been obtained with high yields, iron content in nanocomposite samples varies in ranges 1.0-12.0%, the transition metals’ contents are 0.5-24.0% they are depending from synthesis conditions. Composition and structure of the materials described previously [1, 2].

The spectral properties of transition metal oxides nanocomposites systematically were studied. The IR spectra recorded signals indicative the organic template and the bond O-metal presence. On the contrary, arabinogalactan is transparent both in UV and Vis ranges, and electronic absorption spectroscopy can be successfully used to study arabinogalactan assisted nanoparticles. The refraction of nanocomposites aqueous solutions were measured. Specific refractive index arabinogalactan is close to that of other polyols and amounted to 1.476. Specific refractive index of metal oxides nanocomposites was lower and varies in the range of 1.432 - 1.470. Nanocomposites polarizability was proportional for specific and molecular refraction. It is due to the nature of the metal oxides and its content in the samples.

The study of the magnetic properties of the new composites on the basis of oxides encapsulated into arabinogalactan matrix has been performed by the SQUID magnetometer. The temperature dependence (5 – 320 K) of magnetization of the sample in an applied magnetic field of 15 kOe, and the field dependence of the magnetization at 5 and 320 K were measured. It has been shown that the field dependencies of samples magnetizations have the appearance of a hysteresis loop whose coercive force are 40- 80 Oe at 5 K. The Curie temperature of the samples are ~ 50 K.

The new nanostructured biomagnetics are distinct for their water solubility and moderate magnetization that would be especially important for the development of biocompatible magnetic preparations on their basis. Arabinoganactan and its cobalt oxide nanobiocomposites were selected as immunomodulators to elucidate the possibility of intensification of specific immunity against plague. Nanobiocomposites with target optical characteristics have a great potential to design promising multifunctional nanosized materials with controlled optical properties as well as new optical systems and optical markers in biology and medicine. The work has been carried out at support of RFBR (grant 14-43-04127 r_sibir_a, 14-03-00859_а). [1] G. Aleksandrova, S. Medvedeva, L. Grishchenko, et all, Patent RF № 2260500, 2005. [2]. L. Grishchenko, S. Medvedeva, G. Aleksandrova, et all, Russ. J. Gen. Chem., 2006, vol. 76, no. 7, p. 1111.


136

Pyrimidine Nucleoside Derivatives with Both

Antitubercular and Antiviral Activity

I.L. Karpenko, L.A. Alexandrova

Engelhardt Institute of Molecular Biology RAS, Vavilova 32, Moscow 119991, Russia


Nucleoside analogues play an important role in medicine as antiviral agents. Many 5-substituted

derivatives of 2’-deoxyuridine inhibit the replication of herpes viruses (e.g., HSV and VZV) [1].

Only at the beginning of the XXI century several groups reported a few sets of modified

nucleosides that displayed antimycobacterial activity in vitro [2]. Recently we synthesized 2’-

deoxypyrimidine nucleoside derivatives bearing extended alkyloxymethyl or alkyl(1,2,3-triazol-1-

yl)methyl substituents at С-5 position and demonstrated their effective bacteriostatic activity against

M. tuberculosis strains [3]. However, the nucleosides with large hydrophobic fragments are

insoluble in water thus limiting the biological investigations. The goal of this work was to

synthesize and to study anti-HSV-1 activity and stability of 5’-monophosphates of C-5 modified

nucleosides in human blood serum.

R = C8H17; b: C10H21; c: C12H25

All 5’-monophosphates of C-5 modified nucleosides were not cytotoxic at concentrations up to

200 µM for Vero cells and up to 100 µM for A549 and Jurkat cell lines and stable in human blood

serum over a period of 2 – 12 hours, that allowing their application as potential prodrugs. 5’-

Monophosphates of C-5 modified nucleosides demonstrated moderate activity towards herpes

simplex virus (HSV-1) in the Vero cell culture.

Acknowledgment. This study was supported by the Program of the Presidium of the Russian Academy of Sciences “Molecular and Cell Biology” and by the Russian Foundation for Basic Research (grants № 14-04-00755 and № 15-04-05116).

References 1. E. De Clercq, E. Curr. Opin. Virol. 2012, 572 2 .E.R. Shmalenyuk, S.N. Kochetkov, L.A. Alexandrova et al., Russian Chem. Rev. 2013, 82, 896 3. E.R. Shmalenyuk, S.N. et al., Bioorg. Med. Chem. 2013, 21, 4874


137

Phosphocholine Liposomes as a Tool for Delivery of Anti-Tuberculosis

2’-Deoxypyrimidine Nucleoside Derivatives into Mammalian Cells

L.A. Alexandrova,1 G.M. Sorokoumova,2 M.A Zaretskaya2 and I.L.Karpenko1

1 Engelhardt Institute of Molecular Biology RAS, 119991, Russia, Moscow, Vavilova str., 32 2 Lomonosov State University of Fine Chemical Technologies, 12951, Russia Moscow, Vernadsky

avenue, 86


Development of novel anti-infectious agents is often hampered by low solubility of the compounds which are to block virus/bacteria and their insufficient ability to penetrate into the infected cell. In particular, this is true for the derivatives of 2’-deoxypyrimidine nucleosides bearing extended alkyloxymethyl or alkyl(1,2,3-triazol-1-yl)methyl substituents at С-5 position which target Mycobacterium tuberculosis[1]. We have recently showed that these compounds, although active against wild-type and multiple drug resistant strains of M.tuberculosis, are poorly soluble in aqueous media and are likely to have no active transport system into cells. So the goal of the present work was to develop an approach for delivery of this group of nucleosides into mammalian cells in order to increase their antimycobacterial activity. Our rational was to use liposomal vesicles composed of phospholipids as one of the most popular system for drug/gene delivery. An additional argument for choice of such carrier was that some of the phospholipids also suppress growth of M.tuberculosis[3].

As a model we used two cell lines: murine macrophages (J774) and human epithelial lung carcinoma (A549) since they are widely used for propagation of mycobacteria[4,5]. To ensure easy tracking of liposome penetration into the cells two alternative strategies were employed. One was based on a fluorescent phospholipid derivative - 1-oleoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (NBD-PC) which in combination with unmodified phosphocholine at 1:19 ratio assembled into liposomes. An alternative strategy was based on encapsidation of a reporter fluoresceine into phosphocholine (PC) liposomes. Visualization of the fluorescence by microscopy revealed extranuclear localization of the fluorescent compounds. Noteworthy, these approaches also allowed simple monitoring of delivery process during incubation of the cells.

Phosphocholine was found to be non-toxic for J774 and A549 cells at concentrations up to 300 µg/ml (LD50> 300 µg/ml). At lower concentrations (10-60 µg/ml) it enhanced cell proliferation, whereas presence of the nucleoside in these liposomes decreased effective concentration and/or increased its range. Moreover, encapsidation of the nucleosides with anti-mycobacterial activity allowed to substantially decrease their cytotoxicity.

Altogether, fluorescent compounds allowed us to determine an effective range of concentrations of the liposomes required for optimal delivery of anti-infectious agents into the cells as well as to prevent their toxic side-effects. This study was supported by the Program of the Presidium of the Russian Academy of Sciences “Molecular and Cell Biology” and by the Russian Foundation for Basic Research (grants № 15-04-05116 and № 14-04-00755)

References [1]. E.R. Shmalenyuk, S.N. Kochetkov, L.A. Alexandrova et al., Bioorg. Med. Chem. 2013, 21, 4874-48-84 [2] A.A. Mokhtarieh, S. Cheong, S. Kim, B.H. Chung, M. K. Lee, BBA, 2012, 1818, 1633–1641 [3] Sorokoumova G. M., Andreevskaya S. M., Smirnova T. G., et al., Bull. Exp. Biol. Med., 2009, 148(5),797-799 [4] D. Forge, D. Cappoen, J. Laurent, et al., Eur. J. Med. Chem., 2012, 49, 95-101. [5] Greco E., Santucci M.B., Sali M., et al., Immunology, 2010, 129, 125–132.


138

Antioxidant Activity of Rare-Earth Carboxylates with

2,6-Di-tert-butylphenol Moiety

T.A. Antonenko,1 D.B. Shpakovsky,1 Yu.A. Gracheva,1 T.V. Balashova,2

M.N. Bochkarev,2 E.R. Milaeva1

1 Organic Chemistry Department, Moscow State Lomonosov University, 119991, Russia, Moscow, Lenin Hill, 1/3

2 Razuvaev Institute of Organometallic Chemistry, 603950, Russia, Nizhny Novgorod, Tropinina, 49


The hindered 2,6-dialkylphenols are widely used antioxidants in industry and medicine as

vitamin E mimetics. The introduction of rare-earth elements into the molecules of phenolic

antioxidants enhances its activity and thermal stability. A series of rare-earth metal complexes

M(RCOO)3, (where M = La (1), Eu (2), Gd (3), Tb (4), Er (5); R = 3,5-di-tert-butyl-4-

hydroxyphenyl) were synthesized and characterized by elemental analysis, UV-vis and IR

spectroscopy. Compounds 1-5 are stable covalent monomers with bidentate coordination of

carboxylate groups with the metal center.

COOH

OH

But But

RCOOH

M(N(SiMe3)2)3 3 HN(SiMe3)2 M(RCOO)3

M = La (1), Eu (2), Gd (3), Tb (4), Er (5)

3 +20 oC

+

The radical scavenging activity of compounds was monitored spectrophotometrically in reaction

with stable 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and in the enzymatic process of linoleic

acid peroxidation by lipoxygenase (LOX). The high antiradical activity of all complexes was

demonstrated. The results of DPPH test show that the activity depends upon the nature of metal, the

antiradical activity was found the highest one for complex 5 (EC50 = 60 µM). The most effective

LOX inhibition activity was found for the complex 1 (EC50 = 32 µM). These results open up the

possibility for the design of novel physiologically active compounds.

Kinetic curves of enzymatic linoleic acid hydroperoxides formation in the presence of

different concentration of compound 1: 1 – 33.3 µM; 2 – 16.6 µM; 3 – 8.3 µM; 4 – 3.3 µM; 5 – 0.8 µM,

control LOX-1B, (λmax 234 nm, pH 9.0).

Acknowledgements - The financial support of RFBR (grant № 14-03-00611) is gratefully acknowledged.


139

Interaction of Glycyrrhizic Acid with Lipid Membrane: Perspectives

for Drug Delivery

I.E. Apanasenko,1,2 O.Yu. Selyutina,1,2 N.E. Polyakov,1 N.P. Isaev,1 A.V. Kim1

1Institute of chemical kinetics and combustion SB RAS,630090, Russia, Novosibirsk, Institutskaya, 3

2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova, 2


Glycyrrhizic acid (GA) is a natural oligosaccharide extracted from Glycyrrhiza Glabra (liquorice

root). Due to its amphiphilic properties GA can form supramolecular complexes with a wide range

of hydrophobic drugs resulting in increase of drug solubility, stability and bioavailability. The

hypothesis has been put forward that bioavailability enhancement may be caused not only by the

increase of the drug solubility, but also by the GA influence on the properties of lipid membranes.

In the present study we've made an attempt to clarify the mechanism of GA influence on drugs

bioavailability by investigation the mechanism of GA interaction with lipid membrane. The

unilammelar liposomes were used as model membranes. We’ve studied the GA influence on the

mobility of lipid molecules using nuclear magnetic resonance (NMR) relaxation technique with

shift-reagents addition. Shift-reagent allows to separate the inner and the outer part of the lipid

bilayer. Due to high sensitivity of spin-spin relaxation time to the mobility of measured groups of

protons, using NMR technique one can obtain the data on the mobility of different functional

groups of lipid molecule and make an assumptions about GA localization in membrane. To confirm

these assumptions electronic spin resonance (ESR) experiments and molecular dynamics (MD)

simulation of GA interaction with lipid bilayer were carried out.

It was established that GA interaction with liposome leads to significant changes in lipid

mobility. It could mean that GA is able to integrate into the lipid bilayer. Obtained results point that

GA is integrated only in the external half-layer, and its hydrophilic part remains outside the bilayer.

The fact of GA integration into bilayer is also confirmed by MD simulation. Also we've discovered

that combined action of GA and cholesterol results in the formation of rigid structures and pores in

the bilayer.

This comprehensive study sheds some light on the mechanism of the drugs bioavailability

enhancement in the presence of GA and offers the new possibilities of the GA application as a drug

carrier or drug delivery system.


140

Prospects of Plant Essential Oils Components Study for the Development of

Original Drug Substances

G.А. Atazhanova

JSC “International Research and Production Holding “Phytochemistry” 100009, Republic of Kazakhstan, Karaganda, fax: 8 (7212) 43 31 27


Among secondary metabolite of plants, essential oils, that are rich of biologically active

isoprenoids, are of substantial interest. We have studied chemical composition of essential oils of 51 plant species. First blend

composition of essential oils of 35 plant species of Kazakhstan flora were studied with

chromatography-mass spectrometry method, i.e., Ajania tenuifolia (Jacq.) Tzvel., Artemisia

aralensis Krasch, Artemisia arenaria DC., Artemisia viridis L., Artemisia ferganensis Krasch. ex.

Poljak, Artemisia halophila Krasch, Artemisia Кelleri Krasch, Artemisia monogyna Waldst. еt Kit.,

Artemisia saissanica (Krasch.) Poljak ex Filat., Artemisia tianschanica Krasch. et Poljak, Artemisia

valida Krasch et Poljak., Dracocephalum grandiflorum L, Dracocephalum karataviense Pavl. et

Roldug., Ferula ceratophylla Regel et Shmalh, Мicrocephala subglobosa Krasch, Tanacetum

kittaryanum C.A. Mey Tzvel, Tanacetum saxicola Krasch Tzvel, Thymus mugodzaricus Klok. et

Schost, Thymus karatavicus A. Dmitr. ex Gamajun, Salvia macrociphon L., Schrenkia congesta

Koro, Scutellaria immaculata Lam и т.д., from which 1647 compounds were identified. From

essential oils for chemical modification carvone, pulegone, limonene, cis-tongaosu, sabinene,

buddledine С, thymol, menthol were developed. Quantum-chemical calculations of electronic and

space structure of above mentioned terpenoids were conducted by means of Hartree-Fock ab initio

method in valent-sptitted bases 6-31G(d) and 6-31G(d,p) using GAUSSIAN program.

Furthermore biological screening of obtained essential oils, basic components and its derivatives

was conducted in regard of antibacterial, anti-inflammatory, anti-TB, phagocytosis stimulating,

analgesic, antiviral, cytotoxic activity. It was established, that studied essential oils and its

components have certain biological activity.


141

Study of Camphecin Influence on Mice Behavior in the Open Field Test

A. Babina,1 V. Lavrinenko,1 O. Yarovaya,1,2 N. Salakhutdinov1,2

1 Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova Str, 2

2 N.N. Vorozhtsov Institute of Organic Chemistry, 630090, Russia, Novosibirsk, Lavrentieva Str, 9


Behavior is the possibility of animals and humans to react to the influence of the inner and outer

factors, which provide the survival of individuals or species. According to the modern conception

the brain integrative activity, which underlies a behavior, consists in the consecutive order of

functional systems activation. Psychophysiological mechanism of motivations and emotions has

difficult neurotransmitter character, which depends on the influence of the outer factors.

The aim of the present study was the analysis of Camphecin (1,7,7-

trimethylbicyclo[2.2.1]heptan-2-ylidene-aminoethanol) influence on the mice behavior in the open

field test. Camphecin possesses the antiviral features, and inhibits influenza replication, but not

enough is known about its influence on the nervous system activity.

Experiments are carried out on the nubilous mice of several genetic strains. The preparation was

injected intraperitonealy in the dose of 100 mg/1000 g body weight. The influence of the acute

(single injection) and chronical (5 times per 2 weeks) injections was studied.

One-way variance analysis shows, that single injection of Camphecin doesn’t affect the

locomotive activity, but significant increase of the freezing behavior in the experimental group has

been revealed. This fact is considered as a heightened anxiety. The decrease of the vegetative

activity is found under single injection, which may indirectly indicate the increased activity of the

parasympathetic centers. Thus under the single Camphecin administration reveals no material

change in the behavior patterns. The chronic Camphecin solution introduction to the mice of the

different strains doesn’t show the significant influence of the preparation on the behavior patterns,

which have been studied in the open field test (locomotive, research activity, anxiety, emotionality

and vegetative state of the organism).

The received results are discussed from the position of the neutral influence of Camphecin on the

behavioral patterns in animals.

This work was supported by the Russian Science Foundation (15-13-17)


142

Creation Medications on the Basis of Extractives from Wood of Siberian Larch Species

V.A. Babkin, L.A. Ostroukhova

A.E. Favorsky Institute of Chemistry Russian Academy of Sciences Siberian Branch, 664033, Russia, Irkutsk ,Favorsky St., #1


Theoretical foundations of the integral processing of conifer wood of Siberia and Far East biomasses to develop new technologies for innovative medical and veterinary preparations.The work involves at fundamental and applied researches of flavonoids and polysaccharides of wood and its polyphenol components as well as bark pectines of larch. Its relevance proved to be important to elaborate innovative technologies for deep integrate processing of the larch biomass, to have prospects for processing industry located immediately in the region of the raw material growth (the biorefinary principle) to produce science intensive and value-added products.

Currently, the innovative medicines having no analogues and based on bioactive extractives from larch biomass are under developments in the Laboratory.

The main concept of this investigation is the progress at domestic pharmaceuticals, based on highly active non-toxic, produced by the deep chemical processing of Siberian and Gmelin larch wood, the natural compounds: biflavonoids, polysaccharides, phenolic acids, proanthocyanidines, lignans.

The reason for use of Siberian and Gmelin larch biomasses is specified by peculiar characteristics of their extractives, unique component compositions of the latter, with a difference the other. species of the Larix genus, growing in the Russian Federation, they differ on quantitative content of DHQ, arabinogalactan (AG), spirobiflavonoids, proanthocyanidins, etc.

Dihydroquercetin (DHQ) is a natural flavonoid isolated from Siberian larch (Larix sibirica Ledeb.) and Gmelin larch (Larix gmelinii Rupr (Rupr.)), which content is up to 4.0% by weight of dry wood. The DHQ molecule contains two asymmetric carbon atoms C-2 and C-3, therefore, it can exist as four enantiomers (two pairs of diastereoisomers). The main natural isomer is trans-(+)-(2R3R)-dihydroquercetin. Technology for DHQ production is developed.

The technique for several analysis of DHQ isomers in lumped product, obtained by the technology, has been worked out. The lumped substance consists of no less than 96 % of bioactive trans-(+)-(2R3R) isomer, and so, it has a high antioxidant activity (on scale ORAC 32742,95 µmol TE/g. These studies were performed by ABC Testing laboratory (Advanced Botanical Consulting & Testing, INC.), USA).

Products based on developments of the laboratory, are commercial ones, registered and approved for use by the Ministry of Health of Russian Federation (ROSPOTREBNADZOR).

One of the higher-priority investigation of the Laboratory is the technology for extraction of arabinogalactan (AG), i.e. natural polysaccharide containing in wood and bark of the larch, having unique biological and physicochemical properties. Its natural macromolecular structure, water-solubility, membranotropic properties, ability to receptor-induced endocytosis have aroused the interest for its soluble dietary fibers, and not for that, but it is the carrier for pharmacological molecules to bring biologically active substances to tissues that prolongs the latter action. AG matrix is used as carrier for poor soluble and toxic medications to create new soluble medicines. For example, clathrates of AG with arbidol, remantadine and DHQ are developed by solid-phase mechanochemical synthesis. Bioavailability (solubility) of the drugs consisting of clathrates is increased, so, the therapeutic doses can be substantially decreased at the same adequate level of activity of unmodified preparations.

Another high-priority area of the Laboratory research is the technological synthesis of complex compounds on the basis of available antioxidants (flavonoids) and biogenic metals, the study of their pharmacological properties and the elaboration of import-substituting domestic medicines and veterinary drugs. Thus, a lot of socially hazardous diseases, which etiology and pathogenesis caused by cellular redox-status affection, can be treated by new-developed effective high-antioxidative drugs.


143

Computer Simulation of 11-Phenylundeca-5Z,9Z-dienoic Acid Human

Topoisomerase I Inhibitory Activity

D. Baev,1 V. D'yakonov,2 and T. Tolstikova1

1N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, Siberian Branch, Russian Academy

of Sciences, 630090, Russian Federation, Novosibirsk, Lavrentiev Avenue, 9

2Institute of Petrochemistry and Catalysis, Russian Academy of Science,, 450075, Russian Federation, Ufa, Prospekt Oktyabrya, 141


The search for new efficient and low-toxicity antitumor agents is a highly important task of

modern medicinal chemistry. Numerous studies in this field demonstrated that topoisomerase I

(hTop1) is among the key molecular targets for the development of modern antitumor agents [1].

The molecular docking of 11-phenylundeca-5Z,9Z-dienoic acid was performed using the refined

computer model of the compound in interaction with topoisomerase I active site obtained by

crystallographic methods [2].

A comparison of the location of 11-phenylundeca-5Z,9Z-dienoic acid (2) and its initial (5Z,9Z)-

5,9-eicosadienoic acid (1) [3], which also exhibited a high hTop1 inhibiting activity, in the

topoisomerase I active site, indicates that the lipophilic moiety of acid 2, like acid 1, forms

favorable hydrophobic contacts with Arg364 (Fig. 1A and B). The replacement of the alkyl chain in

the initial acid 1 molecule by the lipophilic phenyl moiety brings about active involvement of the

phenyl π-system into the stacking (π–π interaction) with the aromatic structure of nucleotides

(adenine). Stacking is known to be the principal type of noncovalent interaction in the DNA

structure stabilizing the helical configuration.

Figure 1. Docking of acid 2 (A) and acid 1 (B) in the topoisomerase I active site.

A B

References [1] Y. Pommier, ACS Chem. Biol. 2013, 8 (1), 82-95 [2] V. D'yakonov at al., Bioorg. Med. Chem. Lett. 2015, 25 (11), 2405-2408 [3] V. D'yakonov at al., Chem. Commun. 2013, 49, 8401-8403


144

The Influence of Organotin Compounds on Lipid Peroxidation in Liver of the

Russian Sturgeon in vitro

N. Berberova,1,2 E. Mukhatova,1 V. Osipova,2 M. Kolyada,2 D. Shpakovsky,3 E. Milaeva3

1Astrakhan State Technical University,414056, RUSSIA, Astrakhan, Tatischeva, 16e

2Southern Scientific Centre of the RAS, 344006, RUSSIA, Rostov-on-Don, prosp. Chekhova, 41 3Moscow State Lomonosov University, Moscow, Russia


The combination in one molecule the prooxidant and antioxidant groups relevant to the search and establishment of pharmacological agents with no adverse side effects. The reduction of prooxidant activity of organotin chlorides, as it was shown in the model peroxidation of (Z)-octadec-9-enic (oleic) acid, may be achieved by introducing a 2,6-di-tert-butyl-4-mercaptophenol antioxidant fragment in the molecule [1].

The influence of methyl- and phenyltin(IV) (1-4) containing 2,6-di-tert-butyl-4-mercaptophenol pendant [2] on the accumulation of thiobarbituric acid reactive substances (TBARS) – the secondary products of the lipid peroxidation (LP) that can react with thiobarbituric acid at different stages of lipid peroxidation of the Russian sturgeon liver homogenate in vitro for 48 h.

HO

But

But

S Sn(CH3)22

HO

But

But

S Sn(CH3)3

HO

But

But

S SnPh3 HO

But

But

S SnPh2

2

2

4

1

3

Fig. Variation of TBARS accumulation in Russian sturgeon liver in vitro in presence of compounds 1-4 (C = 10-4M). Differences from control of a - (p <0.01); b- (p <0.05).

It is found that phenyltin thiolates significantly reduce the accumulation of TBARS level at all stages of oxidative process in contrast to the initial organotin chlorides. The inhibitory activity of the methyl derivatives was significantly lower in the initial phase (1 h) for Me2Sn(SR)2 TBARS levels decreased by 18%, and for Me3SnSR (3-24 h) accumulation of lipid peroxidation products was similar to the control (Fig.). The greatist antioxidant effect found for 4 at 24 h period of oxidation, the level of TBARS was reduced by 26% when compared with the control. It should be noted that all compounds under study do not demonstrate an inversion of antioxidant action.

Thus, the introduction of 2,6-di-tert-butyl-4-mercaptophenol fragment in organotin complexes leads to the inversion of prooxidant properties of starting organotin clorides. The obtained data allow one to consider the organotin compounds with thiophenolate ligands as potential antitumor agents. The financial support of RFBR (grants № 13-03-00487, 14-03-00578, 15-03-03057) is gratefully acknowledged.

References [1] E. Mukhatova, V. Osipova, M. Kolyada vet. al., Doklady Chemistry, 2013, 1, 177-180. [2] D. Shpakovsky, С. Banti, E. Mukhatova vet. al., Dalton Trans., 2014, 43, 6880-6890.


145

Antiulcerative Activity of New Omeprazole Analogues

M. Biryukova,1 N. Zhukova,1 T. Tolstikova,1,2 O. Yarovaya,1,2

D. Sokolov1 and N. Salakhutdinov1,2

1 N.N. Vorozhtsov Institute of Organic Chemistryof the Siberian Branch of Russian Academy of

Science, 630090, Russia, Novosibirsk, Lavrentieva Ave, 9 2 Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova St, 2


A benzimidazole derivative omeprazole is a proton pump inhibitor used in the treatment of

peptic ulcer disease and some other gastrointestinal pathologies. It is known as one of the most

important and widely prescribed medication in the basic health system. In order to obtain new

pharmacological substances revealing gastroprotective activity 22 new benzimidazole derivatives

were synthesized.

The experiment was performed on 210 male Wistar rats. The derivatives of benzimidazole under

investigation were administered intragastrically in the dose 100 mg/kg while omeprazole was

administered at the same dose as the reference substance. The antiulcerative activity of these new

compounds was investigated in vivo on the indomethacin-induced peptic ulcer model.

The results of the experiment show that 4 of 22 new benzimidazole derivatives demonstrate

significant antiulcerative activity which is comparable to the one omeprazole has. Moreover, two of

these four compounds had twice the pharmacological effect compared to the reference substance.

Thus, the researched compounds may be considered as a promising basis in future investigations

in the field of ulceration therapy.


146

One-pot ‘on-solvent’ Multicomponent Protocol for the Synthesis of Medicinally

Relevant Pyrano[3,2-c]quinolone Scaffold

S.I. Bobrovsky, M.N. Elinson

N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Science, 119991, Russian

Federation, Moscow, Leninsky prospect, 47


Pyrano[3,2-c]quinolone scaffold is of particular interest due to the fact that this scaffold was

found in many natural alkaloids with anti-inflammatory and cancer cell growth-inhibitory activity.

Recently pyrano[3,2-c]quinolone scaffold was found to display low nanomolar antiproliferative

activity to induce apoptosis in human cancer cell lines and to be microtubletargeting agent [1,2].

The role of multicomponent reactions with the formation of biologically active pharmacophores,

which do not require further isolation and purification, should get much more attention in the future,

because they satisfy for the modern environmental requirements of ‘green chemistry’.

We reported our results on the novel efficient multicomponent assembling of salicylaldehydes

1a-g, malononitrile and 4-hydroxy-1-methylquinolin-2(1H)-one into earlier unknown pyrano[3,2-

c]quinolones 2a-g by catalyzing NaOAc in ethanol with 88–98% yields.

The developed multicomponent procedure requires simple and reasonable starting materials. It is

easily carried out, and the reaction products are crystallized directly from the reaction mixture.

References [1] G. Brader, M. Bacher et al., Phytochemistry. 1996, 42, 881-884 [2] I.V. Magedov, M. Manpadi et al., J.Med.Chem. 2008, 51, 2561-2570


147

Degradation of B-domain Deleted Factor VIII in Serum Containing Medium

V. Bodyagin, D. Milekhin, and J. Krasotkina

Biochemical Production Department, CJSC ”GENERIUM”, 6001125, Russia, Volginsky, 17


B-domain deleted factor VIII (BDD-FVIII) is a novel drug for replacement therapy of

hemophilia A. It is the active pharmaceutical ingredient of Refacto/Xynta, manufactured by Pfizer.

Recombinant FVIII has been engineered extensively throughout the years to increase the low

production yields that were obtained from mammalian cell cultures. It was shown that deletion of

bulky, heavily glycosylated B-domain significantly improves expression level of desired protein

without reduction of its coagulation activity. Although the functional role of native FVIII B-domain

is not finally clarified, it is reliably known that this domain does not contribute either to the intrinsic

tenase formation or to the activation of factor X. Yet, the recombinant production of B-domainless

FVIII is complicated because of its interaction with the membrane of the producing cells in serum

free medium. In 2010 Kolind et al found that 90% of rFVIII is attached to the cell membrane of the

producing cell when the rFVIII variant contains a short part of B-domain (21 aa). By increasing the

length of the B-domain the membrane attached fraction can be reduced only to 50% of the total

expressed rFVIII. In serum containing medium BDD-FVIII does not attach to host cell. Instead of

this it rapidly degrades. The heavy chain of BDD-FVIII undergoes proteolytic cleavage by activated

coagulation factors: thrombin, Factor Xa and Factor IXa.

The scope of this work was to investigate the role of serine proteases in BDD-FVIII degradation

in serum containing medium. The use of recently launched in market affinity chromatography resin

VIIIselect (GE Healthcare) enabled to characterize integrity of BDD-FVIII directly in culture

media.

The results showed that BDD-FVIII is stable in buffer solutions or regular growth media like

DMEM or RPMI for at least 24 hours at 37C. At the same conditions but in the presence of serum

(from 1 to 10 % v/v) BDD-FVIII retained less than 10% of its initial activity and its heavy chain

was almost completely degraded. Addition of benzamidine up to 1 mM did not prevent degradation

of the heavy chain and the loss of coagulation activity. Probably the intrinsic instability of BDD-

FVIII attributed to the weak interaction between the A2 domain and the metal ion-linked A1/A3-

C1-C2 dimer has to be considered in further investigation.


148

1-Phenyl-1,4-dihydrophosphinolines – New Type of Tertiary Phosphines

A. Bogachenkov,1 A. Dogadina,2 and A. Vasilyev1

1 Institute of Chemistry, Saint-Petersburg State University, Petrodvorets, Saint Petersburg, 198504,

Russian Federation 2 Saint- Petersburg State Technological Institute (technical university), Saint Petersburg, 190013,

Russian Federation


Tertiary phosphines are important organic derivatives of phosphorus. They are of interest, on the

one hand, as ligands in organometallic chemistry due to their ability to complexation, and on the

other hand, as compounds with potential (and some explicit) biological activity, both in free form

and in the form of phosphonium salts and complexes [1].

This work presents a method of obtaining previously unknown tertiary phosphines, 1-phenyl-1,4-

dihydrophosphinolines, based on reduction of the corresponding 1-phenyl-1,4-

dihydrophosphinoline 1-oxides with trichlorosilane in the presence of triphenylphosphine. 1-

Phenyl-1,4-dihydrophosphinoline 1-oxides were obtained via intramolecular cyclization of

diphenylphosphorylallenes in the presence of Brønsted superacids (CF3SO3H, FSO3H) and Lewis

acids (AlCl3, AlBr3) [2].

R2

R2

R1

PPh2OP

R2 R2

O PhR1

HSiCl3

R1= H, Br, Alk, Ph; R2= Alk

P

R2 R2

Ph

R1

R1= H, Br; R2= Me

Acid

References

[1] Ramachandran, E., Raja, D.S., Rath , N.P., Natarajan K., Inorg. Chem. 2013, 52, 1504-1514. [2] Bogachenkov, A.S., Dogadina, A.V., Boyarskiy, V. P., Vasilyev, A.V. Org. Biomol. Chem. 2015, 5, 1333-1338.


149

Design Salbutamol and Budesonide Solid

Dispersion for Pulmonary Drug Delivery

E. Bogdanova,1,2 A. Ogienko,1 E. Boldyreva,2,3 S.A. Myz,3 A. Ogienko,4 B. Kolesov,1,2

V. Drebushchak,2 A. Manakov,1,2 Yu. Kovalenko,5 N. Trofimov,5 V. Boldyrev2,3

1Nikolaev Institute of Inorganic Chemistry SB RAS, 630090, Russia, Novosibirsk, Lavrenteva pr., 3.

2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova st., 2. 3Institute of Solid State Chemistry and Mechanochemistry SB RAS, 630128, Russia, Novosibirsk,

Kutateladze, 18. 4Institute of Cytology and Genetics SB RAS, 630090, Russia, Novosibirsk, pr. Lavrenteva, 10.

5JSC “Nativa”, 123001, Russia, Moscow, Ermolaevsky per., 25. E-mail: katerinazevak@gmail.

Recently, we proposed an alternative method of producing novel improved forms of active

pharmaceutical ingredients (APIs) based on spray-freeze drying of API solutions in “volatile

organic liquid – water” systems giving clathrate hydrates [1]. Salbutamol and budesonide are APIs

used in asthma treatment. They are poor water soluble drugs, so their manufacture is limited. Herein

we present a composition design using molecular-level encapsulation of an API in a large porous

composite matrix.

Compositions were prepared by spray-freeze drying a salbutamol (budesonide)/glycine

tetrahydrofuran (THF) – water solution of the composition, corresponding to the clathrate hydrate

existing in the THF-water system. The prepared formulations were light fluffy powders that

demonstrated easy flow and very low bulk density. According to the SEM images, the prepared

samples were porous spherical agglomerates with sizes 30-70 µm, consisting of plane particles 1-

10 µm in length and less than 100 nm in thickness. In contrast to micronized and nanosized

powders, the formulations obtained in this work did not suffer agglomeration on storage. Raman

spectroscopy, DSC and XRPD experiments shows the following main features for compositions:

glycine was present as the metastable β-modification, API were not seen in the crystal structure of

either compositions, API molecules did not form an individual crystalline phase, and, probably were

uniformly distributed between nanosized glycine, thus resulting in a solid dispersion. In vitro tests

by Anderson Cascade Impactor indicated that our samples exhibited significantly higher fine

particle fractions (amount of the substance allegedly penetrating into human lungs) of 59±2 %

compared to the known behavior of nano-salbutamol sulphate blends (45.2%) and micronized

salbutamol sulfate blends (31.3%). For budesonide based formulations - 51.2 % (obtained in this

work) and 20 % (commercial sample).

References [1]. A. Ogienko et. al. Pharm. Res., 2011, V 28 (12), 3116-3127


150

The Comparison of Pharmacological Activity of Ibuprofen and Aspirin after

Complexation with Arabinogalactans of Different Descent

S.A. Borisov,1 T.G. Tolstikova,1 M.V. Khvostov,1 A.V. Dushkin2 and J.V. Chistyachenko2

1 Institute of Organic Chemistry of the Siberian Branch of Russian Academy of Science, 630090,

Russia, Novosibirsk, Lavrentieva st., 9 2Institute of Solid State Chemistry and Mechanochemistry, Siberian Branch, Russian Academy of

Sciences, 630128, Russia, Novosibirsk, Kutateladze st., 18


Non-steroidal anti-inflammatory drugs (NSAIDS) aspirin (AS) and ibuprofen (IB) are among the

most popular pharmaceutical agents currently available on the market. Yet they have some serious

adverse effects the most significant of them being the irritation of the gastrointestinal mucosa. A

promising technique to avoid these side effects is the formation of complexes with a larch

polysaccharide arabinogalactan. In this study we compare the anti-inflammatory and analgesic

effects of the complexes of AS and IB with two different arabinogalactans. One is a highly purified

AG produced from Canadian larch trees (AGC) which is very popular in the western medicine and

the other is obtained from Siberian larch trees (AGS) and is currently under research.

The experiments were performed on albino mice of both genders. Complexes with AGC and

AGS were administered intragastically at doses 200 and 400 mg/kg for ibuprofen and 250 and 500

mg/kg for aspirin (double doses for inflammation test). Pure aspirin and ibuprofen were also

administered orally as a positive control at respective doses (20 and 40 mg/kg for ibuprofen, 25 and

50 mg/kg for aspirin). For the evaluation of analgesic activity of these compounds we used two

standard pharmacologic tests: the hot-plate test and the acetic acid writhing test. As a model of

inflammation, a histamine swelling test was performed. The results of both analgesic tests show that

complexes of AS and IB with AGS have a significant beneficial effect compared to pure AS and IB,

because they were active at doses 200 mg/kg for IB and 250 mg/kg for AS while the pure

substances were not active in the corresponding doses. On the other side, complexes with AGC

didn’t show any difference in analgesic activity in comparison with pure AS and IB. In the

histamine swelling test similar data was obtained showing that complexes with AGS at both doses

significantly decreased the inflammation rate, while the complexes with AGC produced the same

effect only on the higher of these doses, same as pure AS and IB.

To sum up, our study clearly shows that AGS is able to reduce the therapeutic dose of AS and IB

in complexes without losing their pharmacological effects, something that AGC was not capable of

achieving. Thus, we can claim that AGS is superior than AGC in terms of using it as a

complexation drug delivery system.


151

Alternative Desalting Approach for Semi Preparative Oligonucleotide

Manufacturing on Industrial Scale

E. Bulgakova 1Skolkovo Institute of Science and Technology, Dep. of Functional Genomics and Inflectional

Diseases, Skolkovo Innovation Center, Bldg. 5, Moscow, 143026, Russia (at present) 2Alnylam Pharmaceuticals, Cambridge, MA, USA (ex- employee)

At present time a great number of pharmaceutical companies in the world are developing new

therapeutics based on various modified oligonucleotides and their mimics. There are some examples of FDA- approved oligo-compounds as drugs (as Macugen of ISIS Pharm). Other companies, like Alnylam Pharmaceuticals, are actively working on the development of new drug generation based on siRNAs (short 22-mer ribo-oligonucleotides) for down regulation of undesirable genes using so called RNA silencing technologies. So, there is a distinctive trend for oligonucleotide manufacturing to transfer from research to industry scale, and in other words it is manufacturing of oligo- products starting from milligrams to grams and even more.

The main goal of this research was the development of alternative desalting method for solid phase manufacturing of ribo- oligonucleotides, which has been worked out in Medium Scale Synthesis Group of Alnylam Pharmaceuticals. Usually, desalting is the third and final stage of oligonucleotide preparation after solid phase synthesis on automated synthesizers and automated HPLC purification using ion exchange columns. Basically, desalting is an additional chromatography step using preparative gel- filtration columns ( typically with Sephadex G-25).Thus, the desalting stage is necessary due to the immense amount of salt ( up to 10-20 g ) per oligo- sample after every ion exchange purification for semi-preparative(100mg) and preparative( gram) oligo synthesis scale.

Unfortunately, the high amount of salt on final stage turns to be a real manufacture problem as there is a concentration limit of salt amount compatible with gel filtration for each desalting run.

Here, we propose alternative desalting procedure consisting of a) oligo pre-concentration step by using precipitation in organic solvents (ethanol or ethanol/acetonitrile mixture) followed by b) normal gel filtration on preparative Sephadex G-25 columns. During the first step of a procedure, as it has been shown by our results, about 80-90% of salt in grams has been removed and thus the next gel filtration step on Sephadex G-25 column has been facilitated.

So, the developed method allowed us to decrease the amount of salt dramatically before final Sephadex G-25 desalting. This remarkable fact led us for a number of advantages of alternative method over common desalting.

Briefly, alternative 2-step desalting procedure enabled us to: 1)shorten a total number of gel-filtration runs per sample, 2) increase oligonucleotide

concentration limits for gel filtration up to 3-5 times,3)intensify the whole manufacturing cycle up to 30% as an overall oligo-samples throughput has been increased.

Other benefits of the method proposed: Method is simple and robust and proved for scaling up the desalting procedure up to 50-100

mg of final oligonucleotide product Method is compatible with automated desalting procedure on Gilson HPLC system with auto

sampler which has been developed in Alnylam Pharmaceuticals, as well Method has been proved on more than 40 RNA oligo-compounds ( using native or 2’-OMe

modified ribo-oligonucleotides) *All parts of research development have been performed using laboratory equipment of Medium

Scale Synthesis group of Alnylam Pharmaceuticals. **Author greatly thanks her former directors of Alnylam Pharmaceuticals: Dr. M. Maier and

Dr.K. Charisse for their support and interest for the described research.


152

Total Synthesis of Tiasporine B and C

A.S. Bunev, S.A. Sokov, Ya.I. Rudakova, and G.I. Ostapenko

Laboratory of Functional Heterocyclic Compounds, Togliatti State University, 445667, Russia,

Togliatti, Belorusskaya St., 14


In this work will be present results for the total synthesis series thiazole-containing alkaloids,

Tiasporine B and C, which isolated from marine actinomycetes Actinomycetospora Chlora SNC-

032 [1].

The authors are grateful to the Ministry of Education and Science of the Russian Federation (state job No. 426)

References

[1] Fu P., MacMillan J. B. J. Nat. Prod., 2015, DOI: 10.1021/np500929z


153

Synthesis of New Potential Inhibitors of Protein Kinases

A.S. Bunev, Ya.I. Rudakova, A.P. Pavlova and G.I. Ostapenko




In this report will be present the main results of the synthesis bis-thiazolamines and in vitro

investigation their inhibiting activity agains certain tyrosine-kinase (EGFR).



154

Synthesis of Novel Thiazole-containing Amino Acids

A.S. Bunev, Ya.I. Rudakova, S.A. Sokov and G.I. Ostapenko




By now, there is known a comparatively large class of natural compounds possessing biological

activity, which contain imidazole, thiazole, or oxazole moieties. Studies of biological activity and a

total synthesis of cyclic alkoloids isolated from marine microorganisms are very important

directions. In many cases, they are promising antitumor and antibacterial compounds.

Analysis of molecular sctructures of such alkaloids as microcococin P1, promothiocin A,

amithiacin, etc. shows thatthey have a 2-aminomethyl-1,3-thiazole-4-carboxylic acid moiety as one

of the building blocks. Despite that the methods for the synthesis of this compounds derivatives are

well enough described in the literature, we suggest a synthetic approach to the preparation of new

thiazole-containing amino acids, the sctructural analogs of 2-aminomethyl-1,3-thiazole-4-

carboxylix acid.



155

Tetracationic 1,4-Diazabicyclo[2.2.2]octane Derivatives as an Attractive Scaffold

in Antimicrobial Drug Design

E. Burakova, I. Saranina, N. Tikunova, V. Silnikov

Institute of Chemical Biology and Fundamental Medicine, 630090, Russia, Novosibirsk, Lavrent’ev

Ave, 8


Among the potential candidates in search of antibacterial drugs, the attention may be drawn to

quaternary ammonium compounds (QACs), which are used as antimicrobial, disinfectant and

sterilizing agents [1]. The main effects of QACs in bacteria are structural and functional changes in

the cell wall, release of wall components and initiation of autolysis, inhibition of membrane

ATPase, and electrostatic interactions with negatively charged polar head groups of phospholipids.

The antibacterial activity of QACs strongly depends on the nature of the organic groups attached to

the nitrogen, the number of nitrogen atoms present, and the hydrophobicity of alkyl chain.

We designed and synthesised a series of tetracationic QACs, each with two 1,4-

diazabicyclo[2.2.2]octane (DABCO) moieties substituted at the bridge-nitrogen atom with alkyl

chains connected by different linkers. Their antimicrobial activity was evaluated on five gram

negative bacteria (Pseudomonas aeruginosa, Proteus vulgaris, Citrobacter freundii, Salmonella

enterica, Escherichia coli), two gram positive bacteria (Staphylococcus aureus, Enterococcus

faecalis) and one fungi - yeast Candida albicans.

4X-

N+N+H2n+1CnN+ N+ CnH2n+1LINKER

HYDROPHOBIC GROUPS

CATIONIC GROUPS

The experiments in vitro confirmed the relationship between the antimicrobial activity and the

structure of the studied compounds. The some DABCO derivatives showed high inhibitory activity

against both Gram-positive and Gram-negative bacteria as well as fungus. Most compounds

exhibited anti- S. aureus activity better than the common antibacterial drugs ciprofloxacin and

gentamicin. The further studies of these compounds may provide a convenient platform for the

development of new class of antibiotics.

References [1] P. Gilbert, and L.E. Moore, J. Appl. Microbiol. 2005, 99, 703–715


156

Mechanical Disordering of Cell Walls Supramolecular Structure

A.L. Bychkov,1 E.M. Podgorbunskikh,1 E.I. Ryabchikova,2 O.I. Lomovsky1

1Institute of Solid State Chemistry and Mechanochemistry SB RAS,

18 Kutateladze str., Novosibirsk, Russia 2Institute of Chemical Biology and Fundamental Medicine SB RAS,

8 Lavrentiev Avenue, Novosibirsk, Russia


The main obstacle to the development of mechanical methods of lignocellulose pretreatment is

the difficulty of scaling laboratory development and transferring it to the semiindustrial or industrial

equipment. Most of the current research is aimed at achieving the maximum possible reactivity of

cellulose without considering the supramolecular structure of cell walls polymer complex, which is

often not profitable for energy consumption.

In our study the information about fine grinding of plant materials in laboratory and

semiindustrial mills is introduced. It is shown that in the initial period of grinding plant material

forms dense aggregates consisting of crushed particles. In this case grinding lignocellulose is done

by cracking of tissues by cell walls. The mechanism of destruction of cell walls (fig. a-c) strongly

depends on the conditions of mechanochemical experiment [1].

With further grinding is a destruction of aggregates, releasing of particles and a further decrease

in their size takes place. Carried correlation between of reactivity of lignocellulose in further

enzymatic hydrolysis with physicochemical parameters such as specific surface area, total pore

area, the degree of crystallinity of cellulose and morphology of the resulting product. Several

proposals to enable optimize energy consumption at increasing the reactivity of cellulose for the

case of centrifugal-roller mills are expressed in our study [2].

References [1] A.L. Bychkov, E.I. Ryabchikova, K.G. Korolev and O.I. Lomovsky, Biomass and Bioenergy. 2012, 47, 260-267. [2] A.L. Bychkov, V.A. Buchtoyarov and O.I. Lomovsky, Cellulose Chemistry and Technology. 2014, 48, 545-551.


157

Interaction of Tris(1-alkylindol-3-yl)methylium salts with Human Serum

Albumin-the Docking Study

E.E. Bykov,1 N.A. Durandin,2 S.N. Lavrenov,1 M.N. Preobrazhenskaya1

1Gause Institute of New Antibiotics, Russian Academy of Sciences,

11 B. Pirogovskaya Street, Moscow, 119021, Russia; 2Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, 4 Kosygin Street,

Moscow, 119334, Russia E-mail: [email protected]

Recently we have described derivatives of tris(indol-3-yl)methylium salts with the alkyl

substituents at the N-atoms of the indole rings (Fig.1), which were proved to have high cytotoxic

properties at nanomolar-to-submicromolar concentrations for human tumor cell lines1 . Binding of

drugs to serum albumin in biological systems is a key process that can modulate a number of

properties of the carried agent, such as increased solubility in plasma, decreased toxicity, protection

against oxidation or prolongation of the in vivo half-life; hence, binding is essential for

understanding biodistribution. From this point of view it is important to study the interaction of

biologically active compounds with serum albumin (HSA).

R

R RN N

N

N N

NR

RR Figure 1.Tris(indol-3-yl)methylium salts: R =H (antibiotic turbomycin), Me, Et , n-Pr, n-Bu, n-C5H11 , n-C6H13 , n-C7H15 , n-C8H17 , n-C9H19, n-C8H17,C10H21 , C11H23.

The free energy of the interaction of ligands tris(1-alkylindol-3-yl)methylium salts ( Fig.1 ) with

the target (HSA) was calculated by means of the package Molegro Virtual Docker 2. The results

show that free energy value (LgK) depends on the length of R. (Table1)

Table1: The dependent of ∆G (LgK) from the length of R R H Me Et Pr Bu C5H11 C6H13 C7H15 C8H17 C9H19 C10H21 C11H23

∆G, kcal.mol-1 -9.3 -9.4 -11.5 -13.1 -11.9 -12.9 -13.6 -14.1 -11.8 -15.2 -14.6 -13.1

lgK 7.1 7.3 8.8 10.0 9.1 9.9 10.4 10.7 9.0 11.6 11.2 10.0

They almost monotonically increase from R = H to R= n- C7H15; from n- C7H15 to n- C11H23

scatter in the data was observed. This is consistent with experimental data on the binding energy of

the above series of compounds. References

[1]. S. N. Lavrenov, Y. N. Luzikov, E. E. Bykov, M. I. Reznikova,E. V. Stepanova, V. A. Glazunova, Y. L. Volodina, V. V.Tatarsky, Jr., A. A. Shtil, M. N. Preobrazhenskaya, Bioorg.Med. Chem., 2010, 18, 6905.

[2]. http://molegro-virtual-docker.software.informer.com


158

New Approaches to Chemo- and Stereoselective Synthesis of Substituted

Dihydropyrimidines or Dihydropyridines

V. Charushin,1 O. Fedorova,1 Yu. Titova,1 A. Vigorov,1 D. Gruzdev,1 V. Krasnov,1

I. Krivtsov,2 A. Murashkevich,3 G. Rusinov1

1Postovsky Institute of Organic Synthesis of RAS, 620990, Russia, Ekaterinburg,

S. Kovalevskoy/Akademicheskay st., 22/20 2South Ural State University, 454080, Russia, Chelyabinsk, Lenina ave., 76

3Belarusian State Technological University, 220006, Belarus, Minsk, Sverdlova st., 13a, E-mail: [email protected]

New catalytic systems, based on nano-sized metal and silicon oxides, for chemo- and

stereoselective synthesis of substituted 3,4-dihydropyrimidines (DHPMs) and 3,4-dihydropyridines,

showing a high cardiotropic or anticancer activity have been developed. Nowadays, one of the main

problems in DHPMs synthesis is to obtain their enantiopure forms. Indeed, both enantiomers of

DHPMs have been found to show distinct or opposite pharmacological activities [1,2]. We have

shown that the addition of these nanooxides enhances the enantioselective effect of L-proline-

derived chiral inducers in the Biginelli reaction (compound 1). The best results (ее 70-76%) have

been achieved by using chiral inducer 3 and mixed nanooxides ZrO2-SiO2, TiO2-SiO2. For a well

studied anticancer agent, monastrol (2), the presence of the same catalytic system has enabled to

obtain this compound in 45% ее.

NH2

NH2 X NH

NH

XMe

EtO2C

R

OMe

EtO2C CHO

R

NH

N

O CH3

OH

+

1, X = O, R = H; 2, X = S, R = OH

3*

It has been shown that also nifedipine 4 is formed in yields of 92-95% in the presence of nano-

sized oxides SiO2, TiO2-Al2O3 or MgO-SiO2 (GLC data). This method appears to be a good basis to

develop a new technology for nifedipine, which is widely used in medicine.

NH

Me Me

MeO2C CO2Me

NO2

NH3O Me

CO2Me

OMe

MeO2CCOH

NO2

+

4

This work was supported by the Russian Foundation for Basic Research (grants 13-03-12188-ofi_m, 14-03-31357 mol_a,) and by the State Program for the Support of Leading Scientific Schools (grant NSh 3656.2014.3).

References [1] K.S. Atwal, B.N. Swanson, S.E. Unger, B.C. O’Reilly, J. Med. Chem. 1991, 34,806-809. [2] C.O. Kappe, Eur. J. Med. Chem. 2000, 35, 1043-1055.


159

Arylethynyl Nucleosides as Potent Inhibitors of

Tick-borne Encephalitis Virus Reproduction

A.A. Chistov,1,2 A.A. Orlov,3,4 A.V. Guz,1 P.P. Streshnev,1 S.V. Kutyakov,1 A.V. Ustinov,1

V.A. Korshun,1,5 D.I. Osoloskin,3,4 L.I. Kozlovskaya,3 G.G. Karganova3

1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997, Russia, Moscow, Miklukho-

Maklaya st., 16/10 2Institute of Biomedical Chemistry, 119121, Russiwwa, Moscow, Pogodinskaya st., 10 building 8

3Chumakov Institute of Poliomyelitis and Viral Encephalitides, 142782, Russia, Moscow, P.O. Institute of Poliomyelitis

4Lomonosov Moscow State University, Department of Chemistry, 119991, Russia, Moscow, Leninskie Gory 1/3

5Gause Institute of New Antibiotics, 119021, Russia, Moscow, Bolshaya Pirogovskaya st., 11


Novel 5-arylethynyl substituted nucleosides were obtained over the last two decades. Although, some aryl derivatives (e.g. phenylethynyluridine) were considered inactive [1], compounds with bulky perylene fragment showed high activity against herpes viruses [2]. Further investigations discovered similar activity against another enveloped viruses. It was assumed that such compounds (also called RAFIs – rigid amphipathic fusion inhibitors) inhibit viral entry of enveloped viruses by preventing the membrane fusion process [3,4]. RAFIs are believed to inhibit the membrane curvature change, although alternative mechanism is suggested [5].

There is an urgent need for the specific treatment of the infections caused by members of the Flavivirus genus, such as pathogen dengue virus, West Nile virus, tick-borne encephalitis virus (TBEV). The flaviviruses are small enveloped RNA viruses, thus RAFI were suggested to be active against them. We examined two perylene nucleosides (dUY11 and aUY11) on antiviral activity against TBEV. These non-cytotoxic (CC50>250 µM) compounds were found to be efficient reproduction inhibitors of different strains with IC50 values in the nanomolar range. It was found that they could act by the mechanisms mentioned above. This research was supported by Molecular and Cellular Biology Program of the Russian Academy of Sciences, RFBR (project 13-04-01317), Russian President scholarship for young investigators (project SP-2494.2012.4) and Russian Foundation for Basic Research (grants no. 14-03-31566, 15-04-08365). References [1] De Clercq E., Descamps J., Balzarini J., Giziewicz J., Barr P.J., Robins M.J. J. Med. Chem. 1983, 26, 661. [2] Андронова В.Л., Пчелинцева А.А., Устинов А.В., Петрунина А.Л., Коршун В.А., Скоробогатый М.В.,

Галегов Г.А., Вопр. вирусол. 2006, 34. [3] St.Vincent M.R., Colpitts C.C., Ustinov A.V., Muqadas M., Joyce M.A., Barsby N.L., Epand R.F., Epand R.M.,

Khramyshev S.A., Valueva O.A., Korshun V.A., Tyrrell D.L.J., Schang L.M.., Muqadas M., Proc. Natl. Acad. Sci. USA. 2010, 107,17339.

[4] Colpitts C.C., Ustinov A.V., Epand R.F., Epand R.M., Korshun V.A., Schang L.M., J. Virol, 2013, 87, 3640. [5] Vigant F., Hollmann A., Lee J., Santos N.C., Jung M.E., Lee B., J. Virol, 2014, 88, 1849.

NH

O

ONO

OH

HO

R

R = H (dUY11), OH (aUY11)


160

Physicochemical Properties and Anti-opisthorchosis Effect of

Mechanochemically Synthesized Supramolecular Complexes of Albendazole

with Polysaccharide Arabinogalactan from Larch Wood

Yu.S. Chistyachenko,1 A.V. Dushkin,1 E.S. Meteleva,1 M.Y. Pakharukova,2 A.V. Katokhin,2

V.A. Mordvinov,2 N.E. Polyakov,3 M.V. Khvostov,4 T.G. Tolstikova4

1 Institute of Solid State Chemistry and Mechanochemistry of Siberian Branch of Russian Academy

of Sciences, 630128, Russian Federation, Novosibirsk, Kutateladze,18 2 Institute of Cytology and Genetics of Siberian Branch of Russian Academy of Sciences, 630090,

Russian Federation, Novosibirsk, Lavrentieva pr.,10 3 Institute of Chemical Kinetics and Combustion of Siberian Branch of Russian Academy of

Sciences, 630090, Russian Federation, Novosibirsk, Institutskaya, 3 4 N.N. Vorozhtsov Institute of Organic Chemistry of Siberian Branch of Russian Academy of

Sciences, 630090, Russian Federation, Novosibirsk, Lavrentieva pr., 9


Opisthorchosis provoked by trematodes Opisthorchis felineus is one of the most widely spread

helminthiasis in Russia, especially in Western Siberia. Large action spectrum drugs, such as

praziquantel and albendazole are used to treat opisthorchosis. Albendazole’s action is limited by

poor absorption from the intestinal lumen after oral administration because of very low water

solubility (~1 mg/l). Therefore it is necessary to use higher doses of this medication and repeated

dosing. With this approach, the risk of adverse effects of albendazole seems to increase. Therefore,

the increase in efficacy of existing anthelmintic agents is a challenging problem.

In the present study we obtained supramolecular complexes of albendazole with natural

polysaccharide arabinogalactan isolated from the wood of larch Larix sibirica and Larix gmelinii.

For synthesis of these complexes, we used the solid-state mechanochemical method. The properties

of these complexes were evaluated in solid state and in solution. Activity of the complexes against

Opisthorchis felineus was studied in vivo and in vitro. Comparative activity of albendazole and its

complex in vitro was studied on adult European liver flukes O. felineus. Albendazole complex

showed much higher activity in comparison to free albendazole. Efficacy of albendazole and its

complex in vivo was studied on hamsters Mesocricetus auratus that were previously infected with

metacercariae of the European liver fluke O. felineus. Albendazole complex showed enhanced

efficacy in comparison to free albendazole. We also evaluated the histomorphological parameters of

liver samples of hamsters. The data obtained allow us to conclude that the tested doses of

albendazole complex cause no hepatocyte toxicity, unlike free albendazole. These results open the

perspective of new anthelmintic agents’ creation on the basis of albendazole complex with

arabinogalactan.


161

Palladium(II) Complexes of S-methylcysteine and Methionine Derivatives:

Synthesis and Biological Activity

S.G. Churusova,1 D.V. Aleksanyan,1 E.Yu. Rybalkina,2 and V.A. Kozlov1

1A. N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences,

119991, Russia, Moscow, Vavilov Street 28 2Institute of Carcinogenesis, N. N. Blokhin Russian Cancer Research Center, Russian Academy of

Sciences, 115478, Russia, Moscow, Kashirskoe sh. 24


Successful introduction of cisplatin into clinical therapy promoted investigation of anticancer

activity of metal-containing compounds, including various types of palladium complexes [1].

Despite substantial similarity in structure and coordination behavior of Pt(II) and Pd(II) compounds,

the main drawback of the latter appeared to be higher reactivity, leading to deactivation in

biological systems. The application of multidentate chelating ligands can provide higher

thermodynamic and kinetic stability, which is important to improve selectivity and efficiency of

potential drugs. In this respect, a series of novel ligands have been obtained by the interaction of S-

methylcysteine and methionine esters with functionalized carboxylic acids bearing ancillary S-, N-

or P-donor groups. The ligands obtained readily undergo direct cyclopalladation under action of

PdCl2(NCPh)2 to afford κ3-X,N,S-complexes (X = S, N, P) I, II, III and IV. Preliminary

investigations on cytotoxicity of the resulting complexes against three cancer cell lines (HCT116

(colon cancer), MCF7 (breast cancer) and PC3 (prostate cancer)) showed that the most promising

type of metallocycles are palladium(II) complexes III based on picolinamide derivatives.

N

N ∗

Pd

Cl

SMe

COOMe

( )n

O

S

N

O

Pd∗

COOMe

SMe

ClMe

( )n

P

N

O

Pd∗

COOMe

SMe

ClPh

( )n

Ph

PS

N

O

Pd∗

COOMe

SMe

ClPh

Ph

I II III IV

X

X = H, Cln = 1, 2

( )n

Cytotoxicity of palladium(II) complexes III, IC50/µM

Cancer cell line

X = H X = Cl Cisplatin

n = 1 n = 2

(L/D-isomers) n = 1

n = 2 (L/D-isomers)

HCT116 1.8 6.2/7.0 1.2 0.5/3.5 400 MCF7 22 18/15 10 5.1/11.5 25 PC3 >30 13/6.5 16.5 3.6/5.0 120

References [1] A. Garoufis, S. K. Hadjikakou, N. Hadjiliadis, Coord. Chem. Rev. 2009, 253, 1384–1397


162

Synthesis of Pyrazolo[1,5-a]pyrimidines by Recyclization of the Pyrimidine Ring

G. Danagulyan1,2 and A. Tumanyan 2

1Russian-Armenian (Slavonic) State University, 0051, Armenia, Yerevan, H. Emin St., 123

2Scientific-Technological Center of Organic and Pharmaceutical Chemistry of the National Academy of Sciences of the Republic of Armenia, 0014, Yerevan, Azatutyan Ave, 26


Fused systems consisting of a pyrimidine ring and azole have high biological activity. Synthesis

of similar systems, in particular, of various azolo[1,5-a]pyrimidines can be carried out in two ways:

proceeding from the pyrimidine or five-membered cycles by adding the second, missing ring to the

existing heterocycle. We have developed a method for the synthesis of pyrazolo[1,5-a]pyrimidines

by recyclization of 1,4,6-trimethyl-2-(ethoxycarbonyl)methyl-pyrimidinium iodide. Iodide 1 under

the action of various carboxylic acid hydrazides containing biogenic heterocycles (pyrimidine,

pyridine, pyrazol, triazole) forms derivatives of 3-ethoxycarbonylpyrazolo[1,5-a]pyrimidine

containing a fragment of the initial hydrazide with heterocycle (pyrimidine, pyridine, pyrazol,

triazole) in the pyrazol ring.

The reaction proceeds through substitution of the nitrogen atom of the pyrimidine ring by

triatomic N-N-C hydrazide fragment.

N

N

Me

MeMe NHNHCO-Het

COOEtHet-CONHNH2

NH

N

Me

Me

Me N-NHCO-HetCOOEt -MeNH2 N

N

Me

MeNH

O Het

COOEt

N

N

Me

MeNH

O Het

COOEtNN

NCOOEt

HetMe

Me

1 +

-H2O

The work has been accomplished within the framework of the joint Russian-Armenian grant number 13RF-087 of the Committee on Science of the Ministry of Education and Science of the Republic of Armenia and Russian Foundation for Basic Research (RFBR) as well as with financial support from the Committee on Science of Armenia (theme 13-1D334).


163

Аdamantylation of 1,2,4-Triazolo[5,1-c][1,2,4]triazines as Approach for

Synthesis of Antiviral Agents

S.L. Deev,1,2 T.S. Shestakova,1 I.A. Khalymbadzha,1 V.L. Rusinov,1 O.N. Chupakhin1,2

1 Ural Federal University, 19 Mira Street, 620002 Ekaterinburg, 620002, Russia,

Еkateriburg, 19 Mira street 2 Institute of Organic Synthesis, Ural Branch of the Russian Academy of Sciences,

22 S. Kovalevskoy Street, 620219 Ekaterinburg, Russia E-mail: [email protected]

The incorporation of the lipophilic adamantyl moiety in small molecules appears to be an

important approach towards obtaining of compound with diverse pharmacological properties, such

as antiviral, anticancer and antimicrobial activity [1]. The synthesis of heterocycles based on

adamantane leads to a modification of their biological action, often enhancing the effect or

imparting a new kind of activity. For example N-adamantyl tetrazoles show low toxicity and potent

activity against the influenza A virus [2].

Herein, we report an efficient method for incorporation adamatyl moiety in structure of 1,2,4-

triazolo[5,1-c][1,2,4]triazin-7-ones 1a-f by use 1-adamantanole 2. The reactions of compounds 1a-c

with 2 in the sulfuric acid afford derivatives 4a,d and 5a-f. It should be noted that compounds 5a,d

formed by isomerisation triazolotriazines 4a,d.

The structures of compounds 4-5 were confirmed by two-dimensional correlation analysis (2D 1H-13C HSQC, 1H-13C HMBC and 1H-1H NOESY spectroscopy).

The study of the antiviral effect respiratory syncytial virus showed that structure 3a possesses

moderate activity.

This work was supported by the RFBR (projects 13-04-02059, 12-03-33144, 12-03-31476 and 12-04-00581).

References [1] L. Wanka, K. Iqbal, P.R. Schreiner, Chem. Rev. 2013, 113, 3516-3604. [2] V.V. Zarubaev, E.L. Golod, P. M. Anfimov, A. A. Shtro, V.V. Saraev, A.S. Gavrilov, A.V. Logvinov, O.I. Kiselev, Bioorg. Med. Chem. 2010, 18, 839-848.


164

Design, Synthesis and Pharmacological Evaluation of Dipeptide Analogs of

Alpidem, Potential Ligands of TSPO Receptor

O.A. Deeva,1 S.A. Yarkov,2 M.A. Yarkova,2 T.A. Gudasheva,1 S.B. Seredenin2

1Chemistry department, Federal State Budgetary Scientific Institution “Research Zakusov Institute

of Pharmacology”, 125315, Russia, Moscow, Baltiyskaya str., 8 2Department of Pharmacogenetics, Federal State Budgetary Scientific Institution “Research

Zakusov Institute of Pharmacology”, 125315, Russia, Moscow, Baltiyskaya str., 8


The translocator protein (TSPO, 18 kDa) plays an important role for the synthesis of

neurosteroids by promoting the transport of cholesterol from the outer to the inner mitochondrial

membrane, which is the rate-limiting step in neurosteroidogenesis [1]. Neurosteroids interact with

non benzodiazepine site of GABAa receptor causing an anxiolytic effect without the side effects of

benzodiazepine anxyolytics. Using drug-based peptide design method [2] and structure-functional

analysis of well-known synthetic TSPO ligands the dipeptide N-carbobenzoxy-L-tryptophanyl-L-

isoleucine amide (GD-23) was designed as putative ligand. We used Alpidem as non-peptide

prototype[3]. This structure contains all the necessary TSPO ligands pharmacophore elements. This

compound was synthesized by classical methods of peptide synthesis in solution. The anxiolytic

activities were investigated in Balb/C mice using the open-field test (OFT) and elevated plus-maze

test (EPMT) in CD-1 mice. In this study we used the technique of illuminated open field, where the

movement of the animal from the dark into the brightly illuminated arena creates additional

stressing factors[4]. GD-23 in the dosage range 0.05-1.0 mg/kg significant increased total

locomotor activity of mice compared with control groups. GD-23 significantly (p <0.001) increased

the percentage of time spent in the open arms of the maze in the dose range of 0.1-1.0 mg/kg. It was

shown that anxiolytic effect of GD-23 is stereoselective: its D,L and L,D diastereomers abolished

activity in OFT. Anxiolytic and nootropic effects of GD-23 were antagonized by PK11195, a TSPO

antagonist. These results indicate that GD-23 produces anti-anxiety effect that are mediated by

TSPO, but does not cause the side effects normally associated with conventional benzodiazepines.

Hence, GD-23 shows potential for the treatment of stress-related disorders including anxiety.

References [1] C. Nothdurfter, G. Rammes, T.C. Baghai, C. Schüle, M. Schumacher, V. Papadopoulos, R. Rupprecht, J

Neuroendocrinol, 2012, 24 (1), 82-92 [2] T.A. Gudasheva, T.A. Voronina, R.U. Ostrovskaya, N.I. Zaitseva, N.A. Bondarenko, V.K. Briling, L.S. Asmakova,

G.G. Rozantsev, S.B. Seredenin, J. Med. Chem., 1998, 41(3), 284-290 [3] A. Cappelli, G. Giuliani, S. Valenti, M. Anzini, S. Vomero, G. Giorgi, G. Sogliano, E. Maciocco, Bioorg. & Med.

Chem., 2008, 16, 3428–3437 [4] S.B. Seredenin, A.A. Vedernikov, Biull .Eksp. Biol. Med. (Russia), 1979, 88, №1, 714-716


165

One-pot Natural Monoterpene Alcohol Amination to Biologically Valuable

Compounds Over Gold Catalysts

Yu.S. Demidova,1,2 I.L. Simakova,1,2 M. Estrada,3 S.A. Beloshapkin,4 E.V. Suslov,5

K.P. Volcho,2,5 N.F. Salakhutdinov,2,5 A.V. Simakov,6 D.Yu. Murzin7

1Boreskov Institute of Catalysis, 630090, Russia, Novosibirsk, pr. Lavrentieva, 5

2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova, 2 3Posgrado en Física de Materiales, CICESE, 22860, México, Ensenada, B.C

4Materials & Surface Science Institute, University of Limerick, Limerick, Ireland 5Novosibirsk Institute of Organic Chemistry, 630090, Russia, Novosibirsk, pr. Lavrentieva, 9

6Centro de Nanociencias y Nanotecnología, UNAM, 22860, México, Ensenada, B.C 7Process Chemistry Centre, Åbo Akademi University, FI-20500, Finland, Turku/Åbo


The one-pot alcohol amination is a perspective approach to the synthesis of complicated amines

with the different structure that are widely used as pharmacophores in numerous biologically active

compounds, important chemicals for fine organic synthesis and functionalized materials. One of the

challenges in catalytic fine organic chemistry is to develop selective amination of alcohols to yield

amines of a certain structure having specific physiological properties. Complicated monoterpene

amines, synthesized from renewable raw materials, were recently shown to exhibit specific

physiological properties and can be used as intermediates of potential drugs for neurological

diseases [1]. The main objective of the present work was to study general regularities of one-pot

natural monoterpene alcohol amination in the presence of gold containing catalysts over different

metal oxides and to tune selectivity to desired product through optimization of catalyst

nanostructure, active component composition and support nature.

A series of nanosized gold catalysts including Au, AuPd and Pd (for comparison) nanoparticles

supported on different metal oxides (ZrO2, MgO, Al2O3, CeO2, La2O3) were synthesized and tested

in one-pot natural monoterpene alcohol amination with equimolar amounts of the substrates without

any additives at 140-180°C under nitrogen pressure. Myrtenol, which represents a natural terpene

alcohol with primary –OH group, as well as aniline were selected as model substrates in this work.

The main peculiarities of one-pot monoterpene alcohol amination were studied. The products

distribution as well as alcohol conversion was shown to strongly depend on support nature, active

metal component composition and conditions of catalysts pretreatment [2]. A nearly complete

myrtenol conversion as well as the highest selectivity to corresponding secondary amine was

reached over pre-oxidized Au/ZrO2 catalyst.

References [1] I.G. Kapitsa, E.V. Suslov, G.V. Teplov, D.V. Korchagina, N.I. Komarova, K.P. Volcho, T.A. Voronina,

A.I. Shevela,. N.F. Salakhutdinov, Pharm. Chem. J. 2012, 46(5), 263-265. [2] Yu.S. Demidova, I.L. Simakova, M. Estrada, S. Beloshapkin, E.V. Suslov, D.V. Korchagina, K.P. Volcho,

N.F. Salakhutdinov, A.V. Simakov, D.Yu. Murzin, Appl. Catal. A 2013, 464-465, 348-356.


166

MNPs Modified with APS and PMIDA for MRI in vivo

A. Demin,1 A. Pershina,2,5 V. Ivanov,2 O. Shevelev,3 K. Nevskaya,2 N. Shchegoleva,4 A. Minin,4

M. Uimin,4 A. Sazonov,2 V. Krasnov1

1Postovsky Institute of Organic Synthesis of RAS (Ural Branch), 620041, Russia, Yekaterinburg, S.

Kovalevskoy St., 22 2Siberian State Medical University, 634050, Russia, Tomsk, Moskowsky trakt, 2

3Institute of Cytology and Genetics SB RAS, 630090, Russia, Novosibirsk, Lavrentiev Ave., 10 4M.N. Miheev Institute of Metal Physics of RAS (Ural Branch), 620990, Russia, Yekaterinburg, S.

Kovalevskoy St., 18 5Russia National Research Tomsk Polytechnic University, 634050, Russia, Tomsk, Lenina ave., 30


The nanomaterials based on magnetic nanoparticles (MNPs) are intensively developed and used

as therapeutic and visualizing diagnostic agents, including application for MRI. The aim of this

work is modification of MNPs based on Fe3O4 nanoparticles (obtained by a chemical co-

precipitation method from a solution of Fe3+ and Fe2+ salts mixture (MNPs(1), up to 20 nm) and by

method of mechanochemical synthesis (MNPs(2), 12 nm)) with 3-amiopropilsilane (APS)

(according to [1]) or N-(phosphonomethyl)iminodiacetic acid (PMIDA), their comparative

characteristic and study of their ability to serve as MRI contrast agent.

Fe3O4

HOP

HOO

N

O

HO O

OH

OP

O

O

N

O

HO O

OH

NH2SiOMe

MeOMeO

OH

OH

OH

EtOH (95%) O

O

O

NH2SiMNPs(1)-NH2, 1

Fe3O4

Fe3O4MNPs(2)-NH2, 2

MNPs(1)-COOH, 3MNPs(2)-COOH, 4

H2O

Co-precipitation method

(MNPs(1), 20 nm)

(MNPs(2), 12 nm)Mechanochemical synthesis

In vivo hamster liver MR images before (left) and

40 min after (right) administration of MNPs (1)

It has been shown that MNPs (1) possess highest T2-contrast properties in MR-imaging in

experiments in vivo. The working dose of 0.6 mg/kg was lower then the recommended dose of

injection for the contrast agent approved by FDA (USA) Lumirem (2 mg/kg). The MNPs(1) and

MNPs(2) showed no cytotoxicity effect in the working concentration in experiments in vitro. Thus

MNPs (1) are perspective as a liver MRI contrast agents.

This work was partially supported by RFBR, research project No 14-03-00146 (synthesis of nanoparticles) and by RSF research project No 14-15-00247 (in vivo and in vitro experiments). References [1] A.M. Demin, V.P. Krasnov and V.N. Charushin, Mendeleev Commun. 2013, 23, 14-16.


167

Development of Drug Delivery System for Anticoagulant Warfarin on the Basis

of Mesoporous Silica Particles: Adsorption as a Route of Preparation of the

Drug Composite

E.S. Dolinina, E.V. Parfenyuk

G.A. Krestov Institute of Solution Chemistry of Russian Academy of Sciences, Ivanovo, Russian Federation

In spite of a wide spectrum of drugs meant for treatment of various illnesses, many of the drugs

suffer from grave shortcomings such as a low bioavailability, unsatisfactory pharmacokinetics,

photo- and thermal instability, etc. One of efficient ways of improvement of pharmacological and

consumer properties of such drugs is development of optimal drug delivery systems. In the present

time much attention has been paid to porous silica due to its biological, physical chemical

properties, structural and morphological diversification and possibility of preparation under mild

conditions by sol-gel synthesis. Amorphous silica has been recognized as a safe food additive in the

whole world. Numerous studies testify that formation of drug-silica composites can essentially

change properties and behavior of the drug. Adsorption is one of the most widely used and the

simplest routes to prepare the silica-drug composites. The present work is a part of development of

the silica-based delivery system for anticoagulant warfarin.

Warfarin is one of the most commonly used oral anticoagulants in the clinic. Although effective,

warfarin has limitations that complicate its use. These include unpredictable pharmacokinetics and

pharmacodynamics related to variations in dietary vitamin K intake and numerous drug-drug

interactions. It is possible that adsorption of warfarin in porous silica matrix can prevent the drug-

drug interactions and help to develop new silica-based formulation with predictable kinetic behavior

in biological medium.

To develop the efficient drug delivery system, it is necessary to know what silica material can

adsorb the highest amount of the drug and retain it strongly in the silica matrix. For this purpose a

series of mesoporous silica materials (unmodified, phenyl, methyl, mercaptopropyl modified) was

synthesized by sol-gel method. The adsorption of warfarin onto the synthesized silica materials was

studied at different temperatures and medium pH. The effects of surface chemistry of the silica

materials, temperature and pH on the loaded amount of warfarin in the composites as well as energy

of warfarin – silica interactions were elucidated. It was found that warfarin adsorbs only under

acidic conditions. The methyl modified silica showed the highest warferin loading but the energy of

interaction in the composite is the lowest. The effects were explained by pH dependent structural

diversity of the drug and ionization state of the silica surfaces.

The work is supported by RFBR grant 14-03-00022


168

Structure-Function Relationships in Cobra Cardiotoxins: Antibacterial Activity

P.V. Dubovskii,1 A.A. Ignatova,1 Y.N. Utkin,1 and A.V. Feofanov 1,2

1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Russia, Moscow, Miklukho-

Maklaya str., 16/10 2 Biological Faculty, M.V.Lomonosov Moscow State University, 119234 Russia, Moscow, Lenin

Hill, 1/12


In an effort to overcome broadening resistance to conventional antibiotics, novel antibacterial

compounds are being looked for. The polycationic peptides, isolated from insect and snake venoms

attract the attention [1,2]. Among them linear, i.e. cysteine-free peptides, e.g. spider venom latarcins

have been found to feature broad-spectrum antibacterial activity [1]. The structurally rigid

cardiotoxins (or cytotoxins, CTs), containing multiple disulphide bonds, exhibit activity only

against some Gram-positive bacteria, in particular M. luteus [3]. To continue unraveling the

structural determinants of such activity, in the present work we tested antibacterial properties of

seven cardiotoxins (cytotoxin 1, isolated from the venom of cobra Naja melanoleuca, or CT1Nm;

cytotoxin 1 from Naja haje, CT1Nh; cytotoxin 2 from Naja haje, CT2Nh; cytotoxin 1 from Naja

kaouthia, CT1Nk; cytotoxin 2 from Naja kaouthia, CT2Nk; cytotoxin 1 from Naja oxiana, CT1No;

cytotoxin 2 from Naja oxiana, CT2No) against Gram-positive bacterium B. subtilis. Using

techniques, described earlier [3], we found that only two CTs (CT3Nk and CT2No) exhibited

activity in the low micromolar concentration range. Among the CTs tested only these two feature a

net electric charge above 8. CT2No exhibits higher activity, compared to CT3Nk, which is in

accord with different hydrophobicities of these CTs, reflected in their HTL-scores [4]. Thus,

previously made conclusion, that antibacterial activity of CTs depends on the net positive charge of

the molecule and its HTL-score, receive strong support with the present data.

The work was supported by the Russian Foundation for Basic Research (grant 13-04-02128).

References [1]. Kozlov SA, Vassilevski AA, Feofanov AV, Surovoy AY, Karpunin DV, and E.V. Grishin, J. Biol, Chem. 2006,

281 (30), 20983-20992. [2]. Dubovskii PV, and Y.N. Utkin, Acta Naturae 2014, 6 (3), 11-18. [3]. Dubovskii PV, Vorontsova OV, Utkin YN, Arseniev AS, Efremov RG, and A.V. Feofanov, Mendeleev Commun.

2015, 21 (1), 70-71. [4]. Dubovskii PV, Konshina AG, and R.G. Efremov, Curr. Med. Chem. 2014, 21 (3), 270-28.


169

Improvement of Dissolution Properties of Valsartan by co-crystallization with

Vitamin C via Mechanochemical Method

L. Du, A.V. Dushkin and W. Su

Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou 310014, China

Valsartan is a BCS class II angiotensin-II receptor antagonist. Its clinical application is limited

by the poor solubility and low bioavailability.1 Pharmaceutical co-crystal is great potential value

with modifying the unfavorable physicochemical properties, without changing the molecular

structures of drugs.2Thus in this study, a new pharmaceutical co-crystal of valsartan was

synthesized with co-crystal former, vitamin C, to improve its water solubility by mechanochemical

method. A high intensity oscillating mill was applied for producing valsartan-vitamin C co-crystal.

Multi-instrumental characterizations confirmed the formation of the new co-crystal prepared by

high mechanical process, as following: nuclear magnetic resonance, the optical activity test, Fourier

transform infrared spectroscopy, differential scanning calorimetry and powder X-ray

diffractometry. Meanwhile, the ratio of co-crystal components, which determined by quantitative

nuclear magnetic resonance (qNMR), was found to be 1:1 (valsartan to vitamin C). Eventually,

dissolution test were carried in enzyme-free simulated fluid (pH 6.8). It showed that co-crystal of

valsartan and vitamin C significantly increased the dissolution rate (nearly 18 times than free

valsartan). It was concluded that valsartan-vitamin C co-crystal was successfully synthesized by

mechanochemical method and behaved better in modifying physicochemical properties and

pharmaceutical properties.

References 1. G.Flesch,M.P. Lloyd, Absolute bioavailability and pharmacokinetics of valsartan, an angiotensin II receptor

antagonist in man, Eur. J.Clin.Pharmacol.1997,52, 115-120. 2. P.Mura, M.Cirri, M. T. Faucci, J. M.Gines-Dorado, G. P.Bettinetti, Investigation of effects of grinding and co-

grinding on physicochemical properties of glisentide, J.Pharmaceut. Biomed.2002,30, 227-237.


170

Natural and Synthetic nZ,(n+4)Z-Dienoic Fatty Acids: A New Method for the

Synthesis and Inhibitory Action on Topoisomerase I and IIα

V.A. D'yakonov,1 L.U. Dzhemileva,2 A.A. Makarov,1 A.R. Mulyukova,1 U.M. Dzhemilev1

1Institute of Petrochemistry and Catalysis of Russian Academy of Sciences, 450075, Russian

Federation, Ufa, Prospekt Oktyabrya, 141 2Department of Immunology and Human Reproductive Health, Bashkir State Medical University,

450003, Russian Federation, Ufa, Lenin Street, 3


Relying on the earlier results on cross-cyclomagnesiation of O-containing and terminal aliphatic

1,2-dienes [1,2], we developed an efficient versatile method for the synthesis of dienoic acid

containing a 1Z,5Z-diene group.

According to the developed strategy of the synthesis of nZ,(n+4)Z-dienoic fatty acids, the first

step is Ti-catalyzed cross-cyclomagnesiation of tetrahydropyran ethers of alkadien-1-ols 1 with

terminal aliphatic allenes 2 induced by EtMgBr to afford 2,5-dialkylidenemagnesacyclopentanes,

which are then hydrolyzed to give the tetrahydropyran ethers of alkadienols 3. The Jones oxidation

of these products furnished the target dienoic acids 4a-l with the specified 1Z,5Z position of the

diene group with respect to the carboxy group in 61-75% yields and with stereoselectivity of >98%.

.THPO

.R

HO2C RTHPO R

MgRTHPO

+~90%

4a-l

1

2

()n

()n-1

(a): EtMgBr, Mg, [Ti]; (b): H3O+; (c) Jones oxidation. [Ti] = Cp2TiCl2(R = Me) n = 2: m = 11 (a); n = 4: m = 5 (b), 9 (c), 11 (d), 13 (e), 17 (f); n = 5: m = 8 (g); n = 6: m = 7 (h); n = 10: m = 3 (i), 11 (k). (R = Ph) n = 4: m = 1 (l)

(a)

()m

()m

~80%3a-l

()n

(c)()m

(b)()n ()m

The synthesized acids exhibiting high inhibitory activity with respect to human topoisomerase I

and IIα.

This work was performed under financial support from the Russian Science Foundation (Grant 14-13-00263). References [1] V.A. D’yakonov, A.A. Makarov, L.U. Dzhemileva, E.Kh. Makarova, E.K. Khusnutdinova, U.M. Dzhemilev, Chem. Commun. 2013, 49, 8401–8403. [2] V.A. D’yakonov, A.A. Makarov, E.Kh. Makarova, U.M. Dzhemilev, Tetrahedron 2013, 69, 8516–8526.


171

Stereoselective Synthesis of 11-Phenylundeca-5Z,9Z-dienoic Acid and

Investigation of Its Human Topoisomerase I and IIα Inhibitory Activity

V.A. D'yakonov,1 L.U. Dzhemileva,2 A.A. Makarov,1 A.R. Mulyukova,1 U.M. Dzhemilev1

1Institute of Petrochemistry and Catalysis of Russian Academy of Sciences, 450075, Russian

Federation, Ufa, Prospekt Oktyabrya, 141 2Department of Immunology and Human Reproductive Health, Bashkir State Medical University,

450003, Russian Federation, Ufa, Lenin Street, 3


The search for new efficient and low-toxicity antitumor agents is a highly important task of

modern medicinal chemistry. Generally, a solution of this problem is reduced to the search for new

compounds affecting molecular targets that play an important role in carcinogenesis.

Numerous studies in this field demonstrated that topoisomerase I and topoisomerase II are

among the key molecular targets for the development of modern antitumor agents.

(5Z,9Z)-11-Phenylundeca-5,9-dienoic acid was stereoselectively synthesized, based on original

cross-cyclomagnesiation of 2-(hepta-5,6-dien-1-yloxy)tetrahydro-2H-pyran and buta-2,3-dien-1-

ylbenzene with EtMgBr in the presence of Cp2TiCl2 catalyst giving 2,5-

dialkylidenemagnesacyclopentane in 86% yield. The acid hydrolysis of the product and the Jones

oxidation of the resulting 2-{[(5Z,9Z)-11-phenylundeca-5,9-dien-1-yl]oxy}tetrahydro-2Н-pyran

afforded (5Z,9Z)-11-phenylundeca-5,9-dienoic acid in an overall yield of 75%.

.THPO

.Ph

HO2C PhTHPO Ph+

()4

()3

(a,b) (c)()4

1

23 4

A high inhibitory activity of the synthesized acid with respect to human topoisomerase I (hTop1)

and II (hTop2α) was determined.

This work was performed under financial support from the Russian Science Foundation (Grant 14-13-00263).


172

Cytotoxic Activity of Salicylic Acid-containing Ionic Liquids

K.S. Egorova,1 M.M. Seitkalieva,1 A.V. Posvyatenko2 and V.P. Ananikov1

1N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 119991, Russia,

Moscow, Leninsky prospect 47 2Institute of Gene Biology, Russian Academy of Sciences, 119334, Russia, Moscow, Vavilova str.

34/5


Ionic liquids (ILs) are unique systems employed in various scientific fields, such as synthesis,

catalysis, biomass conversion, etc. Initially ILs had been suggested ‘green’ chemicals due to their

non-flammability and non-volatility; however, biological properties of ILs have been attracting

much attention recently.[1] Thus, ILs have been demonstrated to possess anticancer activity, whereas

their micelles are used as drug delivery platforms. ILs containing active pharmaceutical ingredients

are subjects of active studies. In our previous work we demonstrated that introduction of an amino

acid into ILs increased their cytotoxicity.[2] In this study, we synthesized ILs containing salicylic

acid (SA-ILs) in the cation (1-(2-((2-hydroxybenzoyl)oxy)ethyl)-3-methylimidazolium

tetrafluoroborate, 1-(2-((2-hydroxybenzoyl)oxy)ethyl)-3-methylimidazolium chloride, 1-(3-((2-

hydroxybenzoyl)oxy)propyl)-3-methylimidazolium chloride) or anion (1-ethyl-3-

methylimidazolium salicylate, 1-butyl-3-methylimidazolium salicylate, 1-hexyl-3-

methylimidazolium salicylate) and investigated their cytotoxic activity towards the human cell lines

CaCo-2 (colorectal adenocarcinoma) and 3215 LS (fibroblasts) in comparison with cytotoxicity of

conventional imidazolium ILs and salicylic acid. Cytotoxicity of SA-ILs was significantly higher

than that of conventional ILs and was close to that of SA; however, SA-ILs displayed much higher

water solubility than SA, which is a remarkable feature since low solubility is one of major factors

impeding administration of many drugs. Thus, introduction of salicylic acid into an IL does not

disturb the biological activity of the former, but increases its bioavailability. The results confirm the

strategy of obtaining new prodrug ILs to be a promising tool for pharmaceutical research.

The work was supported by the Russian Foundation for Basic Research (project 14-03-31478) and by the Grant of the President of the Russian Federation (project MK-5053.2015.3).

References [1] K.S. Egorova and V.P. Ananikov, ChemSusChem 2014, 7, 336-360. [2] K.S. Egorova, M.M. Seitkalieva, A.V. Posvyatenko, and V.P. Ananikov, Toxicol. Res. 2015, 4, 152-159.


173

New Water-Soluble Antifungal Agents Based on

2-Amino-6-Methylpyrimidin-4(3H)-thione

A. Erkin,1 V. Krutikov1 and V. Tets2

1Department of Chemistry and Technology of Synthetic Biologically Active Substances,

Saint Petersburg State Institute of Technology (Technical University), 190013, Russia, Saint Petersburg, Moskovsky ave., 26

2Zabolotny Department of Microbiology, Virusology and Immunology, Pavlov First Saint Petersburg State Medical University, 197022, Russia, Saint Petersburg, L’va Tolstogo str., 6-8


Searching for new biologically active substances in the series of 2-aminopyrimidine derivatives

[1] some water-soluble 6-benzyl substituted 2-amino-4-methylpyrimidines 1-5 [R1 = H2N, R2 = H,

X = S (1); R1 = H2N, R2 = Br, X = S (2); R1 = R2 = H (3), X = S; R1 = H2N, R2 = H, X = O (4); R1 =

H2N, R2 = H, X = NH (5)] were synthesized and tested as potential inhibitors of growth of

pathogenic bacteria and fungi.

Among the compounds obtained, only thioether 1 was found to possess a significant activity

against Candida albicans (strain 15) with a value of IC100 0.093 mM [2].

Introduction of halogen atoms into phenyl ring of thioether 1 resulted in some decrease of IC100

values for compounds 6 and 7 [R = R1 = H, R2 = F (6); R = H; R1 = R2 = Cl (7)] up to 6 and 25%,

respectively. At the same time, solubility of benzyl ether 7 in water was about 100-fold lower than

that of benzyl ether 6. Furthermore, bromination of compounds 6 and 7 at the pyrimidine nuclei

caused a total loss of antifungal effect of polyhalogenated benzyl ethers 8 and 9 [R = Br; R1 = H, R2

= F (8); R = Br; R1 = R2 = Cl (9)].

In conclusion, benzyl ethers of 2-amino-6-methylpyrimidin-4(3H)-thione appear to be suitable

objects to develop new antifungal agents.

References [1] E. Koroleva, K. Gusak, and Zh. Ignatovich, Russ. Chem. Rev. 2010, 79, 655 [2] A. Erkin, V. Gurzhii, and V. Krutikov, Russ. J. Gen. Chem. 2015, 85, 79


174

The Scallop Patinopecten yessoensis Enzyme Acting Hydrolysis of Fucoidans

M.S. Avtushenko, A.M. Zakharenko, S.P. Ermakova, T.N. Zvyagintseva

1 G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of Russian Academy of Sciences, Laboratory of Enzyme Chemistry, 159 100-Let Vladivostok Ave., 690022 Vladivostok,

Russian Federation.


The scallop Patinopecten yessoensis is commercial species has also been introduced from Japan to France and Canada. The complex of enzymes with active fucoidanase has been found in the mollusk liver, which is often a waste at scallop’s processing.

Fucoidanases are enzymes which catalyze hydrolysis of O-glycosidic bonds in the main chain of the fucoidans. Today we know about twenty sources of such enzymes. They were found only in marine invertebrates and bacteria [1-5]. Fucoidanases remain under-investigated, and only a few enzymes are isolated in a homogeneous state. The major interest to these enzymes stems from the high biological activity of their substrates – fucoidans.

Fucoidans are sulfated homo- and heteropolysaccharides producing by brown algae. The analogues of fucoidans have not been found in terrestrial organisms. The structures of these polysaccharides are very differ and vary on the type and stage of development of algae, growth place, harvest season and other factors [10, 11]. Unfortunately the structure of fucoidans is studied less intensively in compare with their biological activity.

Fucoidanases that are produced fucooligosaccharides may be useful for the study of the structure of a native fucoidans, and to determine biological activity. The products of enzymatic hydrolysis of fucoidan, which have more simple structure, possess the same or higher biological activity than the native polysaccharide.

We have developed the scheme of purification of fucoidanase from the liver of mollusk Patinopecten yessoensis, defined some properties of this enzyme. The fucoidanase has an unusually broad specificity, showing activity in structurally different fucoidan from Undaria pinnatifida, Saccharina cichorioides, Saccharina gurjanovae and Fucus evanescens. For more detailed research we have chosen fucoidan from U. pinnatifida with complex structure. The Km values of the enzyme in the reaction with fucoidan from Undaria pinnatifida is ≈ 3 mg/ml. Also we have compared degree of hydrolysis of fucoidan from F. evanescens by studied bacterial fucoidanase and fucoidanase from mollusk. MS data has shown that degree of fucoidan hydrolysis by fucoidanase from P. yessoensis was higher than bacterial enzyme has.

Thus applying of enzymes to produce new substances is a topical task of modern enzymology. The results of this study will not only expand the existing knowledge about the specificity and mechanism of action of fucoidanases from marine organisms, and the structure of fucoidan, but also create a scientific foundation for the development of new biologically active compounds from natural fucoidans.

This work was supported by FEB RAS Grant # 15-II-5-009. References [1]. N.M. Thanassi, H.I. Nakada, Arch. Biochem. Biophys. 1967, 118, 172–177. [2]. K. Kitamura, M. Matsuo, T.Yasui, Biosci. Biotechnol. Biochem. 1992, 56, 490–494. [3]. R. Daniel, O. Berteau, J. Jozefonvicz, N. Goasdoue, Carbohydr. Res. 1999, 322, 291–297. [4]. V. Descamps, S. Colin, M. Lahaye, M. Jam, C. Richard, P. Potin, T. Barbeyron, J.C. Yvin, B. Kloareg, Mar.

Biotechnol. (N. Y.). 2006, 8, 27–39. [5]. A.S. Silchenko, M.I. Kusaykin, V.V. Kurilenko, A.M. Zakharenko, V.V. Isakov, T.S. Zaporozhets, A.K.

Gazha, and T.N. Zvyagintseva, Mar. Drugs. 2013, 7, 2413–2430. [10]. A.V. Skriptsova, N.M. Shevchenko, T.N. Zvyagintseva, T.I. Imbs, J. Appl. Phycol. 2010, 22, 79-86. [11]. W. Mak, N. Hamid, T. Liu, J. Lu, W.L. White, Carbohydr. Polym. 2013, 95, 606-614.


175

In vitro Anticancer Activity of Laminarans from Far Eastern Brown Seaweeds

and Their Sulfated Derivatives

O. Malyarenko (Vishchuk), S. Ermakova, N. Shevchenko, T. Zvyagintseva

G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of Russian Academy of Sciences, Laboratory of Enzyme Chemistry, 159 100-Let Vladivostok Ave., 690022 Vladivostok,

Russian Federation. E-mail: [email protected]

Brown seaweeds have drawn worldwide attention due to their involvement in many industrial

applications. They are potentially prolific sources of highly bioactive polysaccharides, namely

laminarans, alginic acids and fucoidans. Many of these active compounds have been found to be

useful functional ingredients in many applications such as pharmaceutical, cosmeceutical, and

functional food. In recent years, the interest in sulfated polysaccharides from brown seaweeds

(fucoidans), its structure and biological activities has increased. However the study of biological

effects and molecular mechanisms of neutral polysaccharides, laminarans, are very limited.

Laminarans are low-molecular weight (5-10 kDa) β-D-glucans, which consisted of β-D-(1→3)-

and in less degree (1→6)-linked glucose residues. They differ from each other with regard to their

length and branching structure [1]. β-D-glucans possess antimicrobial and anticancer activities by

enhancing the host immune [2].

Metastasis is the final stage in tumor progression and is found to be responsible for up to 90% of

deaths associated with solid [3]. The invasion and metastasis of tumor cells has been shown to

require proteolytic activity in order to degrade components of the extracellular matrix (ECM) [4].

The matrix metalloproteinases MMP-2 and MMP-9 (also known as gelatinase A and gelatinase B,

respectively), play an important role in the degradation of basement membrane type IV collagen,

which is associated with tumor cell invasion and metastasis [5]. Therefore, the development of

anticancer drugs, which regulate expression of MMP-2 and MMP-9 is crucial in inhibiting tumor

invasion and metastasis.

In the present work we determined that laminarans isolated from Far Eastern brown seaweeds

Saccharina cichorioides, Saccharina japonica, and Fucus evanescens and their sulfated derivatives

inhibited proliferation, colony formation and migration of human melanoma, breast cancer and

colorectal adenocarcinoma cells, as well as expression of MMP-2 and MMP-9 in vitro.

This work was supported by RFBR Grant # 14-04-93003. References: [1] S. Ermakova, R. Menshova, O. Vishchuk, S. Kim, and et.al., Algal Res. 2013, 2, 51–58. [2] G. Chan, W. Chan, D. Sze., J. Hematol. Oncol. 2009, 2, 25. [3] D. Hanahan and R. Weinberg, Cell. 2011, 14, 646-674. [4] L. Liotta, P. Steeg, and W. Stetler-Stevenson, Cell. 1991, 64, 327-336. [5] J. Rundhaug, J. Cell. Mol. Med. 2005, 9, 267-85.


176

The Fucoidan, a Marine Sulfated Polysaccharide from Brown Alga Fucus

evanescens, Prevents Colorectal Cancer Growth by Targeting PDZ-Binding

Kinase/T-LAK Cell-Originated Protein Kinase

O. Vishchuk,1,2 D. Qiuhong,2 S. Ermakova,1 F. Zhu,2 T. Zvyagintseva1

1 G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of Russian Academy of Sciences, Laboratory of Enzyme Chemistry, 159 100-Let Vladivostok Ave., 690022 Vladivostok,

Russian Federation. 2 Tongji Medical College of Huazhong University of Science and Technology, Laboratory of

Biochemistry, No 13 Hangkong Road, 430030, Wuhan, Hubei, PR China


Carcinogenesis is a multistage process in which numerous genes important in the regulation of

cellular functions may be prime targets for cancer preventive agents. Prevention and therapeutic

intervention by natural compounds is a newer dimension in cancer management. Administration of

nontoxic natural compounds was shown to prevent initiation, promotion, and progression events

associated with carcinogenesis in different animal models, and has been suggested to effectively

reduce cancer mortality and morbidity [1].

Fucoidans, sulfated polysaccharides, are constituents of brown algae and some marine

invertebrates (such as sea urchins and sea cucumbers) [2], [3]. Nowadays they are known to be a

topic of numerous studies as nontoxic compounds, possessing wide spectrum of biological activities

[4], [5]. While the development of research efforts involving the structure of the fucoidans and their

biological activities are advancing, the understanding of the molecular mechanisms of their action

and identification of target proteins for cancer prevention are still incomplete. Moreover, a direct

anticancer target of fucoidan from brown algae has not yet been identified in vitro or in vivo.

In the present study the sulfated and partially acetylated (1→3);(1→4)-α-L-fucan from brown

alga Fucus evanescens was shown to attenuate the mitogen-activated protein kinases downstream

signaling in a colorectal cancer cells with different expression level PDZ-Binding Kinase/T-LAK

Cell-Originated Protein Kinase (TOPK), resulting in cells growth inhibition. Moreover, the results

suggest that fucoidan exerts its effects by directly interacting with (TOPK) in vitro and ex vivo and

inhibits its kinase activity. Taken together, our findings support the cancer-preventive efficacy of

fucoidan through its targeting of TOPK for the prevention of progression of colorectal carcinomas.

This work was supported by NSFC Foundation and FEB RAS Grant # 15-II-5-004. References: [1] A. Bode and Z. Dong., Nutrition. 2004, 20, 89-94. [2] T. Zvyagintseva, N. Shevchenko, E. Nazarenko, and et.al. J. Exp. Mar. Biol. Ecol. 2003, 294, 1-13. [3] B. Li, F. Lu, X. Wei, and R. Zhao. Molecules. 2008, 13, 1671-1695. [4] I. Wijesekara, R. Pangestuti, S. Kim. Carbohydr. Polym. 2011, 84, 14-21. [5] G. Jiao, G. Yu, J. Zhang, and H. Ewart. Mar. Drugs. 2011, 9, 196-223.


177

Brown Algae Fucus evanescens as Sourse of Inhibitors of O-glycoside hydrolases

A.S. Silchenko, T.I. Imbs, M.S. Avtushenko, S.P. Ermakova, T.N. Zvyagintseva

G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian

Academy of Sciences, 690022, Vladivostok, Russia E-mail: [email protected]

Investigation of enzyme inhibitors is an urgent task of modern enzymology. Enzyme inhibitors have a great practical importance in various fields: biotechnology, pharmacology and the study of biochemical processes occurring in the organisms. For example, terrestrial plants are protected from insects and herbivores by biosynthesis of inhibitors of digestive enzymes - amylase and/or protease. Details about function of enzymes inhibitors in marine organisms are scarce.

The aim of this study is to determine the presence of inhibitors of O-glycoside hydrolase among metabolites of the brown alga Fucus evanescens. For the experiment were used four different enzymes involved in the catabolism of polysaccharides of brown algae: highly purified fucoidanases FFA from the marine bacterium Formosa algae KMM 3553T and PPF from marine mollusk Patinopecten yessoensis, as well as recombinant fucosidase FucFa from F. algae and glucosidase G-II from the marine mollusk Littorina sitkana. Fucoidan from F. evanescens was used as substrate for evaluation of fucoidanases activity. Substrates for glycosidase and fucosidase were p-Np-α-L-fucopyranoside and p-Np-β-D-glucopyranoside, respectively. F. evanescens metabolites were recovered by successive extraction with organic solvents. The first, fresh samples of F. evanescens were extracted with ethanol to give ethanol extract (Eth). Then, crude Eth extract was successively treated with chloroform and ethyl acetate to give chloroform soluble (Chl), ethyl acetate soluble (EthAc) and water soluble extracts (Wt2) (Table). The metabolites of the EthAc extract inhibited the action of the o-glycoside hydrolase under study. Further separation on a silica gel column EthAc extract using different eluents gave rise to 13 fractions (Fr1-Fr13), that have different inhibitory effects on enzymes under study. So, the Fr6 and Fr9 eluted with ethyl acetate and chloroform-methanol (30:1), respectively, possessed inhibitory activity against fucoidanases FFA and PPF as well as fucosidase FucFa only. The nature of these metabolites is currently set. The Fr11 with highest inhibitory effect on investigated enzymes was separated on the silica gel column eluting with chloroform-methanol (1:1), then additionally separated on hydrophobic column to give 5 subfractions (Fr11.1-11.5). The Fr11.1-Fr11.4 did not inhibit the action of fucoidanases FFA and PPF, however, possessed inhibitory activity against glycosidases FucFa and G-II. This indicates the specificity of the inhibition of metabolites fractions. The subfractions 11.2 and 11.3 were purified over ultrafiltration (cut of 1kDa) to afford phlorotannins (polyphenolic), based on NMR analysis, with strong inhibitory effect toward FucFa (IC50 = 0.025 mg/ml) and G-II (IC50 = 0.025 mg/ml). In conclusion, as shown present study, the metabolites of ethyl acetate soluble extract of the brown algae F. evanescens possessed inhibitory activity against o-glycoside hydrolase under study. Further separation of the extract resulted phlorotannins that inhibited the action of oligosaccharide hydrolases FucFa only and G-II as well as metabolites Fr6 and Fr9 that inhibited the action of fucosidase FucFa and fucoidanases FFA and PPF. This may indicate that metabolites of the brown alga Fucus evanescens exhibit specificity inhibitory activity against different types of enzymes.

This work was supported by RFBR Grant # 15-04-01004_а.

Table. Inhibitory effect of the fractions obtained after separation of F. evanescens extract Inhibition ratio was expressed as a percentage of the original activity without adding of preparation: - not inhibited; + - <30 %; ++ - 30-60 %; +++ - > 70 ( at final concentration 0.1 mg/ml), n.d. – not determined.

Enzymes Fractions Eth Chl Wt1 EthAc Wt2 Fr.6 Fr.9 Fr.10 Fr.11 Fr.12 Fr.11.1 Fr.11.2 Fr.11.3 Fr.11.4

FFA + n.d. + +++ - +++ +++ + +++ ++ - - - - PPF + n.d. - +++ - +++ +++ + +++ + - - - - Fuc + n.d. + +++ - +++ +++ +++ +++ + ++ +++ ++ + G-II + n.d. + +++ - - - + +++ + + ++ +++ ++.


178

Activity of Cytisine Derivatives Against Human Parainfluenza

Virus Type 3 in vitro Experiments.

V.A. Fedorova,1 I.P. Tsipisheva,2 A.V. Kovalskaya,2 A.N. Lobov,2

E.A. Nikolaeva,2 M.S. Yunusov,2 V.V. Zarubaev1

1Research Institute of Influenza, Saint – Petersburg, Russia.

2Institute of Organic chemistry, Ufa science center, Ufa, Russia.

Parainfluenza viruses (HPIVs) can cause upper respiratory tract infection disease in individuals

of all age groups, although young children between 6 months and 3 years present more severe

diseases. HPIVs account for approximately 33% of lower respiratory tract infections in children

younger than 5 years. HPIVs have been associated with laryngotracheobronchitis or croup,

bronchiolitis and pneumonia. Furthermore, immunocompromised children and adults appear to be

particularly susceptible to developing severe and fatal lower respiratory tract infection. There are no

antivirals approved for the treatment of HPIV infection, although several compounds have

demonstrated in vitro and in vivo activity against these viruses. Ribavirin exhibits in vitro antiviral

activity against HPIVs, but it is expensive and has numerous side effects. Thus, its use is limited to

high risk or severely ill infants. In our study, we have evaluated the inhibitory effects of 16

synthetic cytisine derivatives in comparison with those of ribavirin on the in vitro replication of

human parainfluenza virus type 3 (HPIV3). Cytotoxicity of each compound and level of viral

replication were evaluated by MTT-test. The selectivity index was calculated for each analyzed

compound. According to the received data, 9 of 16 synthetic cytisine derivatives have antiviral

effect against HPIV3 and 1 compound was more efficient than ribavirin. Obtained data serve as the

basis for further development of the compounds of this group as a means for the treatment of

parainfluenza infection.


179

Methyl 3-benzoyl-4-oxo-1.4-dihydroquinoline-2-carboxylate: an in vitro

Assessment of Biomembrane Permeability and Hydrolytic Stability

I. Fefilova, A. Trefilova, A. Boteva, A. Mayorov, O. Krasnykh

Research and Educational Center of Applied Chemical and Biological Research

Perm National Research Polytechnic University 29 Komsomolsky Pr., Perm 614990, Russia,


ADME (absorption, distribution, metabolism, and elimination) studies of drug candidates is an

essential part of drug discovery process [1]. The success rates of drug development programs

heavily depend on the properly and timely evaluated pharmacokinetic (PK) profile of lead series

including permeability and stability both in gastrointestinal tract (GIT) and blood plasma.

Compounds containing 4-quinolone fragment possess a wide range of biological activities

including antibacterial, antimalarial, antitumor, etc [2]. In the process of development of a novel 4-

quinolone series we undertook an in vitro evaluation of the key pharmacokinetic (PK) parameters.

Methyl 3-benzoyl-4-oxo-1.4-dihydroquinoline-2-carboxylate was chosen as a model substrate for

the initial PK studies.

Permeability of compounds across biological membranes is the essential factor for in vivo

absorption and distribution. The ability of the 4-quinolone model substrate to permeate through

membranes was assessed in vitro by PAMPA (Parallel Artificial Membrane Permeability Assay)

experiment at different pH. The obtained data demonstrates high permeability of the tested 4-

quinolone, with its permeability being the highest at the pH range of 5.0-6.2.

Methyl 3-benzoyl-4-oxo-1.4-dihydroquinoline-2-carboxylate contains an ester functionality,

which can be hydrolyzed in aqueous media. To determine the predisposition of this compound to

hydrolytic degradation, we have studied its stability in phosphate-saline buffer (PBS). As

determined by HPLC the substrate concentration declined by approximately 40% (from 21µM to

12.2 µM) in the first two hours of the experiment. The product of the hydrolysis was identified by

LC-MS as 3-benzoyl-4-oxo-1.4-dihydroquinoline-2-carboxylic acid. Studies of the in vivo stability

and excretion parameters of both compounds are currently underway.

Acknowlegements: This work was funded in part by The PNRPU - EPFL Collaboration on Metabolism and Diabetes Program supported by the Neva Foundation.

References [1] J. Hodgson, Nat Biotech. 2001, 19, 722-726. [2] Quinolone antimicrobial agents, Eds D. C. Hooper, E. Rubinstein, Washingtin, ASM Press, 2003.


180

Virtual and Experimental Screening of the Maillard Reactions Inhibitors in

Class of Diphenyl Oxide Derivatives

V. Frantseva,1 P. Vassiliev,1 A. Spasov,1 V. Kuznetsova,1 O. Solovieva,1

Yu. Popov,2 V. Lobasenko,2 T. Korchagina2 and O. Efremova2

1Pharmacology Department, Volgograd State Medical University, 400131, Russia, Volgograd,

Pavshikh Bortsov Sq., 1 2Organic and Petrochemical Technology Department, Volgograd State Technical University,

400060, Russia, Volgograd, Lenin avenue, 28


Non-enzymatic glycation of proteins is enhanced in diabetes mellitus due to chronic

hyperglycemia and causes severe complications, such as nephropathy, encephalopathy, and others.

Therefore the search for the Maillard reactions inhibitors is promising for development of new type

of antidiabetic drugs [1]. Diphenyl oxide derivatives demonstrated hypoglycemic activity [2]. Thus,

the search for novel antidiabetic substances with antiglycation effect among

diphenyl oxide derivatives is very promising.

Virtual screening of antiglycation activity of six diphenyl derivatives with

general formula I was carried out by two methods: the method of similarity with standards in IT

Microcosm [3], based on the calculation of the modified Tanimoto similarity coefficient; and in

PASS system, which estimates the probability of the presence of a given activity [4]. The substance

is considered active if at least one prediction system has received a positive evaluation.

According to the prediction results, the probability of presence of the Maillard reaction inhibitor

activity for the two structures was high, for the two was moderate, and for the two was low. Thus,

all six compounds were promising and were tested.

In vitro testing was performed as described in [5]. Experiments have shown that the three

substances have high activity, two have low activity, and one substance is inactive. Therefore, the

accuracy of virtual screening amounted to 83.3%. The results of these studies demonstrate the

prospects of finding of novel antiglycation substances among diphenyl oxide derivatives.

The study was performed in VSMU by RSF grant № 14-25-00139.

References [1] N. A. Ansari, and Z. Rasheed, Biomedical chemistry. 2010, 56, 168-178 [2] Yu. V. Popov, T. K. Korchagina, and M. V. Smirnova, News VSTU. 2009, 6, 62-65 [3] P. M. Vassiliev, and A. N. Kochetkov, IT Microcosm. State Registration Certificate for software program 2011618547 (Russian). 2011 [4] D. A. Filimonov, V. A. Poroikov, T. A. Gloriozova et al., PASS (Prediction of Activity Spectra for Substances). State Registration Certificate for software program 2006613275 (Russian). 2006 [5] A. Jedsadayanmata, Naresuan University Journal. 2005, 13, 35-41

O R1

I


181

Cytotoxicity of Pomolic Acid Isoplated from Chamaenerion angustifolium

Against Human Cancer Cells

T.S. Frolova,1,2,3 T.P. Kukina,2 N.V. Gubanova,3 O.I. Sinitsyna3 1Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogov Street, 2

2Novosibirsk Institute of Organic Chemistry, 63090, Russia, Novosibirsk, Lavrentyev Avenue, 9 3Institute of Cytology and Genetics, 63090, Russia, Novosibirsk, Lavrentyev Avenue, 10


Previously, we have studied the composition of the lipophilic acid fraction obtained from rose

bay willow-herb (Chamaenerion angustifolium). It has been shown that plant raw material from

rose bay willow-herb is enriched by pomolic acid and could be industrial source of triterpene acids.

Pomolic acid is a very promising prospective compound for pharmacology, in vivo and in vitro

models have been shown its strong cytotoxic effect against cancer cells, and weak effect against

normal.

Pomolic acid was found to play positive role in overcoming multiple drug resistance syndrome.

Significant antioxidant effect of pomolic acid was observed, known to correlate with anticancer

properties by the bacterial test systems.

Cytotoxic properties of pomolic acid from willow-herb on several malignant cell lines were

investigated. These lines are characterized by the presence or absence of key apoptosis gene p53.

Human fibroblasts were used as a control cell line. It was shown that cytotoxicity of pomolic acid is

decreased for the lines in which p53 gene is absent. Normal cells are resistant to the toxic effects.

Microscopic analysis of the cell cultures treated pomolic acid was conducted. Significant

morphological changes exhibit after 24 h of exposure to the drug.

References 1. T. S. Frolova, O. I. Sal’nikova, T. A. Dudareva, T. P, Kukina, O. I. Sinitsyna. Russian Journal of Bioorganic

Chemistry, 2014, 40, 82–88. 2. European Patent № 1549330 «Pomolic acid for treating multidrug resistant tumours».


182

Stereospecific 7α-alkylation and 7,7-bis (alkynation) 20 hydroxyecdysone. The

first example spirocyclization of ecdysteroids

I.V. Galyautdinov, Z.R. Khairullina, V.P. Sametov, G.G. Gibadullina, V.N. Odinokov

Institute of Petrochemistry and Catalysis Russian Academy of Science, 450075, Ufa, pr. Oktyabrya, 141,


7α-Alkylsteroids constitute an important class of pharmacologically active compounds. The

stereoselective introduction of an alkyl group into the 7-position is a key transformation of steroids,

which is usually accomplished by conjugated addition of lithium alkyl cuprates to 4,6-unsaturated

3-ketosteroids, resulting in the formation of a mixture pharmacologically significant 7α- and 7β-

alkylsteroids [1]. The alkylation of ecdysteroids have not been studied. Recently, stereospecific 7α-

alkylation of 20-hydroxyecdysone has been described. The reductive alkylation of α,β-unsaturated

ketones in lithium-ammonia solution could be a convenient method for direct introduction of alkyl

groups in the 7α-position ecdysteroids.

We have found that by treating the lithium-ammonia solution 20-hydroxyecdysone (1) a solution

of methyl-, ethyl iodide or allyl bromide in THF are formed in high yield 7α-methyl- (2), 7α-ethyl

(3), and 7α-allyl-14-deoxy-∆8(14) -20-hydroxyecdysone (4), respectively.

When used as the electrophilic reagent propargyl bromide the reaction product is 7,7-bis (2-

propyn-1-yl) derivative of 5. The catalytic hydrogenation leads to the first representative of the

spiro compounds among ecdysteroid – spiro compound 6.

O

OH

OHH

OH

OH

H

OH

H OH

O

OH

OHH

OH

OHOH

H

H

R1

R

Li / liq. NH3

RHal H2 / Pd - C

O

OH

OHH

OH

OHOH

H

H

Me

Me1 2-5

7

123

45

6

8

9

10

11 1314 15

1617

18

19

2021

2223

24

25

26

2712

6

1'

2'3'

4'5'

6 '

7 '

2: R=Me, R1=H; 3: R=Et, R1=H; 4: R=CH2CH=CH2, R

1=H; 5: R=R1=CH2C≡CH.

References [1] Joyce F, Harvey D, J.O'Neal Johnston, Petrow V. Antiprogestational agents. The synthesis of 7-alkyl steroidal

ketones with anti-implantational and antidecidual activity. Steroids. 1976; 27, 759-771.


183

Antioxidative Activity of Silibum Marianum Cultivated in Mongolia

J. Ganbaatar,1 E. Lkhagvamaa,1 E.E. Shults2 1Institute of Chemistry and Chemical Technology, Mongolian Academy of Sciences,

Ulaanbaatar, Mongolia 2Novosibirsk Institue of Organic Chemistry, Russia

Silybum marianum (L.) Gaerth (Milk thistle) (Asteraceae family) is a medical plant widely used

in a traditional medicine. The fruit of this plant is used in the form of liquid extracts for treatment of

liver and gall bladder disorders, including hepatitis and cirrhosis, and to protect the liver against

poisoning from wild mushroom, alcohol, chemical, and environmental toxins. The efficiency of the

extracts from the Milk thistle fruit for a treatment of liver deseases motivated their examination in a

chemical and pharmacological sense. Nowadays, standardized mixture of flavonoids, known as

silymarin or silymarin complex, is used as a hepatoprotective pharmaceutical preparation. Silibum

marianum (Milk thistle) contains high amount of phenolic compounds with antioxidant properties.

This plant was successfully cultivated in Chuluunkhoroot soum, Dornod province, Mongolia, by

“Ereentsav”CoLtd. The aim of this study is to evaluate the antioxidtive activity of methanolic

extracts of the seeds and leaves of S. marianum cultivated in Mongolia. The antioxidant properties

of the leaves and seeds of milk thistle were examined by determining its ability to scavenge the 2,2-

diphenyl-1-picrylhydrazyl (DPPH) free radical.

Table 1. The IC50 of theleaves and seeds of S. marianum

Sample, µg/ml

DPPH scavenging activity, %

IC50 (sample concentration required to scavenging 50% of the DPPH radicals),

µg/ml

Sample, µg/ml

DPPH scavenging activity, %

IC50 (sample concentration required to scavenging 50% of the DPPH radicals),

µg/ml Leaves Seeds

1600 43.7

1800

800 86.4

268 800 21.2 400 71.2 400 9.8 200 40.7 200 5.5 100 23.2

The results obtained from the DPPH assay confirm that Milk thistle extracts have high

antioxidative activity. The IC50 of leaves of Milk thistle was higher (1800 µg/ml) than seeds (268

µg/ml). The antioxidant activity of the seeds prevails activity of leaves more than 6 times.


184

Determining Oxidized Proteins in Human Serum by Micromethod

A.P. Godovalov and N.V. Vavilov

Department of immunology, Acad. E.A. Wagner Perm State Medical University, 614000, Russian

Federation, Perm, Ekaterininskaya str. 85

E-mail: [email protected], [email protected]

In recent years, free-radical processes and their impact on intra- and extracellular metabolism are

actively studied. Native conformation of the protein molecules changes under the effect of oxidative

stress, up to fragmentation that affect their function.

Aim of reserch was to determine the level of protein’s oxidative modification (POM) in human

serum using micromethod.

Materials and methods. We investigated serum samples of peripheral blood of 34 healthy donors

in the age of 21.1±0.3 years. The evaluation of POM was performed by our modification of the

method of Reznick A.Z. et al. [1] the essence of which was in proportional decrease in the volume

of the reactants. The principle of the reaction is based on the interaction of oxidized amino acid

residues of proteins with 2,4-denitrofenilgidrazin to form 2,4-dinitrophenylhydrazone derivatives,

whose absorption density was evaluated spectrophotometrically at 360 nm. The concentration of

oxidized proteins expressed in nmol/mg of total serum protein, which was determined with micro-

biuret method. Statistical analysis used the Student t-test, for significance threshold value accepted

p<0.05.

Results and discussion. The level of POM in serum from healthy donors was 1.84±0.06 nmol/mg

protein, which corresponds to the results obtained in a classic embodiment reactions (p>0.05), and

the data in the literature [2, 3].

Conclusion. Thus, the proposed option of micromethod can be used to determine the serum POM

and it significantly reduces the amount of reagents used.

References [1] A.Z. Reznick, and L. Packer, Methods in enzymology. 1994, 233, 357-363 [2] I. Dalle-Donne, R. Rossi, D. Giustarini et al., Clinica Chimica Acta. 2003, 329, 23-38 [3] J.J. Sheth, U.J. Shah, F.J. Sheth et al., Kamla-Raj Enterprises. 2011, 11, 83-88


185

New Copper-catalyzed Synthesis and Acetylcholinesterase Inhibitory Activity of

N-substituted 2-Aminoisonicotinic Acid

N.E. Golantsov, A.V. Varlamov, and L.G. Voskressensky

Faculty of Science, Peoples’ Friendship University of Russia, 117198, Russia, Moscow, Miklukho-

Maklaya St., 6


Different physiologically active compounds contain in their molecules isonicotinic acid fragment

[1,2]. N-Substituted 2-aminoisonicotinic acids are important reagents in synthesis of such

compounds [2]. Unfortunately, a convenient preparative synthetic method to obtain simple N-

substituted 2-aminoisonicotinic acid has not been published yet. We propose an effective

preparative procedure, based on copper-catalyzed substitution of chlorine in 2-chloroisonicotinic

acid by amines in the presence of sodium hydroxide.

R1 R2 Yield, % Percentage Inhibition, 10-5 mol/l 3 H CH3 83 15±7 4 CH3 CH3 90 ~ 0 5 H C6H5 95 ~ 0 6 H CH2CH2CH3 72 58±4 7 H CH2CH=CH2 68 43±7

The reaction proceeds in water under increased temperature in a sealed tube. The presence of

sodium hydroxide is important to prevent amide formation. Thus obtained N-substituted 2-

aminoisonicotinic acids were screened for acetylcholinesterase inhibitory activity in order to our

ongoing program of seeking new acetylcholinesterase inhibitors. Tests were conducted by Ellman’s

method [3] using multifunctional microplate reader Tecan Infinite M1000 Pro. Two compounds

showed noticeable activity and may serve as a starting point for the future investigation.

This work was performed using scientific equipment of Shared Research and Educational Center for Рhysico-chemical studies of new materials, substances and catalytic systems PFUR. References [1] J. Y. Chung, F. A. Pasha1, S. J. Cho, M. Won, J. J. Lee, K. Lee, Arch. Pharm. Res. 2008, 32, 317-323. [2] C. A. Lipinski, J. L. LaMattina, A. Hohnke, J. Med. Chem. 1985, 28, 1628-1636. [3] G.L. Ellman, K.D. Courtney, V. Andres jr., R.M. Featherstone, Biochem. Pharmacol. 1961, 7, 88-95.


186

Novel Hydroxylaminoisoxazoles as Perspective Lipoxygenase Inhibitors

Yu. Gracheva,1 D. Vasilenko,1 E. Averina,1,2 M. Neganova,2 O. Redkozubova,2

E. Shevtsova,2 E. Milaeva1,2

1 Moscow State Lomonosov University, 119991, Russia, Moscow, Lenin Hill, 1-3

2 Institute of Physiologically Active Compounds of RAS,142432, Russia, Chernogolovka, Moscow Region, Severny proezd, 1


The novel approach to 5-[hydroxy(tetrahydrofuran-2-yl)amino]isoxazoles 1 and functionalized

5-nitroisoxazoles 2 [2] was used for the design of lipoxygenase (LOX) inhibitors which appear to

be a certain structural analogous of the known drug Zileuton.

The synthesis, physico-chemical properties and biological activity of a series of novel

compounds 1 that contain adamantane fragment as R’ will be discussed. The biological screening

was performed using LOX inhibition assay, lipid peroxidation, DPPH-test, metal chelating method,

mitochondrial potential study. The compounds show a high activity as LOX inhibitor with IC50 in

micro- and nanomolar region and non-toxicity in cerebrial granual neurons MTT test.

Neurons survivability control

Kinetic curves of enzymatic linoleic acid hydroperoxides formation in the presence of different concentration of 1

Cerebrial granual neurons (MTT test) for compound 1

Acknowledgments: The financial support of RFBR (grants 15-03-03057, 14-03-00469, 13-03-01151), Presidium RAS (program №8P) is gratefully acknowledged, the equipment of Center for Collective Use IPAC RAS (agreement N14.621.21.0008, ID RFMEFI62114X0008) were used. References [1] E.B. Averina, D.A. Vasilenko, Y.V.Samoilichenko, Yu.K. Grishin, V.B.Rybakov, T.S. Kuznetsova, N.S. Zefirov. Synthesis, 2014, 46(8), 1107. [2] (a) Y.A. Volkova, E.B. Averina, Yu.K. Grishin, P. Bruheim, T.S. Kuznetsova, N.S. Zefirov J. Org. Chem., 2010, 75, 3047. (b) E.B. Averina, Y.A. Volkova, Y.V.Samoilichenko, Yu.K. Grishin, V.B.Rybakov, Kutateladze A.G., Elyashberg M.E., T.S. Kuznetsova, N.S. Zefirov. Tetrahedron Lett., 2012, 53, 1472.


187

New Chlorin E6 Based IR Photosensitizers for Photodynamic

and Antimicrobial Therapy

O.I. Gushchina,1 E.A. Larkina,1 T.A. Nikolskaya,2 A.F. Mironov1

1M.V. Lomonosov Moscow State University of Fine Chemical Technologies, Moscow, Russia. 2Emanuel Institute of Biochemical Physics of Russian Academy of Sciences, Moscow, Russia.


Photosensitizers (PS) became popular drugs in the last decades. It can be explained by simplicity

and noninvasive characteristics of the process of PDT (photodynamic therapy). PDT is a promising

new treatment modality for several diseases, especially cancer. In PDT, light, O2, and a

photosensitizing drug are combined to produce a selective therapeutic effect. The antimicrobial

therapy is provided in the same manner. The PS in this case should be positively charged because

the membrane of bacteria is charged negatively. Studies are underway to create new photosensitizer

candidates, because the most commonly used porphyrin photosensitizers are far from ideal with

respect to PDT and antimicrobial therapy. Finding a suitable photosensitizer is crucial in

improving the efficiency of PDT. Chlorins seem to be promising candidates for PDT owing to their

photophysical properties.

In this work, amide derivatives of chlorin e6 are proposed as promising tetrapyrrole

photosensitizers due to hydrophobic substituents which is believed determines the affinity of the

photosensitizer to the cell membrane and contributes an embedding the molecules in the membrane

systems and therefore the efficiency of photocatalytic processes increases. Given the above, the

purpose of this study was to obtain derivatives of chlorin e6 and to investigate their biological

activity. To synthesize new photosensitizing drugs we carried out the disclosure of exocycle E of

pheophorbide a and its methyl ester with primary unbranched aliphatic mono- and di- amines with

the formation of the corresponding amide derivatives of chlorin e6. The reactions yielded 50-75%.

In case of preparation of cationic PS we performed reaction of methylation of the compounds

obtained 2a,b by adding excess of methyl iodide in chloroform at room temperature.

The purity and the structure of all the compounds obtained were confirmed by methods of TLC,

electronic, IR, 1H-NMR spectroscopy and mass-spectrometry.

HNN

NNH

ONH

MeOOCR1OOC

R3COOMeR1OOC

HNN

NNH

O

CHCl3

a: R2 = CH2CH2NH2, R1 = CH3; b: R2 = CH2CH2N(CH3)2, R1 = CH3; c: R2 = (CH2)3N(CH3)2, d:(CH2)6NH2, R1 = CH3; e: R2 = (CH2)2O(CH2)2O(CH2)2NH2, R1 = CH3;f: R2 = (CH2)6NH2, R1 = CH3; g: R2 = (CH2)3, R1=H; h: R2 = (CH2)3CH3, R1=H;i: R2 = (CH2)4CH3, R1=H; j:R2 = (CH2)5CH3, R1=H; k:R2 = (CH2)6CH3, R1=H; l:R2 = (CH2)7CH3, R1=H; m:R2 = (CH2)8CH3, R1=H; p:R2 = (CH2)9CH3, R1=H.

1

a: R 1 = CH3 b: R1 = H

2 a-p

CH2

N

HNN

NNH

ONH

CH 3OOCCH3OOC n

CH3I

CHCl3

R2 NH2

a,b 3 b,c

b: n=2c: n=3


188

Synthesis and Analgesic Activity of Monoterpenoid Derived 2-Alkyl-4,7-

dimethyl-3,4,4a,5,8,8a-hexahydro-2H-chromene-4,8-diols

I. Il’ina,1,2 A. Pavlova,1 D. Korchagina,1 T. Tolstikova,1 K. Volcho1,2, and N. Salakhutdinov1,2

1 Novosibirsk Institute of Organic Chemistry, Siberian Branch, Russian Academy of Sciences,

Lavrentjev av., 9, 630090 Novosibirsk, Russian Federation 2 Novosibirsk State University, Pirogova St. 2, 630090 Novosibirsk, Russian Federation


Recently, we have found a new series of heterocyclic compounds with chiral hexahydro-2H-

chromene skeleton, which possess a considerable analgesic activity in in vivo tests [1]. These

compounds were synthesized in one preparative step by the reaction of verbenol epoxide 1 or diol 2

(the product of isomerization of epoxide 1) with aromatic aldehydes in the presence of

montmorillonite clay K10.

In this work, we have continued to search for new compounds with high analgesic activity

combined with low toxicity. Therefore the aim of the work was synthesis and analgesic activity

study of 2-alkyl-4,7-dimethyl-3,4,4a,5,8,8a-hexahydro-2H-chromene-4,8-diols 3, derived from

aliphatic aldehydes and diol 2.

2-Alkyl-substituted compounds 3a-n with a hexahydro-

2H-chromene framework were synthesized with yields 36%

- 80%. The analgesic activity of compounds 3a-n was

studied in the acetic acid-induced writhing test and hot plate

test. The majority of the compounds under study exhibited

pronounced analgesic activity. Thus we have found a series

analgesics which are very promising for further studies. References: [1] I. Il’ina, O. Mikhalchenko, A. Pavlova, D. Korchagina, T. Tolstikova, E. Pokushalov, K. Volcho, N. Salakhutdinov. Med Chem Res 2014, 23, 5063–5073.

Aldehyde Yields of 3,a %

4a acrolein 76 4b crotonaldehyde 70 4c α-methacrolein 62 4d trans-2-penten-1-al 80 4e trans-2-hexen-1-al 78 4f trans-2-octen-1-al 54 4g 2,4-hexandien-1-al 57 4h butanal 60 4i isobutyraldehyde 36 4j hexanal 59 4k heptanal 36 4l octanal 77 4m decanal 60 4n cyclohexanecarboxaldehyde 68


189

Anticancer Activity of Some Plant Extracts on U87MG

Human Glioblastoma Cell

V.A. Isidorov,1 S. Bakier,1 J. Moskwa2 and M.H. Borawska2

1Forest Faculty, Bialystok Technical University, 17-200 Hajnowka, Poland

2Faculty of Pharmacy, Medical University of Bialystok, 15-089 Bialystok, Poland E-mail: [email protected]

Glioblastoma is a common primary brain tumor which demonstrates a high proliferation rate and

an aggressive growth pattern and is largely resistant to chemotherapy. One of the most promising drugs for brain tumor therapy is temozolomide (TMZ), an oral alkylating agent belonging to imidazotertrazines. However, the average survival of patients with glioblastoma after TMZ treatment is 22 months, which is still not enough to be satisfied with the therapy. The aim of our study was to examine anti-cancer activity of selected plant extracts in U87MG human glioblastoma cell line expressed as changes in cell viability. Objects of study were supercritical fluid CO2 extracts (SFE) of buds of plants which are common on the Northern Hemisphere: two species of white birch and aspen. Developed technology (CO2-SFE) ensures high resin recovery efficiency of over 95%. Besides, diethyl ether extract of aspen twigs was examined.

We have found that all extracts had a dose- and time-dependent inhibitory effect on the U87MG cell line growth, which was manifested by gradual reduction of cell viability (Figure 1). After 24 h of incubation, all extracts at concentrations 50 µg/ml did not significantly reduce (with exception of aspen buds) the viability of the U87MG cell line. The marked reduction of viable cell number (17.0-45.1% of control) at that time point was observed only when extracts were used in concentration of 100 µg/ml. After 48 h of exposure, the viability of glioma cells decreased gradually with increasing extract concentrations reaching 20.0-46.4% of control for the lowest concentration and 12.0-40.0% for the highest. A strong reduction in the number of living cells in relation to control was observed after 72 h of treatment with concentrations 50 µg/ml (viability from 8.6 to.25.3%) and 100 µg/ml (viability from 2.3 to 12.0%).

The observed anti-tumor effect can be addresed to the high amounts of flavonoids (Betula

pubescens buds), phenyl propanoids (Populus tremula buds) and triterpenoids (B. pendula buds) in the extracts under investigation. In spite of the great differences in chemical composition, anti-cancer activity of the extracts of buds was observed. Extract of aspen (P. tremula) twigs is characterized by the high content of glycosides with different phenolics as aglicone moeties. As can be seen, this extract demonstrates some lower antitumor activity.

0102030405060708090

100

50 µg/ml 100 µg/ml 50 µg/ml 100 µg/ml 50 µg/ml 100 µg/ml

24 h 48 h 72 h

Via

bil

ity

of

HT

B 1

4 (%

of

con

tro

l)

Control 100% B. pubescens buds B. pendula buds P. tremula buds P. tremula twigs


190

Study on Effects of Plant Extracts on Paenibacillus larvae, a Causative Agent of

American Foulbrood: Preliminary Results

V.A. Isidorov,1 K. Buczek,2 M. Zambrzycka3 and I. Swiecicka3

1Forest Department, Bialystok Technical University, 17-200 Hajnowka, Poland

2Veterenary Medicine Department, Lublin University of Life Science, 20-950 Lublin, Poland 3Department of Microbiology, Bialystok University, 15-950 Bialystok, Poland


The managed honeybees are the only important productive “livestock” used for pollination of crop and fruit monocultures. The economic effect of that activity is estimated in billions of $US in the USA only. Hence, pollination through honeybees is extremely important for the reproduction of many crops, fruits as well as wild plants. Due to honey bee’s role in pollination, factors affecting bee health affect natural ecosystems, as well as sustainable agriculture. Because of this, the worsening of honeybee health, vitality and longevity is a serious concern.

The serious problems in beekeeping worldwide are connected with honeybee diseases, American foulbrood (AFB), caused by the Gram-positive spore forming bacteria Paenibacillus larvae, which is extremely pathogenic and aggressive microorganism attacking honeybee larvae. A common strategy for treatment of infected colonies is the use of antibiotics; however, several serious problems are associated with their extensive use: development of antibiotic-resistant strains of P.

larvae, as well as the contamination of bee products by residues of antibiotics. Literature of the last decade confirms an interest in finding substitution of chemical drugs with

natural substances for the control of AFB. In recent investigations antibacterial action of propolis on P. larvae was shown. However, propolis is highly valued bee product and the demand for it is still growing. Moreover, it plays an extremely important role in bees “social immunity”. Because of that, it is desired to replace propolis with a plant material which is common on the Northern Hemisphere.

Table shows the results of preliminary microbiological search on buds and young twigs extracts. These in vitro experiments are very promising: all tested plant extracts inhibit the growth P. larvae. Some of these extracts are more effective than propolis. According to our GC-MS analysis, these extracts contain high amounts of flavonoids, phenyl propanoids and other phenolic compounds. A promising technology of isolation from plant material of resins with high anti-microbial activity is supercritical fluid CO2 extraction.

Table. Minimal inhibitory concentration (MIC) demonstrated by extracts from selected parts of different plants and „poplar-type” propolis on five „wild” strains of P. larvae

Material/extract MIC (µg·ml-1) KB3 KB6 KB8 KB9 KB13

Aspen P. tremula (buds) 19.5 39.0 39.0 78.0 39.0 Litwinow birch B. litwinowii (buds) 39.0 39.0 156.0 78.0 19.5 Silver birch B. pendula (twigs) 313.0 19.5 625.0 19.5 19.5 Downy birch B. pubescens (twigs) 156.0 39.0 156.0 78.0 39.0 “Poplar-type” propolis (70% ethanol extract)* 19.5 39.0 78.0 19.5 39.0 * The MIC values are comparable with literature data for four samples of “poplar-type” propolis against the reference strain of P. larvae (MIC values from 31.2 to 250 µg·ml-1) (Bilikova et al., 2013) This work was supported by National Science Centre (Poland) grant 2014/13/B/NZ7/02280


191

High Temperature High Resolution Gas Chromatography Coupled with Mass

Spectrometry for Analysis of Biologically and Environmentally Important

Organic Compounds

V.A. Isidorov

Forest Faculty, Bialystok Technical University, 17-200 Hajnówka, Poland e-mail [email protected]

Presently high resolution gas chromatography coupled with low-resolution mass spectrometry

(HRGC-MS) has been the most effective and widespread method for investigation of chemical compositions of organic compounds of different origins. The area of application of this technique covers metabolomic profiling, phytochemical and pharmacological investigations, medicinal chemistry, forensic medicine and criminology, food chemistry and cosmetology, chemical ecology and ecotoxicology, as well as environmental chemistry.

For a long time the impossibility of transformation to a gas phase without decomposition or isomerization of labile or non-volatile compounds involving many plant metabolites has been considered a main limitation in gas chromatography. In contemporary gas chromatography, this limitation is overcome by derivatization of analytes, i.e. transformation of them into volatile and stable at high temperatures derivatives and decreasing their polarity. To this end different reagents are used, with the most widely applied method being trimethylsilylation (TMS) due to high reaction rate and low labor intensiveness of a derivatization procedure.

Success of such kind of research is largely determined by the availability and sufficiency of databases of analytical parameters which, in case of GC-MS, are mass spectra and chromatographic retention parameters in their most reproducible form of retention indices (RI). Although mass spectra are the main identification parameters in such investigations, one cannot underestimate importance of retention indices. They serve not only for confirmation of mass spectrometric identification results: a special need for them arise in analysis of isomers, frequently giving essentially indistinguishable mass spectra, as well as in case of minor mixture components when one fails to obtain mass spectra of a sufficiently high quality. Due to this an important role of retention indices in studying the composition of complex mixtures by GC-MS is generally recognized. Unfortunately, the available databases contain only a small number of both mass spectra and retention indices of TMS derivatives of biologically and environmentally important compounds.

Many years of investigating the composition of various mixtures of organic compounds (essential oils, extracts from plant tissues, food products, environmental and industrial samples, etc.) resulted in compiling data on both MS and RI of TMS derivatives [1]. This collection contains linear temperature programmed retention indices (from 650 to 4500 i.u.) on the chromatographic column with HP-5ms stationary phase and mass spectra more than 1150 organic compounds. A point of special interest has analytical parameters of previously none investigated substances detected by author in plant tissues such as phenylpropenoids of sesquiterpenols, glycerides of cinnamic acids, flavonoids and their glycosides. More than 60% of presented in [1] compounds were not previously characterized by their mass spectra and more than 70% by RI values.

Experimental factors influencing the accuracy of retention indices and possible artifacts and complications of mass spectral identification are discussed.

[1] V A. Isidorov (2015) GC/MS Identification of Biologically and Environmentally Important Organic Compounds as their Trimethylsilyl Derivatives. 400 p., A4, hardcover, PWN, Warsaw


192

Mechanochemical Technology of Obtaining Amino Acid Containing Products

I.V. Ivanov,1 A.L. Bychkov,2 O.I. Lomovsky2

1Faculty of Natural Science, Novosibirsk State University, 630090, Russia,

Novosibirsk, Pirogova Str., 2 2Institute of Solid State Chemistry and Mechanochemistry, 630128, Russia, Novosibirsk,

Kutateladze Str., 18


Modern health products are basеd on bioavailable oligopeptides and amino acids [1]. Control of

technological process is important to obtain a required hydrolysis product. The final product should

have required chemical composition, namely, it should contain the correct amount of amino acids,

oligopeptides with a predetermined chain length. The product should not contain harmful aromatic

molecules.

Mechanochemical activation of enzymatic hydrolysis is a difficult but promising method to

control the reactivity of solid-phase proteins in the raw material.

The aim of this work is to study the influence of mechanical activation on the rate and depth of

the subsequent hydrolysis of solid-phase proteins included in the composition of plant raw

materials for further amino acid containing products.

As a result of this work, the influence of particle size on the yield of water-soluble substances

was investigated, the molecular weight of proteins was determined by gel electrophoresis. For

isolation of the solid peptides from the solution, obtained after extraction of soluble substances,

methods such as freeze-and spray-drying were used. Size of the particles and fibers of water-

soluble substances were determined by scanning electron microscopy.

Preliminary experiments in the effect of mechanical treatment on the yield of extractives showed,

that, it is possible to extract at most 30%, without enzymatic hydrolysis. Enzymatic hydrolysis

enables to increase the yield (60 % and more), but it requires detailed study of the influence of

mechanical activation on the reactivity of proteins. Freeze-drying method is most suitable for

obtaining the products in solid phase.

[1] Самошкин С.П., Бычкова Е.С., Бычков А.Л. Супы пюре лечебно-профилактического назначения с питательными веществами в легкоусвояемой форме// Пищевая промышленность. – 2013 - №8 – стр. 26-27


193

Anti-osteoporosis Activity of Artemisia capillaris Extract and its Major

Compound Scoparone via Regulation of Osteoclastogenesis and Bone

Resorption Activity

Sang-Hyun Lee, Yong-Beom Kwon, Dae-Kun Lee, Fang-Fang Wang, Myung-Hee Kang,

Young-In Kwon, Young-Ho Kim1, and Hae-Dong Jang

Department of Food and Nutrition, Hannam University,

Daejeon, 305-811, Korea 1College of Pharmacy, Chungnam National University, Daejeon, 305-764, Korea


Bone is dynamic tissue that is constantly destroyed or resorbed by osteoclasts and then replaced

by osteoblasts in physiological process referred to as bone remodeling. The regulation of formation

and function of bone-resorbing osteoclasts is a key to understanding the pathogenesis of skeletal

disorders. The inhibitory effect of Artemisia capillaris extract (ACE) on osteoporosis was

investigated by analyzing bone mass, biomarkers of bone formation and resorption in

ovariectomized (OVA) SD rats as a postmenopausal bone loss model and the underlying

mechanism was elucidated by examining the suppressive effect of scoparone as a major compound

of ACE on RANKL-induced osteoclast differentiation. ACE inhibited the decrease in total BMD

and BMC of the right femur induced by OVA, which was accompanied by a significant change in

biomarkers of bone remodeling. The bone turnover markers in bone formation such as BALP,

PICP, and OPG were markedly enhanced by ACE and those in bone resorption as RANKL, TRAP,

and ICTP were dose-dependently reduced by ACE treatment. The TRAP and bone resorption

activity were dose-dependently alleviated by scoparone in RANKL-induced osteoclast

differentiation. In addition, the suppressive effect of scoparone on RANKL-induced osteoclast

differentiation was executed by down-regulating ROS production through NADPH oxidase 1 and

mitochondria and by scavenging generated ROS. In conclusion, the protective effect of ACE on

osteoporosis can be accomplished by attenuating RANKL-induced osteoclast differentiation and

bone resorption activity. These results imply that ACE may utilized as a therapeutic agent for the

prevention of bone metabolism-related diseases.

References [1] Na Kyung Lee, Young Geum Choi, Ji Youn Baik, Song Yi Han, Dae Won Jeong, Yun Soo Bae, NackSung Kim, and Soo Young Lee, Blood. 2005, 106(3),852-859


194

Turmeric Extract Fermented with Bifidobacterium Induces Apoptosis via

Caspase-Dependent Pathway and Cell Cycle Arrest in HCT116 Human

Colorectal Cancer Cells

Ji-Eun Park, Sang-Hyun Lee, Min-Ji Seo, P. Belousova, Myung-Hee Kang, Young-In Kwon

and Hae-Dong Jang

Department of Food and Nutrition, Hannam University, 1646 Yuseongdaero,Yuseonggu

Daejeon, 305-811, Korea


Turmeric (Curcuma longa L.) has been used in traditional Indian medicine for many centuries for

its anti-inflammatory, anti-oxidant and anti-carcinogenic properties. Several studies have showed

that the fermentation with microorganism can enhance the beneficial components and effects of

tumeric. This study investigated anti-proliferative and apoptotic effects of turmeric extract

fermented with Bifidobacterium in HCT116 human colorectal cancer cells. There was no significant

different in curcumin content profile between fermented turmeric (FTE) and turmeric extract (TE).

Cell viability assay demonstrated that FTE and TE similarly inhibited the growth of HCT116 cells.

However, annexin V/PI staining showed that FTE between 300~600 µg/mL obviously induced the

apoptosis of HCT116 cells compared to TE. The stronger apoptotic effect of FTE than that of TE

was observed in activation of caspase 3 and PARP cleavage, which is the hallmark of caspase-

dependent apoptosis. Furthermore, we found that FTE was more potent than TE as an anti-

carcinogenic agent which potentially suppresses TNF-α-induced IKK and NF-κB activation. In

addition, it was conformed that FTE remarkably alleviated the expression of cyclin D1 by inducing

the expression of cyclin-dependent kinase inhibitor p21 compared to TE. Consequently, our study

implies that the enhanced apoptotic effect of FTE through NF-κB inactivation and cell cycle arrest

may be due to the transformation of bioactive compounds or the newly produced bioactive

compounds by fermentation with Bifidobacterium.

References [1] Jayaraj Ravindran, Sahdeo Prasad, and Bharat B. Aggarwal, The AAPS Journal. 2009,11(3),1550-7416


195

Solvent-Free Hofmann Rearrangement under High-Vibration

Ball-Milling Conditions

Zhijiang Jiang, Zhenhua Li, Weike Su

Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou 310014, P. R. China.

Hofmann Rearrangement is one of the most important organic reactions in drug synthesis

transforming amide to amine. During the past decades, efforts have been paid to make the reaction

greener. High efficient green synthetic methods with high functional group compatibility and

excellent yields have been widely studied usingN-haloamides1 and hypervalent iodine reagents2 as

oxidants. However, in order to avoid over oxidation of amines, methanol was used as solvent giving

methyl carbamates in most cases, which need additional deprotection to give free amines. To best of

our knowledge, there haven't been any cases of solvent-free Hofmann Rearrangement reported.

Besides, the cost brought by N-haloamides and hypervalent iodine reagents also limited the

application of the methods. Thus, it’s still challenging to develop anefficient Hofmann

rearrangement using inexpensive oxidant under solvent-free conditions.

Recently, a solvent-free Hofmann rearrangement promoted by high-vibration ball milling was

found in our lab. Inexpensive and eco-friendly trichloroisocyanuric acid (TCCA) was chosen as

oxidant, giving free amines directly after 80 minutes grinding with high yields. The reaction

mechanism, functional group tolerance and synthetic application were under investigation.

CONH2

t-BuOH / K2CO3

Ball-Mil ling, 30 Hz, 80 min

Cl

Cl

NH2

Cl

Cl

N

N

N

O

Cl

O O

Cl

Cl

+

95% yield

TCCA • solvent-free• low cost and eco-friendly oxidant• forming free amine directly• high yields with short reaction time

References 1. a) Borah, A. J.; Phukan, P. Tetrahedron Lett., 2012, 53, 3035; b) Huang, X.; Seid, M.; Keillor, J. W. J. Org. Chem.,

1997, 62, 7495; c) Crane, Z. D.; Nichols, P. J.; Sammakia, T.; Stengel, P. J. J. Org. Chem., 2010, 76, 277. 2. a) Zagulyaeva, A. A.; Banek, C. T.; Yusubov, M. S.; Zhdankin, V. V. Org. Lett. 2010, 12, 4644; b) Yoshimura, A.;

Middleton, K. R.; Luedtke, M. W.; Zhu, C.; Zhdankin, V. V. J. Org. Chem., 2012, 77, 11399.


196

A New Thiazolidine-4-carboxylic Acids as a Potent Antitumor Substances.

Syntheses and in vitro Investigations

D.S. Khachatryan,1 K.R. Matevosyan,2 S.Y. Frolova,1 A.V. Kolotaev,1 A.N. Balaev,1

M.A. Baryshnikova3 and V.N. Osipov1

1FSUE «IREA», 107076, Russia, Moscow, Bogorodsky shaft, 3

2D.I. Mendeleev University of Chemical Technology of Russia, Moscow, Miusskaya sq. 9 3FSSI "Russian Cancer Research Center named. NN Blokhin", Moscow, Kashirskoye d. 23


The recent interest in the derivatives of thiazolidine-4-carboxylic acid is due to the fact that some

of the substances are as urease inhibitors [1], as anticancer agents [2], influenza neuraminidase

inhibitors [3]. 2-ariltiazolidin-4-carboxylic acids demonstrated a significant antiproliferative activity

and selectivity for tumor cells [4, 5]. In this regard, we synthesized a series of analogs of such acids

and studied their inhibitory activity depending on the nature of the introduced substituent.

Condensation reaction of L-cysteine hydrochloride monohydrate with the corresponding

aldehyde was used to construct a pharmacophore center – thiazolidine cycle [5]:

O

O

Cl

RH / K2CO3

O

O

R

R =S

NSH

NH

OH

ClF

; ; ;

OH

O ;OH

FCl

F

OH

NH

NO2N

NH

N

O2N

O

O

CH2O / HCl

-KCl O

R

NH

S

OH

O

SHNH2

OH

O

N NH

O

N NHF;

;

; ;

; ;

;

;

Starting 4-methoxy aromatic aldehydes were prepared by following the above procedure

according to [6]. Prepared compounds were tested for cytotoxic activity in vitro. Some compounds

were active against human tumor cell lines: Jurkat, A549, MCF-7, PC-3, HCT-116.

This work was financially supported by the Ministry of Education and Science of the Russian Federation (grant agreement № 14.576.21.0044 from 05.08.14, the FTP "Research and development on priority directions of scientific technological complex of Russia for 2014-2020", a unique identifier RFMEFI57614X0044).

References [1] K. Khan, Z-Ullah, M. Lodhi, M. et al., Molecular Diversity 2006, 10, 223-231 [2] Y.Lu, Zh.Wang, C-M.Li, et al., Bioorganic and Medicinal. Chemistry. 2010, 18, 477-495 [3] Yu Liu, Fanbo Jing, Yingying Xu, et al., Bioorg. Bioorganic and Medicinal. Chemistry. 2011, 19, 2342 -2348 [4] D.D.Miller, L. Wei, J.Chen, et al., US patent 2014/0213623 A1 [5] Y.Mi Ha, Y.J.Park, J.Y.Lee, et al., Biochimie.94. P. 533-540. [6] D.S. Khachatryan, A.L. Razinov, A.V. Kolotaev, et al., Russian. Chemical. Bulletin., 2015. (2), 395 (russ).


197

Innovative Technologies in the Creation of Plant Protection Products

S.S. Khalikov,1 N.G. Vlasenko,2 O.I. Teplyakova2 1Federal State Institution of Science Nesmeyanov Institute of Organoelement Compounds Russian

Academy of Sciences; 28, Vavilov str., Moscow, Russia, , 2 Federal State Budgetary Scientific Institution “Siberian Research Institute of Soil Management

and Chemicalization of Agriculture; Krasnoobsk, Novosibirsk region, Russia


Modern agricultural production couldn't be imagined without efficient and effective use of crop protection chemicals (fungicides, herbicides, insecticides, etc.). Fungicides, in particular, from the class of triazoles tebuconazole, has a wide range of systemic effects, namely, along with protective, treating and eradicating properties, preparation has and growth-regulatory action that may go into retardant under adverse conditions (lack of moisture, soil moisture, too deep placement of seeds, their low germination energy and germination). As a result, tebuconazole may cause some damage to crops of cultivated plants [1].

To eliminate these negative effects of fungicide resistance inductors are used to crop phytotoxicants - natural stimulants that stimulate the adaptive capacity of cultivated plants. The use of natural stimulants in a mixture with the fungicide seems preferable not only for economic reasons (increasing grain yield), but also the biological effects (reduced toxicity of the fungicide action on crop plants and rizosphere layer of soil.

The present work is devoted to the creation of multi-component compositions comprising in its composition fungicides (tebuconazole) and natural antidepressants (polysaccharides). To accomplish this, use the original solid state technology mechano-chemical modification of the properties of tebuconazole due to the synthesis of supramolecular complexes such as "Guest (tebuconazole) - Host (polysaccharide)" [2].

The synthesized complexes of tebuconazole with several polymers (Na-CMC, hydroxyethyl starch, PVP, arabinogalactan et al.) have high biological activity:

complexes positive effect on the germination of seeds soft spring wheat "Novosibirskaya 29"; effectively inhibit the growth of pathogenic microorganisms and therefore treatment with this

composition, gave 100% normally developed seedlings, whereas in the control (no drug treatment) and standard (Raxil SC 60) was observed only 73.4% and 91.2% of healthy grains respectively.

positively influenced the growth process, namely, the seed treatment composition of the proposed increase in the number of roots (+ 4.7%), their length (+ 32%) and air-dried biomass (+ 23.9%);

showed a high level of health-improving effect on the root system of spring wheat grown on soil with a high density of spores Bipolaris sorokiniana;

was effective in the field experiment for destruction of plant pathogens common root rot of wheat.

Continuation of our research will enable a better understanding of the processes occurring in the processing plant compositions, as well as mechanisms to protect the crop from the negative effects of fungicide residues in the soil. The financial support of RFBR (grants №15-29-05835) is gratefully acknowledged.

References: [1]. Vlasenko N.G., Teplyakova O.I. Effect of plant growth regulators on the formation of grain quality of middle-late

soft spring wheat cultivars.– Agricult. Chem., 2012, №1, P.56-64. [2]. Khalikov S.S., Dushkin A.V., Khalikov M.S., Guskov S.A., Davletov R.D., Sapozhnikov Yu. E., Kolbin A.M.

Fungicide composition and method for its preparation. Pat. RF N2469536 (2012).


198

Complex Environmentally Friendly Products for the Protection of Potatoes

S.S. Khalikov,1 A.A. Malyuga,2 N.S. Chulikova2

1Federal State Institution of Science Nesmeyanov Institute of Organoelement Compounds Russian Academy of Sciences; 28, Vavilov str., Moscow, Russia

2 Federal State Budgetary Scientific Institution “Siberian Research Institute of Soil Management and Chemicalization of Agriculture; Krasnoobsk, Novosibirsk region, Russia.

E-mail: [email protected] [email protected]

World potato crop losses from fungal, viral, bacterial, and many other types of diseases account

for about 90 million tons or 17% of croppage. To control the level of fungal, bacterial, and other

infections in crops of potatoes and the period of storage after harvest must be rational use of

chemical and biological methods of protection [1]. And so, should be considered highly relevant

research on the development of efficient, environmentally friendly multi-purpose protection of

potato pests and diseases during the growing season and period of storage [2].

We propose a method of producing such a promising crop protection product in the form of

multi-component compositions based on the use of mechanochemistry methods, namely, by

machining the joint acting formulations known substances with water-soluble polymers [3]. As

preparations were selected tebuconazole carbendazim, thiuram and others. The auxiliary substances

for modifying the properties of preparations were used carboxymethyl cellulose derivatives,

arabinogalactan, hydroxyethyl starch, PVP et al. These compositions thus have a high biological

activity, namely:

- developed by us on the basis of Carbendazim preparations showed high effectiveness against

dry rot in the period of storage and black scab (Rhizoctonia solani) during the vegetation period;

- compositions reduced weight percentage of tubers with dry rot in 3,0-14,5 times compared to

the control. As compared with the standard drug (Colfugo-Super, SC) biological efficiency was

higher on average by 25-26%.

- preparations had a positive effects on the biometric characteristics of plants and crop yields.

The highest productivity was observed in potato plants by using a silica compositions Carbendazim

and sodium carboxymethylcellulose. Thus potato productivity increased by 35% when the tubers

were treated in the autumn and 57% - in the spring.

The financial support of RFBR (grants №15-29-05792) is gratefully acknowledged.

References: [1]. Pshechenkov K.A., Zeyruk V.N., Elansky S.N., Maltsev S.V. Storage technology of potatoes.- Moscow:”Potato”,

2007, 192 p. [2]. Melnikov N.N., Novozhilov K.V., Belan S.R. Directory. Pesticides and plant growth regulators.- Moscow:

“Chemistry”, 1995, 384 p. [3]. Khalikov S.S., Dushkin A.V., Khalikov M.S., Meteleva E.S., Evseenko V.I., Buranbaev V.S., Fazlaev R.G.,

Galimova V.Z., Galiulina A.M. Mechanochemical modification of the properties of anthelmintic drugs. - Chemistry for Sustainable Development, 2011. T.19, №6, S.705-710.


199

Regioselective Au(Ш)-Catalyzed Cycloisomerization of Propargylaminomethyl-

or Propargyloxymethyl 15,16-Disubstituted Methyllambertianate

Yu.V. Kharitonov, E.E. Shults

N. N. Vorozhtzov Institute of Organic Chemistry, Siberian Branch of the Russian Academy of

Sciences, 630090, Russia, Novosibirsk, pr. Lavrentieva 9


Recently we have found that 16-propargylaminomethyl- or 16-propargyloxymethyl- substituted

furanolabdanoids under the action of AuCl3 in acetonitrile underwent cycloizomerization with the

formation of 7-hydroxyisoindolines or 7-hydroxydihydroisobenzofurans substituted by a terpenoid

fragment [1].

Herein we have synthesized dialkyne substituted methyl 15,16-epoxy-8(9),13(16),14-

labdatrienoates 1-4 and found that Au(Ш)-catalyzed cycloisomerizations proceeds selectively with

the formation of the corresponding 6-propargylaminomethyl- or 6-propargyloxymethyl-7-hydroxy-

1,3-dihydroisobenzofurans 5, 6 and 6-propargylaminomethyl- or 6-propargyloxymethyl-7-hydroxy-

isoindolines 7, 8.

The synthetic route to dialkynyl derivatives 1-4 from methyllambertianate 9 through the

formation of 16-propargyloxymethyl- or 16-[N-(tosyl)propargylaminomethyl]-substituted

furanolabdanoids 10, 11 are listed in the Scheme.

This work was supported by RFFI (grant 15-03-06546) References [1] Yu. V. Kharitonov, E.E. Shults, M.M. Shakirov, I.Y. Bagryanskaya, G.A. Tolstikov, Russian Journal of Organic Chemistry, 2012, V. 48, N. 6, 1081–1089


200

Modified Fluorine-containing Indazolones: Synthesis and Anticancer Activity

T.S. Khlebnicova,1 Yu.A. Piven,1 F.A. Lakhvich,1 M.B. Golubeva,1

B.B. Kuzmitsky,1 M. Turks,2 V. Kumpins2

1Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, Acad. Kuprevicha str. 5/2, 220141 Minsk, Belarus

2Faculty of Material Science and Applied Chemistry, Riga Technical University, P. Valdena Str. 3, Riga, LV-1048, Latvia


Indazole and indazolone derivatives are attractive structural scaffolds because of their various

biological activities such as anti-inflammatory, antibacterial, and anticancer activities. In particular,

tetrahydroindazolone was recently recognized as an important pharmacophore because of its

excellent biological activity as a potent inhibitor of heat-shock protein 90 (HSP90) [1]. We have

synthesized 3-polyfluoroalkyl-6,7-dihydro-1H-indazol-4(5H)-ones (I) [2] which can be used as

starting compounds for the preparation of a number of biologically active polyfluoroalkyl-

substituted heterocyclic systems.

NN

RO

NN

RNHO

NN

RNR1O

NN

NH2R2

O

X

X

X

X

NN

ROO

NN

ROHON

III R1=C2H5,IV R1=3F-C6H4CO

II

I

VII

V VI

X X

R = CF3, C2F5, C3F7R1 = C2H5, 3F-C6H4COR2 = CF3, C2F5X = H, F By chemical transformations of 3-polyfluoroalkyl-6,7-dihydro-1H-indazol-4(5H)-ones (I) we

synthesized (E)-oximes (II), O-alkyl- and O-acylderivatives of the oximes (III, IV), 6,7-dihydro-

1(H)-indazole-4,5-diones (V), monooximes of the indazole-4,5-diones (VI) and 4-amino-1(H)-

indazoles (VII). The cytotoxicity of prepared compounds (II-VI) against SK-Hep-1, A549, MCF-7,

M-Hela 11, PA-1, Caco-2 tumor cell lines was evaluated by MTT assay. Some of the investigated

compounds showed a strong inhibition against SK-Hep-1 cell line (84-97%).

The work was financially supported by grant GKNT-BRFFR X14LAT-002 References [1] S. Shandrarlapaty [et al.]. Clin. Cancer Res. 2011,14, 240-248 [2] T.S. Khlebnikova, V.G. Isakova, A.V. Baranovskii; F.A. Lakhvich. Russ. J. Gen. Chem. 2008, 78, 1954-1963


201

New Biological Active Selenium-Containing Antioxidants Based on Alkylated

Phenols and Catechols

S. Kholshin,1 S. Yagunov,1 V. Cheblukova,1 N. Kandalintseva1,2 and A. Prosenko1,2

1Novosibirsk state pedagogical university

630126, Russia, Novosibirsk, Viluiskaya str, 28 2Novosibirsk institute of antioxidants

630091, Russia, Novosibirsk, Krasnyi prospekt str, 54а


Glutathione peroxidase (GPx), a selenoenzyme, plays a key role in protecting the organism from

oxidative damage by catalyzing the reduction of harmful hydroperoxides with thiol cofactors [1].

Moreover shown that wide range of selenorganic compounds mimics the catalytic activity of GPx

both in vitro and in vivo [2]. On the other hand, alkylated phenols derivatives are frequently used in

the therapy or prevention of wide range of oxidative stress-derived diseases [3, 4].

In this work, we are present new synthetic selenium-containing derivatives of alkylated phenols

and catechols 1-7, like efficient antioxidants with low toxicity and hypoglycemic, anti-

inflammatory and chondroprotective agents.

R1 R2

OH

Se

R1 R2

OH

Se Se

R1

OH

R2 R1

OH

R1R1

OH

R2

SeSO3Na

R1

OH

R2

SeAlk

R1

OH

R2

Se(CH2)nCOOH(Na)

OH

R1 R2

SeC12H25

R1, R2 = H, Me, t-Bu; Alk = n-Bu, C12H25; n = 1,2

1 2 3

5 6 7

R1

OH

OH

SeAlk 4

References [1] J. Köhrl, R. Brigelius-Flohé, A. Böck, R. Gärtner, O. Meyer, L. Flohé, Biol. Chem. 2000, 381, 849-864 [2] G. Mugesh, W.W. du Mont, H. Sies, Chem. Rev. 2001, 101, 2125-2179 [3] M. D. Mashkovskyi, Lekarstvennye sredstva, Novaya volna, Moscow, 2011, 1216 p. [4] E. B. Men`shikova, N. K. Zenkov, V. Z. Lankin, I. A. Bondar`, V. A. Trufanin, Okislitelny stress: Patologicheskye sostoyanya i zabolevanya, ARTA, Novosibirsk, 2008, 284 p.


202

Antibacterial Property of 2-Aminoethylphosphonates

N. Khusainova and I. Galkina

Kazan Federal University, A.M.Butlerov Chemistry Institute, 420008, Russia, Kazan, Kremlevskaya str., 18


In recent years 2-aminoalkylphosphonates have attracted much attention as compounds with a

broad spectrum of biological activities. With the aim to extend the number of these compounds we

have obtained the series of 2-aminoethylphosphonates on the basis of the reactions of

vinylphosphonates with secondary aliphatic amines. Then we have investigated a sensitiveness of

different kinds pathogenic microorganisms towards synthesized 2-aminoethylphosphonates:

It have been shoved the 100% ruin of the colony of Esherichia coli for compounds (1- 4), and

the 100% ruin of Pseudomonas aeruginosa (compound 1-3) have been established. It have been

showed also the high antibacterial activity (100%) of the compounds (1 -4) against Staphylococcus

aureus, and high sensitiveness of the Esherichia coli, Proteus, Pseudomonas aeruginosa,

Staphylococcus aureus and Candida albicans to the solution of hydroxiaminophosphonate

in DMSO in cons. 1-0.1 %.

This results show that the 2-aminoethylphosphonates are perspective enough compounds for the

deep investigation of theirs biological activity.

This work was funded by the subsidy allocated to Kazan Federal University for the project part of the state assignment in the sphere of scientific activities.

(BuO)2P(O)CH2CH2NH(CH2)2OH


203

Synthesis and Inhibition of Mycobacterium tuberculosis by

A-homoaminotriterpenoids

N.I. Medvedeva, E.F. Khusnutdinova, I.E. Smirnova, O.B. Kazakova

Ufa Institute of chemistry of Russian Academy of Sciences, 71, pr. Oktyabrya, 450054 Ufa, Russian Federation


Tuberculosis is the leading cause of mortality among all infectious diseases worldwide and is responsible for two million deaths annually. The main problem of antitubercular teraphy is high resistance development of M. tuberculosis against new drugs. So, searching of new antitubercular substance is actual. In this context, the class of triterpenoids are of particular interest. Known, that triterpenoids and their derivatives possess cytotoxic, antiproliferative, hepatoprotective, antidiabetic activity.

In this case we synthesized several derivatives of olean, ursan and lupan types of triterpenes and investigated its antituberculosis activity (scheme).

Antitubercular screening showed that compounds 6, 10, 11, 14, 17-19, 22, 24, 25 were identified

as a potent inhibitors against M. tuberculosis. And derivative 9 was the most active with MIC value 1.1 mg/ml. The work was supported financically by the Russian Foundation for Basic Research (project No. 15-03-04058 А) and grant for young scientists of Bashkortostan Republic (No. 28, 13.03.2015).


204

Cycloaddition Reaction of 3-substituted Benzo[b]furans in Synthesis of

Amaryllidaceae alkaloids and Their Derivatives

A.S. Kilmetiev, E.E. Shults

Laboratory of Medicinal Chemistry, Novosibirsk Institute of Organic Chemistry by N.N. Vorozhtsov of the Siberian Branch of Russian Academy of Science, 630090, Russian Federation, Novosibirsk,

Lavrentiev avenue, 9


Phenolic compounds are known as useful intermediates for synthesis of valuable natural

products, including the substances with dibenzofuran core, e. g. galantamine and related

Amaryllidaceae alkaloids [1]. Most synthetic approaches to galantamine are based on two reactions:

phenolic oxidative coupling in the presence of metal oxidants and the intramolecular Heck reaction

of halogenated phenols [1]. We present a novel methodology to synthesize galantamine and its

derivatives. Key step of this total synthesis is Diels-Alder reaction between 3-substitued

benzo[b]furans 2 and 3-trimethylsiloxy-1,3-butadienes 3. This reaction is known to proceed

regiospecifically and allow to obtain cycloadducts 4 with good yield [2, 3]. Required

benzo[b]furan-3-carboxylates and benzo[b]furan-3-carbaldehydes were synthesized from

appropriate 2-methoxyphenols 1 by known convenient procedures [3, 4, 5].

This work was supported by the Russian Science Foundation (project No 14-13-00822). References [1] J. Marco-Contelles, M. do Carmo Carreiras, C. Rodriguez, M. Villarroya and A. G. Garcia, Chem. Rev. 2006, 106, 116-133 [2] N. Chopin, H. Gerard, C. Rodriguez, I. Chataigner and S. R. Piettre, J. Org. Chem. 2009, 74, 1237-1246 [3] A. S. Kil’met’ev, E. E. Shul’ts, M. M. Shakirov, T. V. Rybalova and G. A. Tolstikov, Rus. J. Org. Chem. 2013, 49, 872-885 [4] M. E. Dudley, M. M. Morshed and M. M. Hossain, Org. Synth. 2009, 86, 172-180 [5] K. Ando, Y. Kawamura, Y. Akai, J. Kunitomo, T. Yokomizo, M. Yamashita, S. Ohta, T. Ohishid and Y. Ohishi, Org. Biomol. Chem. 2008, 6, 296-307


205

Aziridinederivative of Grosheimin

A.S. Kishkentaeva,1 О.А. Nazarova,1 B.А. Abdigalymova,1 Ju.V. Gatilov,2,3

G.A. Atazhanova,1 S.M. Adekenov1

1JSC “International Research and Production Holding “Phytochemistry”100009, Republic of Kazakhstan, Karaganda, fax: 8 (7212) 43 31 27

2N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry Of the Siberian Branch of Russian Academy of Science, 630090, Novosibirsk, 9 Lavrentev avenue

3Novosibirsk State University,630090, Novosibirsk, 2 Pirogov street E-mail: [email protected], [email protected]

Grosheimin (1) is sesquiterpene lactone of guaine group, represents itself as recyclable chemical

compound, which are a lot in Chartolepis species. For the synthesis of a new derivative of

grosheimin (1) a reaction with sodium nitrite and ammonium chloride in aqueous - ethyl alcohol

was conducted by hard conditions, revealed a new derivative (2) with melting point of 176-178 °С

and yield 17%.

O

O

+O OH NaN3; NH4Cl

C2H5OH; H2O;H

H

T=60-70oCO

O

O

H

H

OH

NH

(1) (2)

The structure (2) was established on the basis of physical-chemical constants and spectral data

(IR-, UV-, NMR 1Н and 13

С, two-dimensional spectra NMR 1Н-1

Н (COSY, NOESY) and 13С-1Н

(COSY,COLOC), mass spectrometry. The stereochemistry of crystal structure (2) is studied by the

X-ray diffraction method (see picture 1).

Picture 1 – The spatial structure of the molecule (2)

Thus, when conducting grosheimin azidation reaction (1) 3-oxo-8α-hydroxy-1,5,7α,4,6,8β(Н)-

guaia-10(14)-en-13,11-azaspiro-12,6-olide (2) was synthesized.


206

Reliable Immunomodulatory Evaluation for Substances of Natural Origin

E. Kmoníčková,1,2 P. Drašar,3 J. Harmatha,3 M. Kverka,4 Z. Zídek1

1 Dept. of Pharmacology, Institute of Experimental Medicine, Academy of Sciences of the Czech

Republic, CZ-14220, Czech Republic, Prague, Vídeňská 1083 2 Institute of Pharmacology and Toxicology and Biomedical Centre, Faculty of Medicine in Pilsen,

Charles University, CZ-30100, Czech Republic, Pilsen, alej Svobody 76 3 Dept. of Natural Products, Faculty of Food and Biochemical Technology, University of Chemistry

and Technology, CZ-16628, Czech Republic, Prague, Technická 5 4 Institute of Microbiology, Academy of Sciences of the Czech Republic,CZ-14220, Czech Republic,

Prague, Vídeňská 1083 E-mail: [email protected]

In process of pharmacological and toxicological screening, results might be biased by possible

presence of bacterial contaminants in samples of both natural and synthetic origin. It is well established that bacterial macromolecules lipopolysaccharide (LPS), lipoteichoic acid (LTA) and peptidoglycan (PGN) activate immune innate system. Similar to LPS, both LTA and PGN are potent inducers of nitric oxide (NO) and cytokines, and synergize with LPS in this respect. As far as plant-derived substances and xenobiotics may contain bacterial contaminants, a care should be taken to detect and then eliminate them from test samples. LPS can be neutralized with polymyxin B. On the other hand, removal of LTA and PGN has not been available, so far. The aim of our studies was: 1) to develop a convenient protocol for the elimination of these agents, 2) to investigate potential immunostimulatory effects of probiotics, and 3) thapsigargin-related sesquiterpene lactones (SL) isolated from plant sources.

Methods: Detection and elimination of LPS, LTA and PGN, the method of Limulus amebocyte lysate assay was compared to the microfiltration assay. Amicon® Ultra 0.5 mL centrifugal devices and filters with molecular weight cutoffs of 3 kDa and 100 kDa were used for this purpose. The fractions of probiotic strain (Lactobacillus casei probiotic strain DN-114001) were prepared by microfiltration of the original lysate using the 3- to 100-kDa cutoff microfilters. Immunobiological activities (in vitro production of NO, cytokines secretion) were investigated in rat peritoneal macrophages. SL were isolated from Laser trilobum (L.) Borkh. and Thapsia garganica L.

Results: 1) For academic labs we have developed a convenient protocol for the elimination of polymeric substances of bacterial origin. The 100 kDa filters have been found sufficient to completely remove all LPS, LTA and PGN from test samples. 2) Stimulatory activity (NO production, IL-6, TNF-α secretion) of the original lysate (Lactobacillus casei strain DN-114001) and its microfiltrates were observed in peritoneal macrophages. Interestingly, the immunobiological effects of probiotic bacteria are contributed to by substances characterized by the molecular weight of ≤ 3 kDa. 3) Contamination of SL with bacterial components was excluded. In a group of guaianolide type of SL we found trilobolide (and thapsigargin), which induced IFN-γ and IL-2. Targeted trilobolide molecules were newly prepared and also showed the structure-related immunostimulatory properties.

At present, the immunomodulation is recognized as a prospective strategy for the treatment of many autoimmune, inflammatory and cancer diseases. Immunomodulatory potential of our microfiltrates of probiotic lysates and targeted trilobolide molecules are promising for further pre-clinical studies. This work was supported by Czech Science Foundation 303/12/0535 and 303/14-04329S.


207

A New Approach to the Automated Identification of Metabolites in

Multi-vendor Datasets.

D. Hardy,1 V. Lashin,2 R. Kubilius,3 J. Blanz,4 M. Trunzer4 and E. Kolovanov2

1 ACD/Labs, Bracknell, UK 2 ACD/Labs, Moscow, Russia

3 ACD/Labs, Vilnius, Lithuania 4 Novartis Institutes for Biomedical Research, Basel, Switzerland.


A challenge in the study of drug metabolism with the aim of rapid and accurate identification of

metabolites are the variety of software packages. Most instrument vendors offer software to support

the identification of metabolites. However, in multi-vendor laboratories or large companies a tool is

required that avoids big investments in customisations for data evaluation, data storage and

reporting.

Here we present a new approach for the automated identification of metabolites, which allows

data from nearly all the mass spectrometry vendors to be processed, reviewed and databased. The

automated file capture and processing capabilities make it suitable for a high-throughput

environment whilst the biotransformation scientist can still check the results and make any changes

such as modifying a structure assignment. Batch processing allows multiple time point samples to

be processed, the calculation of pharmacokinetic parameters such as area under the curve (AUC) or

multi-sample analysis for e.g. interspecies or interassay metabolism comparison.

A structure based prediction approach is used to identify potential metabolites. The site of

biotransformation is evaluated using MS/MS spectra from data-dependent acquisition by LC/MS.

Mass shifts of fragment ions and the common collision-induced fragmentation rules are applied to

localise the site of biotransformation. In the cases where there is not sufficient evidence to support a

single site of biotransformation, metabolite structures can be represented using the Markush

notation. Unexpected metabolites are also identified using control sample comparison in

combination with the time course of the formation of new components.

The automatically generated metabolic scheme, mass spectra, chromatograms, semi-quantitative

results based on peak area as well as study metadata can be stored in a database for future use and

for reporting of the results including a web-based interface. The software allows to speed up

metabolism studies without compromising the quality of the interpretation. Both, discovery and

development departments could benefit and use the potential to save a huge amount of time and

effort.


208

ACD/Percepta – a Software Platform for ADME/TOX Property Prediction,

Compound Profiling and Lead Optimization

P. Japertas,1 K. Lanevskij,1 A. Sazonovas,1 E. Kolovanov2

1 ACD/Labs, Vilnius, Lithuania 2 ACD/Labs, Moscow, Russia.


One of the most significant challenges facing medicinal chemists in general is transforming an

active molecule into a viable drug. Lead optimization efforts are guided by a combination of

factors, such as potency, ease of synthesis, patentability concerns, specific synthetic constrains of

the interaction with the target, as well as lead’s toxicity and ADME properties. Balancing numerous

inter-related molecular properties often becomes a major issue.

Despite constant advances in the field of in silico modelling and huge number of available

predictive models, computational approaches still fail to meet high initial expectations, as the

majority of existing models are ‘statistics driven’ and have been developed with the only pursuit of

the best possible fit in mind. It is usually not accounted that many biological properties, from

permeation of biological membranes (intestinal absorption, BBB) to protein-ligand interactions, are

to a certain extent governed by basic physicochemical parameters, such as lipophilicity or

ionization. Changing their values may have distinct, even opposite effects on different ADME

properties, and these effects are inter-related. Only the aggregated output of different models makes

it possible to account for numerous effects and identify the physicochemical space regions most

likely occupied by analogs with favorable ADME profiles.

In this work we present a novel software platform – ACD/Percepta – to aid medicinal chemistry

projects by providing reliable prediction modules for a multitude of physicochemical, ADME, and

drug safety-related properties, along with a ‘Chemical Spreadsheet’ workspace that implements

powerful data mining, visualization, compound profiling and ranking capabilities, as well as

ACD/Structure Design Engine for generating libraries of virtual analogs compatible with the

desired biological characteristics.

ACD/Percepta predictors are specifically designed to comply with general physicochemical

trends even for the complex properties involving protein-ligand interactions (P-gp, hERG). This

gives a key advantage of the models being clearly interpretable. Not only they produce an estimate

of the property value for a compound, but also provide a mechanistic insight that has the potential to

guide lead optimization in the desired direction. Also, many models are trainable, enabling the

researcher to adapt them to ‘in-house’ chemical space without model rebuilding. In our opinion, a

rational strategy of lead optimization should aim at achieving balanced physicochemical profiles of

candidate compounds that would translate into the desired ADME properties.


209

Endosorb - New and Effective Tool for Purification of Proteins

Contaminated with LPS

A. Morozov,1 M. Kopitsyna,1 A. Nuzhdina2 and I. Bessonov1

1 Bauman Moscow State Technical University, 105005, Russia, Moscow, 2nd Baumanskaya str. 5.

2 D. Mendeleev University of Chemical Technology of Russia, Moscow, 125047, Russia, Miusskaya sq. 9.


Bacterial endotoxin which is structurally lipopolysaccharide (LPS) was first discovered by

Richard Pfeiffer in 1892 as a stable toxic compound released upon the destruction of bacterial cells.

It causes pyrogenic effect and other inflammatory symptoms in mammals and human. The LPS

content is strictly regulated in injectable drugs because even trace amounts of bacterial endotoxin

can cause life-threatening complications in patients.

Many purification methods have been developed for endotoxin removal, including LPS affinity

interactions, two phase extractions, ultrafiltration, affinity chromatography and anion exchange

chromatography. We developed very simple and extremely effective technique for protein

purification. Beads of Endosorb are placed in protein solution and stirred for 30-60 min at room

temperature. Consequently all amounts of LPS are removed from solution while concentration of

protein remains almost unchanged. The principle of high selectivity of Endosorb to LPS is

electrostatic interaction between negatively charged LPS and positively charged ligand covalently

bonded to macroporous polymeric matrix of Endosorb. Endosorb was tested on 3 different proteins

on the following conditions: protein concentration 0.125 mg/ml, LPS concentration – 30 EU/ml,

Endosorb content – 15 mg/ml. Protein concentration was determined spectrophotometrically and

LPS content was determined using Limulus Amebocyte Lysate (LAL) test. Results are summarized

in table 1.

Table 1. LPS removal from protein solutions using Endosorb.

Protein pI Buffer solution

Initial LPS

content,

EU/ml

LPS content after

purification,

EU/ml

Protein

recovery, %

Cytochrome C 9.6 PBS (0.1M, pH 7.0) 30 <0.3 92

Hemoglobin 6.8 PBS (0.1M, pH 7.0) 30 <0.3 82

BSA 4.7 Tris-borate buffer

(0.1M, pH 8.3) 30 <0.3 98


210

Information-analytical system "IR-EXPERT"

T.A. Kornakova, K.S. Chmutina

N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry SB RAS, 630090, Novosibirsk,

Lavrenteva ave.,9

Databases and computer information systems working with them are widely used by researchers

in the medical industry, pharmacology, ecology, forensic examination and synthetic laboratories.

The chemists use both large universal and a small problem-oriented database. Expanding of the

database is often faced the problem of confidentiality of substances under investigation and new

synthesized compounds.

System “IR-EXPERT” allows you to form peculiar your "spectrum - structure - property"

database and to add to the existing database. The information-analytical system IR- EXPERT [1]

created in the NIOC SB RAS permits solving of the various problems in analytical chemistry, such

as:

• to search for spectral analogs of the compound tested

• to perform taxonomy structures on the base of structure fragments and IR spectra selected

• to create the list of the most probable fragments of the compound tested

• to check the "spectrum-structure" accordance

• to define the spectral features of structure fragment

• to simulate the IR spectrum for a compound with the given structure

• to determine the structural similarity

• to select compounds including and/or excluding reliable structure fragments

• to perform a taxonomy of text information

The structures of chemical compounds in IR-EXPERT are presented both as graphs (relations

"atom" - "bond") and as the exhaustive set of connected non-isomorphic 2-7 node structure

fragments [2]. Such unique representation enables to perform substructure search, to compare sets

of structural fragments with each others, to distribute structural and spectral data by classes, to form

descriptors for the treatment of chemical information.

In each particular case, the result of search or of establishing "spectrum-structure-property"

relationships is determined primarily by the presence of the close spectral and structural analogues

in the database used. It was shown in [3] that the efficiency of a database usage depends on the

wideness of different chemical class representation, as well as on the completeness of a particular

test chemical class presentation.

The analytical procedures of IR-EXPERT can be applied to the databases of various physical

nature [4], to establish the structure of new synthetic compounds for their spectrum and to use in the


211

study of biologically active compounds. IR-EXPERT database includes about 80 thousands records

for chemical compounds, including biochemical, toxic, pharmaceutical, agricultural substances and

other. The database contains about 140 thousands different k- node (k=2-7) structure fragments. [1]. V.N. Piottukh-Peletsky, K.S. Chmutina, M.V. Korolevich, J. Struct. Chem. 44 (2003) 835-84 [2]. V.N. Piottukh-Peletsky, I.K. Korobeinicheva, T.F. Bogdanova, S.G. Molodtsov, B.G. Derendyaev, Anal. Chim.

Acta 409 (2000) 181-195 [3]. B.G. Derendyaev, V.N. Piottukh-Peletsky, T.A. Kornakova, Chemistry for Sustainable Development, 9 (2001) 17-

26 [4]. T.A. Kornakova, T.F. Bogdanova, B.G. Derendyaev, V.N. Piottukh-Peletsky Anal. Chim. Acta 543 (2005) 177-

180


212

Alkaloids of Papaver kuvajevii Schaulo et Sonnikova

M.O. Korotkikh, N.A. Pankrushina, and O.I. Salnikova

N.N. Vorozhtsov Institute of Organic Chemistry, 630090, Russia, Novosibirsk, pr. Lavrentieva, 9


A new poppy species was discovered in 2003 in the Sayano-Shushensk Biosphere Reserve and

described as Papaver kuvajevii Schaulo et Sonnikova [1].

We studied the alkaloid composition of Papaver kuvajevii aerial part. There were identified both

main alkaloids amurensinine, mecambridine and minor α-allocryptopine, amurensine,

tetrahydrocoptisine, norsanguinarine, oxysanguinarine, norhelerythrine, oxoberberine, alpinine

using GC-MS. The structures of amurensinine and mecambridine were confirmed by 1H NMR

spectra.

Amurensinin belongs to isopavines and possesses important pharmacological properties for

nerve system treatment. It can be used against Alzheimer’s disease, Hungtington’s chorea,

amyotrophic lateral sclerosis, and Parkinson’s and Down’s syndromes [2].

Mecambridine has isoquinoline structure and inhibits individual CYP enzymes. In [3] was

indicated high-affinity interaction of several isoquinoline alkaloids, particularly mecambridine, to

specific CYP enzymes.

We continue the study to improve of alkaloid separation scheme.

References [1] Shaulo D., Sonnikova A. Turczaninowia. 2003, 6 (4), 5-6.

[2] Carrillo L., Badia D., Dominguez E., Vicario Jose L., Tellitu Imanol A. J. Org. Chem. 1997, 62

(20), 6716–6721.

[3] Kaisa A., Achim Meyerb, Lenka Jerabkovaa, Laura E. Korhonena, Minna Rahnastoa, Risto

O. Juvonena, Peter Immingb, Hannu Raunio Salminena. Achim Phytomedicine. 2011, 18 (6), 533 –

538.


213

Synthesis of Novel Coumarin-substituted Fluoroquinolones

S. Kotovskaya,1,2 N. Mochulskaya,1 Z. Baskakova,1 E. Nosova1 and V. Charushin1,2

1 Department of Organic Chemistry, Ural Federal University named after the first President

of Russia B.N. Yeltsin, 620002, Russia, Ekaterinburg, Mira St., 19 2 Postovsky Institute of Organic Synthesis, the Ural Branch of the Russian Academy of Sciences,

620041, Russia, Ekaterinburg, S. Kovalevskaya/Academicheskaya St. 22/20


Fluoroquiolones represent a very important class of highly active antibacterial drugs, which are

widely used in chemotherapy. Many compounds of this family are also capable of exhibiting other

types of activity, including antitubercular, antiviral and antineoplastic ones. Derivatives of

coumarins (2H-chromen-2-one) proved to possess antispasmodic, photosensitizing, antineoplastic,

anticoagulant and other types of pharmacological activity. Therefore, incorporation of coumarin

fragments into the structure of fluoroquinolones is of a great interest for medicinal chemistry.

A new synthetic approach to cyanomethyl fluoroquinolones 3a,b, based on nucleophilic

substitution of fluorine atom in quinolones 1a,b by action of cyanoacetates, followed by acidic

hydrolysis of 2a,b and decarboxylation, has been suggested. A series of new (2H-chromen-2-one)-

substituted bi- and tricyclic fluoroquinolones 4a,b have been obtained by reacting 3а,b with

derivatives of salicylic aldehyde.

N

F

O

R1

O

O

F

R2

N

F

O

R1

O

O

O

N

O

R

R2

N

F

O

R1

OH

O

N

R2

N

F

O

R1

OH

O

O OR

2

OH

OR

3

O

N

O

R

O

R3

H2SO4conc/AcOH/H2O (0.5:10:10)

DMSO, K2CO3

R= Et, tBu

R1=Et, R2= H (a); R1,R2= (b)

1a,b 2a,b

3a,b 4a,b

R3= H, Br, Cl, OEt, tBu

The evidence for the structures of 2-4 is provided by their spectral data, including the data of the 1Н and 19F NMR-spectroscopy.

The research was supported by the Ministry of Education of Russian Federation (Project 2458).


214

4-Quinolones: the Old Friends in a New Role

A. Boteva, I. Fefilova, A. Trefilova, S. Solodnikov, G. Ljushina, V. Maslova, O. Krasnykh

Research and Educational Center of Applied Chemical and Biological Research at the Perm

National Research Polytechnic University, 614990, Russia, Perm, Komsomolsky Av. 29. E-mail: [email protected]

Scaffolds containing the 4-quinolone (4-Q) fragment are regarded as privileged. In addition to

typical for 4-Q interactions which include H-bonding and various π-interactions, functionalized

substituents can provide extra binding points and significantly facilitate diverse and multiple

interactions with biological targets of 4-Q. Such multifunctionality leads to noticeable diversity of

pharmacological effects of 4-quinolones. Besides the well known antimicrobial activity,

representatives of the 4-Q class possess anti-viral and anti-cancer properties, inhibit PDE-5,

modulate CR type 2 and inhibit JNKs [1-3].

We have chosen 2,3-diacyl-4-quinolone fragment as a starting point for a “structure driven” drug

discovery project aiming to a new effective and non-toxic agent to treat inflammatory conditions.

Beginning with the known methodology based on the decarbonylation of substituted pyrrolodiones

we have synthesized a series of 2,3-diacyl-4-quinolones which were used for further modifications

leading to several types of heterocyclic systems containing a 4-Q fragment [4,5].

A selected set of synthesized compounds was evaluated for ability to pass through the biological

membranes, as well as for their potency as anti-nociceptive and anti-inflammatory agents. In

summary, a lead series of compounds is identified. The compounds are characterized by an

acceptable “drug-like” profile, anti-inflammatory and analgesic effects of some representatives is

almost equal or superior to the one of diclofenac, and LD50 values are higher than 1000 mg/kg.

Acknowledgements: This work was partially funded by the Program «The PNRPU - EPFL collaboration on Metabolism and Diabetes» supported by «Neva» foundation. References [1] M. Welsch, S. Snyder, and B. Stockwell, Curr Opin Chem Biol. 2010, 14(3), 347–361 [2] C. Mugnaini, S. Pasquini , F. Corelli F, Curr Med Chem. 2009, 16(14), 1746-67. [3] A. Boteva, O. Krasnykh, I. Fefilova, E. Babushkina and P. Slepukhin, Russ.Chem.Bull., 2014, 63, 3, 731-738[4] A. Boteva, O. Krasnykh, S. Solodnikov, RU 2381229 C2 20100210, 2010


215

Complex Study of Phycobiliproteins and Sulfated Polysaccharides from Sterile

and Reproductive Forms of Red Alga Ahnfeltiopsis flabelliformis

A. Kravchenko,1 S. Anastyuk,1 V. Glazunov,1 V. Isakov,1 W. Helbert,2 I. Yermak1

1G.B. Elyakov Pacific Institute of Bioorganic Chemistry, 690022, Russian Federation, Vladivostok, Prospect 100 let Vladivostoku, 159

2Centre de recherches sur les macromolécules végétales, BP 53, 38041, France, Grenoble cedex, 9


Cell walls of red algae are mostly composed of sulfated galactans such as agar or carrageenan which are not found in land plants, and pigments such as phycobiliproteins (PBPs). PBPs (phycoerythrin (PE), phycocyanin (PC), allophycocyanin (APC)) have bright color and very intense fluorescence, so they are used in immunofluorescence diagnostics, cytometry, cosmetic and food industries. In addition, these pigments exhibit a variety of biological activities. The importance of polysaccharides in pharmaceutical development in last years has been shown. A comprehensive study of PBPs and polysaccharides from red algae is one of the important problems today.

In our research PBPs and polysaccharides were extracted sequentially from seaweed A.

flabelliformis which forms extensive populations in the Russian Far Eastern seas. PBPs were extracted by two methods with 0,1 M phosphate buffer and 1,5% NaNO3 solution. The most effective method for PBPs isolation is its extraction with 1,5% NaNO3. It was shown PE is predominant phycobiliprotein. After pigments isolation the polysaccharides were extracted from algae with hot water. Sequential extraction of PBPs and polysaccharides from A. flabelliformis makes to increase the yield of polysaccharides. It was shown that polysaccharides represent carrageenans. It is known that κ-, ι- and λ-carrageenans are regular polysaccharides which are widely used in various fields of industry and medicine. Natural polysaccharides often have hybrid structure containing disaccharide units of several types of carrageenans. We studied the structure of the polysaccharides from the sterile and reproductive forms of A. flabelliformis. Polysaccharides under study contain gelling fractions predominately, in which galactose and 3,6-anhydrogalactose are main monosaccharides. According to IR and NMR spectroscopy, and mass spectrometry, gelling polysaccharide from sterile algal form has a hybrid κ/β structure with κ- and β-units ratio 3:1. The polymer chain of this polysaccharide is built from κ-carrabiose, κ-carratetraose, κ-carrahexaose, β-carrabiose, β-carratetraose, hybrid tetra- and hexa-oligosaccharides. A gelling polysaccharide from reproductive form of A. flabelliformis represents ι/κ-carrageenan with a predominance of ι-type units, contains β-carrageenan and minor amounts of ν-carrageenan. According to the enzymatic hydrolysis of the polysaccharide with ι-carrageenase from Microbulbifer thermotolerans and analysis of the fractions by ESI MS/MS, main components of ι/κ-carrageenan is ι-carrabiose, ι-carratetraose and hybrid tetra- and hexasaccharides. According ESI MS/MS and methylation analysis, xylose is substituent of one of the hydroxyl groups of the galactose in carrageenans. The anticoagulation activity of carrageenans has been shown.


216

Simulation of Bioavailability and Pharmacokinetics in Preclinical Studies

I. Yakovlev,1 V. Yuskovets,1 I. Narkevitch,1 A. Radilov2 and D. Krivorotov 2

1Organic chemistry department, St. Petersburg State Chemical-Pharmaceutical Academy,

Professora Popova str., 14-A, Saint Petersburg, 197376, Russian Federation 2Chemical modeling laboratory, Research Institute of Hygiene, Occupational Pathology and



4-[(3-Oxo-3-ethoxypropanoyl)amino]benzoic acid has been developed in St. Petersburg State

Chemical-Pharmaceutical Academy. This pharmacological substance (PhS) has antioxidant and

anti-ischemic effect on myocardium. The preclinical pharmacokinetic studies of this substance have

been performed within program «Pharma-2020» at the RIHOPHE.

Nowadays prediction of pharmacokinetic profile in drug discovery has become very useful. This

method optimizes the labor costs and contributes to the humanization of research. Thus, comparison

of the pharmacokinetics data and absolute bioavailability in in vivo and in silico experiments is of

practical interest for many researchers engaged in drug discovery.

In order to predict the pharmacokinetic parameters ACD/Percepta (Canada, Lithuania, Russia)

software was used. While calculating this computer program considers solubility, stability in GIT

media, absorption, metabolism, active transport, P-gp specificity and a dose of a substance. The

calculated data were compared with those obtained from intravenous and oral administration of

equivalent doses of PhS to rabbits.

OH

O

NH

O

O

O

CH3

Results of preclinical pharmacokinetic studies

Results of pharmacokinetic parameters modeling

Experimental model Rabbit (6 animals) ACD/Percepta PhS 26 mg/kg 7,8 mg/kg (550 mg per a 70 kg human) Tmax, min 56±20 min 152 min Cmax 7.5±1.6 µg/mL 7,9 µg/mL

AUC0-24 iv - 32 µg*h/mL or - 22 µg*h/mL

iv - 63 µg*h/mL or - 44 µg*h/mL

Bioavailability F 73% 70%

Pharmacokinetic curves of blood plazma levels of PhS

Calculated and found bioavailability of the substance is almost the same. The data obtained show

the oral bioavailability of PhS to be less than 100%, due to, inter alia, the active metabolism of the

substance. The results demonstrate the use of computer modeling to optimize preclinical studies.


217

Synthesis of Mimetics of Known GPCR Modulators with Peripheral Action

V. Kuznetsov,1 D. Krivorotov 1




Modulators of GPCRs based on quaternary salts of semi-synthetic alkaloids, e.g. Relistor (2011),

are used in the treatment of drug-induced intestinal disorders due to their peripheral action towards

the nervous system of mammals. Thus, it makes it challenging to conduct target-oriented synthesis

of new quaternary ammonium salts of related scaffolds.

We have synthesized ammonium salt 1 from papaverine biogenetically related to Papaver

somniferum alkaloids. It has a similar structure to Relistor and thus can have similar biological

activity.

N+ CH3

O

CH3

O CH3OCH3 O

CH3

Br-

N+

CH3

OCH3

O

CH3

O

CH3

OCH3

Br-

N CH3

OCH3

O

CH3

O

CH3

OCH3

N

OCH3

O

CH3

O

CH3

OCH3

Papaverine 1 1

O

N+

OH

OOH

H CH3

Br-

Relistor

Using benzhydrylpiperazine as in the synthesis of DOR agonist BW373U86, we have

synthesized analogous quaternary salts. Changing the order of introduction of substituents allows

obtaining salts 2 and 3 with various anions. These salts are crucially different in terms of solubility.

N

N+

CH3

Cl

N N

Cl

N N CH3

Cl

N

NH

Cl

CH3I

X-

2 X-=I-

3 X-=Br-

Br

CH3I

N

NCH2

CH3

CH3

OH

O N

CH3

CH3

BW373U86

Br

Thus, a set of chemical compounds, mimetics of GPCR modulators with peripheral action, has

been synthesized. The salts obtained are potentially active with respect to the peripheral nervous

system and optimized regarding a pharmacologically important solubility number.


218

The Search for Novel Adaptogens Among Thiourea Amino Acid Derivatives

S. Ramsh,1 V. Kuznetsov,2 S. Petunov,2 D. Bobkov,2 A. Radilov2 and D. Krivorotov 2

1Dept. of Chemical Technology of Organic Colors and Phototropic Сompounds, Saint Petersburg State Technology Institute (Technical University), Moskovsky Pr., 26, Saint Petersburg, 190013,

Russian Federation 2Chemical modeling laboratory, Research Institute of Hygiene, Occupational Pathology and



Prolonged physical activity is accompanied by hypoxia progression, violation of energy

production in the mitochondria and depletion of adaptive reserves of the body. The lack of synthetic

adaptogens available (Bemithylum et al.) stimulates the search for the drugs that have directed

actions [1].

We have synthesized new cyclic thioureas (type 1 and 2) by means of subsequent

aminomethylation by amino acids and alkylation.

(CH2)nCOOHNH

NN

SR

+

HI-

NH2 NH2

SNH2(CH2)nCOOH

NH

NNH

S (CH2)nCOOHRHal

CH2O

type 1n=1, 2, 3 RHal = MeI, EtI type 2 Their influence on the performance of the animals after exhausting loads in test “Swimming with

maximum loads” has been studied. The effectiveness of performance recovery has been determined

by calculating a recovery index (RI) and its comparison with a control group index. This index is a

ratio of the duration of execution of swimming load in 40 min after complete exhaustion to the

duration of the 1st swimming. Compound Recovery index RI change versus control (water)

Acute experiment Subacute experiment RS-4, type 1 0,72 0.96 – RS-5, type 1 0,73 1.15 0.92 RS-6, type 1 0,99 1.15 1.19 RS-7, type 2 0,83 1.25 1.29 RS-8, type 2 0,73 1.10 – RS-9, type 2 0,72 0.94 – Bemithylum 0,97 1.30 1.31

The most active pharmaceuticals RS-6 and RS-7 may serve as a basis for the development of

novel effective Russian drugs with adaptogenic activity.

References [1] E. Studentsov, S. Ramsh, N. Kazurova, O. Neporozhneva, A. Garabadzhiu, T. Kochina, M. Voronkov, V. Kuznetsov, D. Krivorotov, Obz. Klin. Farmakol. Lek. Ter. 2013, 11, 3-43.


219

Scots Pine Phloem Reaction to the Anthropogenic and Biotic Stress

T.P.Kukina,1 G.G. Polyakova,2 T.S. Frolova,1, 3 O.I. Sal’nikova1

1Novosibirsk Institute of Organic Chemistry, Siberian Branch of the Russian Academy of Sciences 9 prosp. Acad. Lavrent’ev, 630090 Novosibirsk, Russian Federation

2Sukachev Institute of Forest, Siberian Branch of the Russian Academy of Sciences, Akademgorodok, 660036 Krasnoyarsk, Russian Federation

3Novosibirsk State University, 2 Pirogov Street, 630090, Novosibirsk, Russian Federation E-mail: [email protected]

The interactions between trees and pathogenic fungi are investigated actively since the 1960s in

relation to the usage of the method of conifer tree stem inoculation with the mycelium of

ophiostomatoid fungi spread by insects and causing wood blueness. For inoculation, a small hole is

cut in the living stem inner bark to the sapwood, and fungal mycelium is placed into it. Studies of

the stem phloem response to the inoculation of living trees with pathogenic fungi are complicated

by the risks of fungal epiphytotic diseases. The application of fungal extracts instead of the living

mycelium excludes the risk of decease spreading in the forest. Our previous results and published

data confirm the validity of mycelial extract as an alternative to viable mycelium in studies of

interactions between conifers and ophiostomatoid fungi under natural conditions. The contents of

lignin, bound PA, and resins were found to increase, while those of free PA, low-molecular-weight

carbohydrates, and the cellulose components decreased.

The aim of the present study was to elucidate the role of secondary metabolites in the

enhancement of phloem necrosis induced by compounds extracted from ophiostomatoid fungi as

compared with wounding-induced necroses. To this end, the content of lipophilic constituents—

aliphatic and terpenoic compounds were analyzed in pine trees in the zone of phloem response to

mechanical injury (control treatment 1) and to injury combined with inoculation of high-molecular-

weight compounds extracted from mycelia of the ophiostomatoid fungus Ceratocystis laricicola

(treatment 2). In the experiment with Scots pine trees, significant changes were detected in all

measured biochemical characteristics of the bark, irrespective of the type of injury (treatments 1 and

2). The chemical compositions of lipophilic fractions from Scots pine phloem were determined by

gas chromatography-mass spectrometry (GC-MS). As a result, more than 50 acid and neutral

constituents were identified. Some components such as resin acids demonstrated multifold

increasing in comparison with intact phloem whereas aliphatic acid content decreased. When the

mycelial extract was introduced into holes in the treatment 2, biochemical changes were more

pronounced than in the control treatment 1. This effect is especially impressive in the case of

oxygenated resin acids and dehydroabietic acid. The increase of level is from 5-10 (treatment 1) to

7-15 (treatment 2).


220

Human Recombinant Factor VII For Replacement Therapy

P. Kustov, A. Orlov and J. Krasotkina

Biochemical Production Department, CJSC ”GENERIUM”, 6001125, Russia, Volginsky, 17


Coagulation factor VII (FVII) is a key component of normal plasma hemostasis. This single–

chain 50-kDa protein is synthesized in liver and secreted in blood vessel. Deficiency of FVII, either

inherited or acquired, results in bleeding disorders of different severity. Management of associated

hemorrhages primarily consists of FVII replacement therapy to stop bleeding. Nowadays treatment

of FVII-deficient patients includes the use of fresh frozen plasma, prothrombin complex

concentrates, plasma derived FVII concentrates and the use of recombinant FVIIa (rFVIIa). Yet,

there is no preparation of recombinant FVII that would provide virus safety as compared with

plasma derived drugs and low risk of thrombosis as against activated FVIIa. The main problem in

FVII production is its autoactivation during purification process. The use of serine protease

inhibitors, benzamidine, for instance, helps to overcome this problem. All methods, reported to

date, employ serine protease inhibitors up to final stages of FVII purification and, thus, prevent

usage of such FVII product as a pharmaceutical substance.

Here we report a new method for recombinant FVII production without use of any protease

inhibitor. Method includes protein expression in mammalian cell line followed by anion exchange

and affinity chromatography purification steps. The resulting FVII exhibits more than 95%

homogeneity and coagulation activity of native protein. Comparative pharmacokinetics of rFVII

and rFVIIa in rabbits showed that after single injection plasma levels of FVII exceeded that of

rFVIIa probably due to rapid inactivation of rFVIIa with antithrombin III. On this evidence we

conclude that highly purified recombinant FVII has definite advantages on its activated form in the

replacement therapy of FVII deficiency.


221

Synthesis and Studying of Biological Activity of 2-[2-(3-R-adamanthane-1-yl)-2-

oxoethylidenhydrazino]-4-(het)aryl-4-oxobut-2-enoic Acids

A. Kuznetsov,1 N. Pulina,1 A. Krasnova1

1Perm State Pharmaceutical Academy, 614990, Russia, Perm, Polevaya st., 2


The previous studies have shown that derivatives of 4-(het)aryl-2,4-dioxobutanoic acids have

hypoglycemic activity [1]. Our further research involves interaction of 1-(3-R1-adamanthane-1-yl)-

2-(triphenylphosphoranylidenhydrazono)ethanons (1) with 4-(het)aryl-2,4-dioxobutanoic acids (2)

via diaza-Wittig reaction, forming 2-[2-(3-R-adamanthane-1-yl)-2-oxoethylidenhydrazino]-4-

(het)aryl-4-oxobut-2-enoic acids (3):

Screening for hypoglycemic activity of synthesized acids 3 has not revealed high active

compounds. However, in view of the revealed changes of motivated behaviour of the animals

tested, it was of interest to study their influence on behavioral responses of rats with alloxan-

induced diabetes mellitus (DM) by “open field” test. DM was modeled by oral administration of

alloxan trihydrate at the dose of 160 mg/kg with followed by registration of physical activity

parameters and emotional status in the “open field” test. Administration of test compounds was

found not to have eliminated oppressive behavior under alloxan-induced DM, though improved

locomotor activity was observed. Antidiabetic reference drug (metformin) also did not have a

positive impact on the motivation of rats with alloxan-induced DM behavior disorders.

The results obtained indicate the feasibility of further synthesis and study of neurotropic activity

of derivatives 3.

References: [1] N. Pulina, F. Sobin, A. Krasnova, et al, Pharmaceutical Chemistry Journal. 2011, 45 (5), 18-21.


222

Synthesis and Biological Evaluation of

2,2,2-tris(1H-indol-3-yl)acetic acid Derivatives

S.N. Lavrenov, Ya.A. Solomatin

Gause Institute of New Antibiotics, Russian Academy of Sciences,

119021, Russia, Moscow, Pirogovskaya Street, 11 B.


Some of 2,2,2-trisarylacetic acid and derivatives of trisindolylmethanes are known to posses

interesting biological activity.1,2 We have made first synthesis and described some derivatives of

2,2,2-tris(1H-indol-3-yl)acetic acid, some of which were proved to have high cytotoxic properties

for human tumor cell lines (best activity were shown with compounds of type 8-9, IC50 0.1-0.8 µM

on HCT-116 and K-562 tumor cell lines).

References: [1] R. Dothager, K. S. Putt, B. J. Alien, B. J. Leslie, V. Nesterenko, P. J. Hergenrother, J. Am. Chem. Soc., 2005, 127,

8686 [2] Stepanova E. V., Shtil’ A. A., Lavrenov S. N., Bukhman V. M., Inshakov A. N., Mirchink E. P., Trenin A. S.,

Galatenko O. A., Isakova E. B., Glazunova V. A., Dezhenkova L. G., Solomko E. Sh., Bykov E. E., Preobrazhenskaya M. N., Russian Chemical Bulletin. 2010, 59, N. 12, 2259-2267.


223

Synthesis and Anti-inflammatory Activity of Derivatives of

2-Hydroxy-4-oxo-4-thienyl-2-butenoic Acid

K. Lipatnikov, N. Pulina, A. Krasnova, T. Yushkova and F. Sobin

Perm State Pharmaceutical Academy, 614990, Russia, Perm, Polevaya st., 2


The previous studies have shown that 4-(het)aryl-2-hydroxy-4-oxo-2-butenoic acids possess anti-

inflammatory, hypoglycemic, anticoagulant, antimicrobial, analgesic effects [1]. Fragments of

thiophene, benzo[d]thiazole and 1,3,4-thidiazole are pharmacophores and they are contained in

drugs like pyrantel, riluzole, pramipexole, acetazolamide. In continuation of our research amides

and water-soluble derivatives of 2-hydroxy-4-oxo-4-thienyl-2-butenoic acid 2,4 were synthesized.

Screening for anti-inflammatory activity of the synthesized compounds 2,4 was performed on

acute exudative inflammation model. Active compounds acting at the level of reference drugs

nimesulide and diclofenac, but at the same time having less acute toxicity have been found.

References [1] A. Kuznetsov, F. Sobin, V. Kozhukhar’, and N. Pulina, Global Scientific Potential. 2013, 4 (25), 8-11.


224

Novel Moronic Acid Semisynthetic Derivatives

I.Ya. Mainagashev,1 O.V. Salomatina,1 and N.F. Salakhutdinov1,2




Moronic and morolic acids are naturally occurring pentacyclic triterpenes, containing double

bond in E-ring. These compounds were isolated from Acridocarpus vivy and Phoradendron

reichenbachianum (its extracts have been extensively used in traditional Mexican and African

medicine); they possess various physiological activities such as cytotoxic, anti-HIV, anti-HSV, and

anti-inflammatory [1]. But there are only few papers about semisyntetic derivatives of moronic

acid; therefore we decided to use moronic acid as a scaffold for the synthesis of new compounds.

Moronic acid has been prepared based on the available triterpene betulin according to the literature

technique [2]

In this work, new semisynthetic moronic acid derivatives were synthesized by modification of

carboxylic group and triterpene skeleton ring A. Amides (1-2) were synthesized via formation of

chloroanhydride and its reaction with corresponding amines. Compound 3 was synthesized by

introducing of pharmacophore (2-cyano-1-ene-3-one in the ring A); this fragment is essential

structural element that causes increasing of biological activity of semisynthetic medicinal agents

based on different triterpene acids [3].

Structures of all new compounds were confirmed by 1H and 13C NMR and high resolution mass

spectrometry. The anti-cancer and anti-inflammatory activities of the target semisynthetic products

are going to be evaluated in vitro.

References [1]. J. Ito, F.-R. Chang, H.-K. Wang, Y. K. Park, M. Ikegaki, N. Kilgore, K.-H. Lee, J. Nat. Prod. 2001, 64, 1278–1281 [2]. O. Flekhter, N. Medvedeva, G. Tolstikov, F. Galin, M. Yunusov, H. Thi, L. Tien, O. Savinova, E. Boreko, L. Titov,

and I. Glukhov, Biorgan. Khim. 2009, 35(2), 253-259. [3]. T. Honda, T. Janosik et al., Journal of Medicinal Chemistry. 2004, 47(20), 4923-4932; E. Logashenko, O.

Salomatina, A. Markov, D. Korchagina, N. Salakhutdinov, G. Tolstikov, V. Vlasov, and M. Zenkova, ChemBioChem 2009, 12, 784-794


225

Biofilm Probiotics

V. Melnikov,1 V. Chistyakov2

1Department of Molecular Diagnostics and Epidemiology, Central Research Institute of

Epidemiology, 111123, Russia, Moscow, Novogireevskaya street, 3a 2Rostov Research Institute of Biotechnology, Academy of Biology and Biotechnology,

Southern Federal University, 3444006, Russia, Rostov-na-Donu, B. Sadovaya street, 105/42


On the market there is a large number of probiotics, which contain live beneficial

microorganisms intended for the restoration of violated intestinal biocenosis. Current probiotics are

obtained by laboratory cultivation on nutrient media. However, bacteria living in nature are greatly

different by the properties from the cultures grown in ‘‘classic’’ laboratory conditions. The

dominant lifestyle of bacteria in the natural environment is biofilm. Biofilm is a stress-tolerant

community of microorganisms held together by intercellular junctions and a self-produced

extracellular matrix, which forms on the surface of objects of the environment and the tissues of

living organisms. We proposed that biofilm probiotics had advantage over the marketed planktonic

ones. One can suppose that biofilm-driven gene expression might make the probiotics bacteria

harmful for health. Three responses can be given as a counterclaim. First, the resident microbiota of

the human and animal body grows in a biofilm. Second, according to our study the traditional

health-giving foods Natto (similar to Pepoke of Myanmar, Chungkukjang and Doenjang of Korea,

Sufu or Furu of China and Thua nao of Thailand, soya beans fermented by terrestrial bacilli), which

Japanese consume for hundreds of years, include fermenting bacteria in a biofilm. Third, our

experiments on animals (mice, dogs, poultry, aquaculture fish) and human volunteers show that the

biofilm bacteria are not only harmless but health beneficial. So the results of our studies indicate the

possibility of bacterial biofilm growing technology application for the development of

veterinary/medical probiotic preparations and products of functional nutrition.


226

The Stereoselective Transformation of Eudesmanolide of (-)-α-Santonin

N. Merkhatuly, A.T. Omarova, A.K. Ibrayeva, D.D. Naushabekova, Zh.E. Suleimenova,

L.T. Balmagambetova

Ye.A.Buketov Karaganda State University,

100028, The Republic of Kazakhstan, Karaganda, str.Universitetskaya,28 Е-mail: [email protected]

The chemical transformations of plant’s metabolites, leading to the physiological active

derivatives, became the important direction in synthetic and medical chemistry. Continuing the

researches in the field of diene eudesmanolides chemistry [1-3] we investigated the reaction of

alkaline transesterification of (-)-α-santonin (1). The reaction (1) with ethanol at sodium ethylate

presence leads stereoselectively to formation of a new cis-condensed 10β(CH3),5β(H), 4β (CH3) - 6-

keto-eudesman ester (4) with yield 94% instead of 6-hydroxy-ester (3).

O

OO

O

OH CO2Et

4 5

10

(1)

6

O

OH CO2Et

O

O CO2EtH

64 510

O

OH CO2Et

6

(3)

EtO EtOH

(2)

2. EnolizationTautomerization

enol (4)

1. Allylic shift

x

Obviously, the initial stage of formation of an anion (2) initiates the following stages of the

electrophilic rearrangement, which are proceed stereoseleсtively, giving as a result ketо - cis-

eudesman ester (4), with the quantitative yield.

The formation of "key" enol is confirmed with synthesis of isosantonin (5) according to the

following scheme:

O

OO

56

t-BuO-

OO

O

(5)O

O

O

(1)

RT

t-BuOH · DMSO

K +H3O+

The structures of the synthesized compounds (4) and (5) are established by the UV-, IR -, NMR - 1H,13C – spectroscopy, mass spectrometry and X-ray analysis.

References [1] N. Merkhatuly, S.K. Zhokizhanova, L.T. Balmagambetova, Rus. J. Gen. Chem. 2006, 76, 1347. [2] N. Merkhatuly, S. K. Zhokizhanova, L. T. Balmagambetova, Rus. J. Org. Chem . 2007, 43, 150. [3] N. Merkhatuly, A.N. Iskanderov, L.T. Balmagambetova, Rus. J. Org. Chem. 2013, 49, 1405.


227

Liquid Phase Oxidation of Halo-vinyl Compounds on the Catalyst Ag/SiO2:

Extremely Stable Water-Soluble AgxCly Cluster

N. Asatryan, A. Mikaelyan

Department of Chemical Technologies and Environmental Engineering, National Polytechnic

University of Armenia, 0009(Zip code), Republic of Armenia, Yerevan, Teryan Str., 105


Liquid phase oxidation (LPO) processes functional groups of organic molecules with air or

molecular oxygen, is one of the important areas of organic chemistry and are widely used in

laboratory and industrial syntheses [1]. In this report presents the process of catalytic LPO of 1,1,5-

trichloro-1-pentene and 1,1-dichloro-5-bromo-1-pentene. As a promising catalytic system we have

chosen Ag/SiO2 (100-200 µm, 5% Ag) and packed column bubbling reactor [2]. The interaction can

be represented by the following scheme:

The process is characterized by a pronounced induction period of 2.5-3 h. At the same time, after

1.5-2 h of interaction, there is complete homogenization of the metallic silver into the organic phase

was observed. It is noteworthy that after the induction period the reaction rate increases

dramatically, and after 8-10 hours there is a selective formation of desired products with a yield of

70-80%. Although, for this reactions as end product was observed formation of appropriate

carboxylic acid chloride, that was isolated from reaction mixture by means of ethanolysis. After

this transformation and removal of rest of ethanol the formation of crystals was observed. This

compound is high photo-stabile, stable by heating up to 200 °C, decomposes by melting to AgCl

and gas (preliminary Cl2), solution in ethanol-water is stable in acidic media.

This work was supported by SCS MES RA, research project № SCS 13Ap_2e021.

References [1] A.K. Suresh, M.M. Sharma, T. Sridhar, Ind. Eng. Chem. Res. 2000, 39, 3958-3997 [2] N.L. Asatryan, Bulletin of SEUA (Polytechnic), 2010, 2, 473-478


228

Water-soluble Metal Complexes of Pectin

Polysaccharides with Antianemic Effect

S. Minzanova, V. Mironov, A. Vyshtakalyuk, L. Mironova, V. Milyukov

A.E. Arbuzov Institute of Organic and Physical Chemistry of Kazan Scientific Center

of Russian Academy of Sciences, 420088, Russia, Kazan, Arbuzov str., 8


New water-soluble complexes on the basis of natural biopolymers – pectin polysaccharides,

containing sodium and readily available bivalent ions Fe2+, Ca2+, Co2+, Cu2+, are synthesized. These

compounds are promising, since they contain macro-and microelements in the biologically

accessible form, that is a necessary condition for many vital processes in organism. They are

synthesized on the basis of pectin biopolymer matrix, which possesses a wide spectrum of

physiological activity, enlarged therapeutic effect and low toxicity. The procedure of metal-

complex synthesis is rather simple, environmentally safe and does not require expensive equipment

and expenses. The obtained substances are water-soluble, what makes them the perspective

preparations for application in medicine and pharmacology [1].

The target products are low-toxic compounds. They do not have irritant, skin-absorption and

ulcerogenic action, do not effect negatively on exercise performance of laboratory animals. LD50 for

mice at intraperitoneal introduction is 500-1200 mg/kg, as for peroral introduction, the substances

don’t manifest any toxic effect even at the doze of 25000 mg/kg. The metal complexes of pectin

polysaccharides improve substantially the hematological indices/ Thus, after introduction of the Na,

Co- and Na, Fe, Ca-pectates the hemoglobin concentration is increased by 10%, and the

erythrocytes number - by 20%. The introduction of Na, Cu- and Na, Fe-pectates increases the

hemoglobin concentration by 12.5% and 10%, respectively. The maximum effect is reached by

application of polymetallic complexes, containing Fe2+, Co2+, Cu2+ simultaneously (the hemoglobin

concentration is increased by 13-23%, and the erythrocytes number - by 21-38%). The antianemic

activity of the obtained complexes is higher than the activity of Ferroplex, Tot’hema, Sorbifer,

Actiferrin, Ferroglobin B12 preparations. Therewith, the iron doze demonstrating the strong effect

on hemogenesis is 2-3 times less than therapeutic doses recommended for the investigated

medicines.

All the results are covered by the patents of RF.

This work was supported by Russian Foundation for Basic Research (project no. 13–03–00046)

References: [1] S.T. Minzanova, V.F. Mironov, A.I. Konovalov, A.B. Vyshtakalyuk, O.V. Tsepaeva, A.Z. Mindubaev, L.G. Mironova and V.V. Zobov. Pectins from non-traditional raw materials: technology, properties and biological activity, Kazan. 2011, 226.


229

Phlomisoic Acid in the Synthesis of Labdane Type Naphthoquinones and

Biphenyl Substituted Spiroundecanes

М.Е. Mironov,1 М.A. Pоkrovsky,2 A.G. Pоkrovsky,2 E.E. Shults1

1NIOCH SB RАS, 630090, Russia, Novosibirsk, Lavrentiev’s av., 9

2Novosibirsk State University,Russia


Furanolabdanes, which include the phlomisoic acid 1, possess diverse biological activity [1]. We

have developed a selective approaches to the synthesis of furanolabdanes containing an additional

naphthoquinone or 7-biphenylspiro[5,5]endecane-1,5,9-trione moiety in the furan ring. As the main

synthetic approach chosen Diels-Alder rection of terpenoid siloxydienes 2, 3 and appropriate

dienophiles.

The cytotoxic activity against several cancer lines of the resulting compounds in the

conventional MTT assay have been determined. The biological results for the modified

furanolabdanoids, reported herein, shown that compounds exhibited the activity with respect to

model cancer cell lines (within micromolecular range) and the structural modification with the

introduction of napthoquinone or 7-biphenyl-spiro[5,5]undecane-1,5,9-trione substituent in the

furan ring proved of great importance to obtain cytotoxic anti-cancer agents.

This work was supported by the Russian Foundation for Basic Research (project № 15-03-06546).

References [1] Shults Е.E., M. E. Mironov, and Yu. V. Kharitonov, Chemistry of Natural Compounds, 2014. Vol. 50, No. 1, p. 2-21.


230

Chemical Modification of Cirsilineol Molecule

G.K. Mukusheva, G.M. Baisarov, S.M. Adekenov

JSC “International Research and Production Holding ”Phytochemistry”,

100009, Republic of Kazakhstan, Karaganda, fax: 8 (7212) 43 31 27,


Plant polyphenols are one of the perspective directions in the field of phytochemical studies.

Research in this field is mainly dedicated to studies of plant flavonoid aglycones, anthraquinones

and coumarines, and also obtaining on its basis the new biologically active derivatives.

To the number of available flavons belongs 5,4’-dihidroxy-6,7,3’- trimetoxyflavon (1), isolated

from Artemisia glabella Kar.et Kir.

On the basis of cirsilineol molecule (1) we have synthesized its relevant oxime (2) with yield of

65%.

Bromation of cirsilineol with NBS in methanol results in formation of relevant bromoderivative

(3) with yield of 59%.

By acylation of cirsilineol Ас2О/ТsОН acylderivative is formed (4) with yield of 67 %.

O

OOH

OH

OCH 3

H3CO

H3CO

NH2 OH*HCl

O

N OHOH

OH

OCH3

H 3CO

H 3CO

12

O

OOH

OH

OC H3

H 3CO

H 3CO

NBS

Br

3

O

OOAc

OH

OC H3

H3CO

H3CO

Ac

2 O/T

sOH

4 The composition and the structure of new synthesized compounds are proved by the results of

IR-, UV-, NMR 1Н, 13С – spectroscopy, and also by data of elemental analysis, mass-spectrometry.


231

Synthesis of Novel Purine and 2-Aminopurine

Conjugates with Heterocyclic Amines

V.V. Musiyak, D.A. Gruzdev, E.N. Chulakov, A.Yu. Vigorov, G.L. Levit, V.P. Krasnov

Postovsky Institute of Organic Synthesis, Russian Academy of Sciences (Ural Branch), Ekaterinburg, Russia


Purine derivatives with C-6-amino substituents have attracted considerable attention due to their physiological and pharmacological properties. The purpose of this work was to obtain a series of new 2-aminopurin-6-yl and purin-6-yl conjugates with chiral heterocyclic amines 1-6 and achiral amines 7 and 8.

ONH

Me1

ONH

Me2

F

FS

NH

Me3

NH

Me4

NH

Me5

F

NH

Me6

NH

7

NH

8 To prepare 2-aminopurine conjugates linked with fragments of the above amines via 6-

aminohexanoyl moiety, we carried out the nucleophilic substitution of chlorine in 2-acetamido-6-chloropurine by enantio pure N-(6-aminohexanoyl) derivatives of heterocyclic amines 1–5. The same approach was applied for the synthesis of racemic and achiral 6-purinyl derivatives bearing the fragments of amines 2, 6, 7.

YZ

N

X

Me

(CH2)5O NPhth

YY

N

X

R'

(CH2)5O NPhth

YY

N

X

R'

(CH2)5O NH

N

N

NH

N

YZ

N

X

Me

(CH2)5O NH

N

N

NH

N

NHR

i. N2H4, EtOH, ∆ii. 2-acetamido-6-chloropurine,

TEA, DMA, 100 oC

R = Ac

1N NaOH, rtR = H

X = CH2, O, SY = H, FZ = H, F

i. N2H4, EtOH, ∆

ii. 6-chloropurine, TEA, n-BuOH, 90 oC

X = CH2, OCH2, (CH2)2Y = H, FR' = Me, H

62-85%

40-68%

55-98%

In another approach, we performed the coupling of N-(6-purinyl)aminohexanoic acid with

racemic amine 6 and achiral amines 7, 8 under the action of TBTU in the presence of DIEA. However, the reaction of amines 6 and 7 afforded the target compounds in low yields (28 and ~7%, respectively), whereas the attempts to perform the reaction with amines 2 and 4 failed.

To prepare conjugates of purine (2-aminopurine) linked directly with racemic heterocyclic

amines 2, 4, 6 we carried out the nucleophilic substitution of chlorine atom under various conditions. The highest yields were observed when the reaction was carried out in boiling water in the presence of sulfuric acid.

NH

X

Me

Y

ZN

N NH

N

Cl

R

N

X

Me

Z

N

N

NH

N

R

Y

+H2SO4, H2O, X = CH2, (CH2)2, OCH2

Y = Z = H, F

R = H, NHAc

2, 4, 6 Thus, a series of novel conjugates of purine and 2-aminopurine linked with the fragments of

heterocyclic amines via 6-aminohexanoyl fragment or directly were obtained for the first time. The presence of free 9-NH group in the purine system allows further transformation of these compounds aimed at preparation of modified nucleosides, promising biologically active compounds. This work was financially supported by the Russian Scientific Foundation (grant 14-03-01077).


232

Highly Stereoselective Synthesis of (2SR, 3aSR, 8aSR)-3a-

methyloctahydrocyclohepta[b]pyrrol-4(1H)-one Derivatives

I.N. Myasnyanko, E.R. Lukyanenko, A.V. Kurkin

Lomonosov Moscow State University, Chemistry Department, 119991, Moscow, GSP-2, Leninskie Gory, 1/3

E–mail: [email protected] Fax: +7-495-939-17-14

Recently, most players in the pharmaceutical industry working on small molecule drug discovery

have focused their efforts on enriching the newly designed drug prototype with sp3-hybridized

carbon atoms as their goal was to bring the chemo-types closer to natural products and to boost

chemo types’ three-dimensional properties.1

However, the reaction portfolio currently available to the pharmaceutical industry lacks solid

synthetic methodologies to produce compound libraries of natural product-like molecules for

screening purposes. Furthermore, the synthetic approaches commonly used for small molecule

compound libraries often miss a stereo controlled bond formation. Conversely, most natural

products do have well stereo-defined structural features.

Herein we report the highly stereoselective synthesis of (2SR, 3aSR, 8aSR)-3a-

methyloctahydrocyclohepta[b]pyrrol-4(1H)-one derivatives (6-30) via the aza-Cope-Mannich

reaction (Scheme) with several stereocenters.

This study was supported by the Russian Foundation for Basic Research (RFBR), Russia (Projects No. 14-03-01114 [1] Ritchie, T. J., Macdonald S. J. F., Young, R. J., Pickett S. D. Drug Discovery Today, 2011, 16, 164-171.


233

Multicomponent Efficient Transformation of Salicylaldehydes, Dimedone and

Coumarin into Substituted Tetrahydro-1H-xanthen-1-ones

R.F. Nasybullin,1 O.O. Sokolova2 and M.N. Elinson 1

1N. D. Zelinsky Institute of Organic Chemistry RAS, Russia, 119991 Moscow, 47 Leninsky prosp.

2Higher Chemical College RAS, Russia, 125047, Moscow, 9 Miusskaya square


Xantenes (dibenzopyranes) are one of the most widely distributed classes of natural compounds

possessing diverse pharmacological activities such as antiviral, anti-inflammatory and anti-cancer

properties [1].

Coumarin (2H-chromen-2-one) has attracted much attention as it found naturally in many plants

and due to its remarkable biomedical applications [2].

Thus, the 9-(4-hydroxy-2-oxo-2H-chromen-3-yl)-3,3-dimethyl-2,3,4,9-tetrahydro-1H-xanthen-1-

one system appears to be of the interest as it incorporates a tetrahydro-1H-xanthen-1-one and

coumarin heterocyclic rings, which are both promising with respect to biological responses.

We herein report a fast and efficient multicomponent transformation of salicylaldehydes,

dimedone and 4-hydroxycoumarin into substituted 9-(4-hydroxy-2-oxo-2H-chromen-3-yl)-3,3-

dimethyl-2,3,4,9-tetrahydro-1H-xanthen-1-one in ethanol in the presence of 10 mol.% of NaOAc:

The developed procedure utilizes simple equipment, reasonable starting materials and is valuable

from the viewpoint of environmentally benign diversity-oriented large-scale processes.

Acknowledgements

The authors gratefully acknowledge the financial support of the Russian Foundation for Basic

Research (Project No. 14-03-31918).

References [1] Giri R., Goodell J.R., Xing C., Benoit A., Kaur, H., Hiasa H.; Ferguson D.M. Bioorg. Med. Chem., 2010, 18, 1456-1463. [2] Sandhu S., Bansal Y., Silakari O., Bansal G. Bioorg. Med. Chem., 2014, 22, 3806-3814.


234

5,5’-Modified 2’,5’-Dideoxyuridines: Synthesis and Biological Properties

S.D. Negria, I.L. Karpenko, S.N. Kochetkov, L.A. Alexandrova

Engelhardt Institute of Molecular Biology RAS, Vavilova 32, Moscow 119991, Russia


Tuberculosis is currently the worlds second leading infectious killer after AIDS and causes 1.4 -

2 million deaths annually [1]. The emergence of new multidrug resistant (MDR) and extensively

drugresistant (XDR) strains of Mycobacterium tuberculosis, the causative agent of tuberculosis,

means that there is an urgent need for new antituberculosis drugs. at present, no new

antituberculosis drugs were introduced into clinical practice since the 1970s. Recently we

synthesized 2’-deoxypyrimidine nucleoside derivatives bearing extended alkyloxymethyl or

alkyl(1,2,3-triazol-1-yl)methyl substituents at С-5 position and demonstrated their effective

bacteriostatic activity against M. tuberculosis strains [2]. We found that introduction of azido- or

iodo substituents in position 5’ of the carbohydrate moiety does not cause the loss of

antimycobacterial activity. The goal of this work was to synthesize and to study biological

properties of 5, 5’-modified 2’-deoxyuridine derivatives bearing extended substituents.

The 1,3-dipolar cycloaddition of azide (1) with the corresponding olefins in a binary phase

system of methylene chloride-water under the catalysis of Cu(I) obtained in situ from copper

sulphate and sodium ascorbinate according to method [3] resulted in 5’-[4-substituted (1,2,3-triazol-

1-yl]-5-[4-(1-decyl)-1,2,3-triazol-1-yl]methyl-2’,3’-dideoxyuridines (2,3). The treatment of

nucleoside (3) with NH3aq or 0.5M KOH gave compounds (4 and 5, respectively). All compounds

were not cytotoxic at concentrations up to 200 µM for Vero and up to 100 µM for A549 and Jurkat

cell lines. The anti-TB-activity will be reported.

Acknowledgment. This study was supported by the Russian Scientific Foundation, project № 14-50-00060 References 1. WHO, Global tuberculosis control. WHO report 2014, 115 2. E.R. Shmalenyuk, Kochetkov S.N., Alexandrova L.A. et.al. , Bioorg. Med. Chem. 2013, 21, 4874 3. Lee B.-Y.; Park S.R.; Jeon H.B.; Kim R.S., Tetrahedron Lett. 2006, 47, 5105


235

Menthyl Ester of Gamma-aminobutyric Acid as Potential Anticonvulsant Agent

M. Nesterkina, I. Kravchenko

Pharmaceutical Chemistry Department, Odessa National University, 65026, Ukraine, Odessa,

Dvorjanskaya st., 2


Menthol is well known for cooling effect due to its ability to chemically activate the cold-

sensitive transient receptor potential cation channel (TRPM8) [1]. In addition to this peripheral

action, a number of studies have demonstrated that menthol has actions within the CNS such as

nootropic and anticonvulsant activity [2]. Moreover, menthol was found to act as a potent positive

allosteric modulator of GABAA receptors [3]. These promising data served as precondition for

present study aimed to obtain menthyl ester of gamma-aminobutyric acid (GABA) with subsequent

investigation of its anticonvulsant activity. Acyl derivative of (–)-menthol with amino acid moiety

was synthesized by reaction with N-Boc-protected GABA in the presence of

dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) followed by deprotection

of the amino groups in the HCl/CH3COOH medium.

O

O

NH

O

OOH

ONH

O

OOH

O

O

NH2 ClHDCC, DMAP

ClH

CH3COOH.

The anticonvulsant activity of menthyl ester was examined in model of acute generalized

seizures induced by intravenous infusion of 1% pentylenetetrazole solution (PTZ). Dose-response

relationship for the compound has been investigated over a wide range of doses: 87-1350 mg/kg.

Considering the possible prolonged action of menthyl ester, pharmacological tests were carried out

at large time period: 0.5-96 hours after oral administration. Maximum anticonvulsant effect was

observed at 18 hours: DCTC and DTE values are 191% and 196%, accordingly. Synthesized

compound was shown to demonstrate prolonged anticonvulsant activity: DCTC and DTE values are

133% and 142%, respectively, at 96 hours after administration (175 mg/kg). The acute oral LD50 of

menthyl ester to adult white mice was measured and estimated at 2700 mg/kg; noteworthy is the

fact that experimental animals show signs of convulsions and respiratory rhythm inhibition before

dying.

References [1] G. Kamatou, I. Vermaak, A. Viljoen and B. Lawrence, Phytochemistry. 2013, 96, 15-25 [2] X-B. Zhang, P. Jiang, N. Gong et al., PLoS ONE. 2008, 3, 1-11 [3] A. Hall, C. Turcotte, B. Betts et al., Eur. J. Pharmacol. 2004, 506, 9-16


236

Hemocoagulating Activity of Sulfur-Containing Monoterpenoids

S.V. Kiselev,1 L.E. Nikitina,1 V.A. Startseva,1 N.P. Artemova,1 A.V. Bodrov,1

А.А. Rakhmatullina,2 G.A. Yakubovskaya1 and V.V. Klochkov3

1Department of General and Organic Chemistry, Kazan State Medical University, 420012, Russian

Federation, Kazan, ul. Butlerova, 49 2Republican Center of Blood, 420140, Russian Federation, Kazan, ul. Pobedy, 85

3Institute of Physics, Kazan Federal University, 420008, Russian Federation, Kazan, ul. Kremlevskaya, 18


We previously showed that sulfur-containing monoterpenoids possessed antifungal, anti-

inflammatory, anti-helicobacter, and other types of activity [1]. Herein the hemocoagulating activity

of sulfides, sulfoxides and sulfones synthesized on the basis of (−)-β-pinene and (+)-camphene [2-

4] was studied using human blood plasma in vitro.

The most water-soluble sulfoxide of pinane series and sulfone of camphene structure inhibit

completely the spontaneous and induced by collagen and arachidonic acid aggregation of platelets;

they also reduce the coagulating activity of human blood plasma. Interaction of sulfones with model

of biological membranes (SDS, DPC) have been studied using 2D DOSY and 2D NOESY spectra.

Taking into account the low toxicity of thioterpenoids the compounds obtained can be considered as

potential medicinal agents for treatment and prevention of trombophilia, and as a stabilizer of blood

specimen in transfusiology.

The work was performed using a subsidy under state support of K(VR)FU for increasing the competitiveness of leading global science-education centers.

References [1] L.E. Nikitina, N.P. Artemova, and V.A. Startseva, Natural and thiomodified monoterpenoids, LAP LAMBERT, Germany. 2011, 167 p. [2] L.E. Nikitina, V.A. Startseva, S.A. Dieva, I.A. Vakulenko, and G.A. Shamov, Chem. Nat. Compd. 2006, 42, 178-181 [3] V.V. Gavrilov, V.A. Startseva, L.E. Nikitina, O.A. Lodochnikova, O.I. Gnezdilov, S.A. Lisovskaya, N.I. Glushko, and E.N. Klimovitskii, Pharm. Chem. J. 2010, 44, 126-129 [4] A.V. Bodrov, L.E. Nikitina, V.A. Startseva, O.A. Lodochnikova, R.Z. Musin, and O.I. Gnezdilov, Russ. J. Gen. Chem. 2013, 83, 80-86


237

Synthesis of Compound of Ceftriaxone with L-α-Alanine

G.V. Novikova, N.A. Staloverova

Institute of Non-Ferrous Metals and Materials Science, Siberian Federal University, 660041

Russian Federation, Krasnoyarsk, Svobodny Prospekt, 79


Cephalosporins are semisynthetic broad-spectrum antibiotics. Nowadays they are the most

widespread antimicrobial drugs exhibiting mechanisms of action, resistance and some other

properties similar to the ones of penicillins. Ceftriaxone antibiotic belongs to the third generation of

cephalosporins. Ceftriaxone has proven in the treatment of gram-negative microorganisms, but at

regular use, the bacteria increase the defense reaction, and efficiency of antibiotic becomes reduces.

The amino acid alanine (НАla) belongs to the nonessential amino acids, it is easily synthesized in

the organism of animals and humans. L-α-НАla is raw material for the synthesis of glucose in the

organism. Therefore, it is important to design and produce new drugs, which are based on the

cephalosporin compounds. The current research is focused on the synthesis of ceftriaxone

compound with L-α-alanine and study of antibacterial properties.

The compound (Na2CefTriaН)Ala·(H2O)2 was synthesized by mixing Na2CefTria·3,5H2O with

L-α-НАla at the molar ratio 1:1 in minimum water volume. Then, 70 ml of ethanol was added to the

solution to precipitate a white powder (pH = 7,1). The compound was characterized by chemical

and spectrophotometric analysis, TGA and IR spectroscopy. The antibacterial activities of the

compound was tested in vitro against Gram-positive bacterium Staphylococcus aureus 25923,

Gram-negative bacteria Escherichia coli 25922 and Klebsiella pneumoniae 13883. The effects of

ceftriaxone, L-α-alanine and (Na2CefTriaH)Ala·(H2O)2 on the bacteria were investigated using the

paper disk diffusion method. The bacteria were cultured in incubator for 18–24 h at 36 °C. Standard

disks were impregnated with the solutions of the compounds in phosphate buffer (pH = 6). The

biological activities of (Na2CefTriaН)Ala·(H2O)2 against Gram-positive and Gram-negative bacteria

were investigated at the (Na2CefTriaН)Ala·(H2O)2 concentrations of 0.4, 0.8, and 1.0 mg/mL−1.

Antibacterial activity (Na2CefTriaН)Ala·(H2O)2 is increased by 30% relative Na2CefTria against

Escherichia coli 25922 and 5% relative to Staphylococcus aureus 25923. There is no antibacterial

activity observed against Klebsiella pneumoniae. The initial amino acid does not exhibit

antibacterial properties against all bacteria.

The research has been funded and carried out in terms of state contract of Ministry of Education and Science of Russian Federation.


238

Synthesis of 1-Methyl-9-phenyl-3,11-dioxatricyclo[5.3.1.02,4]undecane

A.M. Novoselov, E.R. Lukyanenko, D.S. Belov, A.V. Kurkin

Lomonosov Moscow State University, Chemistry Department, 119991, Moscow, GSP-2, Leninskie Gory, 1/3

E–mail: [email protected] Fax: +7-495-939-17-14

In modern world the most important purpose in medicinal chemistry is the search of new drugs

against cancer. There are some natural substances which have a biological activity against different

cancer cells. One of this natural substances is derivatives of Englerin A. Englerin A is a newly

discovered guaiane sesquiterpene from the stem bark of Phyllanthus engleri collected in Tanzania 1

The key intermediate in synthesis of these cores is the macrocyclic ketones which we have

obtain by the oxy-Cope rearrangement from the respectively 1,5-dien-1-ols. This way is shortest

and simplest compared with all known methods in synthesis derivatives of natural Englerin A.

Herein we report the synthesis of 1-methyl-9-phenyl-3,11-dioxatricyclo[5.3.1.02,4]undecane 10

via the oxy-Cope reaction (Scheme).

This study was supported by the Russian Foundation for Basic Research (RFBR), Russia (Projects No. 14-03-31685, 14-03-31709, 14-03-01114) [1]. Ratnayake, R.; Covell, D.; Ransom, T. T.; Gustafson, K. R.; Beutler, J. A. Org. Lett. 2009, 11, 57–60.


239

Synthesis of Novel Monoterpenoid and Methoxyphenyl Substituted Coumarins

T. Odarchenko,1,2 T. Khomenko,1,2 D. Korchagina,1 K. Volcho,1,2 N. Salakhutdinov1,2

1N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, 630090, Russia, Novosibirsk, pr. Lavrentieva, 9

2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova st., 2


Coumarin derivatives are of great interest due to their diverse bioactivity. In particular, their

antiviral activities make the compounds attractive for further derivatization and screening as novel

therapeutic agents [1]. We have synthesized novel coumarin compounds for testing their inhibitory

anti-influenza virus A (H1N1) activity. Substituted coumarins were prepared by the reactions of

monoterpenoid and methoxyphenyl bromides with coumarins synthesized by the Pechmann

condensation of resorcinol with β-keto esters.

This work was supported by the Russian Science Foundation (15-13-17). References [1] Chia-Lin Lee et al., J. Nat. Prod. 2009, 72, 1568–1572.


240

The Effect of Lithium, Cerium (III), Copper (I) Chlorides on the Synthesis of

1-(2-Chloro-3-quinolyl)-4-dimethylamino-2-(1-naphthyl)-1-phenyl-butan-2-ols

A. Omelkov,1 V. Fedorov2

1CJSC «Pharm-Sintez», 115419, Russia, Moscow, Vtoroy Roschinskiy blvd., 8


Tuberculosis (TB) is a major health problem worldwide. In 2013, an estimated 9.0 million developed TB and 1.5 million people died, 0.36 of whom were HIV-positive [1]. The development of effective chemotherapy for the treatment of tuberculosis began in 1940s and has been reinvigorated recently due to concern regarding the emergence of highly drug-resistant TB strains [2]. The new anti-TB drug Bedaquiline (Sirturo®) was introduced in 2013. Diarylquinolines show new type activity against sensitive and resistant Mycobacterium tuberculosis strains [3].

CJSC «Pharm-Sintez» developed and explored new derivatives of Bedaquiline. One of them shows a good activity against Mycobacterium tuberculosis resistant strains and can be produced in industry. It is necessary to note, that production of 1-(2-chloro-3-quinolyl)-4-dimethylamino-2-(1-naphthyl)-1-phenyl-butan-2-ols is very complicated and requires application of expensive reagents. Our work’s purpose was to search simple methods to get the alcohols 3 with good yield. We decided to accomplish the goal using cerium (III) chloride and lithium chloride. Under optimized conditions it was observed that the yield of reaction increases from 42% to 70%. It was subsequently found that (1R*, 2R*)- and (1S*, 2S*)-isomers of 1-(2-chloro-3-quinolyl)-4-dimethylamino-2-(1-naphthyl)-1-phenyl-butan-2-ols obtained more than (1R*, 2S*)- and (1S*, 2R*)-isomers. The ratio between R*R*, S*S* and R*S*,S*R* diastereomers was 9/1.

N

Cl CHCeCl2

R3,R2,R1

O

N

CH3

H3C

N

Cl

R1,R2,R3

OH

N+H

CH3

H3C

Cl-

1 2

1.) -78°C

2.) HCl

3

+

**

Another way to increase the yield of the reaction is activation of the ketone group. The methods

of activating the ketone group at the 3-dimethylamino-1-(1-naphthyl)-propan-1-one have been developed. This effect is achieved by forming a complex of copper (I) chloride and 3-dimethylamino-1-(1-naphthyl)-propan-1-one. Reaction between 3-benzyl-6-bromo-2-chlorquinoline and copper complex gives the yield of products 81%. The ratio between R*R*, S*S* and R*S*,S*R* diastereomers was 9/1.

In conclusion, we have demonstrated that the nucleophilic addition with cerium (III) chloride and lithium chloride has stereoselectivity and gives good yield. Our work has successfully demonstrated the first application of the copper complex method for synthesis of 1-(2-chloro-3-quinolyl)-4-dimethylamino-2-(1-naphthyl)-1-phenyl-butan-2-ols which has a stereoselectivity and gives a good yield. References [1] Organization W. H. Global tuberculosis control: WHO report 2014. [2] Elliott R. L. Third World Diseases / Ed. by Springer. 2011. [3] Baptiste V., Crauste C., Flipo M. et al. Eur. J. Med. Chem., 2012, 51, 1 – 16.


241

Solubility of Powder Preparations from Quercetine Containing Plant Raw

D.V. Orlov, O.I. Lomovsky

Institute of Solid State Chemistry and Mechanochemistry SB RAS, 630128

Russia, Novosibirsk, Kutateladze 18


One of the main criteria of biological availability of pharmaceutical substances and preparations

is the solubility in water and physiological solutions. Powder preparations produced from plant

materials by grinding of plant parts are used in modern medicine practice and traditional medicine

as a biologically active additives and components of functional food. The content of soluble

substance in plant vary between some percents for roots and wooded parts and 30 - 40% for special

case as for green tea or flowers petals.

Mechanochemical treatment of plant raw materials with special solid reagents was applied. The

chemical reactions with formation of soluble forms of biologically active minor components and

depolymerization of the main components of plant tissues occurs. Formation of soluble salt forms

of biologically active substances of acids or phenol nature is carried up under mechanical treatment

of raw with solid alkalis. Reactions of alkaline hydrolysis of polymer components are also possible

during mechanochemical treatment. The aim of our work is to define the influence of the main

components content – cellulose, hemicellulose, lignin – upon the effect of mechanochemical

enhancing of preparation solubility. The comparative range of plant raw was chosen with high

content of flavonoids and quercetin especially.

The range of powered preparations with different solubility (in brackets, mass%) after

mechanochemical treatment is presented: brank, grain husk (5); blueberry, press cake (17); larch,

timber (18); onion, husk (25); sophora, root (32), dill, herb (43).

Content of cellulose, hemicellulose and lignin in initial raw is carried out. The solubility of

mechanochemicaly produced preparations decrease with the increase of cellulose content, the

variation of lignin content in 10 mass% interval doesn’t influence really upon solubility preparation,

there are no any correlation of hemicellulose content upon solubility.

The reason of cellulose content effect is based upon the mechanochemical reaction of flavonoid

quercetin with cellulose. It was shown that chemical reaction in cellulose and qurcetin solid mixture

lead to the adsorption of quercetin and decrease of its bioavailability.


242

Antioxidative Activity of Novel Organogold(I) Complexes Based on

2,6-di-tert-butyl-4-mercaptophenol

V. Osipova,1 M. Polovinkina,2 N. Movchan,1 D. Shpakovsky,3 E. Milaeva3

1Southern Scientific Centre of the RAS, 344006, Russia, Rostov-on-Don, st. Chehova, 41

2Astrakhan State Technical University, 414056, Russia, Astrakhan. st. Tatishcheva 16

3Moscow State Lomonosov University, 119991, Russia, Moscow, Lenin Hill 1-3


Despite the clinical success of known drugs based on organometallic compounds, many tumors

are inherently resistant to them, and some of them can cause serious side effects. Consequently,

there is a growing demand for new metal-based pharmaceuticals with a mechanism of action

different from the known anticancer drugs. Substances based on Au(I) are known for their anti-

arthritic properties and have a clinical interest as potential antitumor agents.

The effect of gold complexes 1-2 on the lipid peroxidation (LPO) level in rat and sturgeon liver

was studied in comparison with the effect of 2,6-di-tert-butyl-4-mercaptophenol (RSH). The

concentration of secondary products of lipid peroxidation, which react with thiobarbituric acid

(TBARS) was determined at different stages of oxidation (3, 24 and 48 h). In a control experiment

the level of TBARS in rat and sturgeon liver after 48 h increased 1.3 times and 9 times in

comparison with 1 h (p<0.01). The addition of RSH to liver leads to the reduction of TBARS

accumulation of at all stages of oxidation (p<0.01). The addition of Au complexes-1 and 2

significantly decrease TBARS’ level of LPO in rat liver. The level of TBARS in sturgeon liver in

the presence of gold complexes is comparable with control experiment.

1

2

a) b) Effect of the compounds on TBARS level in rat (a) and sturgeon (b) liver.

The addition of Au compounds results in more effective inhibition of LPO in rat liver

homogenate, when compared with sturgeon liver homogenate. Moreover the effect of 1 and 2 on rat

liver LPO was more pronounced than the influence of the known organic antioxidant RSH.

Acknowledgements - The financial support of RFBR (grants № 13-03-00487, 14-03-00578, 15-03-03057) and RSF (14-13-00483) is gratefully acknowledged.


243

Certain Short Peptide Analogues of Somatostatin

V.N. Osipov,1 A.N. Balaev,1 K.A. Okmanovich,1 A.V. Kolotaev2 and D.S. Khachatryan2

1CJSC «Pharm-Sintez», 115419, Russia, Moscow, Vtoroy Roschinskiy blvd., 8

2FSUE «IREA», 107076, Russia, Moscow, Bogorodsky shaft, 3


Somatostatin (SST) is a cyclic peptide consisting of 14 amino acids and inhibiting the release of

growth hormone [1]. Since native somatostatin is short-acting and of limited usefulness in clinical

treatment, synthetic somatostatin analogues were developed [2]. Peptide analogues of somatostatin,

such as octreotide, vapreotide (RC-160), and lanreotide have a high resistance to enzymatic

degradation and different activity profiles and now are used in the clinic for the treatment of

neuroendocrine tumors.

It was found that the amino acid residues Phe, Trp, Lys and Thr in the native molecule SST are

necessary for biological activity [3]. So, we have previously obtained short peptides possessed as

cytotoxic activity [4] and antitumor activity in mice [5]. At present research a next series of short

peptides, which are fragments of octreotide with various functional groups on the N- and C-

terminal, were synthesized.

NH

SR1

CO-

(R1 = H, Ar)X-Cys = Boc-Cys(Acm)-,

Y = OH, OAlk, NH2, NHOH; R = Boc, Z;

X-Cys-Phe-D-Trp-Lys(R)-Thr-Y

Prepared peptides were tested for cytotoxic activity in vitro. High inhibitory activity has been

demonstrated in tumor cells with high density somatostatin receptors.

Work was carried out at financial of Ministry of Education and Science of the Russian Federation. (agreement № 14.576.21.0044 from 05.08.14)

References [1] C.B. Srikant, Somatostatine. 2004, Kluwer Academic Publishers, Boston [2] O. Ovadia, S. Greenberg, B. Laufer, et al., Expert Opinion on Drug Discovery. 2010, 5(7), 655-671 [3] S.W. Lamberts, A.J. van der Lely, and L.J. Hofland, European Journal of Endocrinology. 2002, 146(5), 701-705 [4] A.N. Balaev, V.N. Osipov, V.E. Fedorov, et al., Rossiiskii Bioterapevticheskii Zhurnal. 2012, 11(4), 47-53 [5] A.N. Balaev, V.N. Osipov, V.E. Fedorov, et al., Rossiiskii Bioterapevticheskii Zhurnal. 2013, 12(3), 57-60


244

Mechanochemical, Microwave, Ultrasound assisted

Isolation of Natural Alkaloids

N. Pankrushina,1 M. Korotkih,1 and A. Mikheev2,3

1N.N. Vorozhtsov Institute of Organic Chemistry, 630090, Russia, Novosibirsk, Lavrentjev ave., 9

2 Nikolaev Institute of Inorganic Chemistry, 630090, Russia, Novosibirsk, Lavrentjev ave., 3 3Novosibirsk State University, Pirogov Str. 2, Novosibirsk, 630090, Russia,


Alkaloids possess a broad spectrum of biological activity and are the base of many remedies. It

was proved that alkaloids pharmacological properties were used for treatment of humans, animals

and plants [1, 2]. The aim of our research was to develop alternative effective methods to isolate

alkaloids from plant materials. We have studied the mechanochemical treatment of Aconitum

septentrionale Koelle roots as well as Sophora flavescens Ait. roots with alumina and silica. The

alkaloids yield rose up to 2 times and the procedure was more simple and ecological as it was

shown. The MW assisted extraction improved the yield of alkaloids up to 2- 4 times with high

selectivity using even water as the solvent. We have studied the ultrasound assisted extraction

conditions of alkaloids from Sophora flavescens Ait. vegetative organs (roots, herb, seeds and

pericarps). It was found that seeds and pericarps of this plant are perspective resource of alkaloids.

The alkaloid composition of the new species Papaver kuvajevii Schaulo et Sonnikova has been

investigated by GC-MS method both after conventional and MW assisted extraction. The main

alkaloid components amurensinine, mecambridine and minor alkaloids α-allokriptopin, amurensin,

β-homochelidonine, tetrahydrocoptisine, sanguinarine, norsanguinarine, oxysanguinarine, biflorine,

oxyhydrostinine, cryptopalmatine, homochelidonine, norchelerythrine, 8,9-dimethoxy-2,3-

(methylenedioxy) isopavinane were identified.

References [1] M.A. Turabekova, B.F. Rasulev, F.N. Dzhakhangirov, et al., J. Environ. Sci. Health, Part C, 2014, 32, 213–238. [2] Peng-Fei Yue, Hai-Long Yuan, et al., Int. J. Pharm., 2010, 387, 139–146.


245

A New Method for Synthesis of Substituted Diazepines[1,4] Annelated with

Indole Nuclei

A.A Panov, S.A. Lakatosh

Gause Institute of New Antibiotics ul. Bol'shaya Pirogovskaya 11, Moscow, 119021 Russia

HN

OO

NH

N

N

N

HN OO

NH

HN

II

IIIa

HN

OO

N

N

NR'2R'1

IIIb

Compounds (I) are new promising analogues of various known biologically active compounds,

such as an pigment arcyriacyanin A (II), posessing the antitumor properties, and the potent PIM1

kinase inhibitors (III). We report a synthesis of novel compounds (VIa,b) as precursors for (Ia,b)

The method employs sequential nucleophilic substitution of the bromine atoms in 3,4-

dibromomaleimide and indoline or indoline-3-alkanol with subsequent cyclisation in trifluoroacetic

acid to produce diazepine annealed with two indoline nuclei. The oxidation of the diazepines (V)

with DDQ with subsequent acidification of reaction mixture, allowed to transform an undescribed

unknown intermediate products (presumably with an open maleimide cycle) into the desired

diazepines (VIa,b).

HN OO

NN

Va

VIa

OPG( )n

HN OO

NN

OPG( )n

HN OO

NN

OPG

TFA

( )nIVa

DDQ


246

Diversity of Solid Forms of Felodipine: Structure and

Physicochemical Properties

A.O. Surov, G.L. Perlovich

G.A. Krestov Institute of Solution Chemistry of RAS, 153045, Russia, Ivanovo, Academicheskaya St.,1


In many cases, active pharmaceutical ingredients (APIs) are established to exist in different

crystal forms such as polymorphs or solvates/hydrates. Different polymorphs of a given crystalline

drug substance may have different physicochemical properties such as melting temperature,

solubility, stability, dissolution rate, and bioavailability which may affect their usefulness in drug

formulations. Solvates in general also play a significant part in drug development, and solid-form

screening to identify solvates/hydrates is a crucial prerequisite for consistent manufacturing and

processing of drugs. Accidental solvate formation is highly undesirable during processing of drugs

due to lack of predictability and, as a consequence, potentially

uncontrollable changes of the physicochemical properties of a

product. Systematic investigation of the the well-known calcium-

channel blocking drug felodipine (figure 1) revealed that it has at

least four distinct polymorphic modifications, and two of which are

practically isoenergetic [1]. It was found that the separate

crystallization of the different polymorphs can be effectively controlled by adjustment of the pH

values of the respective crystallization media. In addition, felodipine exhibits a strong propensity

for solvate formation with various solvents. So far, nine solvates of the drug, including structures

with common organic solvents such as dimethyl sulfoxide, dimethylformamide and acetone, were

isolated and characterized. [2] The diversity of the crystal forms were investigated by a wide

spectrum of experimental methods and approaches: X-ray diffraction, thermal analysis, solution

calorimetry and dissolution. In this work, we consider all experimental data obtained so far in a

systematic manner, with an aim to rationalize the relationships between the crystal structures of the

felodipine solid forms and their observed physicochemical properties, which is an important issue in

a pharmaceutical development.

This work was supported by a Grant from the President of the Russian Federation no. МК-

67.2014.3 and Russian Foundation for Basic Research (project № 14-03-31001).

References [1] A.O. Surov, K.A. Solanko, A. D. Bond, G.L. Perlovich, A. Bauer-Brandl, Cryst. Growth Des. 2012, 12, 4022–4030; [2] A.O. Surov, K.A. Solanko, A. D. Bond, A. Bauer-Brandl, G.L. Perlovich, CrystEngComm. 2015, DOI: 10.1039/C5CE00350D.

N

O

OO

OCH3

CH3

CH3CH3

H

Cl

Cl Figure 1. Molecular

structures of felodipine


247

Using of Modern Preparative Chromatography Methods in Biotechnology

O.A. Pisarev

Institute of Macromolecular Compounds RAS , St.-Petersburg,V.O., Bolshoi pr.31, Russia


Because their technological flexibility preparative chromatography on sorbents of various

structure become in the present time the most effective method of isolation, purification and

superpurification of biologically active substances (BAS). Superpure BAS are very necessary for

biotechnology and for pharmaceutical industry and their adjacent fields.

The report deals with the structure and properties of highly selective crosslinked polymeric

sorbents synthesized mainly by radical copolymerization of acrylic acids and various polyvinyl

crosslinking agents and with the application of these polymeric sorbents for low – pressure effective

preparative chromatography of biologically active substances.

The two-stage chromatographic method of preparative processes realization, which permit the

degree of content of basis substance in final products, was elaborated. By this method and by the

method of frontal-displacement chromatography in super state antitumor antibiotics rubomycin,

doxorubicin, carminomycin were obtained.

Organic anion – fusidic acid – was purified from the native solution by the method of coion-

hydrophobic chromatography when the formation of sharp boundaries was determined by the more

rapid kinetics of coions sorption in comparison with counterions.

The displacement chromatography of human serum albumin by using of low-molecular

displacement substances was investigated and was shown that these substances possesses by the

ability of selective displacement of protein. By definite concentration of displacement substance the

aim protein appears from the column by displacement mechanism. Another components with the

more high and the more low sorption ability may be elute or desorbed by the suitable solvent.

By using of kinetic selectivity effect the methods of purification of bee venom components

(melittin, phospholipase A2), hydrolytic enzymes ribonuclease, desoxyribonuclease, trypsin and

chymotrypsin was elaborated. The effect of kinetic selectivity allow essentially to increase (with

comparison of traditional methods of preparative chromatography) the velocity and effectivity of

substances separation. This method opens additional possibilities for isolation and purification of

labile BAS with close values of distribution coefficients between mobile and immovable phases

and low velocity of diffusion. The effect of kinetic selectivity deal not with concrete mechanism of

sorption and, consequently, may be realized in equal degree in ion- exchange, hydrophobic,

distribution and another kinds of chromatography.


248

Mechanochemistry in Technology Bioconversion of Plant Raw Materials

E.M. Podgorbunskikh, A.L. Bychkov, O.I. Lomovsky

Institute of Solid State Chemistry and Mechanochemistry SB RAS, 630128, Russia, Novosibirsk,

Kutateladze str., 18


Enzymatic hydrolysis of lingocellulosic biomass into monosaccharides with subsequent

fermentation to produce bioethanol is an important branch of biotechnology. Agricultural wastes are

the most promising feedstock for production of renewable biofuels [1]. Among the modern methods

of bioconversion of lingocellulosic biomass into soluble sugars is the most effective solid-phase

techniques associated with mechanical pretreatment and subsequent heterogeneous enzymatic

hydrolysis of substrate. Mechanical activation is helpful to increase the reactivity of lignocellulosic

materials by changing the morphology, structural disordering, surface area and crystallinity of

cellulose [2].

The purpose of this research work was to study processes occurring during mechanical activation

difficult convertible plant raw materials and leading to the production of high reactivity products

suitable for further enzyme conversion into low molecular weight carbohydrates.

We used the following “model” conditions of mechanical activation of plant materials: at -

196°C, when the components are in the vitreous state, and 180°C, when lignin and hemicellulose are

in highly elastic state. Mechanical activation at optimal conditions leads to increase yield of the

enzymatic hydrolysis, which can be explained by an increasing of surface area, reducing the degree

of crystallinity of cellulose, structural disordering of lignocellulose.

Mechanical pretreatment of rice husk in a semi-industrial centrifugal roller mill leads to

increasing in the specific surface area in 4 times, reducing the degree of crystallinity of cellulose in

2 times and increasing yield of low molecular weight sugars in 7 times, which is a significant

contribution to the implementation of biocatalytic conversion of lignocellulose using industrial

machines [3].

References [1] REH. Sims et al., Bioresource Technology. 2010, 101, 1570-1580. [2] AL. Bychkov et al., Cellulose Chemistry and Technology, 2014, 48, 545 – 551. [3] EM. Podgorbunskikh et al., Catalysis in Industry (in press).


249

Synthesis and Cytotoxicity Evaluation of Benzene-fluorinated 2-Arylindoles

L. Politanskaya,1 V. Shteingarts,1* O. Zakharova2 and G. Nevinsky2


Novosibirsk, Lavrentiev av., 9 2 Institute of Chemical Biology and Fundamental Medicine, SB RAS, 630090, Russia, Novosibirsk,

Lavrentiev av., 8


The polyfluorinated indole nucleus is a structural component in a vast number of biologically

active natural and synthesized compounds and pharmaceuticals [1]. We have demonstrated an

efficient method for the synthesis of polyfluorinated 2-arylindoles including the Sonogashira cross-

coupling of fluorinated ortho-iodanilines with ethynylarenes and the following cyclization of

prepared ortho-alkynylanilines in alcoholic solvents catalyzed by p-toluenesulfonic acid.

X1 X2 X3 X4 Alk

1 F H H H Me 67%

2 F F H H Et 56%

3 CF3 F H H n-Bu 64%

4 F H F NH2 Et 78%

5 F F F NH2 Et 55%

6 CF3 F F NH2 n-Bu 58%

Pd(PPh3)2Cl2,CuI, Et3N PTSA

MeCN AlkOH

Alk = Me, Et, n-Bun = 3 _ 4

NH2

I

Fn

R = H, p -NH2, m-F

RNH

F

X1

X2

F1_ 6

X3

X4

Compounds 1─6 were examined for their ability to

inhibit the growth of two mammalian cell lines: tumor

cell lines from human mammary adenocarcinoma

(MCF-7) and human myeloma (RPMI 8226) as well as

normal Chinese hamster Ag 17 cells (AG). It has been

revealed that the polyfluorinated compound 3

demonstrates the best cytotoxicity in all tested tumor

cell lines, the average IC50 value towards tumor cell

lines being approximately 5-fold lower than that towards normal cells.

The work is supported by the RFBR (grant № 14-03-00108) and by the interdisciplinary grant from SB RAS № 98. References [1] L. Bjeldanes, H. Le, G. Firestone, U.S. Patent 2005/58600, (A1) 2005 [2] J. Frost, M. Dart, K. Tietje, T. Garrison, G. Grayson, A. Daza, O. El-Kouhen, L. Miller, L. Li, B. Yao, G. Hsieh, M. Pai, C. Zhu, P. Chandran, M. Meyer, J. Med. Chem. 2008, 51, 1904–1912. *Deceased

Compound

IC50

MCF-7 RPMI AG

1 27.1 14.5 38.9

2 7.7 21.0 22.5

3 5.6 6.0 26.7

4 38.4 8.0 27.0

5 35.1 10.0 19.4

6 18.8 9.5 19.3


250

Molecularly Imprinted Polymers as New Generation of Sorbents with Properties

of Artificial Receptors to Biologically Active Substances

I. Polyakova,1 N. Ezhova,1 A. Leshchinskaya,1 I. Garkushina,1 L. Borovikova,1

A. Osipenko,2 O. Pisarev1,2

1Institute of Macromolecular Compounds of Russian Academy of Sciences, 199004, Russia, Saint-

Petersburg, VO Bolshoy pr.,31 2Departmen of Medical Physics, Peter the Great Saint-Petersburg Polytechnic University, 195251

Russia, Saint-Petersburg, Polytechnicheskaya str., 29,


Molecular imprinting is a technique of making molecularly imprinted polymers (MIPs), the

artificial materials of new generation with properties of artificial enzyme-like receptor. MIPs attract

great attention in down-stream processes such as isolation and purification of medicinal substances

in biotechnology, or removal of toxic compounds from blood in hemoperfusion treatment due to

their advantages such as pre-determined selectivity and chemical stability. In this method, a target

molecule as a template (T) is introduced into polymerization bulk. After polymerization, the T is

removed from the matrix leaving cavities (imprint-sites) complementary in shape and size to target

molecule. In our work, molecularly imprinted polymers (MIPs) were prepared via bulk

polymerization and microemulsion polymerization. L-lysine (Lys), erythromycin (Erm), uric acid

(Ua) and cholesterol (Ch) have been used as the T to prepare imprinted polymers LysMIP,

ErmMIP, UaMIP and ChMIP, correspondently. Methacrylic acid and hydroxyethyleneglycol

methacrylate were used as functional monomers (M) and ethyleneglycol dimethylacrylate was used

as a crosslinker (X) to prepare the MIPs. To improve the binding performance of the MIPs and their

surface homogeneity and to shift toward higher values the energy of the different types of sites, the

synthesis method was optimized for the effect of different the X/M/T ratio both on the polymers

structural stability in wide the pH range and on the MIPs electrochemical properties. Investigation

of the nature of intermolecular interactions in the studied sorption systems was carried out on the

sorption isotherms. The main parameters (the distribution coefficients) and thermodynamic

functions (∆G, ∆H and ∆S) of the templates as the aim molecules sorption on the MIPs were

compared with those obtained on the reference non-imprinted polymers (NIP). This allowed

studying the influence of recognition conditions and estimating the contribution of specific and non-

specific binding into the sorption selectivity of the templates on the MIPs. The sorption conditions

for shifting the distribution energies toward higher affinity sites and finding the mobile-phase

composition to improve peak shape on the MIPs were found.

Acknowledgements This research was supported by Russian Foundation for Basic Research (project № 15-03-07968).


251

Analgesic Activity of New Diazaadamantanes Containing Pinene Fragment

K. Ponomarev,1 E. Suslov,1 A. Pavlova,1 O. Ardashov,1,2 D. Korchagina,1 A. Nefedov,1,2

T. Tolstikova,1,2 K. Volcho,1,2 N. Salakhutdinov1,2

1N.N. Vorozhtsov Novosibirsk Institute Of Organic Chemistry, 630090, Russia, Novosibirsk, Acad.

Lavrentyev Avenue 9 2Novosibirsk State University, 630090, Russia, Novosibirsk, Pirogova Str. 2


New diazaadamantanes 3a-d containing pinene fragment were synthesized by the interaction

between dimethylbispidinone 1 and aldehydes 2a-d for the first time. The analgesic activity of the

compounds 3a-d was studied in vivo in the hot plate and acetic acid-induced writhing tests.

It revealed that compound 3a in dose 20 mg/kg demonstrated good analgesic activity with a

higher efficacy then the reference drug, diclofenac sodium.

A decrease in the analgesic effect in the acetic acid-induced writhing test and the complete

disappearance of the activity in the hot plate test was observed in animals administrated with

enantiomeric compound 3b. Therefore, the absolute configuration of the compound has a key

influence on the manifestation of its analgesic activity. Addition of the CH2-group during the

transition from 3a to 3c or the keto group during the transition to 3d led to the complete

disappearance of analgesic activity by these compounds.

Thus, we have shown that the structure of the (-)-pinane fragment in the compound 3a is

important for analgesic activity.

We found that compound 3a exhibits a low acute toxicity (LD50 >1000 mg/kg), does not cause

damage to the gastric mucosa, and its analgesic effect is, at least partially, mediated by the

cannabinoid system involving CB1 receptors.

The reported study was supported by RFBR, research project No. 15-03-01092a.


252

Novel Synthetic Approaches to Organochalcogen Compounds with Potential

Biological Activity

V.A. Potapov, M.V. Musalov, E.O. Kurkutov, R.S. Ishigeev, A.S. Filippov, M.V. Musalova,

A.G. Khabibulina, and S.V. Amosova

A. E. Favorsky Irkutsk Institute of Chemistry, Siberian Branch of the Russian Academy of Sciences,

664033, Russian Federation, Irkutsk, Favorsky Str., 1


A number of novel synthetic approaches to organochalcogen compounds with potential

biological activity have been developed based on chalcogen halides.

Selenium dichloride and dibromide exist in equilibrium and can not be isolated in a pure form.

However, we have found that freshly prepared in situ selenium dihalides in solution can take part in

various reactions to give useful products with potential biological activity in high yields.

Another approach is based on the reactions of pyridylchalcogenyl halides with unsaturated

compounds leading to condensed heterocycles. For example, the reaction of 2-pyridylsulfenyl

chloride with 4-vinylpyridine afforded 3-(4-pyridyl)-2H,3H-[1,3]thiazolo[3,2-a]pyridin-4-ium

chloride. Annulation of the hexahydrobenzothiazole cycle to the pyridine ring occurred in the

reaction of 2-pyridylsulfenyl chloride with cyclohexene.

SeHal2

O

SeCl

Se

S HalHal

OO

Se

S HalHalOO

SOO

OH

O

O

Se

+

N S

N SCl

Cl

N S

Cl

N

N

Se

HalHal

O O

OHHO

Hal = Cl, Br

X

SeHalHal

OPh O

HO

X

X = O, S, Se

Efficient synthesis of tetraalkylammonium trichloro(dioxoethylene-O,O')tellurates has been

developed. These compounds represent analogs of known medicament AS-101, which is used for

treatment of clinical immunosuppression conditions involving AIDS.


253

Eudesmanolides of Inula helenium and Its

Aminoderivatives with Cytotoxic Activity

S. Pukhov, L. Anikina, S. Afanasyeva and S. Klochkov

Laboratory of Natural Compounds, Institute of Physiologically Active Compounds Russian

Academy of Sciences, 142432, Russian Federation, Chernogolovka, Severniy proyezd, 1


Natural sesquiterpene eudesmane type lactones are considered as compounds with a wide range

of biological activities [1]. Alantolactone (1), isoalantolactone (2) and epoxyalantolactone (3) are

the secondary metabolites of I. helenium L. and we have shown that this plant origin natural

products inhibit the growth of various cancer cell lines: MCF7, MS and HCT116. In addition the

modification of natural lactones by amines gives the some derivatives (4-15) with an activity

exceeding the starting compounds.

IC50 values of tested compounds

0

10

20

30

40

50

1 4 2 5 6 7 8 3 9 10 11 12 13 14 15Compound

IC50

, uM

MCF7MSHCT-116

The synthesized derivatives and natural sesquiterpene lactones have been tested against the panel

of cancer cell lines by MTT-assay. Therefore, the representatives of derivatives attract interest by

way of promising biologically active compounds for further investigation of mechanism of action

on cancer cells and developing a potent new anti-neoplastic agents.

References [1] I. Merfort, Curr. Drug Targets. 2011, 12(11), 1560-1573 [2] S. Pukhov, M. Neganova, L. Anikina, E. Shevtsova, S. Afanasyeva and S. Klochkov, Fund. Res. 2014, 9(9), 1988-1992


254

Anti-virus Activity Evaluation of Camphor Derivatives Using Pseudoviruses

Yu. Razumova,1 A. Sokolova,1,2 O. Yarovaya,1,2 A. Pokrovsky,1 N. Salakhutdinov1,2

1Novosibirsk State University, 630090, Russia, Novosibirsk Pirogova St. 2.

2N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, 630090 Russia, Novosibirsk, Lavrentjev Avenue 9


Pseudovirus containing envelope proteins from influenza A virus (HA and NA) on a background

of HIV core proteins are useful surrogates for infectious virus. Previously, our groups have shown

that camphor derivatives display antiviral activity against the pandemic 2009 H1N1 influenza

virus1,2. In this study, a novel series of camphor derivatives were synthesized and tested for their

inhibiting activity against H5N1 pseudovirus. In parallel these compounds were tested for their

inhibiting activity against pseudovirus containing an envelope G-protein from vesicular stomatitis

virus (VSV-G pseudovirus). The compounds designed and synthesized are listed in Figure 1 and

their inhibiting activity are presented in Table 1. All compounds showed low toxicity. All

compounds possess inhibitory activities against pseudoviruses. Among them, compound 4 is the

most potent inhibitor. Compounds 3, 4 and 15 inhibit H5N1 pseudovirus with a similar potency to

that of VSV-G pseudovirus, whereas compound 1 inhibit H5N1 pseudovirus better than VSV-G

pseudovirus. Thus H5N1 pseudovirus can be used for anti-influenza activity screening and

structure–activity relationship study of camphor derivatives.

Table 1. Inhibitory activities of camphor derivatives against pseudoviruses.

Compound Against H5N1 pseudovirus, ED50, мМ Against VSV-G pseudovirus, ED50, мМ 1 1,070 ± 0,058 1,815 ± 0,125 2 1,292 ± 0,088 1,528 ± 0,090 3 0,986 ± 0,050 0,974 ± 0,048 4 0,047 ± 0,002 0, 040 ± 0,002 5 0,365 ± 0,021 0,376 ± 0,023

This work was supported by the RFBR grants 15-03-00193_a, 14-04-01461_a.

References 1. Sokolova et al. Bioorg. Med. Chem 2013;21: 6690–8. 2. Sokolova et al. Bioorg Med Chem. 2014;22(7):2141-8

Figure 1. Chemical structures of camphor derivatives


255

Virtual Screening for Novel Atg5-Atg16 Complex

Inhibitors for Autophagy Modulation

J. Reynisson,1 A. Richardson,2 E. Robinson2, E. Leung,3 A. Matuszek,1 N. Krogsgaard-

Larsenn,1 D. Furkert,1 and M. Brimble1

1School of Chemical Sciences, University of Auckland, New Zealand 2Institute for Science and Technology in Medicine, Keele University, Guy Hilton Research Centre,

Stoke-on-Trent, United Kingdom 3Auckland Cancer Society Research Centre, University of Auckland, New Zealand

Cellular homeostasis is maintained by a balance between the synthesis and break down various

cytosolic components. Most of the selective degradation of these structures occurs in the cytosol

through the ubiquitin/proteasome system.1,2 The autophagy process is initiated by appearance of

isolation membranes (phagophores) in the cytoplasm. These membrane structures assemble to

generate an autophagosome that truncates a small part of the cytosol containing the components

determined for degradation. The autophagy process is initiated by a complex cascade of Atg

(autophagy) proteins of which 18 have been identified to date. It is hypothesised that inhibition of

this Atg5-Atg16 complex can modulate the autophagy process, which is of great interest, since

autophagic dysfunction is associated with a number of severe diseases, in particular cancer.1 A host

of small molecule compounds are known to induce autophagy, however no compounds have been

reported that directly inhibit it.3 The x-ray crystals of many of the proteins involved in the

autophagy cascade are available allowing for a virtual high throughput screen (vHTS) to be

conducted to identify small molecules, which are able to modulate the autophagy formation.2, 4, 5

The binding pocket of the ATG5 protein where the natural polypeptide ligand binds (Atg16) was

used for the vHTS campaign to block this interaction. From an initial collection of 5×104 candidates

three clear hits were identified using the A2780 ovarian cancer cell line, i.e., they stopped the

progression of autophagy. Furthermore, preliminary structural activity relationship (SAR)

investigations for the hits were performed. These hit compounds are the first to be reported to be

able to directly modulate autophagy formation and therefore have the potential to be developed into

anticancer therapeutic agents.

1 N.N. Noda, Y. Ohsumi, F. Inagaki, Chem. Rev., 109 (2009) 1587-1598. 2 Y. Fujioka, N.N. Noda, H. Nakatogawa, Y. Ohsumi, F. Inagaki, J. Biol. Chem., 285 (2010) 1508-1515. 3 A. Fleming, T. Noda, T. Yoshimori, D.C. Rubinsztein, Nature Chem. Biol., 7 (2011) 9-17. 4 M. Matsushita, N.N. Suzuki, K. Obara, Y. Fujioka, Y. Ohsumi, F. Inagaki, J. Biol. Chem., 282 (2007) 6763. 5 J. Reynisson, W. Court, C. O’Neill, J. Day, L. Patterson, E. McDonald, P. Workman, M. Katan, S.A. Eccles, Bioorg.

Med. Chem., 17 (2009) 3169-3176.


256

The Nature of Known Drug Space

J. Reynisson

School of Chemical Sciences, University of Auckland, New Zealand

Known drug space (KDS) is defined as all molecules that are used in the clinic [1]. It can be used

as a metric in exploring ideas within the framework of drug discovery programmes, i.e., the

presence of certain undesirable molecular moieties [2]. This begs the question what is the nature of

KDS? It was analysed for the occurrence of natural products and their derivatives. It was found that

10% of the drugs on the market are unaltered natural products, 29% are their derivatives (semi-

synthetics) and the rest (61%) have a synthetic origin [3]. Also, KDS occupies a larger volume in

chemical space with respect to drug-like chemicals in respect to mainstream molecular descriptors

(see figure) [3]. Furthermore, the size of chemical space is estimated to be 1.6×109 compounds

based on the power growth ((f(x) = AxB, Carbon=x)) of known organic compounds. The statistical

distribution of KDS and drugs with good

oral-bioavailability (drug-like) based on

their carbon atom count was used to

deduce the size of these sub-regions

within chemical space. KDS is predicted

to be 2.0×106 molecules and drug-like

chemical space is calculated to be 1.1×106

compounds [4]. Finally, it was found that

serendipity has played a crucial part in

populating KDS [5].

[1] A. Mirza, R. Desai, J. Reynisson, Eur. J. Med. Chem., 44 (2009) 5006-5011. [2] P. Axerio-Cilies, I.P. Castañeda, A. Mirza, J. Reynisson, Eur. J. Med. Chem., 44 (2009) 1128-1134. [3] R. Bade, H.-F. Chan, J. Reynisson, Eur. J. Med. Chem., 45 (2010) 5646-5652. [4] K.L.M. Drew, H. Baiman, P. Khwaounjoo, B. Yu, J. Reynisson, J. Pharm. Pharmaco., 64 (2012) 490–495. [5] E. Hargrave-Thomas, B. Yu, J. Reynisson, W. J. Clin. Oncol., 3 (2012) 1-6.


257

5′-(1,2,3-Triazolyl)substituted Lappaconitine Derivatives

V.E. Romanov, E.E. Shults

Novosibirsk Institute of Organic Chemistry, SB RAS,

630090 Novosibirsk, Lavrentjev Ave. 9, Russia, E-mail [email protected]

Huisgen’s dipolar cycloaddition of organic azides and alkynes is the most direct route to 1,2,3-

triazoles [1]. The discovery that Cu(I) efficiently and regiospecifically unites terminal alkynes and

azides, providing 1,4-disubstituted 1,2,3-triazoles under mild conditions, was a welcome advance

[2]. Our laboratory has had a long-term focus on the chemistry of plant metabolites, in particular, on

the use of metal-catalyzed reactions for transformations of natural compounds including diterpenoid

alkaloids [3,4]. The purpose of this communication is to study the CuAAC reaction of 5'-

ethynyllappaconitine 1 with several organic azides 2-8. New derivatives of lappaconitine 9-15 are

obtained by treatment 5'-ethynyllappaconitine 1 with azides 2-8 in the presence of copper sulfate

(II) and sodium ascorbate in DMF under heating. The isolated yield of 1,4-disubstituted 1,2,3-

triazoles 10,12-15 in all cases was good; an exception were reactions with azides 2 and 11, which

are heat-labile.

C O

NHAc

OCH3

OCH3

HOH3CO

N

O

OH

15

5

14

7

8

1

3

16

17

13

20

12

1'2'5'HC

1

2-8

CuSO4.5H2O

AscNaDMF, 70 оС

C O

NHAc

OCH3

OCH3

HOH3CO

N

OOH

NN

N

R

19

10

18

9 (40%), 10 (65%), 11 (46%), 12 (73%), 13 (70%), 14 (63%), 15 (68%)

R N3

2, 9: R = Ph; 3, 10: R = Bn; 4, 11: R = 4-OH-C6H4; 5, 12: R = 4-OH-C6H4CH2;6, 13: R = 4-OH-C6H4(CH2)2; 7, 14: R = 4-OH-C6H4(CH2)3; 8, 15: R = 4-OH-C6H4(CH2)4

1''4''

5''

This work was supported by the Russian Science Foundation (project. No 14-13-00822). [1]. Huisgen, R. In 1,3-Dipolar Cycloaddition Chemistry; Padwa, A., Ed.; Wiley: New York, 1984. [2]. (a) Rostovtsev, V. V.; Green, L. G.; Fokin, V. V.; Sharpless, K. B. Angew. Chem., Int. Ed. 2002, 41, 2596. (b)

Tornoe, C. W.; Christensen, C.; Meldal, M.J. Org. Chem.2002, 67, 3057 [3]. Osadchii, S.A., Shul’ts, E.E., Polukhina, E.V., Shakirov, M.M., Tolstikov, G.A. Rus. Chem. Bull., Int. Ed. 2006,

55, 1077. [4]. Romanov V.E., Shults E.E., Shakirov M.M., Tolstikov G.A., Russ. J. Org. Chem. 2011. V. 47. P. 581-588.


258

Solvent-free Multicomponent Synthesis of Pyrano[2,3-dpyrimidines]

F. Ryzhkov, M. Elinson

N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 119991, Russia,

Moscow, Leninsky prospect 47, +7-499-137-38-42


Pyrano[2,3-d]pyrimidines have received considerable attention owing to their wide range of

diverse pharmacological activity. They are nicotinic acid receptor (NAR) agonists [1] and show

antitumor, hepatoprotective, antibronchitic, and anti-AIDS activity [2], among others.

The development of solvent-free organic synthesis has become an important research area. This

is not only due to the need for the more efficient and less labour-intense methodologies for the

synthesis of organic compounds, but also because of the increasing importance of the environmental

considerations in chemistry. The elimination of volatile organic solvents in organic synthesis is also

the most important goal in ‘green chemistry’.

We were prompted to use a convenient and facile solvent-free cascade methodology for the

synthesis of pyrano[2,3-d]pyrimidines scaffold from benzaldehydes, N,N’-dialkyl barbiturates and

malononitrile. We have found that potassium fluoride as catalyst can produce, under solvent-free

conditions, approach to substituted pyrano[2,3-d]pyrimidines in 89-95% yields.

The catalytic procedure utilizes simple equipment; it is easily carried out and is valuable from

the viewpoint of environmentally benign diversity-oriented large-scale processes. This new process

opens an efficient and convenient solvent-free multicomponent way to synthesize substituted

pyrano[2,3-d]pyrimidines – the promising compounds for a variety of biomedical applications.

References [1] Huang, X.; Su, J.; Rao, A. U.; Tang, et al. Biorg. Med. Chem. Lett. 2012, 22, 854–858 [2] Furuay, S.; Ohtaki, T. Eur. Pat. Appl. EP 608565, 1994


259

Synthesis and Investigation of 1,2,4-Triazolo[5,1-c]-1,2,4-triazines

Antiviral Action

I.M. Sapozhnikova, N.R. Medvedeva, E.N. Ulomskii, and V.L. Rusinov

Department of Organic Chemistry, Ural Federal University, 620002, Russian Federation,

Ekaterinburg, Mira Str., 19


Antiviral drug "Triazavirin" (1) refers to a class of triazolo[5,1-c]-triazine. Its characteristic

feature is high efficiency in vivo, in contrast to the experiments in vitro, that suggests its possible

metabolic transformations. In present report we examine the nucleophilic substitution of the nitro

group, which on the one hand, simulates possible metabolic transformations, on the other - allows to

expand a number of biologically active compounds.

Due to the fact that the substitution of the nitro group in the "Triazavirin" is hindered, derivatives

having an alkyl substituent on the nitrogen atom were synthesized, in which a nitro group is readily

replaced by various biogenic S- and N-nucleophiles. As a model of S-nucleophiles were selected L-

cysteine and L-cysteine-containing tripeptide glutathione; as N-nucleophiles - various amines, and

the ethyl ester of L-lysine.

N

N

N

NN

O

MeS

NO2

R

N

N

N

NN

O

MeSR

SR'N

N

N

NN

O

MeS

SR'

NH4+

N

N

N

NN

O

MeS

N

R

N

N

N

NH

N

O

MeS

N

NH

NH

N

N

N

NH

N

O

MeS

NO2

R= Me, Et, Bn, t-Bu, CH2OC(O)C(CH3)3, (CH2)4OC(O)CH3, CH2O(CH2)2OC(O)CH3

HS-R'

HS-R' = L-cysteine, L-gluthatione

= BuNH2, s-BuNH2, HO(CH2)2NH2, pyrrolidine, morpholine, L-lysine ethyl ester

R= Me, Et, Bn, t-Bu, (CH2)4OC(O)CH3, CH2O(CH2)2OC(O)CH3, CH(CH3)OCH2CH3

-

1

2

3 4

5 6

(Na)

Investigation of the antiviral action of the compounds in in vitro experiments against influenza

virus (strains A/S.-Petersburg/16/09 A(H3N2), A/Victoria/35/72 A(H3N2), A/S.-Petersburg / 5/09

A(H1N1 )) showed a decrease of viral titer from 0.5 to 3.0 lg.


260

Thiadiazolo[1,5-a]pyrimidines as Perspective Precursors of Heterocycles with

Potentially High Biological Activity

K. Savateev, E. Ulomsky, V. Rusinov

1Ural Federal University, department of Organic chemistry, Ekaterinburg, street Mira, 19,

Ekaterinburg, 620002, Russian Federation


Regiospecific synthesis of biologically active 7-substituted purines is an important problem in

the synthesis of drugs belonging to the group of unnatural nucleosides. We propose an approach

that involves the synthesis of 7-R-purines (II) through the corresponding azolo[1,5-a]pyrimidines

(I).

XY

N

N

R1

R2K N

N

N

N

R3

X = Y = K = N X = N; Y = CH; K = SR1, R2 = nitrogen-containing groups

(I) (II)

The synthetic route from compounds (I) to purines (II) lies through conversion of oxoderivatives

(1) to the corresponding chloroheterocycles (2) with further amination (3) the latter. In this case,

one of the advantages of thiadiazolopyrimidines over tetrazole-analogues is the high stability of the

chloro-derivative (2): it can be isolated from the reaction mixture and analyzed. Accordingly, fewer

byproducts occurs in amination step and that simplifies the isolation of the desired products (3) and

increases its yield.

N N

NS OH

O

NO2 N N

NS Cl

O

NO2 N N

NS NH

O

NO2

R

O O

CH2

R = Bu; t-Bu; i-Bu; Pr; AcO(CH2)2-4;

(1) (2) (3)

POCl3

Py

RNH2

Furthermore, 5-alkylamino-6-nitrothiadiazolo[1,5-a]pyrimidines (3) are close structural

analogues of triazolotriazines which are adenosine receptor antagonists [1], as nitro group is a good

model of the cyclic pyridine N-atom, while the thiadiazole ring is an alternative to the triazole

cycle.

Thus, we synthesized a series of 5-R-amino-6-nitro-7-oksothiadiazolo[1,5-a]pyrimidines which

are a good precursors to the synthesis of a wide range of potentially biologically active

heterocycles. Selection of the substituents at the amino group based on literature data which

indicated that 8-alkyltriazolopurines and 7-R-purines exhibit a high biological activity [2].

We thank the Russian Scientific Foundation grant № 14-13-01301. References [1]: Jorg M., Shonberg J., Mak F., Miller N., Bioorg. And Med. Chem. Let., 2013, vol. 23, p. 3427-3433; [2]: Biamonte M.; Boehm M.; Fan J.; Hong K., J. Med. Chem., 2006, vol. 49, #17, p. 5352 – 5362.


261

ω-Oxo-functionalized 20-Hydroxyecdysone Derivatives in the

Synthesis of Aminoecdysteroids

R.G. Savchenko, S.A. Kostyleva, V.N. Odinokov

Institute of Petrochemistry and Catalysis Russian Academy of Science,

450075, Ufa, pr. Oktyabrya, 141


Ecdysteroids (polyhydroxylated sterols) are hormones of molting and metamorphosis of insects

and other arthropods. 20-Hydroxyecdysone (20-E 1) isolated by us from the plant Serratula

coronata L is the most widely studied and available representative of this class of compounds. The

structure and physiological properties of 20-E (is involved in protein, glucose, lipide metabolism)

are very attractive for the synthesis of target oriented derivative for medicine and agriculture.

We studied the reductive amination of ω-carbonyl derivatives of ecdysteroids. ω-Oxo-

functionalized ecdysteroids are practically important precursors in the synthesis of amino

derivatives which can to act as a neuromuscular block like known commercial aminosteroids.

Each of the aldehyde 3 and corresponding ketone 4 were prepared as a mixture (~2:1) by

ozonolysis of ω-anhydro derivative 2 and isolated by CC. Thus, the earlier unknown secondary

aminoecdysteroid 5 was synthesized using a reductive amination [RNH2/NaBH(OAc)3] of an 24-

aldehyde 3. On the other hand, the new primary amine 6 was obtained by oximation of 25-oxo

derivative 4 and the followed hydrogenation of the resulting -oxime.

The structure of and the gross composition of synthesized compounds were established by 1D

and 2D 1Н and 13С NMR and MS MALDI TOF.

OH

OHH

OH

OHOH

OHH

H

O

AcO

AcOH

OO

OHH

H

O

O

OO

OH

H

OO

OH

H

O

NH

OO

OH

H

RO

O

OH

H

NH2

N

N

O

CH3CH3

Ph

1. Me2CO/TsOH

2. Ac2O-Py/DMAP3. MsCl-Py/DMAP

O3

CH2Cl2-Py

24 25

+

1 23 4

1. RNH2

2. NaBH(OAc)3 yield 40-74%

1. NH2OH*HCl/ Py-Et3N2. H2/ Ni-Ra

yield 67%

5 6R = Pr, Ph,


262

Synthesis of 5-(2,4-Dimethylpyridin-4-yl)-trisnoroleanane

E.V. Salimova, E.V. Tretyakova, S.A. Kostyleva, R.M. Latypova, L.R. Miniyarova

Institute of Petrochemistry and Catalysis, Russian Academy of Sciences,

pr. Oktyabrya 141, Ufa, 450075 Bashkortostan, Russia


Medicinal plants have long been an excellent source of pharmaceutical agents because there are

available and possess potential activity. The main leaders for medical applications among

triterpenoids are glycyrrhizic acid, betulin, ursolic and oleanolic acid. A significant group of

biologically active triterpenoids are nitrogen-containing derivatives. The presence of nitrogen at

different positions of the molecular structure causes various activities [1-4]. 2,4-Dimethylpyridine

derivatives of oleanolic series was synthesized to examine their pharmacological activity.

Interaction of 3,4-secoallobetulin derivative (1) with acetylchloride in presence of AlCl3, followed

by treatment with ammonia lead to pyridine compound (2) formation with a yield of 70%.

Modification of 3,4-seconitrile-28-oxoallobetulone isopropyl group (3), under similar conditions

proceed with hydrolysis of the nitrile function into carboxyl group and compound (4) formed with

a yield of 62%.

O

N

N

O

N

O

N

N

O

OOOH

O

1. AcCl, AlCl3

2. NH3

1. AcCl, AlCl3

2. NH3

1 2 (70%)

3 4 (62%)

This study was performed under financial support by the Russian Foundation for Basic Research (project no. 14-03-97 046 r_Povolzh’e_a). References 1. Lee K.-H. J. Nat. Prod., 2010, Vol. 73, No. 3, P. 500. 2. Wang S.-R., Fang W.-S. Curr. Top. Med. Chem., 2009, Vol. 9, No. 16, P. 1581. 3. Kuo R.-Y., Qian K., Morris-Natschke S. L., Lee K.-H. Nat. Prod. Rep., 2009, Vol. 26, No. 10, P. 1321. 4. Laszczyk M. N. Planta Medica., 2009, Vol. 75, No. 15, P. 1549.


263

Synthesis and anti-TBEV Properties of 4-Aminopyrimidine N-oxides

K.N. Sedenkova,1,2 E.V. Dueva,1 D.I. Osolodkin,1 E.B. Averina,1,2 V.A. Palyulin,1,2

Yu.K. Grishin,1 T.S. Kuznetzova1 and N.S. Zefirov1,2

1 Chemistry Department, Lomonosov Moscow State University, 119991, Russia,

Moscow, Leninskie Gory, 1-3 2Institute of Physiologically Active Compounds, Russian Academy of Sciences,

142432, Russia, Chernogolovka, pr. Severniy, 1


Previously in our group a novel three-component heterocyclization involving gem-

bromofluorocyclopropanes 1, nitrosylating or nitrating agent and organic nitrile yielding previously

unknown 4-fluoropyrimidine 1-oxides 2 was found [1].

F

Br RCNN

N

F

O

R1 2

R1

R2

R1

R2

R1,R2,R= Alk, Ar[NO+]

A series of 4-fluorotetrahydroquinozaline 1-oxides 3 was obtained via this reaction and studied

in SNAr-processes under the treatment with various amines, bearing alkyl, (adamantyl)alkyl and

aryl substituents [2]. Preparative two-step method for the synthesis of 4-aminopyrimidine N-oxides

4 from readily available gem-dihalogenocyclopropanes was elaborated.

N

N

F

R

NHR'R'', DIPEA

N

N

NR'R''

REtOH

O O3 4

N

N

HN

O

2-Ad

N

N

HN

t-Bu

O

1-Ad

Ph

EC50 6.6 µM EC50 5.5 µM

yeilds up to 90%

The activity of tetrahydroquinozaline N-oxides 4 against tick-borne encephalitis virus (TBEV)

reproduction was assessed and a number of compounds were found to inhibit TBEV entry into the

host cells. Bulky hydrophobic substituents R, R’ and R’’ were identified to be important for the

antiviral activity.

We thank the RFBR (Project 14-03-00469-а) and the Presidium of RAS (Program N 8) for financial support. References [1] K.N. Sedenkova, E.B. Averina, Yu.K. Grishin, A.G. Kutateladze, V.B. Rybakov, T.S. Kuznetsova and N.S. Zefirov. J. Org. Chem., 2012, 77, 9893–9899. [2] K.N. Sedenkova, E.V. Dueva, E.B. Averina, Y.K. Grishin, D.I. Osolodkin, L. I. Kozlovskaya, V.A. Palyulin, E.N. Savelyev, B.S. Orlinson, I.A. Novakov, G.M. Butov, T.S. Kuznetsova, G.G. Karganova and N. S. Zefirov. Org. Biomol. Chem. 2015, 13, 3406–3415.


264

Effect of Natural Polysaccharides and Oligosaccharides on Permeability of Cell

Membrane – Implication in Drug Delivery

O.Yu. Selyutina,1,2 I.E. Apanasenko,1,2 N.E. Polyakov,1 A.G. Shilov3

1Institute of chemical kinetics and combustion SB RAS,630090, Russia, Novosibirsk, Institutskaya, 3

2Novosibirsk State University, Russia, Novosibirsk, Pirogova, 2 3Institute of cytology and genetics SB RAS, Lavrenteva, 10


Natural poly- and oligosaccharides are widely used in medical chemistry as a drug carriers or

drug delivery systems. The most well-known examples of such carriers are cyclodextrins (CD), a

family of compounds made up of sugar molecules bound together in a ring. Typically, CD can be

topologically represented as a toroid with hydrophobic interior. Another example examined in

present work is arabinogalactan (AG) which is the polymer consisting of arabinose and galactose

monosaccharides. And the last one is glicyrrhizic acid (GA) which is triterpene glycoside

extractedfromthe liquorice root. All of them demonstrate complex-forming properties with a wide

range of drug molecules resulting in increase of drug solubility, stability and bioavailability.

In the present work we've made an attempt to clarify the mechanism of their action on drugs

bioavailability in terms of molecule penetration through biological membrane. The study of

membrane permeability was carried out in vitro using red blood cells and myeloid leukemia cells.

The method used is nuclear magnetic resonance technique with paramagnetic ions adding. Using

this method one can split the external and internal environment and investigate the diffusion of

molecules through the membrane by measuring the relaxation time of protons.

We’ve shown that the presence of GA and AG reduces exchange time through K562 cell

membrane while there is no significant changes in the presence of CD. Sodium formate (SF) was

usedas a model molecule. These results are important for understanding the mechanism of the drugs

bioavailability enhancement in the presence of complex-forming agents.

SF is found at micromolar concentrations in human erythrocytes and its physiological role in

these cells is very little known. It is assumed that SF participates in the biosynthesis of purine.

Furthermore, it is known that SF is detected in leukocytes in much higher concentrations than in

erythrocytes and perhaps is involved in apoptosis. Therefore the effectof poly- and oligosaccharides

on penetration of SF through leukemia cell membrane could also have an application in cancer

treatment.


265

Synthesis and Anti-acetylcholinesterase Potency of Onium Uracil Derivatives

V. Semenov,1 I. Galyametdinova,1 L. Saifina,1 I. Zueva,1,2 K. Petrov,1,2 V. Zobov,1 V. Reznik1

1A.E. Arbuzov Institute of Organic and Physical Chemistry, 420088, Russia, Kazan, Arbuzov str., 8

2Kazan Institute of Biochemistry and Biophysics, 420111, Russia, Kazan, Lobachevsky str., 2/31


Acetylcholinesterase (AChE) inhibitors are widely used in medicine for pharmacological

correction of cholinergic neurotransmission pathologies. The efficacy of antiAChE drugs is based

on their ability to potentiate the effects of acetylcholine due to a decrease in the rate of hydrolysis

by AChE.

We developed a novel class of AChE inhibitors based on onium uracil and condensed uracil

derivatives with acyclic topology (compounds 1,2) and macrocyclic topology (pyrimidinophanes 3)

[1]. The compounds 1-3 were synthesized starting from 1,3-bis(ω-bromoalkyl)-6-methyluracils or

1,3-bis(5-bromopentyl)quinazoline-2,4-dione, followed by amination with NH2Et or NHEt2,

alkylation of N atoms with substituted benzyl bromide or p-dibromoxylylene(alkane), and finally

alkylation of N atoms of macrocycles with substituted benzyl bromide.

2Br

1

2

34

5

6

1: n=2-4

N

NN

NO

O

n

n

R1

R3

R4R5

R2

R4

R1 R2

R3R5

2Br

2

N

NN

NO

O

3

3

R1

R3

R4R5

R2

R4

R1 R2

R3R5

N

NN

NO

O

3

3

R1

R3

R4R5

R2

R4

R1 R2

R3R5

3: X=(CH2)3,6,9;

X

R1,R2,R3,R4,R5=H, NO2, CN, F, CF3

2Br

The acyclic compouns and their macrocyclic counterparts demonstrate high selectivity against

AChE, and their IC50 (0.5-50 nM) towards butyrylcholinesterase (BuChE) exceed 3-4 order the IC50

towards AChE. The dependence of the toxicity and efficiency against AChE versus BuChE of the

compounds on the topology, number of methylene groups in polymethylene chains, substituent at

benzene ring of the benzyl moieties, structure of the spacer linking 5-(ethyl-substituted-

benzylammonia)pentyl units for pyrimidinophanes 3 was determined. We found that some of the

compounds discussed restored the value of amplitude decrement of integral muscle in animals with

experimental miastenia gravis to the value of healthy animals, and hence the onium uracil

derivatives can be considered as a potential remedy for the treatment of pathological muscle

weakness.

The financial support of RFBR (grant #13-04-40288-Н) is acknowledged.

References [1] V.E. Semenov, R.K. Giniyatullin, S.V. Lushchekina, etc. MedChemComm. 2014, 5, 1729-1735.


266

Synthesis and Biological Properties of Pyrrolylbenzoxadiazoles – Potential Nitric

Oxide (II) Donors

Yu.P. Semenyuk,1 P.G. Morozov,1 E.V. Prazdnova,2 O.N. Burov1

1Department of Chemistry, Southern Federal University, 344090, Russia, Rostov-on-Don, Zorge, 7 2 Academy of Biology and Biotechnology, Southern Federal University, 344090, Russia, Rostov-on-

Don, Stachki, 194/1


Nitric oxide (II) is a regulator of many physiological processes (of the immune and nervous

systems, relaxation of the blood vessels), pathological states (tumor, infectious, inflammatory

disease) in humans [1]. It is known that nitrobenzoxadiazoles are powerful sources of exogenous

nitric oxide (II) [2]. In this work we report about synthesis new dinitrobenzoxadiazole derivatives

and their ability to generate NO.

YO

N

Cl

NO2

O2N NR

YO

N

NO2

O2N

N R

YO

N

NO2

O2N

NR

1 a,b 2 a,b

a Y = N b Y = N+-O-

a R = Bn b R = Me

a Y = N; R = Bn (67%)b Y = N+-O-; R = Me (44%)

3 a,b 4 a,b

a Y = N; R = Bn (11%)b Y = N+-O-; R = Me (33%)

+ +

NO generation in vivo of all synthesized compounds was quantitatively defined using genetic

engineering Lux-biosensors (strain) E. coli MG 1655 pSoxS-lux. The most active inductor 4b is an

order of magnitude more effective than the comparator drug nitroglycerin. The absence of toxicity

for all compounds and the presence of the DNA-protective effect of compound-leader 4b were

found using biosensor E. coli MG 1655 (pXen7-lux) and biosensor E. coli MG 1655 (pRecA-lux)

respectively.

Compound 4b undergoes 1,3-N-oxide tautomerism possibly facilitating the generation of nitric

oxide. Compound structures and mechanism of the 1,3-N-oxide tautomerism were studied by NMR

spectroscopy, X-ray analysis and quantum chemistry.

The study was supported by the Russian Science Foundation, the project № 14-13-00103. References [1] P. G. Wang, T. B. Cai, N. Taniguchi, Nitric Oxide Donors: For Pharmaceutical and Biological Applications, Wiley-VCH, 2005, 407p. [2] V. G. Granik, N. B. Grigoriev, Russ. Chem. Rev. (Engl. Transl.), 2011, 80, 171.


267

Preparation of Solubilized Forms of Meloxicam

S.A. Myz,1,2 A.G. Ogienko,2,3 T.N. Drebushchak,1,2

T.P. Shakhtshneider,1,2

A.A. Ogienko,4 E.V. Boldyreva1,2

1 Institute of Solid State Chemistry and Mechanochemistry SB RAS, Novosibirsk, Russia

2 Novosibirsk State University, Novosibirsk, Russia 3 Nikolaev Institute of Inorganic Chemistry SB RAS, Novosibirsk, Russia

4 Institute of Citology and Genetics SB RAS, Novosibirsk, Russia

The one of the most important problems in the modern pharmaceutical industry is modification of drugs to improve their bioavailability. Meloxicam, 4-hydroxy-2-metyl-N-(5-methyl-2-thiazolyl)-2H-1,2-benzothiazine-3-carboxamide-1,1-dioxide, is a modern non-steroidal anti-inflammatory drug (NSAID). It causes fewer side effects on the gastrointestinal tract, and good tolerability. Due to its low solubility under acidic and neutral conditions, it has low bioavailability. To increase one, various methods have been used and evaluated with respect to aqueous solubility and dissolution rate, resulting in particle size reduction, preparation of solid dispersions, etc. For producing fine powders of pharmaceutical substances, methods based on spray drying or different variants of freeze-drying also are known. Another method of producing fine powders of pharmaceutical substances, a so-called method of obtaining a "glass phase" with the use of high cooling rates (with the formation of amorphous phases), followed by removing the solvent by sublimation has been developed recently [1, 2].

In the last decades, co-crystals, i.e. crystal materials consisting of at least two different components in particular an active pharmaceutical ingredient and a non-toxic partner molecule [3, 4], has extended in pharmaceutical industry as a method to enhance the bioavailability. Various methods such as solid-state grinding or kneading (solvent-drop grinding), classical solution crystallization approach as well as preparation from melt are usually applied to prepare co-crystals. In the present work, a method to prepare meloxicam co-crystals with carboxylic acids based on rapid cooling of solution containing both starting components (meloxicam and partner molecule) in a desired ratio followed by removing the solvent by sublimation has been developed. The current study has been designed to obtain ultrafine forms of meloxicam and to investigate the dissolution rate and solubility of the obtained samples.

Using method of freeze-drying of frozen solutions, the ultra-fine samples of meloxicam were obtained, which exhibited higher dissolution rate in comparison with the initial drug. The co-crystals of meloxicam with carboxylic acids, malonic and succinic, obtained by this method showed much higher rate of dissolution (in comparison with solvent-drop ground co-crystals also) due to small size of the particles and increased specific surface. We suggest that the method based on freeze-drying of frozen solutions using high rates of cooling and formation of amorphous phases can be used for increasing bioavailability of poorly-water soluble drugs as well as decreasing the side effects of the drugs. It can be used for preparation of new forms of different drugs with improved characteristics.

The work was partly supported by the RFBR (grant No. 13-03-00795) and the Council for Grants of the President of RF for Support of Leading Scientific Schools (project no. NSh-279.2014.3). References [1]. K.A. Overhoff, K.P. Johnston, J. Tam, J. Engstrom, R.O. Williams, J. Drug Del. Sci. Tech., 2009, 19, 89. [2]. A.G. Ogienko, E.V. Boldyreva, A.Yu. Manakov, S.A. Myz’, A.A. Ogienko, A.S. Yunoshev, E.G. Zevak, N.V.

Kutaev, A.A. Krasnikov, Dokl. Phys. Chem., 2012, 444, 88-92. [3]. A.B. Bond, CrystEngComm., 2007, 9, 833-834. [4]. N. Schultheiss and A. Newman, Cryst. Growth Des., 2009, 9, 2950.


268

The quantum-chemical DFT-calculating of the thermodynamic properties of

arglabin molecule

S.T. Shamilova,1 L.K. Abulyaisova,2 G.A. Atazhanova,1 S.M. Adekenov1

1"Phytochemistry" International Research and Production Holding, 100009, Kazakhstan,

Karaganda, M.Gazaliev str., 4 2E.A. Buketov Karaganda State University, 100025, Kazakhstan, Karaganda, Mukanov str., 41


The fundamental thermodynamic properties of arglabin molecule such as: enthalpy of formation,

entropy, Gibbs free energy, molar heat capacity, - one calculated by means of quantum-chemical

method of density functional B3LYP / 6-31G (d) in the frames of GAUSSIAN 03W program [1],

taking into the consideration zero-point energy and contributions of translational, vibrational and

rotational motion of the molecule.

The enthalpy of isomers formation was calculated by relatively simple compounds (15С +

9Н2 + 3/2О2) by means of the formula:

∆Н = (Еп+Н298)arg - ∑(Еп+Н298)С, H2, О2,

where пE + Н298 – the sum of the electronic energy and thermal corrections to the arglabin

enthalpy and simple compounds at 298K.

The theoretically calculated physicochemical characteristics of Arglabin molecule С15Н18О3

Factor B3LYP/6-31G(d) method Factor B3LYP/6-31G(d) method *Eп, at. u

-808.0565920

The rotational constants, GHz

0.64397 0.37956 0.25048

**E0, kcal/mole 190.549 , kcal/(mole•K)

123.635 (total) 42.403

∆f , kcal/mole

-2 193.853 translational rotational vibrational

32.929 48.303

∆ , kcal/mole -2 119.001 The dipole moment of D x- component y- component z- component

5.056 3.630 -3.070 -1.721

,J/(mole•K) 23.53

* The total energy of the molecule.

** The zero-point energy of the molecule.

References [1]. Frisch M. J., Trucks G. W., Schlegel H. B., et al. Gaussian 03, Revision C.02, Gaussian, Inc., Wallingford, CT,

2004.

0298S

0298H

0298G

0298pC


269

Determination of the Enantiomeric Purity of Pemetrexed and Levalbuterol on

the Macrocyclic Glycopeptide Bonded Phases

E.N. Shapovalova, I.A. Fedorova, A.A. Priporova, I.A. Ananieva, O.A. Shpigun

Chemistry Department, Moscow State University, GSP-1, Leninskie gory, d. 1, stroenie 3,

119991, Moscow, Russia


Macrocyclic glycopeptides, which are immobilized on the surface of silica, are successfully established as chiral selectors in HPLC for enantioseparation of large range of drugs. These chiral phases have high selectivity due to the presence in their structure of various groups capable to multipoint interaction with shared compounds in polar and nonpolar solvents. The aim of this work was the separation of pemetrexed and albuterol enantiomers on silica gel modified by eremomycin (column Nautilus-E, BioChemMac, Russia) and teicoplanin aglycone (column Chirobiotic TAG, Astec, USA). Pemetrexed is drug that is prescribed for malignant pleural mesothelioma and for non-small cell lung cancer. Active substance of pemetrexed is L-isomer. Active substance of albuterol is R-isomer, so-called levalbuterol, which is used as a bronchodilator.

Separation of the enantiomers was carried out by reverse - phase (RP) and polar organic (PO) modes of chromatography. Mixture of a buffer solution of ammonium dihydrogen phosphate (NH4H2PO4) and organic solvents (methanol, acetonitrile) was used as a mobile phase in the RP-mode. Concentration, pH of buffer and content of organic modifier was varied in the mobile phase. In the PO-mode organic solvents (methanol, acetonitrile) with additive of an acid (acetic or formic) and a base (triethylamine or diethylamine) were used. The better separation of pemetrexed’s enantiomers was obtained on eremomycin-column and albuterol’s enantiomers - on teicoplanin aglycone-column. Pemetrexed’s enantiomers able to separate with a resolution of 2.8-3.3 on column Nautilus-E in RP-mode. The determination of the enantiomeric purity of pharmaceutical preparation of pemetrexed was conducted. Used mobile phase was (55:15:30) MeOH: ACN: NH4H2PO4 (50mM, pH 2.5). Time of analysis was less than 10 minutes. The detection limit of compound was calculated as signal / background ratio = 3: 1. It was 0.3 µg / ml, it correspond 0.12% D-enantiomer of the total amount of the drug. The content of D-pemetrexed is permitted at no greater than 0.3%. Enantiomers of albuterol were separated on Nautilus-E and Chirobiotic TAG columns in PO-mode chromatography. The selectivity was higher on Chirobiotic TAG column (Rs = 1.7). The maximum resolution of enantiomers peaks (1.7) was obtained when mobile phase was (90:10:0.5:0.5) methanol: acetonitrile: triethylamine: CH3COOH. The detection limit of the compound, calculated by the signal / background ratio = 3: 1 was 0.2 µg / ml, it correspond 0.10% S-enantiomer of total composition of drug. The content of S-albuterol is permitted at no greater than 0.2%.

The determination of enantiomeric purity of the pharmaceutical substances such as pemetrexed from one batch and levalbuterol from two batches was carried out.


270

Ortho-dinitrobenzenes in the Synthesis of Biologically Active Benzimidazolones

A. Shchipalkin, D. Krivorotov and V. Kuznetsov

Chemical modeling laboratory, Research Institute of Hygiene, Occupational Pathology and Human Ecology, Federal Medical Biological Agency, p/o Kuz’molovsky, Saint Petersburg, 188663, Russian

Federation


2,3-Dihydro-2-oxo-1H-benzimidazole-1-carboxamides, benzimidazolone derivatives, seem to be

quite attractive. Among these compounds are selective serotonin receptor ligands, inter alia, ligands

of unique 5-НТ4a subtype. The latter is located in the brain region called “pre-Bötzinger complex”.

This region ensures respiratory rhythm generation in humans [1].

The described methods of synthesis of 2,3-dihydro-2-oxo-1H-benzimidazole-1-carboxamide

derivatives are not unified. Therefore, it is difficult to conduct synthesis for creating a compound

library, which can be used ultimately in high-throughput screening of biological activity of

benzimidazolones. We have developed an efficient general approach to the synthesis of 2,3-

dihydro-2-oxo-1H-benzimidazole-1-carboxamide derivatives based on nucleophilic substitution of

ortho-dinitrobenzenes. Variation of substituents in the benzene, imidazole and urethane fragments

has allowed us to obtain a wide range of potentially biologically active and known compounds, such

as BIMU-8 (6a) [1].

N+

O-

O

N+

O-

O

R1

NH2 R2 N

+

O-

O

NH

R1

R2

N

N

R1

R2

O

NHO

R3

1 26a-j

6a: R1=H; R

2=iso-propyl; R

3=tropine;

6b: R1=H; R

2=iso-propyl; R

3=memantine;

6c: R1=H; R

2=iso-propyl; R

3=2-adamantan;

6d: R1=H; R

2=iso-propyl; R

3=pipirazine;

6e: R1=H; R

2=iso-propyl; R

3=N,N-diethyletheleniamine;

6f: R1=CF3; R

2=iso-propyl; R

3=N,N-diethyletheleniamine;

6g: R1=CF3; R

2=iso-propyl; R

3=tropine;

6h: R1=H; R

2=cyclo-propyl; R

3=tropine;

6j: R1=H; R

2=cyclo-propyl; R

3=nipicotineamide.

The structure of the synthesized compounds has been proved using physicochemical methods of

analysis.

References [1] T. Manzke, U. Guenther, E. Ponimaskin, M, Haller, M. Dutschmann, S. Schwarzacher, D. Richter. Science. 2003, 301, 226-229.


271

Determination of Seven Flavonoids in Epimedium by Liquid

Chromatography–Tandem Mass Spectrometry Method

O.A. Shevlyakova,1 K.J. Vasil’yev,1 A.A. Ihalaynen,1 A.M. Antokhin,1 V.F. Taranchenko,1

D.A. Mitrofanov,1 A.V. Aksenov,1 I.A. Shchetinina,1 I.A. Rodin,2 O.A. Shpigun2

1 Federal State Unitary Enterprise “Scientific Center “Signal”,

Russia, 107014, Moscow, Bolshaya Olenya, 8 2 Lomonosov Moscow State University, Chemistry Department,

Russia, 119991, Moscow, Leninskie gory, 1-3

Epimedium - genus of perennial herbaceous plants in the family Berberidaceae, includes more

than 60 species. Prized as a medicinal plant exhibiting androgen, antioxidant, antidepressant,

antitumor activities [1-3]. In medicine are used the dried aerial parts, which contain a wide variety

of physiologically active compounds, the most interesting of which are flavonoids: icaritin,

icarisides I, II, epimedins A, B, C. The contents of these components largely determines the value

of the plant material.

The approach for detection of icariin, icaritin, icarisides I, II, epimedins A, B, C in water- ethanol

extracts from plants was developed on the basis of liquid chromatography/mass spectrometry. The

separation of the compounds was performed using a Hypersil Gold aQ column (3 µm, 2.1×150

mm), «Thermo Scientific» wish gradient elution by 0,1% formic acid in water/acetonitrile (95/5)

and 0.1% formic acid in acetonitrile. Separation conditions were optimized for major components.

Detection of the target compounds was carried out using the high resolution orbital ion trap

hybrid mass spectrometer Q-Exactive (Thermo Scientific, Germany) with electrospray ionization in

the multiple reactions monitoring mode (MRM). In this paper, we report the results of the

parameters optimization for mass-spectrometric detection, mass-fragmentation of active substances

by the high-energy C-trap dissociation (HCD) and the characteristic ion transitions for each test

compound. Limits of detection were 5×10-4 mkg ml-1, error in the determination of the ions masses

was < 2 ppm.

The advantages of this method of analysis are high reliability, accuracy, sensitivity and stability

over a wide range of concentrations.

[1]. H. K. Kang, Y.-H. Choi, H. Kwon, S.-B. Lee, D.-H. Kim, C. K. Sung, Y. I. Park, M.-S. Dong, Food and Chem.

Toxicology, 2012, 50, 2751-2759. [2]. W. Xu, Y. Zhang, M. Yang, Z. Shen, X. Zhang, W. Zhang, H. Li, Journal of Pharm. and Biomed. Analysis, 2007,

45, 667-672. [3]. Y. Pan, L. Kong, X. Xia, W. Zhang, Z. Xia, F. Jiang, Pharmacol. Biochem. Behav., 2005, 82, 686-694.


272

Phosphoramidates of Pyrimidine Nucleoside Analogues: Unexpected Products of

Known Reaction

N.F. Zakirova, A.V. Shipitsyn

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov St.,

Moscow, 119991, Russian Federation. Fax: +7(499)1351405

E.mail: [email protected]

A series phosphoramidate derivatives of β-L-2',3'-dideoxy-3'-thiacytidine (lamivudine, 3TC) and

3’-azido-3’-deoxythimidine (AZT) - the compounds are potentially latent forms of monophosphates

3TC and AZT was prepared. The best option for the synthesis of these compounds was the

acquisition of phosphoramidates from diphenylphosphite followed by treatment with

phenylphosphite of 3TC or AZT carbon tetrachloride and various amines (Atherton–Todd reaction

[1]). Surprisingly obtaining two groups of products - phenylphosphoramidates 3TC (2) or AZT (3)

and bis-3TC-phosphoramidates (4) or bis-AZT-phosphoramidates (5) synthesis of the previously

undescribed structures.

The work was supported by the Russian Foundation for Basic Research, project 13-04-00829-a; Presidium of the Russian Academy of Sciences, project “Molecular and Cellular Biology.”

References [1] S. S. Le Corre, M. Berchel, H. Couthon-Gourvès, J.-P. Haelters and P.-A. Jaffrès, Beilstein J. Org. Chem. 2014, 10, 1166–1196.doi:10.3762/bjoc.10.117


273

Anti-influenza Activity of Usnic Acid Derivatives in vitro and in vivo

A.A. Shtro,1 V.V. Zarubaev,1 O.A. Luzina,2 D.N. Sokolov,2 N.F. Salakhutdinov2

1Influenza Research Institute, Saint-Petersburg, Russia. 2Novosibirsk Institute of Organic Chemistry, Novosibirsk, Russia.

Influenza A virus is a human pathogen of primary importance for health care systems all over the world and represents serious problem for drug development, vaccination and treatment strategies. Only three chemical classes of compounds are currently used as anti-influenza drugs: adamantane derivatives block protein M2 that acts as a proton channel required for acidification of the virion core, and thus prevent virus decapsidation. Second, there are four neuraminidase inhibitors: zanamivir (Relenza®), oseltamivir (Tamiflu®), peramivir (Rapiacta®), and laninamivir (Inavir®). Third, nucleoside analogs ribavirin and favipiravir are antivirals of broad range of activity, which exhibit a suppressive effect against almost all RNA-genome human viruses.

In the present work we showed wide range of activity of some usnic acid derivatives against influenza viruses differing in their antigenic type and subtype, susceptibility to current antivirals and host specificity. We demonstrated that compounds of this group suppress replication of broad panel of influenza viruses.

Derivatives of usnic acid showed moderate activity in experiments in vivo. Protective index of compound 575 (valine enamine of usnic acid) was 60% (1 LD50) and 33% (10 LD50); beside this it reduced the weight loss during the infection. Replicative activity of the virus in the lung tissue of mice treated by compound 575 was also reduced. The inhibitory effect of compound 575 is not due to its direct inactivation of virion particles, as its incubation with virus for 1 hour had no effect on viral infectivity.

Time-of-addition experiments showed that compound 575 has the target of its action at the early stages of the life cycle. The decrease in viral titer was observed when added 1 h prior to infection (prophylactic administration), simultaneously (administration in a period of viral absorption on the cell surface) or immediately after the infection, including virus entry into cells, virus uncoating, release of viral RNP in the cytoplasm and importing of the RNP into the nucleus; however, the largest decrease in viral titer occurred when the compound has been in contact with infected all cells during the experiment. Thus, it can be concluded that the antiviral effect of compound 575 probably is cumulative.

Finally we can say that usnic acid derivatives have a broad spectrum of anti-influenza activity in vitro and one of them, compound 575 also showed moderate activity in vivo. This compound had shown activity on early stages of influenza virus lifecycle and do not stimulate the selection of resistant strains. The hepatotoxicity of compound 575 was reduced comparatively to usnic acid. Our results suggest that valine enamine of usnic acid could be a potential candidate for the development of a new anti-influenza therapy.


274

Nanoemulsions With Biologically Active Substances

E.G. Shubenkova

Omsk state technical university, 644050, Russian Federation, Omsk, Mira Ave., 11.


Delivery of therapeutic preparations by means of nanosystems of various type – the perspective

direction in modern pharmacology and medical cosmetology. It allows to get the preparations

having noticeable advantages before traditional [1].

The purpose of these researches was development of natural nanoemulsion compositions on the

basis of the oil and water extracts of plant raw materials. This compositions for transdermal

application contains a hydrophilic phase, a hydrophobic phase and surface-active substance. As a

hydrophobic phase using oil extracts of plant raw materials on the basis of natural oils. The surface-

active substance is being choosing from group of phospholipids of a natural origin [2].

Plant extracts as components of natural preparations contain the fullest sum of biologically active

substances (BAS), capable to make complex impact on an organism, to show a wide range of

pharmacological activity and they are characterized by low toxicity. The selection of herbal raw

materials for extraction provides variation composition of extracts and thereby increases the

therapeutic effect. Extraction of a complex BAS from plant raw materials was carried out by

methods of repeated two-phase extraction and ultrasonic extraction [3].

The assessment of anti-oxidizing activity of the received extracts according to a technique is

carried out [4]. Stability of nanoemulsions was studied in the conditions of the accelerated storage

at a temperature of 40 °C [5].

As a result the phospholipid nanoparticles in the form of nanoemulsion systems like "water in

oil" which represent natural compositions on the basis of oil and water extracts of plant raw

materials are received [2]. The nanoemulsion is biologically compatible and well transferable, and

also provides the uniform prolonged release of active ingredient and possesses sufficient stability.

References [1] S. Singh, Nanomedicine-nanoscale drugs and delivery systems, J. Nanosci. Nanotechnol, 2010, 10(12), 7906-7918. [2] Pat. 2535022 RUS; Bul. invent., 2014, 34. [3] Shubenkova E.G., Chzhu O.P. "Biochemical developments of extractions of biologically active

substances". Applied and Fundamental Studies : Proceedings of the 2nd International Academic Conference. March 8-10, 2013, St. Louis, USA. Publishing House "Science & Innovation Center", 2013. – Р. 55-59.

[4] Pat. 2170930 RUS; Bul. invent., 2000, 13. [5] O.S. Strekalova, Diss. cand. biol. Sciences, Research Institute of Biomedical Chemistry of V.N. Orekhovich RAMS,

Moscow, 2010, 117 p.


275

Modification of Cyprofloxacine with Indol-1-ylmaleimide Derivatives. Synthesis

and Study of the Antibacterial Activity

A.Yu. Simonov, S.A. Lakatosh, E.P. Mirchink, E.B. Isakova

Gause Institute of new Antibiotics, Russian Federation. Moscow, 119021, B.Pirogovskaya, 11


The development of new antibacterial substances, effective against the resistant strains is avery

important objective. The structural modification of the known antibiotics is one of the possible

ways to obtain new effective compounds.

The resistance to cyprofloxacine and other fluoroquinolone antibiotics is widely spread in

clinical practice and is a big problem in antibacterial therapy. One of the possible of modification

sites of the cyprofloxacine molecule is a piperazine nitrogen atom, since it is not part of

flouroquinolone pharmakophore. The reaction of cyprofloxacine with an 3-bromo-4-(indol-1-

yl)maleimides gave the corresponding conjugates. The new substances were not active against

MRSA, but some of them were more active against the selected strains than the starting

cyprofloxacine. The activity data for the best compound in the series are given in the Table 1.

N

FO

OH

O

HNO O

N

NN

R2

R1

R3

Table 1

MIC values for the derivatives of cyprofloxacine against G+ and G- microbal strains (µg/ml)

Strains

LCTA Stap

hylo

cocc

us

aure

us 2

5923

A

TC

C

Stap

hylo

cocc

us

aure

us 3

798

Stre

ptoc

occu

s pn

eum

onia

e

4961

9 A

TC

C

Ent

eroc

occu

s fa

eciu

m 5

69

E.c

oli 2

5922

A

TC

C

Kle

bsie

lla

pneu

mon

iae

1388

3

AT

CC

Pro

teus

vul

gari

s 13

315

AT

CC

Salm

onel

la

chol

eras

uis

1402

8 A

TC

C

Cipro 0,25 64,0 4,0 4,0 0,13 0,13 0,25 0,25

2478 4,0 32,0 4,0 4,0 1,0 0,13 0,25 0,13

LCTA-2478: R1 = CH3, R2 = H, R3 = H


276

Search of the Bioactivity of Extracts of Thymus serpyllum L. Growing in Yakutia

S.V. Sivtceva,1 Yun Jeong Choe,2 Kyu Lee,2 Ho Young Kang,2

Su In Cho3 and Zh.M. Okhlopkova 1

1Departmen of Biology, North-Eastern Federal University, 677000, Russia, Yakutsk, Belinskogo Str., 58

2Department of Microbiology, Pusan National University, 609-735, Korea 3Department of Pharmacology and Herbology, Pusan National University, 609-735, Korea


Plants of the Thymus genus are presented in the Yakutian flora by 15 species. The most

widespread specie is the Thymus serpyllum L. Its grass contains 0,79-1,0% of essential oil, tanids,

flavonoids, saponins, coumarins, glycosides etc. [1]. The specie is widely used in the Yakut

traditional medicine [2]. However, the extractive substances of the Yakutian Thymus and their

bioactivity hasn't been specifically studied.

The grass of Thymus serpyllum L. was collected in natural populations in the territory of north-

east, central and south Yakutia. The material was composed with observance of state standards

(6077-80) during 2011-2014. Alcoholic and aqueous-alcoholic extracts were obtained by means of

Multi Rotator RS-60. After 72-hour extraction, filtration and drying under vacuum in the rotor

evaporator (3-4 h) and in the cups (15-18 h) were executed. Extracts are used for the phytochemical

analysis of the main groups of biologically active substances (TLC, quality reactions) and for the

screening of cytotoxicity on mice of the balb/c line. In the first case, extracts were entered orally. In

the second case, extracts were entered intramuscularly. During the experiment changes in behavior

of the animals were not observed. The increase in quantity of red blood cells was noticed. Alcoholic

extracts have high activity and suppress growth of Candida 4075, Torulospora delbrueckii 4063,

Debaryomyces hansenii 4059 by 73-75%. Phytochemical research of dry alcoholic and aqueous-

alcoholic extracts shows the existence the glycosides of flavonol, halkons, saponins. It is necessary

to identify the structure of the found substances. The obtained substances may show antifungal

activity.

This work was partially supported by the Program of the Development of NEFU (the Contract No. 2.37.4. of 20.11.2012, third stage). References [1] Makarov A.A. Herbs of Yakutia. Yakutsk. – 1970. – 180p. (in Russian) [2] Makarov A.A. Vegetable remedies of the Yakut traditional medicine. Yakutsk. - 1974. – 64p. (in Russian)


277

Antioxidants as a criterion of EXTRA VIRGIN olive oil quality

N.V. Sizova

Institute of Petroleum Chemistry, SB RAS, 4, Academichesky ave. Tomsk, 634055, Russia,

Phone: (3822) – 507359


Olive oil is a most common oil in the world, which is widely recommend for internal and external use. It is believed that the olive oils owe their high biological and nutritional properties to antioxidants. Every fatty oil contains such antioxidants as tocopherol homologues and specific minor components. According to the literature, the content of vitamin E in olive oils is 12 to 43 mg%, which is twice lower than in sunflower oil and 8 times less than in the wheat germ oil. The specific components of olive oils are polyphenols − hydroxythyrozol, thyrozol, luteolin, and apigenin, which can also act as antioxidants. We measured the total content of antioxidants in EXTRA VIRGIN olive oils, produced by different technologies in different countries.

Objects. We investigated 25 samples of olive oils produced in different countries. The oil samples were purchased from Russian and foreign retail vendors. For some oils, we determined the peroxidation indices.

Method. The measurements were carried out in an MCDP-2 isothermal microcalorimeter, designed and produced at the Institute of Petroleum Chemistry, SB RAS. The model reaction of cumene oxidation with azobisisobutironitrile as initiator was carried out at 60 oC.

The heat evaluation (W) was proportional to the chain continuation (reaction II) or break of peroxyradicals at the inhibitor-antioxidant (reaction VII):

k2 RH + RO•

2 → ROOH + R• (II)

k7 InH + RO•

2 → ROOH + In• (VII)

The concentration of antioxidants can be calculated by the formula: [ ]

iw

AOXfn 0=τ , where the

initial concentration of antioxidants [AOX]0 (mol/l) is related to both the induction period τ and the rate of initiation wi (l/mol·s), whereas f is the stoichiometric coefficient of inhibition, and n is the number of functional groups.

Results. The reactions yielded significantly different amounts of AOXs in olive oils − from 0.7·10-3 mol/kg to 1.6·10-3 mol/kg. This is explicable, given the variety of olive types, their cultivation regions and ripeness. These values, when recalculated with respect to the content of tocopherol are found to be 30 to 75 mg%, which is more than twice higher than the published data (12 to 43 mg%). This is attributed to the contribution of phenolic acids (up to 4.2 mg%) and the manifestation of synergy. We have found no differences in the content of AOXs in the oils from black or green olives, although according to the literature the content of vitamin E in the olives decreases with maturation, while that of polyphenols increases.

For the EXTRA VIRGIN olive oils in question, the acid and peroxide numbers turned out to be high. According to the TR 024/2011 (Technical Regulations on Butter and Fat Products), the acid number admissible for unrefined extra virgin olive oils should be no more than 1.6 mg KOH/g and the peroxide number − as low as 20 mmol O2/kg. The maximum acid and peroxide numbers were observed in the Cretan Mythos oil (Crete, Greece), 1.6 mg KOH/g and 18 mmol O2/kg, respectively, and the minimum acid and peroxide numbers, equal to 0.5 mg KOH/g and 8.6 mmol O2/kg, respectively, were determined in the Celeste oil (Spain).


278

Triphenylantimony(V) Catechlolates Derived from Alkyl Gallates: Synthesis

and Antioxidant Activity

I.V. Smolyaninov,1,2 A.I. Poddel’sky3 and N.T. Berberova1

1Southern Scientific Centre of RAS, 344006, Russia, Rostov-on-Don, Chekchova str., 41

2Astrakhan State Technical University, 414056, Russia, Astrakhan, Tatisheva str., 16 2G. A. Razuvaev Institute of Organometallic Chemistry of RAS, 603137, Russia, Nizhny Novgorod,

Tropinina str., 49


A new triphenylantimony(V) catecholate complexes (1-4) containing gallic acid esters were

obtained. Radical scavenging and antioxidant activity of organoantimony(V) complexes as well free

ligands 5-7 was considered in assays with DPPH radical, model peroxidation of oleic acid.

The values of EC50, nDPPH, AE for complexes 1-4 point out that the compounds can act as an

antiradical agent. All complexes are displayed inhibition effect on the process of oleic acid

oxidation. The compounds 1-4 are played the role of effective LOOH destructors. For the more

active complexes 1,4 and methyl gallate 5, the effect on lipid peroxidation of rat liver homogenate

of male Wistar was investigated, and membrane-protective properties were evaluated on the

example of induced hemolysis of red blood cells. Antioxidant activity in the process of lipid

peroxidation of liver homogenate in the presence of compound 1 and 4 was displayed. It was found

to increase the rate of decomposition of hydrogen peroxide hemolysate of erythrocytes observed in

the membrane-action induced hemolysis of erythrocytes. The complexes 1 and 4 are obviously not

only effective destructors hydroperoxides of oleic acid, and hydrogen peroxide. In conclusion, our

results suggest that the combination organometallic fragment containing of antimony with bioactive

ligand leads to appear uncommon properties, which are varied from properties the initial

triphenyllantimony(V) compounds.

Acknowledgements - the work was financially supported by Russian Foundation for Basic Research (grants № 14-03-00478, 15-03-02967), grant of President RF (MK–445.2014.3).


279

Synthesis and Biological Activity of Conjugates of Substituted Indoles with

Phenothiazine-3,7-Diamine

V.B. Sokolov, A.Yu. Aksinenko, T.V. Goreva, T.A. Epishina, V.V. Grigoriev, A.V. Gabrel’yan,

D.V. Vinogradova, M.E. Neganova, P.N. Shevtsov, E.F. Shevtsova and S.O. Bachurin

Institute of Physiologically Active Compounds, 142432, Russia, Chernogolovka, Severnyi pr. 1


As part of searching of new perspective multitarget drugs for the treatment of neurodegenerative

diseases a synthetic approach to integrating in one molecule a substituted indoles and structural

fragment of methylene blue was proposed.

Conjugates of substituted indoles 3-6 and methylene blue catalytic (CsF) were obtained by

Michael’s alkylation of various substituted indoles 1 with acrylated phenothiazine-3,7-diamine 2.

The influence of bifunctional conjugates 3-6 on binding with neuronal NMDA-receptors,

assembly/disassembly of microtubules and mitochondrial characteristics were studied. The affinity

of conjugates 3 compared to 4 and 5 to NMDA-receptors are significantly more pronounced,

moreover these conjugates also have a greater selectivity for GluN2B-dependent binding. All

studied bifunctional conjugates 3-6 alike methylene blue alone demonstrated the ability to decrease

the rate of tubulin polymerization/depolymerization with normal structure of formed microtubules.

Study of effects of new synthesized compounds on mitochondrial function allow us to identify as

screening hits some fluorine containing compounds in series of 4 which able to block calcium-

induced mitochondrial permeability transition and protect the neurons from calcium overloading.

The study was performed by a grant from the Russian Scientific Foundation (project № 14-23-00160). The equipment of Center for Collective Use IPAC RAS (agreement N14.621.21.0008, ID RFMEFI62114X0008) was used.


280

The Biological Properties of the New Agent for Photodynamic Therapy And

Luminescent Bioimaging Based on Molybdenum Complexes

A.O. Solovieva,1 K.E. Zubareva,1 A.A. Krasilnikova,1,2 L.V. Shestopalova,2

E.V. Vorontsova,3 K.A. Brylev,4 A.F. Poveshchenko,1 M.A. Shestopalov1,4

1 Federal State Budgetary Scientific Institution «Scientific Institute of clinical and experimental lymрhology», 2 Timakova Str., 630060 Novosibirsk, Russia.

2 Novosibirsk State University, 2 Pirogova Str., 630090 Novosibirsk, Russia. 3 The Institute of Molecular Biology and Biophysics

4 Institute of Clinical and Experimental Lymphology SB RAMS, 2 Timakova Str., 630060 Novosibirsk, Russia.


Molybdenum complexes with {Mo6I8}4+-cluster core are considered as attractive candidates for

bioimaging (luminescent diagnostic), biolabeling and photodynamic therapy because the clusters exhibit red phosphorescence with high quantum yields and, as a consequence, may generate the singlet oxygen. Since such complexes are easily hydrolyzed in water and can react in physiological environments, it is necessary to find a method to stabilize them. We propose to use silica nanoparticals (SNPs) as containers protecting the clusters against hydrolysis and other reactions in physiological environments. The synthesized {Mo6I8}@SiO2 SNPs exhibit a good solubility, an intensive luminescence, a photostability, and a lower susceptibility to cellular efflux mechanisms than small drug molecules.

The main factor determining the efficiency of {Mo6I8}@SiO2 for bioimaging and photodynamic action is the ability the drugs to be internalized into cells and lower cellular efflux degree. Thereby, the purpose of our study was to investigate cytotoxicity, cellular internalization, intracellular distribution, cellular uptake and elimination kinetic of {Mo6I8}@SiO2.

We showed that concentrations of {Mo6I8}@SiO2 from 1.5 µg/ml to 1500 µg/ml do not influence on viability and proliferation of cells. It has been identified that cellular uptake kinetic of {Mo6I8}@SiO2 depended on the incubation time. Maximum cell uptake of {Mo6I8}@SiO2 was observed after 2 hours of incubation. Also, the rate of cellular elimination of {Mo6I8}@SiO2 was studied. It has been shown that the drug was actively excreted from cells during the first 2 hours, after that the fluorescence intensity of stained cells was not significantly changed and stayed on a high level. These findings have thus shown that the drug rapidly enters cells and stays there for a long time.

To determine intracellular localization of {Mo6I8}@SiO2 the fluorescent confocal microscopy and transmission electron microscopy (TEM) were applied. Luminescent imaging clearly showed that {Mo6I8}@SiO2 was localized in the cell cytoplasm into endosome-similar structures. TEM microphotographs demonstrated that SNPs persist as in membrane-enclosed vesicles (endosomes) as well as in cytosol (free SNPs). These findings suggest that nanoparticles uptake into cells by endocytosis and are able to endosomal escape. Endosomal escape is often a critical step in intracellular trafficking of SNPs and subsequent targeting to appropriate subcellular compartment. This quality is crucial for drug efficacy as a photosensitizer.

Thus, {Mo6I8}@SiO2 has low cytotoxicity, high degree of cellular uptake, low level of elimination from the cells and is able to endosomal escape. These properties suggest that the compound is highly promising for photodynamic therapy and luminescent bioimaging. This work was supported by the Russian Science Foundation (Grant №14-14-00192).


281

New Acyclic Nucleoside Phosphonates Bearing Unsaturated

Fragments in the Chain. Synthesis and Antiviral Properties

P.N. Solyev, M.V. Jasko and M.K. Kukhanova

Laboratory of the molecular basis of action of physiologically active compounds,

Engelhardt Institute of Molecular Biology, 119991, Russia, Moscow, Vavilov St., 32


Acyclic nucleoside and nucleotide analogues are of great interest. Since Nobel-winning

Acyclovir, the most used acyclic nucleoside against herpes viruses, appeared, many successful

attempts and achievements in this scope has been made [1]. Phosphonate moiety is a useful bypass

of the limiting first phosphorylation step and does not undergo enzymatic hydrolysis as natural

phosphates do. Several acyclic nucleoside phosphonate-based drugs are currently in clinical use for

treatment of viral infections (e.g. Tenofovir, Cidofovir, Adefovir). The flexible acyclic chain

possesses higher range of conformations giving an opportunity to bind more nucleotide-involved

enzymes – thus, a combined activity against more viruses can be expected. It is quite essential since

over 80% of HIV-infected patients suffer from herpes and other virus-induced infections.

In our recent study we have synthesized new acyclic nucleotide analogues with broad antiviral

activity. One of the series is oxime-containing nucleoside phosphonates:9-{2-

[(phosphonomethyl)oximino]ethyl}adenine, -guanine and 9-{2-[(phosphonomethyl)oximino]-

propyl}adenine. Oximes are rarely used in drug design and yet their conformational rigidness and

their stability in hydrolytic tests may be useful for nucleoside side chain drug design [2]. The

compounds were synthesized using modified Mitsunobu procedure for key intermediate diethyl

aminooxymethylphosphonate, which allowed to double the yield of the original synthesis [3]. The

efficient oxime forming “click” reaction was performed to form the target structures. A convenient

procedure for aminooxy group detection was proposed. Products displayed moderate activity

against HIV and herpes viruses in cell cultures and hepatitis virus C in replicon system without any

toxicity up to 1000 µM.

Other series of the compounds will be further discussed.

This study was supported by the Russian Foundation for Basic Research (grant № 14-04-31163-mol_a). References [1] E. De Clercq. Biochemical Pharmacology, 2007, 73, 911-922 [2] J. Kalia, and R. T. Raines. Angewandte Chemie International Edition, 2008; 47, 7523-7526 [3] Lacoste A.-M., Dumora C., Zon J. Enzyme Inhibition, 1993, 7, 231-248


282

Pentacyclic Triterpenoid Bioregulators – a New

Class of Hepatoprotective Agents

I. Sorokina, N. Zhukova, and T. Tolstikova

N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry of Siberian Branch

of Russian Academy of Science, 630090, Russia, Novosibirsk, Acad. Lavrentjev ave., 9


Now more than 50 percent of hepatoprotective pharmaceuticals and additives have a plant origin.

Most of them are native substances extracted from a raw material (such as silymarin, curcumin,

Liv-52) or the galenical of some plants. New approach to development of hepatoprotective drugs

is based on synthetic modification of secondary plant metabolites possessing high basal activity.

Plant-derived pentacyclic triterpenoids of lupane, ursane and oleanane families provide a versatile

structural platform for synthesis derivatives which are able to stimulate cytoprotective signal

pathway Nrf2-Keap1-ARE. These bioregulators demonstrate potent antioxidant, anti-inflammatory

and immunomodulative activity which can be adequate for the treatment of liver diseases.

New derivatives of moronic, betulinic, dihydrobetulinic, betulonic and levopimaric acids were

recently synthesized in Novosibirsk Institute of Organic Chemistry and Institute of Chemical

Kinetics and Combustion SB RAS. The hepatoprotective activity of them was studied in animals

models of acute and chronic hepatitis, induced by toxic agents, drugs or alcohol. It is found

moronic acid and their derivatives possess hypocholesterolemic action and potent antiholestatic

effect which exceed the same of thioctic acid. The amids of betulonic and betulinic acids which are

modified by amino acids residues, have high antioxidant and anti-inflammatory activities and

efficiently protect hepatocytes against degenerative and necrotic lesions. Alanin- derivative of

betulonic acid is shown to be a more potent anti-fibrotic agent compare with admethionin.

Betulinic acid-peptid conjugates containing histidine, alanine, tryptophan and isoleucine amino

acids are found to exibit high anti-inflammatory activity, comparable to that of indomethacin. It was

demonstrated by molecular docking the conjugates are incorporated into binding site of Kelch-

domen of target protein Keap1 by their amino acid residues and form more non-covalent bonds,

than the triterpenoid core. The modification of betulinic acid with acetylenic moieties leads to

derivatives with hepatoprotective and spasmolytic activity which exceeds the same of drotaverin

and tropicamid. It is found morpholin and piperidin derivatives of dihydrobetulonic acid decrease

hepato-toxic effects of cytostatic drugs and do not stimulate tumor growth and dissemination in

carcinoma-bearing mice. The obtained results confirm high potency of pentacyclic triterpenoid

bioregulators as a new class of hepatoprotective agents.


283

Bioflavonoid’s and Carotenoid’s Based Compositions of Enhanced Solubility

and Antioxidant Activity

L.P. Suntsova,1 A.V. Dushkin,1 N.E. Polyakov,2 I.E. Apanasenko,2

Е.S. Меtеlеvа,1 V.I. Evseenko1

1Institute of Solid State Chemistry and Mechanochemistry,630128, Russia, Novosibirsk,

Kutateladze St.,18 2Institute of Chemical Kinetics and Combustion, 630090, Russia, Novosibirsk, Institutskaya St., 3


Currently, special attention is paid to the creation of effective agents that can reduce the damage

from free radical processes in the body. Examples of such agents are the compounds of plant origin

- carotenoids and bioflavonoids. They demonstrate a wide spectrum of activity in the human body.

Namely, antioxidant, capillary protection, increasing the elasticity of red blood cells, and reduce the

risk of cardiovascular disease. Reacting as antioxidants with free radicals and reactive oxygen

species, macular carotenoids protect the retina against peroxidation and photo-damage. On the other

hand, extremely low water solubility of bioflavonoids and carotenoids and their instability restrict

their practical application as components of food or medicinal formulations. To improve the water

solubility we used mechanochemical approach. Formulations based on flavonoids and carotenoids

with auxiliary substances were prepared. Increasing the water solubility is achieved through the

following mechanisms: partial ionization of molecules of flavonoids by alkaline agents - calcium

carbonate and magnesium carbonate; the formation of “host-guest” complexes with water-soluble

larch polysaccharide arabinogalactan (AG) derived from Larix sibirica and Larix gmelinii; inclusion

of molecules of carotenoids in the micelles formed of glycyrrhizic acid (GA) and its sodium salt in

aqueous solution. The solubility of samples was measured by HPLC, complex formation with GA

was demonstrated by NMR relaxation methods, and complexation with AG was analyzed by SPR

techniques.

It was found a significant increase (in ~101-103) of flavonoids and carotenoids solubility in all

samples prepared by mechanochemical way. Inclusion of the “guest” molecule into GA micelle also

allows to increase stability of carotenoids. These results are interesting for the development of

enhanced dietary supplements and medicines.

References 1. Suntsova L.P., Меtеlеvа Е.S., Dushkin А.V. Mechanochemical obtaining watersoluble compositions based on

flavonoids // Fundamental research. 2014. № 11. pp. 2174-2179. 2. I.E. Apanasenko, O.Yu. Selyutina, N.E. Polyakov, L.P. Suntsova, E.S. Meteleva, A.V. Dushkin, P. Vachali, P.S.

Bernstein Solubilization and Stabilization of Macular Carotenoids by Water Soluble Oligosaccharides and Polysaccharides // Archives of Biochemistry and Biophysics. 2014. DOI:10.1016/j.abb.2014.12.010.


284

Chimeric Structures Based on 1-(oxiran-2-ylmethyl)-1H-indole-3-carbaldehyde

as New Antidiabetic Agents

K.F. Suzdalev,1 A.A. Spasov,2 S.A. Tsirulnik,1 N.I. Chepljaeva2 and V.A. Kuznetsova2

1Department of Chemistry, Southern Federal University, 344090, Russia, Rostov-on-Don,

Zorge Street, 7 2Department of Pharmacology, Volgograd Medical University, 400131, Russia, Volgograd,

Pavshikh Bortsov Square, 1


The term "chimeric structure" means the presence in the molecule of several structurally

different fragments, each of which contributes to the biological activity. It is known that the indole

moiety is often part of antidiabetics. As other pharmacophores can act cyclic secondary amines,

isatin and adamantane derivatives. In addition the drug must be soluble in water.

Our goal is to design hybrid molecules or "chimeric structures", combining three basic

fragments: 1. indole moiety as a privileged structure, 2. pharmacophore group and 3. the residue of

the amine hydrochloride, dissolving the substance in water. Desired structures were obtained from

the aldehyde 1 [1], having two electrophilic centers - a carbonyl group and an oxiran ring.

We have implemented a multi-component one-pot synthesis for compounds 2: Reaction of

aldehyde 1, active methylene compounds and amine leads directly to amino alcohols 2. On their

basis the water-soluble hydrochlorides 3 were synthesized.

Biological tests revealed that the compounds 3 inhibit the Maillard reaction and serve glycogen

phosphorylase inhibitors. These properties allow reducing glucose production in the liver and

neutralizing diabetic complications. Thus, the chimeric structures 3 are promising for the

development of highly active drugs to combat diabetes and its consequences.

Acknowledgements. Research was carried out according the state task of the Ministry of Education

and Science of the Russian Federation № 4.129.2014/K.

References [1] K.F. Suzdalev, S.V. Den’kina, A.A. Starikova, V.V. Dvurechensky, M.E. Kletsky, and O.N. Burov, Mendeleev Communications, 2011, 21, 231-233.


285

In Silico And In Vitro Search For Inhibitors Of Dipeptidyl Peptidase-4 Among

Adamantane Derivatives

A. Taran,1 P. Vassiliev,1 A. Spasov,1 N. Cheplyaeva,1 Yu. Popov,2 V. Lobasenko,2

E. Shishkin2 and V. Mokhov2





Modern strategy of treatment for type 2 diabetes mellitus (DM 2) requires correction of chronic

hyperglycemia [1]. Inhibitors of dipeptidyl peptidase type 4 (DPP-4) are a new class of oral drugs

with low risk of adverse effects and high therapeutic efficacy. The mechanism of action is based on

an increase of incretin’s level [2]. One of the representatives of this group is vildagliptin,

adamantane derivative. Thus, the search for substance with DPP-4 inhibitor activity among

adamantane derivatives is reasonable and appropriate. Prediction of DPP-4 inhibitor activity for 1-

substituted (I) and 2-substituted (II) adamantane derivatives is carried out

using a docking program AutoDock Vina [3]. We used three experimental

3D models of DPP-4 taken from PDBe [4], codes 2ogz, 3g0b and 3ccc. The

average value of the calculated energy ∆E of docking in the catalytic sites of

all three models was used as the final evaluation for the ligand affinity. As a

comparison drug was chosen vildagliptin. Substances were considered

«active» if the docking energy was comparable to or greater than the

docking energy of vildagliptin. By this criterion, 19 of the studied

substances, nine (47%) were potentially active. The inhibitory activity was

tested in vitro by spectrophotometric method [5] using multidetector TECAN Infinite 200 Pro.

Experimental results showed that the substances active were 15 (78%). Thus, the efficiency of

virtual screening was 60%. The results of these studies confirm the promising of search for DPP-4

inhibitors in class of adamantane derivatives.

The study was performed in VSMU by RSF grant № 14-25-00139.

References [1] E.V.Bova and E.N.Pacus, Fundamental research. 2009, 10, 6-9 [2] S.H. Tella, and M.S. Rendell, Expert Opinion on Drug Safety. 2015, 14, 127-140 [3] AutoDock Vina, http://vina.scripps.edu [4] PDBe, http://www.ebi.ac.uk/pdbe/ [5] A.S.Taran and N.I.Cheplyaeva, Volgograd medical scientific journal. 2014, 1, 26-29

(CH2)nN

R1 R2

N

O

R1

R2

R3

I

II


286

Polyphosphate Nucleotide Derivatives Functionalized at

Terminal Phosphate Group

Yu. Tarasenko,1 P. Chalova2 and T. Abramova1

1Laboratory of Organic Synthesis, Institute of Chemical Biology and Fundamental Medicine,

630090, Russian Federation, Novosibirsk, Lavrent’ev Ave, 8 2Department of Natural Science, Novosibirsk State University, 630090, Russian Federation,

Novosibirsk, Pirogova Str., 2


Indispensable role of polyphosphorylated nucleotides (e.g. ATP, 5’,5’-dinucleoside

tetraphosphates, terminal cape structures of RNA) in biochemistry is difficult to overestimate.

Rapid development of molecular biology methods in medicine and urgent tasks in searching for

new inhibitors of key enzymes responsible for heavy diseases pose an actual problem of

development of new effective methods for obtaining of conjugates of polyphosphorylated

nucleotides tethered through polyphosphate chain.

We report on the effective synthetic approach affording nucleoside 5’-polyphosphates I bearing

an extended hydrophilic linker at terminal phosphate and functionalized with an alkylamino group.

A number of protective groups for aminoalkyl functionality and different strategies implying

coupling of nucleoside mono, di, and triphosphate were studied. Our approach allows an easy

changing of conjugated reporter groups and a number of phosphate residues in conjugates.

Based on the key compounds I we synthesized two

series of nucleoside 5’-polyphosphates II and III.

Series II is designed as a number of substrates for

DNA/RNA polymerases for NA sequencing [1].

Series II is proposed as a new class of potential

inhibitors for enzymes of the PARP family [2]. The role

of these enzymes in maintaining genome integrity and

carcinogenesis is now well-known. Compounds of the

series II represent analogues of NAD+ - the substrate in

the poly-ADP synthesis catalyzed by the PARP

enzymes.

The work was supported by RFBR (Russia, Grant No 14-04-01018a). References [1] P.A. Sims, W.J. Greenleaf, H. Duan, and X.S. Xie, Nat. methods. 2011, 8, 575-580. [2] G. Papeo, E. Casale, A. Montagnoli, and A. Cirla, Expert Opin. Ther. Pat. 2013, 23, 503-514.

O

H(OH)OH

OPO-

OOPO

O-

O nucleobase

n

OH2NI

O

H(OH)OH

OPO-

OOPO

O-

O Ade

n

ONH

II

- Fluorecent dye

O

OHOH

OPO-

OOPO

O-

O AdeONH

O

R

III

R - NH2, OH, CH3, CH2NH2, NO2, Cl, etc.


287

Chemical Composition of the Essential Oils of the Plant Family Apiaceae from

Buryatian and Mongolian Flora

V. Taraskin,1,2 B. Urbagarova,1 G. Tykheev1 and L. Radnaeva 1,2

1Medical Institute, Buryat State University, 670000, Russia, Ulan-Ude, Smolin ul., 24a.

2 Laboratory of Chemistry of Natural Systems, Baikal Institute of Nature Management SB RAS (BINM SB RAS), 670047, Ulan-Ude, Sakhyanovoy str., 6.


By GC / MS method there was studied the composition of essential oils of Saposhnikovia

divaricata (Turcz.) Schischk. (aerial part and roots), Bupleurum scorzonerifolium Willd. (aerial

part), Phlojodicarpus sibiricus (Steph. Ex Spreng.) Koso-Pol. (rhizomes and roots) of the family

Apiaceae which grow on the territory of Buryatia and Mongolia. The yield of essential oil from the

plants was around 0,35-1,4% from the weight of air dried raw material. The highest yield of

essential oil was observed from rhizomes and roots of Phlojodicarpus sibiricus – 1,4% from the

weight of air dried raw material. The main components of the essential oils, extracted from herbs of

Saposhnikovia divaricata, are the representatives of bicyclic sesquiterpenes group: δ-cadinene

(18,8%), β-akoradien (8,7%), γ-muurolen (8,8%), α-muurolen (4,3%), γ-cadinene (5.3%),

caryophyllene (3.5%), T-murolol (2.4%). The essential oil from the roots of Saposhnikovia

divaricata is full of falcarinol (also termed as panaxynol) (45,5-49,3%), which induces apoptosis

and has anti-cancer activity [1]. The dominate components of the essential oil from the herbs of

Bupleurum scorzonerifolium are monocyclic sesquiterpenes - germacrene D (5,1-36,8%),

monocyclic monoterpenes - limonene (4,3-6,7%), bicyclic sesquiterpenes - caryophyllene (2,5-

11,4%). There were revealed 30 to 110 components in the samples. The essential oil extracted from

the herbs of Saposhnikovia divaricata is characterized by a high content of bicyclic sesquiterpenes.

The essential oil of Phlojodicarpus sibiricus is characterized by a high content of limonene (62%)

and γ-terpinene.

Thus, the essential oils from the plants of Apiaceae family are distinguished by obligatory

presence of limonene, γ-terpinene and caryophyllene. Germacrene D is one of the dominant

components, generally, in the aerial parts of the plants.

References [1] Zhonghong Yan, Ruolin Yang, Yi Jiang, Zhihui Yang, Junrui Yang, Qian Zhao and Yang Lu , Molecules, 2011, 16, 5561-5573.


288

Novel Dosage Forms on the Basis of 1,2,4-Thiadiazole

Derivative and β-Cyclodextrins

O. Silyukov,1 M. Chislov,1 A. Proshin,2 E. Chibunova,1 T. Volkova1 and I. Terekhova 1

1G.A. Krestov Institute of Solution Chemistry of RAS, 153045, Russia, Ivanovo, Akademicheskaya ,1

2Institute of Physiologically Active Compounds of RAS, 142432, Russia, Chernogolovka, Severniy pr., 1


Thiadiazole derivatives display the therapeutic activity in the treatment of Alzheimer’s disease,

which is one of the socially significant chronic neurodegenerative diseases. Low solubility and,

consequently, bioactivity of these compounds can considerably decrease their pharmacological

action. In this connection, development of water-soluble forms of thiadiazole derivatives is the

urgent task of pharmaceutical chemistry. Cyclodextrins are suitable naturally occurring compounds,

which are widely used in pharmacy as encapsulating materials and drug delivery systems. Solubility

as well as stability and other physicochemical properties of drugs can be significantly improved

through the inclusion complex formation with cyclodextrins.

To this end, complex formation of 1-[5-(3-chlorophenylamino)-1,2,4-thiadiazol-3-yl]-propan-2-

ol with β- and hydroxypropyl-β-cyclodextrins in phosphate buffer (pH 7.4) was studied by

solubility and UV-vis spectroscopy methods. It was demonstrated that rather stable inclusion

complexes of 1:1 stoichiometric ratio are formed in the solution. Insertion of the benzene ring into

macrocyclic cavity results in appreciable increase of drug solubility.

Solid inclusion complexes of 1,2,4-thiadiazol derivative with β-cyclodextrins were prepared by

grinding method without and with addition of small amount of ethanol. Individual compounds, their

physical mixture and solid complexes were characterized by differential scanning calorimetry and

IR-spectroscopy. Existence of the inclusion complexes in the solid state was confirmed on the basis

on the obtained results.

The in vitro dissolution study of the prepared dosage forms 1,2,4-thiadiazol derivative and its

solid complexes and physical mixture with β-cyclodextrins performed in phosphate buffer (pH 7.4)

at 37 0C showed that dissolution rate of 1,2,4-thiadiazol derivative is considerably increased by

complex formation with cyclodextrins. Thus, β-cyclodextrins can be used as appropriate

solubilizing agents for the considered drug.

This work was supported by Russian Science Foundation (grant №15-13-10017).


289

Effect of Acid Modification of Montmorillonite Clays on Synthesis of

Octahydro-2H-chromen-4-ol from Vanillin and Isopulegol

M.N. Timofeeva,1,2 K.P. Volcho,3 V.N. Panchenko,1 O.S. Mikhalchenko,3 N.F. Salakhutdinov,3

K.V. Kochegarova,2 A. Gil,4 M.A. Vicente5

1Boreskov Institute of Catalysis SB RAS, 630090, Russia, Novosibirsk, Pr. Ak. Lavrentieva 5

2Novosibirsk State Technical University, 630092, Russia, Novosibirsk, Pr. K. Marksa 20 3Novosibirsk Institute of Organic Chemistry SB RAS, 630090, Russia, Novosibirsk,

Pr. Ak. Lavrentieva 9 4Department of Applied Chemistry, Public University of Navarra, 31006, Spain, Pamplona

5Department of Inorganic Chemistry, University of Salamanca, Spain, Salamanca


Reaction of homoallylic alcohols with aldehydes, variant of the Prins cyclization reaction, is one

of the ways for the synthesis of a wide range of tetrahydropyran derivatives. Recently, acid-

activated montmorillonites due to their unique structural and physicochemical properties have

received much attention for synthesis of biologically active 2H-chromen-4-ols. The aim of this

study was to investigate the effect of the HCl concentration used for the acid-activation on the

surface acidity and the porous structure of the montmorillonite clays, and the subsequent influence

on the catalytic activity of these solids in the Prins cyclization reaction of (-)-isopulegol (I) with

vanillin (II) to octahydro-2H-chromen-4-ol (isomers IIIa and IIIb).

According to experimental data the concentration of HCl used for the acid activation of the clays

strongly affects the conversion of (II) and the yield of isomers (IIIa-b) due to influence on the

amount of Brønsted acid sites (BAS). It was found that the amount of BAS in acid-activated

montmorillonites is a key factor for the adjustment of the reaction rate and the selectivity of the

reaction. An increase of the amount of BAS leads to a decrease in the ratio of (III)/(IV) products.

The nature of the clay affects the chemo- and stereo-selectivity, i.e. molar ratios of (III)/(IV) and

(IIIa)/(IIIb). Reaction rate and isomer selectivity in the presence of clay with large pore diameter

and low microporosity (clay 1) were lower in comparison to clay with low pore diameter and large

microporosity (clay 2). Noteworthy, in the presence of clay 2 and clay 1 the (IIIa)/(IIIb) molar ratios

were 5.3-5.7 and 4.0-4.5, respectively, that can be also related to the differences in the structural

parameters.


290

Efficient Route to 18-[(5-Bromomethyl)oxazol-2-yl]-18-norisopimaradiene and

its Derivatives

M.A. Timoshenko, Yu.V. Kharitonov, E.E. Shults

N. N. Vorozhtzov Institute of Organic Chemistry, Siberian Branch of the Russian Academy of

Sciences, 630090, Russia, Novosibirsk, pr. Lavrentieva, 9


Functionalized isopimaranes [1] and substituted oxazoles [2] from nature are of interest for their

biological activity. Herein, we report the selective transformation of isopimaric acid 1 to a series of

18-(oxazol-2-yl)-18-nor-isopimaranes.

Cycloisomerization of N-propargyl amide 2 in the presence of ZnCl2 led to compound 3 which

was selective transformed into a diterpenoid substituted 5-(bromomethyl)oxazole 4. By

condensation of 4 with amino acid derivatives the corresponding derivatives 5a-d were obtained.

Reaction of compound 4 with sodium azide in DMF led to diterpenoid azide 6. The selective

formation of 4 allowed the further transformations of azide 6. By Cu-catalyzed reaction of

compound 6 with several alkynes biheterocyclic or triheterocyclic ensembles of type 7-9 were

synthesized.

This work was supported by the Russian Foundation for Basic Research (grant 15-03-06546) References [1] J.R. Hanson, Nat. Prod. Rep., 2015, 32, 76-87. [2] G. Nuzzo, M. L. Ciavatta, R. Kiss, V. Mathieu, H. Leclercqz, E. Manzo, G. Villani, E. Mollo, F. Lefranc, L. D'Souza, M. Gavagnin and G. Cimino, Mar. Drugs, 2012, 10, 1799– 1811.


291

Inhibition of Copper and Iron Induced Hydroxyl Radical Formation by Metal

Chelator Deferiprone

V.A. Timoshnikov,1,2 T.V. Kobzeva,1 N.E. Polyakov,1 G.J. Kontoghiorghes3

1Institute of chemical kinetics and combustion SB RAS, Russia, Novosibirsk, Institutskaya, 3

2Novosibirsk State University, Russia, Novosibirsk, Pirogova, 2 3Postgraduate Research Institute of Science, Technology, Environment and Medicine, Cyprus.


Deferiprone (L1) is an active chelator of metal ions. This drug is used to treat diseases associated

with an excess of iron in the body, such as thalassemia. It is known that both copper and iron ions

can produce highly toxic hydroxyl radical through the Fenton reaction. Aqua complexes of iron

may also generate hydroxyl radicals under irradiation. While some studies have reported the

antioxidant activity of L1, however, the detail mechanism of this activity at the molecular level of

deferiprone has not been determined.

Thus, the purpose of this study was, firstly, to establish the ability of L1 to

inhibit the Fenton reaction induced by iron and copper ions, and secondly, to study

the mechanism of inhibition of active oxygen radicals formation in photoinduced

processes involving Fe(III) aqua complexes in the presence of deferiprone. Finally,

we tried to investigate the effect of ascorbic acid on the antioxidant properties of

L1 in redox reactions. It is known that although ascorbic acid has antioxidant

properties, it can act as a pro-oxidant in the presence of other antioxidants and metal ions. EPR spin

trapping technique was applied to answer these questions.

It was demonstrated that interaction of L1 with Fe(III), Fe(II), Cu(II) and Cu(I) ions completely

inhibits the formation of hydroxyl radicals in the presence of hydrogen peroxide (Fenton reaction)

[1]. Irradiation of Fe(III) aqua complexes in the presence of spin traps TMIO and PBN gives rise to

a EPR signal of spin adduct with OH radical. The presence of L1 completely inhibits the formation

of OH radicals in this reaction. The mechanism of formation of the spin adduct with OH radical was

confirmed by detection of the methyl radicals in the presence of dimethyl sulfoxide. Formation of

singlet oxygen was also not detected under irradiation of L1 or its iron complexes. The presence of

ascorbic acid does not affect the antioxidant properties of L1.

References [1]. V.A. Timoshnikov, T.V. Kobzeva, N.E. Polyakov, G.J. Kontoghiorghes. Inhibition of Fe3+ and Fe2+ induced

hydroxyl radical production by the iron-chelating drug deferiprone. Free Radical Biology and Medicine. 78, l (2015) 118–122.

N

CH3

CH3

OH

O


292

The Development of a New Innovative Anticoagulant Drug

D. Tovbin, D. Tarasov

PharmaDiall, Ltd, 105043, Russia, Moscow, 4-th Parkovaya str., 9/21


Factor Xa (FXa) is known to play a central role in a blood coagulation cascade and considered to

be one of the most attractive targets for oral anticoagulants of new generation. We used a new

approach for the development of new highly potent low-molecular weight FXa inhibitors. The

reaction of amid bond formation from appropriate amides and acids was chosen to be underlying

and the compound groups like R1-(CONH)-СН3, R1-(CONH)-R2-(CONH)-СН3 and R1-(CONH)-

R2-(CONH)-R3 were studied in a successive manner. The best inhibition constants of each group

were found by the two-step multi-iterative approach, which was a combination of in silico

calculations based on physical models and in vitro measurements of antithrombotic activity.

Subnanomolar potency of several developed compounds was achieved. Moreover, the concentration

of prothrombin time doubling (PTx2) in human plasma of compd. no 217 (DD217) turned out to be

130 nM, which puts it among the best FXa inhibitors ever reported. The compound efficacy was

proved by in vivo assays including oral administrations in monkeys. DD217 is now under

preclinical trials.

The comparison of the International normalized ratio for monkeys (Rhesus macaques), after

intragastrically administrated DD217 and the same dose of the well known drug Xarelto

(Rivaroxaban). The effect of DD217 is softer and longer.


293

Antiviral Activity of Preparation from Rice Husk and Green Tea

E.G. Trofimova,1 O.I. Lomovsky,1 E.I. Kazachinskai,2 T.V. Teplyakova2

1Institute of Solid State Chemistry and Mechanochemistry SB RAS, 630128

Russia, Novosibirsk, Kutateladze 18 2State Research Center for Virology and Biotechnology "Vector", 630559, Russia, Novosibirsk

region, Koltsovo


Catechines from the green tea showed good activity against some viruses. Their concentrations vary

within the range 10 to 300 mg per one gram of dry tea. It is possible to introduce flavonoids in capsules

and use carrier compounds providing the delivery of biologically active compound to affected

regions. Compounds based on silicon dioxide occupy a special place among many structurally

stable materials that have been studied for drug delivery. It was established that silicon compounds

are concentrated in definite sites – tumors, wounds, affected regions. Rice husk is notable for a high

content of the inorganic component, silicon dioxide, whose concentration can reach 20 % on

average. Furthermore rice husk is a perfect bio-renewable raw material - around 100 million tonnes

of rice husk wastes are produced every year. Increasing attention is drawn to the mechanochemical

approach as a method to obtain the compositions of biologically active substances with inorganic

carriers. Thus, the goal of the present work is the development of a solid-state method for obtaining

preparation based on flavonoids and silica from the plant raw material, and evaluation of the

antiviral activity of them.

It was demonstrated that the interaction of silicon dioxide with gallocatechines can be realized

using the solid-phase mechanochemical method. During the joint mechanical treatment of the

reactant or a mixture of green tea with rice husk, surface complexes of silicon with gallocatechines

are formed.

Results of research of antiviral activity of compositions against Herpes simplex virus 2 and West

Nile virus are obtained. The data are compared with antiviral activity of some chemotherapeutic

agents such as ribavirin, nitroxide radical and extract of shelf fungus.

The promising character of the application of these preparations in the therapy of virus infections

is due to the discovered antiviral action, general corroborative and immunostimulating effect of the

substances of plant origin, relatively simple and cheap technology of medical preparations by solid-

phase methods.


294

A Computation Modeling for Rational Design and Synthesis of Oligomer

Compounds Capable of Arresting the Viral Fusion Machinery

V. Tsvetkov,1,2 B. Bolshchikov2 and A. Serbin1,2

1Biomodulators RC, Health RDF, 117042, Russia, Moscow, Boulevard Adm. Ushakova, 14-209

2A.V. Topchiev Institute of Petrochemical Synthesis RAS, 119991, Russia, Moscow, Leninsky Pr., 29 E-mail: [email protected]

A computation screening of small molecules for antiviral therapy is very relevant approach to

antimetabolitic-kind interventions or inhibiting the equally small size centers of virus specific

receptors and enzymes. But small molecules cannot be fully efficient and drug-resistance preventive

agents against lager macromolecular and nano-organized viral targets such as, for example, the tri-

molecular protein complexes mediated the class 1 fusion between cell membrane(s) and virion's

envelope. This fusion mechanism is widely used to entry in cells by many viral families, including

the retro- (e.g., HIV), orthomixo- (e.g., influenza A/B), paramixo-, flavi- (e.g., Ebola), and other.

The most crucial targets are the three-α-helical [HR1]3 pre-hairpin intermediates (2.5 x 5-10

nm). Improving the docking and molecular dynamics approaches to modeling interactions between

synthetic and biopolymers [1], we studied the targets-binding capabilities among the specific-

anchor-modified oligomer compounds as the nano-competent ligands (ONL). Being able to belt

[HR1]3 targets, the ONLs are found can be many folds stronger inhibitors than small molecules.

These results are in good correlation with data of in vitro evaluations of experimentally synthesized

inhibitors of HIV-1/2 and Influenza A/B viruses. And recent quantum chemistry modeling of

reactions for ONL preparation gives additional guidelines to a controlled synthesis of new ONL-

compounds with predicted anti-fusion activity against other viruses, including the Ebola as well.

ONL + HIV-1, gp41-target + Influenza, HA2-target + Ebola GP2-target

References [1] V.Tsvetkov and A. Serbin, J Comp-Aid Mol Des. 2012. 26(12):1369-1388; 2014. 28(6):647-673


295

Molecular Modeling of New G-quadruplex Inhibitors of HIV Infection

V. Tsvetkov,1,2 A. Varizhuk,2,3 V. Oslovsky3 and G. Pozmogova 1


2 Institute of Physical-Chemical Medicine, 119435, Russia, Moscow, Malaya Pirogovskaya str. 1a 3 Engelhardt Institute of Molecular Biology, 119991, Russia, Moscow, Vavilov str. 32


New anti-HIV DNA aptamers that inhibit an attachment step of the viral entry pathway are

reported. The natural and thiophosphoryl aptamers were obtained on the basis of monomolecular G-

quadruplex (G4) scaffolds, including the well-known G4 thrombin inhibitor TBA. Their supposed

mechanism of action involves binding with the HIV envelope glycoprotein gp120. This was

investigated by using physicochemical methods and molecular modelling (docking and MD

simulation). The results suggest that interactions with gp120 may underlie the observed minor anti-

HIV activity of unmodified TBA (several gp120 binding sites may be involved, including the V3

loop). The thiophosphoryl TBA analogue, which demonstrated antiviral activity in a middle

nanomolar range, has a higher affinity for the gp120 V3 loop, but in general, we attribute its activity

to non-specific interactions with viral and/or cellular proteins.

The results of TBA and F-thio-TBA docking to gp120: The aptamers are green,

phosphodiester groups are red, S atoms ore black. The protein body is pink and V3 loop is blue

MD simulation results: (R/S)-F-thio-TBA complex with gp120 and free binding energies of

TBA/F-thio-TBA complexes with the gp120 V3 loop


296

Parameters for Describing Conformational Rearrangements in G-quadruplexes

V. Tsvetkov,1,2 A. Varizhuk,2,3 G. Pozmogova2


2 Institute of Physical-Chemical Medicine, 119435, Russia, Moscow, Malaya Pirogovskaya str. 1a 3 Engelhardt Institute of Molecular Biology, 119991, Russia, Moscow, Vavilov str. 32


Conformational changes in DNA G-quadruplex-forming regions affect genome function and,

thus, compose an interesting research topic. Computer modelling may yield insight into quadruplex

folding and rearrangement, particularly molecular dynamics simulations. Here, we show that

specific parameters, which are distinct from those commonly used in DNA conformational

analyses, must be introduced for adequate interpretation and, most importantly, convenient visual

representation of the quadruplex modelling results. We report a set of parameters that

comprehensively and systematically describe G-quadruplex geometry in dynamics. The parameters

include those related to quartet planarity, quadruplex twist, and quartet stacking; they are used to

quantitatively characterise various types of quadruplexes and rearrangements, such as quartet

distortion/disruption or deviation/bulging of a single nucleotide from the quartet plane. Our

approach to describing conformational changes in quadruplexes using the new parameters is

exemplified by telomeric quadruplex rearrangement, and the benefits of applying this approach to

analyse other structures are discussed.

Vladimir Tsvetkov, Galina Pozmogova, Anna Varizhuk The Systematic Approach to Describing Conformational Rearrangements in G-quadruplexes. Journal of Biomolecular Structure & Dynamics 2015 DOI:10.1080/07391102.2015.1055303


297

5-Aminotetrazole in the Synthesis of 6-Ar(Het)-3,5-diamino-1,2,4-triazines.

Lamotriginum and Its Analogues

E.N. Ulomsky, V.L. Rusinov, E.K. Voinkov, V.V. Fedotov, K.V. Savateev

Ural Federal University, 19, Mira str., 620002, Ekaterinburg, Russia


A derivatives of 6-aryl-3,5-diamino-1,2,4-triazines occupy a unique place in medicinal chemistry

and possess remarkable biological properties such as anticonvulsants reagents, for example, the

Lamotriginum is an anticonvulsant drug used in the treatment of epilepsy and bipolar disorder. Our

research group has developed new synthetic methodologies – reconstruction of heterocycles, which

included, at first step, construction of fused azaindolizines on the base nitrogen containing

heterocycles and next step destruction of basis heterocyle.

NHNN

N NH2

NHNN

N N2+ N

H

N

NHNN

N

ArN

Ar

LG

NC

NN

NNN

N

NH2Ar

N

NN

NH2Ar

N3 NN

N

NH2

Ar

NH2

AcONa

Ar = Ph, 4-Cl-C6H4, 2-Cl-C6H4, 2,3-Cl-C6H3, 2-C10H7; Het = 2-Py, 4-Py, benzimidazolyl-2; LG = H; LG = COR', CO2H

NO2-, H+

tO

A tetrazolyl hydrazones were prepared by azo coupling of tetazolyl-5-diazonium and α-pyridyl-

or benzimidazolyl acetonitriles with good yields. Aryl derivatives of acetonitrile were activated by

acyl functional before Japp-Klingemann reaction with diazo compound. A tetrazolotriazines were

destruction by copper or PPh3 to 3,5-diamino-1,2,4-triazines.

This method is superior to the reported methods in all aspects such as short reaction times and

excellent yields of 6-aryl-3,5-diamino-1,2,4-triazines. For example, the proposed method of

preparation of Lamotriginum has 45% yields for finished compounds. Also the method of activation

for aryl derivatives of acetonitriles in Japp-Klingemann reaction can be used to synthesized of

several new 6-aryl, -hetaryl and -alkyl derivatives of 3,5-diamino-1,2,4-triazine.

We thank the Russian Science Foundation grant № 14-13-01301


298

Mechanochemical Activation of Brown Coal and modification of Its humic acids

T.S. Urazova, A.L. Bychkov, O.I. Lomovsky

Institute of solid state chemistry and Mechanochemistry SB RAS,

630128, Russia, Novosibirsk, Kutateladze str.18


The humic acids ability to bind various contaminants from environment arouses heightened interest among scientist in the field of environmental protection and reclaim [1]. The structure of humic molecules ensures water insolubility at neutral рН. There are some methods that allow to increase the yield of water-soluble humic acids by decrease of molecular weight or humic acids transfer to the salt form. Besides of the yield increasing there is the ability of humic acids structure modification [2]. For this purpose mechanochemical oxidation, sulfonation or acid hydrolysis are use.

In recent years investigations in the field of mechanochemistry associated with study of directional mechanochemical effects on polymeric structures have allowed to offer approaches for carrying out mechanochemical reactions under control [3]. This fact opens up new opportunities in the chemistry of humic acids.

Heavy metals binding in chelate complexes is the most effective from the viewpoint of the sorbents development. This capability is typical for phenolic and carboxylic groups at aromatic rings [1].

The aim of this work was to modify the humic acid structure by mechanochemical method in order to obtain effective sorbent and product for soil regeneration.

Using mechanochemical treatment of brown coal we have obtained sorbents showing high yields of humic acids (75 %). The treatment of coal with sodium hydroxide leads to the humic acids transfer to the salt form (sodium humate) without a functional group content increasing. The treatment of coal with an oxidizing agent Na2CO3·1.5H2O2 leads to the formation of new phenolic and carboxylic groups [4].

The sorbents efficiency was tested in experiments under laboratory and natural conditions. We have conducted experiments, in which the contaminated medium was water in natural conditions of Ob reservoir. The salts of copper, zinc and cadmium were added into a confined volume of water. We have shown that the sorbents can successfully bind heavy metals in natural conditions.

The sorbent, obtained by using a sodium percarbonate doesn’t initiate a biomass growth. This allows us to recommend these sorbents for removal of heavy metals from polluted water.

Due to the high yield of humic acids brown coal after mechanochemical activation also can be used as a product for the soil regeneration.

[1] E. Ghabbour, M. Shaker, A. El-Toukhy, I. Abid, G. Davies. Chemosphere. 63. 2006. 477–483 [2] A. Proidakov. Solid Fuel Chem. 2009. 43. 1. 9–14. [3] F. Leibfarth. Science. 2011. 333. 1582-1583. [4] Urazova, T. S., Bychkov, A. L., Lomovskii, O. I. Mechanochemical Modification of the Structure of Brown Coal

Humic Acids for Preparing a Sorbent for Heavy Metals // Russian Journal of Applied Chemistry, 2014, Vol. 87, No. 5, pp. 651−655.


299

Electrophilic Fluorination of 1-Ethyl-7-(4-R-piperazin-1-yl)-4-oxo-1,4-

dihydroquinoline-3-carboxylic Acids with NF-Reagents

A.N. Usoltsev,1,2 P.A. Zaikin,1 G.I. Borodkin,1,2 V.G. Shubin1

1 N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry,

630090, Novosibirsk, Russia, Lavrentiev Ave, 9 2 Novosibirsk State University, 630090, Novosibirsk, Russia, Pirogov St., 2

E-mail: usoltс[email protected]

Nowadays there are a lot of publications devoted to the improvements of synthesis of bicyclic

and tricyclic fluoroquinolones, increase of yield and quality of the products. A number of studies

are dedicated to the synthesis of new fluoroquinolones to obtain new medicines with a high

antibacterial activity [1].

We have shown that fluorination of 1-ethyl-7-(4-methylpiperazin-1-yl)-4-oxo-1,4-

dihydroquinoline-3-carboxylic acid (1) and 1-ethyl-7-(4-tert-butoxycarbonylpiperazin-1-yl)-4-oxo-

1,4-dihydroquinoline-3-carboxylic acid (2) with NF-reagents proceeds predominantly at the 8

position of quinoline fragment in acetonitrile, water and under solvent-free conditions. The main

products of the fluorination reaction are presented in Scheme 1. The structures of the products

obtained were established by 1H, 13C, 19F NMR spectroscopy, and high resolution mass-

spectrometry. The influence of the reaction conditions on the distribution of the products is

discussed.

Scheme 1. Fluorination of quinolones 1 and 2 with NF-reagents.

This work was gratefully supported by Division of Chemistry and Material Sciences of RAS (project 5.1.4). References [1] V.N. Charushin, E.V. Nosova, G.N. Lipunova, O.N. Chupakhin, Fluoroquinolones: synthesis and use, Moscow: Fizmatlit, 2013.


300

Synthesis of bi(hetero)arenes Potential Physiologically Active Compounds

I. Utepova,1,2 O. Chupakhin,1,2 M. Trestsova,1 D. Kucheryavaya,1 A. Musikhina1

1Ural Federal University, 620002, Russia, Ekaterinburg, 19 Mira Street

2Institute of Organic Synthesis, the Ural Branch of Russian Academy of Sciences, 620219, Russia, Ekaterinburg, 22 S. Kovalevskoy Street


Organic compounds, bearing pyrrole or indole moieties, appear to be very attractive for

biological screening, since the data of biological tests indicate that derivatives of this family quite

often become lead molecules. Among them are known antibiotics, antiviral and antitumor drugs,

agents for the neurological and cardiovascular diseases therapy [1].

Recently, we have found a new system for SNH oxidative reactions operating in heterogeneous

phase: air oxygen/nanosized TiO2/UV irradiation. In these conditions «azine-indole/pyrrole»

oxidative coupling is carried out in good yields.

On the other hand, the CdS/TiO2 composite material is allowed to obtain SNH reactions products

bi(hetero)aryls by visible light irradiation. Thus, the heterocycles 1a-d coupling with 4,5,6,7-

tetrahydroindole (2a) and 2-phenylpyrrole (2b) has been carried out. The choice of these

nucleophilic reagents due to the fact that fragments of derivatives 2 are being increasingly used for

the synthesis of natural substances analogues, pharmacophores and building blocks of drugs.

The research was financially supported by the Russian Science Foundation (Project No. 14-13-01177), the Council on Grants of the President of the Russian Federation (Program for State Support of Leading Scientific Schools, Grant NSh-3656.2014.3). References [1] M. Giampieri, A. Balbia, M. Mazzeia, P. La Collab, C. Ibbab, R. Loddo, Antiviral Res., 2009, 83, 179; A. J. Kochanowska-Karamyan, M. T. Hamann, Chem. Rev., 2010, 110, 4489; Z. Jin, Nat. Prod. Rep., 2009, 26, 382; I. S. Young, P. D. Thompson, A. Thompson, Nat. Prod. Rep., 2010, 27, 1801.


301

Synthesis of Nopinane-annelated Pyridines

E. Vasilyev,1 A. Agafontsev1,2 and A. Tkachev1,2

1 N.N.Vorozhtsov Novosibirsk Institute of Organic Chemistry of the Siberian Branch of Russian

Academy of Sciences, 630090, Russia, Novosibirsk, Lavrentjev Ave., 9 2Department of Natural Sciences, Novosibirsk State University, 630090, Russia, Novosibirsk,

Pirogova st.,2 E-mail: [email protected]

Investigation of substituted chiral pyridines gets significant attention in last two decades [1, 2].

In our laboratory chiral pinopyridines I were synthesized by reaction of (+)-pinocarvone oxime II

with enamines III promoted by FeCl3 by solvent-free way [3].

Unusual results were obtained in case of enamines IV containing ketone functional group

neighboring to the C-N enamine fragment. It was found that along with the products V annelation

1:1 C2-symmetrical bipyridines VI were formated.

C2-symmetrical products VI prevail in case of 1) oxime II was taken in excess (2:1); 2) reaction

was performed at high temperature (130-150 ºC); 3) reaction was promoted by FeCl3*6H2O. By this

way crystalline products VI could be obtained with 20-25% yields. Product V prepared from

equimolar amounts oxime and enamine, lower temperature (100-110 ºC) and CuCl2*2H2O as Lewis

acid with 8-10% yields.

The reported study was partially supported by RFBR, research project No. 15-03-03828_a.

References 1. L.-E. Perret-Aebi, A. von Zelewsky, C. Dietrich-Buchecker, J.-P. Sauvage, Angew. Chem. Int. Ed. 43 (2004). N

34. P. 4482-4485. 2. B. Rooks, M. Haas, D. Sepúlveda, T. Lu, S. Wheeler, ACS Catal., 5 (2015), N 1, P. 272–280. 3. E. Vasilyev, A. Agafontsev, A. Tkachev, Synth. Comm. 44 (2014) N 12. – P. 1817-1824.


302

Calculation of the Membership Function in the Consensus Prediction of the

Pharmacological Activity of Chemical Compounds

P. Vassiliev

Pharmacology Department, Volgograd State Medical University, 400131, Russia, Volgograd,

Pavshikh Bortsov Sq., 1


The consensus approach has been successfully used in the search for pharmacologically active

substances by means of IT Microcosm [1]. The approach generalizes a spectrum of prediction

estimates, obtained by several different methods of classification. If several calculated estimates

coincide, the accuracy of prediction increases, because the errors compensate one another. When

planning in silico of the experimental screening, assessment in marks of the levels of prospects of

compounds seems the most rational: 3 – very prospective; 2 – prospective; 1 – little prospective; 0 –

not prospective. Assessment in marks is a standardized and allows to generalize the results,

obtained by several significantly different prediction methods. The following formula of calculation

of the membership function of compound to the set of substances with given activity, with use of

several prediction methods and consensus of prediction evaluations is proposed:

,.

.+

++ 0203

010∑1

C

N

i=Ci

m LN

L+Ind

=F

where N – number of prediction methods; Indi – evaluation of prospects of compound by method i;

LC – the level of consensus, equal to the number of non-zero estimates of all Indi minus

one;

0.01, 0.02 – unbiasedness parameters.

For example, value Fm = 0.832 was calculated for diphenyloxide derivative with laboratory code

TONS-0031 in prediction of dipeptidylpeptidase-4 inhibitor activity by means of PASS [2], IT

Microcosm [1], and docking in AutoDock Vina [3]. In the experiment, this compound showed a

sufficiently high activity comparable with the activity of the reference drug. The proposed metric is

used in the direct search for antidiabetic substances with multitarget action.

The study was performed at VSMU and was supported by RSF grant № 14-25-00139.

References [1] P. M. Vassiliev, A. A. Spasov, V. A. Kosolapov et al., Consensus Drug Design Using IT Microcosm. In: L. Gorb,

V. Kuz’min, E. Muratov (eds) Application of Computational Techniques in Pharmacy and Medicine. Challenges and Advances in Computational Chemistry and Physics, vol. 17. Springer Science, Dordrecht. 2014, 369-431

[2] D. A. Filimonov, and V. V. Poroikov, Ross. Khim. Zhurn. (Zhurn. Ross. Khim. ob-va im. D. I. Mendeleeva). 2006, 50(2), 66-75

[3] O. Trott, and A. J. Olson, J. Comp. Chem. 2010, 31(2), 455-461


303

Regulating the NMDA-receptors Activity by Lambertianic Acid Amide

S.O. Vechkapova,1 T.A. Zapara,1 T.G. Tolstikova,2 A.S. Ratushnyak1

1 Design Technological Institute of Digital Techniques, Siberian Branch of the Russian Academy of

Sciences, Russia, Novosibirsk, Rzhanova st., 6 2 N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry of the Siberian Branch of the Russian

Academy of Sciences, Russia, Novosibirsk, Lavrentjev Ave., 9


The most common neurodegenerative disorders, such as Alzheimer’s, Parkinson’s, Huntington’s,

multiple sclerosis, epilepsy, ischemic brain damage, although caused by different mechanisms, may

share a common pathway: anomalies in working of ionotropic glutamate receptors, particularly of

NMDA type [1, 2]. Currently, drugs with glutamatergic mechanisms of action are intensely being

developed for the treatment of cognitive disorders and neurodegenerative processes.

The aim of this work was to study the effect of lambertianic acid amide (AmLA) on the induced

epileptiform activity in hippocampal CA1 field. The experiments were carried out on the

hippocampal slices of ICR male mice using standard electrophysiological techniques. Stimulation

of Schaffer collaterals and registration of induced population spikes of pyramidal neurons in the

CA1 field were made using glass microelectrodes, filled with saline.

The epileptiform activity was induced by magnesium-free medium or by treatment of corazol

(pentylenetetrazol). The application of AmLA in concentration of 170 µМ significantly decreased

the epileptiform activity or fully terminated it. The preincubation of slices with AmLA for an hour

before applying epileptiform conditions prevented the development of epileptiform activity. Also

AmLA did not affect the initiation of NMDA-dependent synaptic potentiation. It exhibited

neuroprotective effects, normalizing the epileptiform activity caused by the hyperactivity of

glutamatergic system or by decreasing the activity of GABAergic mediator system. Thus,

lambertianic acid extracted from needles and galipot of Siberian cedar pine (Pinus sibirica R. Mayr)

and its derivatives are a promising source of drugs for cognitive disorders and neurodegenerative

diseases treatment.

The work was supported by IV 35.1.5 basic project of fundamental researches of RAS and FNM-46 "Fundamental Sciences for medicine" program of Presidium of RAS. References [1] Lipton S.A., Rosenberg P.A. Excitatory amino acids as a final common pathway for neurologic disorders. N Engl J

Med, 1994, V. 330(9), P. 613-622. [2] Mehta A., Prabhakar M., Kumar P., Deshmukh R., Sharma P.L. Excitotoxicity: bridge to various triggers in

neurodegenerative disorders. Eur J Pharmacol. 2013, V.698(1-3). P. 6-18.


304

A New Look at the Synthesis of 5-Halogen Derivatives of Morpholino Uridine

I. Vohtancev,1 Yu. Tarasenko,2 V. Silnikov2 and T. Abramova2

1Novosibirsk State University, Pirogova St. 2, Novosibirsk 630090, Russia

2 Institute of Chemical Biology and Fundamental Medicine, SB RAS, Lavrent’ev Ave, 8, Novosibirsk 630090, Russia


Morpholino derivatives of modified nucleosides are widely spread in biology and medicine as

monomers of DNA or siRNA analogs, they being used in antiviral [1] or anticancer [2] therapy and

bioanalytical applications [3]. Using the halogenated at heterocyclic base nucleoside derivatives

opens an easy way to functionalization of the nucleosides. Previously, the synthesis of morpholino

nucleoside containing 5-iodouracil as heterocyclic base was reported [4]. The method consisted of a

large number of steps including an introduction and removing of transient protective groups. As a

starting material the ribouridine was used. We report here the simple synthesis of the morpholino

derivatives 5-halogenuridine (I, Br, Cl). N-Trityl protected morpholino 5-halogenuridine was

prepared from 5-halogenribouridine in good yield and without chromatographic purification.

Scheme 1. Synthesis of N-trityl protected morpholino 5-halogenuridine.

Reagents and conditions: i) NaIO4, EtOH, rt; ii) (NH4)2B4O7·4H2O, Et3N, rt; iii) NaBH3CN, CF3COOH, rt;

iv) TrCl, Et3N, DMF, rt.

Our approach is based on the Summerton`s protocol [5] with some modification. The key feature

of our procedure is the careful maintaining of pH at Schiff base formation stage. All morpholino

derivatives of 5-halogenuridine (I, Br, Cl) were characterized by 1H, 13C NMR and mass

spectrometry.

The work was supported by Russian Fond for Basic Research (Russia, Grant No 14-04-01018a).

References [1] A. Nazmi, K. Dutta, and A. Basu, PLoS Negl. Trop. Dis. 2010, 4,11, 2–12. [2] Y. Takei, K. Kadomatsu, K. Yuasa, W. Sato, and T. Muramatsu, Int. J. Cancer. 2005, 114, 3, 490–497. [3] M. Hocek, and M. Fojta, Org. Biomol. Chem. 2008, 6, 13, 2233–2241. [4] B. Nandi, S. Pattanayak, S. Paul, and S. Sinha, Eur. J. Org. Chem. 2013, 7, 1271-1286. [5] J. E. Summerton, and D. D. Weller, US Patent 5034506, 1991.

O

OHOH

HO

NH

O

ONO

HO

X

NH

NH

O

ONO

HO

X

NH

NH

O

ONO

HO

X

NTr

NH

O

ON

OHO

X

HO OHO O

i ii iii iv

NH

O

ON

X

1 2 3 4 5

X=I, Br or Cl


305

Ethylnitrocyanacetic Ester as the Latent Form of the Nitroacetonitrile

E.K. Voinkov, K.V. Savateev, V.V. Fedotov, E.N. Ulomskiy, V.L. Rusinov

Department of Organic chemistry, Ural Federal University named after the first President of Russia

Boris Yeltsin, ZIP 620002, Russian Federation, Ekaterinburg, Mira Street, 19


Synthesis of heterocyclic compounds similar in structure to the natural purines traditionally

attracted interest in terms of search receptors inhibitors of natural purine nucleosides, e.g. the

adenosine and guanosine. These inhibitors can be used in the treatment of numerous diseases, from

Parkinson's and Alzheimer’s deseages to malignant tumors. The perspective target in the wide area

of azoloazines is 6-nitro-7-aminoazolo [5,1-c] [1,2,4] triazines (6). It is already known that some of

those compounds possesses of medicinal action. A method of synthesis of compounds (6) consists

in reacting 3-diazoazoles 4 with nitroacetonitrile. Significant limitation of the method is the

instability of compound, which is the obstacle in the synthesis of nitroaminoazolotriazines. Now we

have developed an effective and simply method of synthesis of 6-nitro-7-aminoazolo [5,1-c][1,2,4]

triazines (6) using suitable nitrocyanacetic ester potassium salt (1), prepared by oxidation

isonitrosocyanacetic ester. The salt 1 is decarboxylated by the treatment with an alkali solution to

produce the potassium salt of nitroacetonitrile 2, which is used without isolation in azocoupling

reaction. A direct proof of the existence of stable potassium salt of nitroacetonitrile is the X-ray

analysis of its crystals grown from methanol. Thus one can receive azolylhydrazones of nitriles of

2-nitroglyoxylic acid (5) in moderate yields by azocoupling 3-diazotriazoles or 3-diazopirazoles

with the potassium salt of nitroacetonitrile 2 in aqueous solution. 6-nitro-7-aminoazolo [5,1-c]

[1,2,4] triazines (6) are obtained by reflux the hydrazones (5) in DMF.

NHN

X N2+

R

NHN

X NH

N

NO2NC

R

NN

NN

X

NH2

NO2

R

NO2

NC

Cl-

NHN

X NH2

R

O

NO2

NCOEt

NO2

NC

+

(4)

(5) (6)

a: R=H, X=N; b: R=Me, X=N; c: R=SMe, X=N;d: R=COOEt, X=N; e: R=b-Py, X=N;f: R=SMe, X=CCN; g: R=SMe, X=CCOOEt.

- K+

t0

AcONaNaNO2

HCl

(3) (2)

-K+

+ KOH- K+

(1) (2)

AcONa

Thus, we have upgrade method for the synthesis aminoazolo [5,1-c][1,2,4] triazines as potential

receptors inhibitors of natural purine nucleosides.

We thank the Russian Science Foundation grant № 14-13-01301.


306

Computer-aided Prediction of Bioactivity of Glucose-modified Benzocaine

T.G. Volkova,1 I.O. Talanova,2 A.M. Ryzhakov,3 A.I. Tsarkova1

1Ivanovo State University, 153025, Ivanovo, street Ermak, 39

2Ivanovo State Medical Academy, 153012, Ivanovo, Sheremetév prospekt, 8 3Institute of Physiologically Active compounds Russian Academy of Sciences, Department of

Computer-Aided Molecular Design, Institute of Physiologically Active Compounds RAS, 142432, Moscow region, Chernogolovka, Severnii proezd, 1


The problem of chemical modification of p-amino benzoic acid has no little interest now. It is

due to possibility of varying of some physicochemical properties of medical drugs without changing

of the physiological effect character by addition of the carbohydrate moieties to the drug structure.

The aim of this research was a comparative analysis of lipophilic properties of benzocaine (a)

and ethyl ester of p-(N-the-D-glucopyranosid)aminobenzoic acid (b) and studies on their potential

target-specific activity.

+ -H2O

OCH2OH

OH OH

OHOH

NH2 COOС2Н5

CH2OH

COOС2Н5

O

OH OH

OHNH

а b

The coefficient of lipophilicity logP for a standard system octane-1-ol – water has been

determined using complex software ACD / Labs. It permits to predict the physico-chemical

behavior of different molecules [1].

The potential bioactivity profile of ethyl ester of p-(N-α-D-glucopyranoside)aminobenzoic acid

was obtained based on data base of substances of known biologically active compounds using

software PASS [2].

It was found that ethyl ester of p-(N-α-D-glucopyranoside)aminobenzoic acid will possess

antiviral properties, and will participate in the metabolism of carbohydrates, amino acids and lipids.

However, screening data indicate a rather high risk under using of dry pure substance in view of its

possible toxicity. Also the hyperuricemia is possible during its application. It was shown that

glucose-modified benzocaine should be more soluble in aqueous solution. It makes possibility to

develop new dosage forms of the latter with a higher bioavailability.

Reference [1] http://www.acdlabs.com. [2] http://pharmaexpert.ru/passonline.


307

Influence of Pectins Isolated from Different Raw Materials on

Blood Parameters of Rats

A. Vyshtakalyuk,1 S. Minzanova,1 L. Mironova,1 V. Zobov,1,2 V. Mironov1

1A.E. Arbuzov Institute of Organic and Physical Chemistry of Kazan Scientific Center of Russian

Academy of Sciences, 420088, Russia, Kazan, Arbuzov str., 8 2Kazan Federal University, 420008, Russia, Kazan, Kremlyovskaya str., 18


Pectin, being a unique biologically active product, possessing functional properties, presents

special interest. According to the literature data pectin effects on some parameters of immunity,

specifically T-lymphocytes and phagocytic activity of neutrophils [1, 2]. It was shown the pectin

with low molecular mass and high esterification degree improves bioavailability of iron from iron

sulfate [3, 4].

The objective of our investigations is to study the influence of pectins isolated from amaranth,

daikon, apple pomace [5] as well apple and citrus pectins supplied by “Herbstreith & Fox”

Company Group (Neuenbuerg, Germany) on morphological and immunological parameters of rats

blood. Investigated pectins had been adding to drinking water of experimental rats in the doses 60-

140 mg/kg for 7-12 weeks. The blood parameters were measured by methods of optical microscopy

and spectrophotometry.

It was established that all investigated pectins effect predominantly on the immunological

parameters of blood. The leucocytes level increased as well leukogram normalized under pectin

treatment. Thus, the quantity of medullocells and young forms of granulocytes as well large

lymphocytes decreased. At the same time, the quantity of mature immune cells increased.

Moreover, the phagocytic activity of neutrophil to Escherichia coli and Staphylococcus aureus

improved specifically the number of englobed neutrophil in the groups of rats treated with pectins.

The effect of pectins on the level of hemoglobin and erythrocytes manifested less specifically in the

increase of the erythrocytes number.

Therefore, the study revealed the positive influence of pectins on the investigated parameters of

the rats blood.

References: [1] N.V. Novikova, R.O. Hamzamulin, N.P. Sheluchina et al., Environment and health of human, Новикова, – Byshkek. 1992, 81-85. [2] D.A. Demidov, M.V. Kostiuchenko, P.A. Dvoenosova and O.N. Shevernitskaia, Experimental & clinical gastroenterology. 2009, Is. 2, 65-73. [3] M. Kim and M.T. Atallah, The Journal of Nutrition. 1992, 122, 2298-2305. [4] M. Kim, M.T. Atallah, C. Amarasiriwardena and R. Barnes, The Journal of Nutritional. 1996, 126, 1883-1890. [5] S.T. Minzanova, V.F. Mironov, A.I. Konovalov, A.B. Vyshtakalyuk, O.V. Tsepaeva, A.Z. Mindubaev, L.G. Mironova and V.V. Zobov, Pectins from non-traditional raw materials: technology, properties and biological activity, Kazan. 2011, 226.


308

Mechanochemically-Assisted Extraction of Water Solutable Pectin from

Gardenia Jasminoides

W. Xu, A.V. Dushkin and W. Su

Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou 310014, China

Gardenia fruit is a common Chinese herbal medicine; active compounds gardenia yellow and

geniposide are mainly product extracted from it. However, a large quantity of pectin contained in

Gardenia fruit can’t be fully extracted and utilized. For the purpose of reducing waste and

enhancing economic benefits of Gardenia, the technology of mechanochemistry assisted extraction1,

which has been successfully used to extract active compound from plants2 was applied to extract

pectin. In this study, according to the chemical property of pectin and experimental results, 10%wt

sodium carbonate was chosen as milling aids and 1% wt sodium bisulfite was added so as to avoid

possible Maillard reaction3. Optimal ball-milling condition was obtained in a planetary ball mill at

400 rpm for 5 min. After extracted with boiling water (10:1 liquid-solid ratio) for 5 min and

neutralized by acetic acid, the yield of pectin could reach to 13.2% in 75% purity.

Mechanochemistry assisted extraction not only avoids the application of excess acids that can

significantly reduce environmental pollution, but also shortens the extraction time that can reduce

energy consumption. Consequently, this method is eco-friendly and has a splendid industrial

prospect.

Gardeniawater solutable Pect in

O

H

H

COOH

H

OHHO

n

1. milled at 400 rpm, 5 min

2. water 5 min then neutralized

Less acid and rapidly

References 1. Martinez, T. Understanding mechanochemistry from first principles. Abstracts of Papers, 249th ACS National

Meeting & Exposition, Denver, CO, United States, March 22-26, 2015, PMSE-103. 2. Xie, J., Li, H., Zhu, X., Wang, P., Su, W. Efficient and selective extraction of magnolol from Magnolia officinalis

by mechanochemical extraction technique. Chem. Eng. Process., 2011, 50(3), 325-330. 3. Scaman, C., Nakai, S., Aminlari, M. Effect of pH, temperature and sodium bisulfite or cysteine on the level of

Maillard-based conjugation of lysozyme with dextran, galactomannan and mannan. Food Chem., 2006, 99(2), 368-380.


309

New Vitamin E Analogues: Sulfides and Selenides Based on 5-Hydroxy-2,3-

Dihydrobenzofurans

S. Yagunov,1 S. Kholshin,1 N. Kandalintseva,1,2 and A. Prosenko1,2

1Novosibirsk state pedagogical university

630126, Russia, Novosibirsk, Viluiskaya str, 28 2Novosibirsk institute of antioxidants

630091, Russia, Novosibirsk, Krasnyi prospekt str, 54а


Chromane ring is the basis of wide range of biologically active compounds (tocopherols,

flavonoids, coumarins) and drugs (troglitazone, ormeloxifene, nebivolol). Derivatives of 6-

hydroxychromanes and in particular vitamin E have unique biological properties and high

antioxidant activity. Thus, 6-hydroxychormanes and its homologues is perspective building blocks

for creation new bioantioxidants and drugs.

Recently, we prepared a number of sulfur-containing derivatives of 5-hydroxy-2,3-

dihydrobenzofuranes and 6-hydroxychromanes with far exceeding antioxidant activity of α-

tocopherol [1]. In this work we present a new sulfur- and selenium-containing analogues of

tocopherols: 2-dodecylthiometyl- and 2-dodecylselenometyl-5-hydroxy-2,3-dihydrobenzofurans 1

and 2.

O

HO

SeC12H25

R1

R2

R3

R1 = R2 = R3 = Me (a); R1 = R3 = Me, R2 = H (b); R1 = H, R2 = R3 = Me (c);R1 = R2 = H, R3 = Me (d); R1 = R2 = R3 = H (e)

O

EtO

I

R1

R2

R3

OH

OEt

R1R2

R3i

ii, iv, v

i: I2, MeCN, rt. ii: C12H25SH, NaOH, EtOH, 50oC. iii: C12H25SeSeC12H25, NaBH4, EtOH, ∆. iv: BBr3, DCM, rt. v: PhMe, ∆.

O

HO

SC12H25

R1

R2

R31

2

iii, iv, v

Sulfides 1 and Selenides 2 were obtained based on 2-iodometyl-5-hydroxy-2,3-

dihydrobenzofurans by efficient synthetic protocol including iodocyclization 2-allyl-4-

ethoxyphenols in acetonitrile at room temperature, substitution iodine by dodecanethiol or -selenol

in EtOH and cleavage ethoxy group with BBr3.

In AIBN-initiated autooxidation of styrene compounds 1 and 2 showed exceed antioxidant

activity than tocopherols.

References [1] S.E. Yagunov, S.V. Kholshin, N.V. Kandalintseva and A.E. Prosenko, Russ. Chem. Bull.. 2013, 62, 1395-1400


310

Silver Nanoparticles Stabilized in a Poly(ethylene oxide) Matrix – a Prospective

Antimicrobial Agent for Hydrocolloid Wound Dressings

O.V. Yakutkina, A.V. Kharenko, S.V. Antonov, Yu.M. Korolev

Over the last 20 years great attention has been focused on antimicrobial hydrocolloid wound

dressings which are used for the treatment of chronic wounds and burns. This is a type of dressing

containing gel-forming agent, which absorbs wound exudate, thereby maintaining a moist wound

environment, while keeping microbal activity at low level by releasing an antimicrobial agent into

the.

Among antimicrobial agents silver is the most effective, because in the process of evolution

bacteria failed to develop resistance to silver, since can attack various sites in the cell. Silver in the

ion form has a cauterizing and irritating effect on the skin and mucous because of its high reactivity.

Also it rapidly interacts with different cell components losing activity. Therefore using colloidal

silver became a tendency of the recent years. It has less toxicity than ionic silver and it is in active

state to provide therapeutic and bactericidal effect.

So, the aim of this work was develop a formulation containing silver nanoparticles stabilized in

the poly(ethylene oxide) (PEO) matrix for further use in the hydrocolloid wound dressings as a

complex antimicrobial agent.

The choice of the PEO as a matrix forming polymer is determined by its unique properties. First,

PEO is a biocompatible polymer which is approved for medical applications. Second, PEO is well

soluble in aqueous media, it has the adhesion to moist surfaces, i.e. the release of the exudate in

wounds and burns would promote the establishment of the adhesion contact as well as initiate silver

release from the formulation into the wound. And finally, PEO is able to form a polymer complex

(PC) in the mixtures of PEO and a silver nitrate, which persists in dry films. It worth notice that

PEO is a meltable polymer, so the preparation of the hydrocolloid compositions by hot melt mixing

is possible, which facilitates manufacturing in an industrial scale.

The optimal method of silver nanoparticles preparation and stabilization in the PEO matrix was

developed with formaldehyde used as a reducing agent. The formation of the PC by PEO and silver

nitrate in films was confirmed by infrared spectroscopy and differential scanning calorimetry.

Transmission electron microscopy images demonstrate very iniform size distribution of silver

nanoparticles. The developed PEO-silver nanoparticles formulations were then used for preparation

of hydrocolloid wound dressings samples. The silver release from the prepared samples was tested

and compared with that of the commercially available wound dressings.


311

Consensus screening of new inhibitors of glycogen phosphorylase in class of

adamantane derivatives

L. Yanalieva,1 P. Vassiliev,1 A. Spasov,1 N. Cheplyaeva,1 E. Vorobiev,1 Yu. Popov,2

V. Lobasenko,2 E. Shishkin2 and V. Mokhov2





Liver Glycogen Phosphorylase (PYGL) plays a central role in the maintenance of glucose

homeostasis [1], it is therapeutic target in treatment of diabetes mellitus type 2. An adamantane

derivative is the most studied and interest class of chemicals. The aim is a consensus virtual

screening and experimental study of 1-substituted (I) and 2-substituted (II) adamantane derivatives

as potential inhibitors of PYGL. Consensus screening in silico of nineteen

compounds (I, II) was executed using two prediction systems. PASS

program [2] allows for the structural formula to estimate the probability of

the presence of activity and the probability of its absence. The method of

similarity to the standards of IT Microcosm [3] is based on the calculation

of modified Tanimoto similarity coefficient for predicted substances to the

reference drugs. According to the prediction results was calculated index of

perspectivity and a membership function to the class of PYGL inhibitors.

The consensus prediction estimates served as the criterion for the selection

of compounds. One substance must be showed PYGL inhibitory activity. Nineteen substances were

studied experimentally by the method of tablet screening [4] using multidetector TECAN Infinite

Pro 200. One of nineteen substances show a statistically significant level of activity. Thus, the

accuracy of consensus screening of active structures is about 100%. The results show that

consensus screening is highly effective and an adamantane derivative is novel class of molecules for

design of glycogen phosphorylase inhibitors.

This work was supported by RSF grant № 14-25-00139.

References [1] L. Somsak, K. Czifrak, M. Toth et al., Curr. Med. Chem. 2008, 15, 2933-2983 [2] D. A. Filimonov, V. A. Poroikov, T. A. Gloriozova et al., PASS (Prediction of Activity Spectra for Substances). State Registration Certificate for software program 2006613275 (Russian). 2006 [3] P. M. Vassiliev, and A. N. Kochetkov, IT Microcosm. State Registration Certificate for software program 2011618547 (Russian). 2011 [4] L. Yu, Y. Chen, and J. Treadway, Pharmacol. Exp. Ther. 2006 , 317, 1230-1237

(CH2)nN

R1 R2

N

O

R1

R2

R3

I

II


312

Pyrophosphate Analogs Suppress Phosphorolytic Activity of Wild-type and

AZT-resistant HIV-1 Reverse Transcriptase

M.K. Kukhanova, A.N. Korovina, N.N. Usanov, S.N. Kochetkov, D.V. Yanvarev

Laboratory of molecular basis of physiologically active compounds, Engelhardt Institute of

Molecular Biology RAS, Moscow, Russia

The first drug approved for the clinical treatment of HIV-1 infection was a nucleoside analog 3’-

azido-3’-deoxythymidine (AZT). However, shortly after the introduction of AZT, cases of

resistance were found. This resistance originates in the phosphorolytic excision of AZT by

inorganic pyrophosphate or ATP catalyzed by viral reverse transcriptase (RT). In most cases,

resistance originates from specific RT mutations called thymidine analog mutations (TAM). The

mutations accelerate the excision of AZT so dramatically that AZT becomes almost ineffective.

Specific inhibitors of AZT-excision could have therapeutic utility, since they might restore the

activity of the nucleoside-based drugs against TAM-containing HIV. Optimally, such inhibitors

should not reduce incorporation of AZT, since this would clearly antagonize it anti-retroviral

activity.

We present the results of an investigation of the inhibition of the PPi or ATP-mediated HIV-1

reverse transcriptase catalyzed phosphorolytic cleavage of AZT-terminated DNA primers by a

series of 30 bisphosphonates (BPs). The influence of the structure of Mg2+-coordinating group and

an electron deficiency of the side-chain aromatic substituent of BPs on their activities were studied.

We compared the ability of the most active BPs to suppress the PPi and ATP induced excision

catalyzed by WT or AZT resistant RTs (M41L, D67N, K70R, T215F, K219Q). For all BPs the

correlation between their inhibiting activities in AZT-excision and DNA primer elongation

processes were evaluated. We postulated that where is an almost linear dependence between the

ability of studied BPs to inhibit of both excision and elongation activity of RT and it should be

mentioned that selective BPs are not sensitive to drug resistant RT.

The work was supported by Russian Science Foundation, project No 14-50-00060


313

Synthesis of Novel Non-natural Conformationally Restricted Cyclopropane

Amino Acids of Spiro[2.3]hexane series

N. Yashin,1,2 E. Averina,1,2 T. Kuznetsova,1 N. Zefirov1,2

1 Lomonosov Moscow State University, Chemistry Department, 119991, Russia, Moscow, Leninskie

gory, 1-3 2 Institute of Physiologically Active Compounds of RAS,142432, Russia, Chernogolovka, Moscow

Region, Severny proezd, 1


The novel approach to 5-aminospiro[2.3]hexanecarboxylic acid was developed on the basis of 3-

methylenecyclobutanecarboxylic acid 1. The first step of the synthesis includes modification of

Curtius reaction of 3-methylenecyclobutanecarboxylic acid 1.

CO2H1) ClCO2Et, Et3N

2) NaN3, H2O3) t-BuOH, t oC

NH-Boc

2, 90%1

Reaction of [1+2]-cycloaddition of diazoacetic ester 3 to (methylene)cyclobutylamine 2 was

thoroughly investigated. The best yield of adduct 4 was achieved using dirhodium tetraacetate as a

catalyst and refluxing dichloromethane as a solvent. Pd(OAc)2

NH-Boc N2 CH

CO2Et

1 1.5:

Rh2(OAc)4

CH2Cl2, 40 oC

Rh2(OAc)4CH2Cl2, 40 oC

CO2Et

NH-Boc4, dr=4:5

2 3

34 %

10 %

5 %

CH2ClCH2Cl, 80 oC The study of hydrolysis of amino ester 4 is under way now.

Acknowledgments: The financial support of RFBR (grant 14-03-32103-a) is gratefully acknowledged. References [1] A. V. Chemagin, N. V. Yashin, Yu. K. Grishin, T. S. Kuznetsova, N. S. Zefirov, Synthesis, 2010, 3379. [2] N.V. Yashin, E.B. Averina, K.N. Sedenkova, T.S. Kuznetsova, N.S. Zefirov, Russ.Chem. Bull, 2013, 928. [3] N.V. Yashin, A.V. Chemagin, Yu.K. Grishin, E.B. Averina, T.S. Kuznetsova, N.S. Zefirov, Dokl. Chem., 2013,

450, 156.


314

The trolox ionic conjugates with amines - a new type of functionalized bioactive

compounds

Yu.V. Yushkova, E.I. Chernyak, S.V. Morozov, I.A. Grigorèv

Novosibirsk Institute of Organic Chemistry SB RAS, 630090,

Russia, Novosibirsk, Lavrentyev ave. 9, E-mail: [email protected]

The cause of many disease processes is the weakening of the body's antioxidant defenses, which

often leads to socially significant diseases such as Parkinson's disease and Alzheimer's disease,

multiple sclerosis, cataracts, epilepsy, rheumatoid arthritis, ischemia and cancer. Effective treatment

of diseases such pathologies are widely used drugs with antioxidant properties.

Synthesis of hybrid pharmacologically active compounds based on molecules of antioxidants is

one of the most promising ways to develop new medicines. The group of compounds-leaders

having high antioxidant activity includes substances with chroman core - α-tocopherol, Trolox and

dihydroquercetin. Antioxidant Trolox is widely known among them and used to produce

polyfunctional conjugates, in which its molecule covalently linked to various pharmacologically

active compounds.

Most described covalently linked Trolox conjugates are hydrophobic compounds, which limits

their use in a water soluble form of the drug as often required in medical practice. One effective

way to increase the water solubility of organic compounds with acidic or basic properties, is

obtaining their salt forms.

In this work the Trolox ionic conjugates 2 were obtained by interaction with ammonia, alkyl

amines, amine-containing heterocyclic compounds, including nitroxides and plant alkaloids, which

exhibit a wide spectrum of pharmacological activity.

For all obtained Trolox ionic conjugates were detected significant increase in the water solubility

(1-3 order) with preserving of high antioxidant activity.


315

Synthesis and Biological Evaluation of Novel Tyrosyl-DNA Phosphodiesterase 1

Inhibitors with a Benzopentathiepine Moiety

A.L. Zakharenko,1 T.M. Khomenko,2,3 S.V. Zhukova,4 O.A. Koval,1,3 O.D. Zakharova,1

R.O. Anarbaev,1,3 N.A. Lebedeva,1,3 D.V. Korchagina,2 N.I. Komarova,2 V.N. Vasiliev,2

J. Reynisson,5 K.P. Volcho,2,3 N.F. Salakhutdinov2,3 and O.I. Lavrik1,3

1Novosibirsk Institute of Chemical Biology and Fundamental Medicine, SBRAS, 630090, Russian

Federation, Novosibirsk, Akademika Lavrentieva Ave., 8 2N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SBRAS, 630090, Russian Federation,

Novosibirsk, Akademika Lavrentieva Ave.,9 3Novosibirsk State University, 630090, Russian Federation, Novosibirsk, Pirogova 2

4Altai State University, 656049, Russian Federation, Barnaul, Lenina Ave., 61 5School of Chemical Sciences, University of Auckland, New Zealand


Tyrosyl-DNA phosphodiesterase 1 (TDP1) is a promising target for antitumor therapy based on

Top1 poison-mediated DNA damage [1,2]. TDP1 plays an important role in removal stalled Top1-

DNA covalent complexes, generated by DNA topoisomerase I (Top1) inhibitors, such as

camptothecin and some other anticancer drugs (for review see 2). Hence, inhibiting the activity of

TDP1 can theoretically enhance the therapeutic effect of Top1 modulators for anticancer treatment

[3,4].

Several novel benzopentathiepines were synthesized and tested as inhibitors of TDP1. The

benzopentathiepines have IC50 values in the range of 0.2 to 6.0 µM. According to the molecular

modeling, the conformational flexibility of the dibutylamine group of the most effective inhibitor

allows it to occupy an advantageous position for effective binding compared to its cyclic

counterparts. The study of cytotoxicity of these compounds revealed that all compounds cause an

apoptotic cell death in MCF-7 and Hep G2 cells. Therefore the new class of very effective

inhibitors of TDP1 was elaborated.

This work was supported by the Ministry of Education and Science of Russian Federation, State Agreement №14.604.21.0018, ID RFMEFI60414X0018. References [1] F. Cortes Ledesma, S.F. El Khamisy, M.C. Zuma, K. Osborn, and K.W. Caldecott, Nature 2009, 461, 674-678. [2] E.Q. Comeaux, and R.C. van Waardenburg. Drug Metab. Rev. 2014, 46, 494-507. [3] Y. Pommier, E. Leo, H. Zhang, and C. Marchand, Chem. Biol. 2010, 17, 421-433. [4] Y. Pommier, Nat. Rev. Cancer. 2006, 6, 789-802.

2-(Dibutylamino)-N-(8-(trifluoromethyl) benzo[f][1,2,3,4,5] pentathiepin-6-yl) acetamide, novel

TDP1 inhibitor. IC50 = 0,22 µM


316

Novel Camphor- based Compound of Broad Range of Anti-influenza Activity

V.V. Zarubaev,1 A.V. Garshinina,1 S.A. Boldyrev,1 V.A. Fedorova,1 Y.R. Orshanskaya,1

A.A. Shtro,1 A.S. Sokolova,2,3 O.I. Yarovaya,2,3 N.F. Salakhutdinov2,3

1 Influenza Research Institute, prof. Popova 15/17, 197376, St. Petersburg, Russia 2 N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, Siberian Branch, Russian Academy

of Sciences, Lavrentjev Avenue 9, 630090 Novosibirsk, Russia 3 Novosibirsk State University, Pirogova St. 2, 630090 Novosibirsk, Russia

Influenza virus continues to remain one of the leading human respiratory pathogens causing

significant morbidity and mortality around the globe. Due to short-term life cycle and high rate of

mutations influenza virus is able to rapidly develop resistance to clinically available antivirals. This

makes necessary the search and development of new drugs with different targets and mechanisms of

activity. Previously (1, 2) we identified derivatives of camphor as potent virus-inhibiting group of

compounds. Here we report anti-influenza activity of camphor derivative 1,7,7-

trimethylbicyclo[2.2.1]heptan-2-ylidene-aminoethanol (camphecene). In in vitro experiments it

inhibited influenza viruses A(H1, H1pdm09, H3 and H5 subtypes) and B with EC50's lying in

micromolar range. Due to low cytotoxicity it resulted in high selectivity indices (75-645 depending

on the virus). This effect did not depend on susceptibility or resistance of the viruses to adamantane

derivatives amantadine and rimantadine suggesting that its mechanism of activity differs from the

inhibition of M2 ion channel. The compound appeared the most effective when added at the early

stages of viral life cycle (0-2 hours p.i.). In direct hemagglutinin inhibition tests camphecene was

shown to decrease the activity of HA's of influenza viruses A and B. The activity of camphecene

was further confirmed in experiments with influenza virus-infected mice, in which, being used

orally by therapeutic schedule (once a day, days 1 to 5 p.i.) it decreased specific mortality of

animals infected with both influenza A and B viruses, in dose-dependent manner (index of

protection 55.6 and 88.9 %, respectively). Taken together, these results are encouraging for further

development of camphecene-based drug(s) and for exploration of camphor derivatives as highly

prospective group of potential antivirals.

References 1. Sokolova AS, Yarovaya OI, Shernyukov AV, Pokrovsky MA, Pokrovsky AG, Lavrinenko VA, Zarubaev VV,

Tretiak TS, Anfimov PM, Kiselev OI, Beklemishev AB, Salakhutdinov NF. New quaternary ammonium camphor derivatives and their antiviral activity, genotoxic effects and cytotoxicity. Bioorg. Med. Chem. 2013; 21:6690-6698.

2. Sokolova AS, Yarovaya OI, Korchagina DV, Zarubaev VV, Tretiak TS, Anfimov PM, Kiselev OI, Salakhutdinov NF. Camphor-based symmetric diimines as inhibitors of influenza virus reproduction. Bioorg Med Chem. 22 (2014) 2141–2148.


317

Mechanochemical-assisted extraction of polysaccharides and

alkaloids from Sophora Flavescens Ait

Q. Zhang, J. Yu, W. Su

Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Hangzhou 310014, P.R. China


A mechanochemical-assisted extraction technology (MAET) was proposed for the efficient

extraction of polysaccharides and alkaloids from Sophora Flavescens Ait (SFA). Polysaccharides

from SFA were functional food and alkaloids were biological pesticide. The objective of MAET is

to eco-friendly extract the polysaccharides and alkaloids as well as reduce the cost of production.

The Highest yield of polysaccharides and alkaloids was obtained by grinding the roots with solid

reagent for 15 min, and extracting with water at room temperature. Polysaccharides were retained

through ultrafiltration membrane with certain pore size. Antioxidant activity of polysaccharides was

assessed by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay. Compared with heat

reflux extraction (HRE), MAET showed the advantages of higher yield, selectivity and efficiency,

lower extraction temperature, no organic solvent consumption, low-cost of production, and

environment-friendliness. The mechanism during the extraction process of ball milling was

investigated by the methods of particle size distribution analysis, scanning electron microscopy,

diffuse-reflectance UV–Vis spectrum, and X-ray powder diffraction.

References 1. X. H. Zhang, L. N. Liu, C. W. Lin, Isolation, structural characterization and antioxidant activity of a neutral

polysaccharide from Sisal waste. Food Hydrocolloids, 39 (2014) 10-18. 2. G. T. Chen, X. M. Ma, S. T. Liu, Y. L. Liao, G.Q. Zhao, Isolation, purification and antioxidant activities of

polysaccharides from Grifola frondosa. Carbohydrate Polymers, 89 (2012) 61-66. 3. W. T. Bi, M. L. Tian, K. H. Row, Solid-phase extraction of matrine and oxymatrine from Sophora Flavescens Ait

using amino-imidazolium polymer. Journal of Seperation Science, 33(2010) 1739-1745. 4. N. Funaya, J. Haginaka, Matrine- and oxymatrine-imprinted monodisperse polymers prepared by precipitation

polymerization and their applications for the selective extraction of matrine-type alkaloids from Sophora flavescens Aiton. Journal of Chromatography A, 1248 (2012) 18-23.


318

Synthesis and Cytotoxicity of New Functionalized

Fluorinated 1,4-Naphtoquinones

S. Zhivetyeva,1 L. Goryunov,1* V. Shteingarts,1* O. Zakharova2 and G. Nevinsky2


Novosibirsk, Lavrentiev av., 9 2 Institute of Chemical Biology and Fundamental Medicine, SB RAS, 630090, Russia, Novosibirsk,

Lavrentiev av., 8


Phosphanium betaine derivatives of polyfluorinated 1,4-naphthoquinones are potential inhibitors

of tumoral cells growth and antioxidants protecting cells against spontaneous mutagenesis [1]. It

was one of the reasons that motivated us to prepare and study a new group of betaines by reaction

of quinone (1) with phosphanes PR1R2Ph (R1 = Me, Ph; R2 = Ph, 3,5-, 2,5-F2C6H3).

Compounds 2─5 were examined for their ability

to inhibit the growth of two mammalian cell lines:

tumor cell lines from human mammary

adenocarcinoma (MCF-7) and human myeloma

(RPMI 8226) as well as normal cell lines (LMTK)

and (AG). It has been revealed that all compounds

2─5 demonstrate enhanced cytotoxicity in all tumor cell lines.

The work is supported by the RFBR (grant № 14-03-00108). References [1] G.A. Nevinsky, O.D. Zakharova, L.I. Goryunov, S.I. Zhivetyeva, V.D. Shteingarts, RU Patent 2535676, 2014. * Deceased

№

IC50

MCF-7 RPMI LMTK AG

2 4.6 6.4 7.7 23.0

3 5.5 8.5 10.4 50.0

4 3.6 5.8 7.6 19.8

5 4.5 4.7 6.8 16.8


319

Synthesis and pharmacological activity of trifluoromethylphenyl substituted

imidazo[1,2-a ]benzimidazoles

V.A. Anisimova, O.N. Zhukovskaya

Institute of physical and organic chemistry, Southern Federal University 344090, Rostov-on-Don, prosp. Stachki, 194/2


Develepment of methods for the modification of fluorine-substituted heterocycles is a promising direction for the creation of biologically active substances, since about 25% of drugs are presented in the pharmaceutical market, contain at least one fluorine atom [1]. Selection of priority determined by aesthetic purposes developed in resent years, the concept of preferred structural- “molecular fragments having affinity for a large number of receptors” [2]. Benzimidazole and its derivatives - imidazo[1,2-a]benzimidazoles are part of the preferred structures class.

4-Fluorophenyl substituted imidazo[1,2-a]benzimidazoles (1,2) were prepared and studied before. It was determined that the compounds (1), where NR2= pirrolidino, morfolino, n=2; m=2, Y=Cl

exhibit kappa-opiod agonist activity [3], and action analgesic effect [4]. Compound 2 (n=3,

NR2=N(C2H5)2, m=2, Y=Cl), has a more effective local anesthetic effect compared to dekain and marcain [4]. In order to expand the scope of research and establish the influence of the trifluoromethyl group on pharmacological activity, we obtained 4-trifluoromethylphenyl substituted imidazo[1,2-a]benzimidazole (5). 4-Trifluoromethylphenacylbromide was used for their synthesis. Thus, the reaction 1-dialkylaminoalkyl-2-aminobenzimidazole with 4-trifluoromethylphenacylbromide produce quarternary salt form (4), cyclization of which leads to 9-dialkylaminoalkyl-2-(4-trifluoromethylphenyl)imidazo[1,2-a]benzimidazole(5).

Currently, the pharmacological active compounds (5) is investigated at the department of

pharmacology, Volgograd State Medical University The work funded as part of the design of the public tasks in scientific research, project #4.196.2014/K

References [1] O'Hagan D. Understanding organofluorine chemistry. An introduction in the C-F bond// Chem.Soc.Rev., 2007,

v.37, p. 308-319. [2] DeSimone R.W. et all. Privileged Structures: Applications in Drug Discovery//Combinatorial chemistry &High

throughput screening, 2004, v.7, p. 473-493 [3] RF pat. 2413512; Bull. № 7 ( 2011). RF pat. 2412187; Bull. №5 (2011) [4] RF pat. 2311416; Bull. № 33 (2007). RF pat. 2313341; Bull. № 36 (2007)


320

Synthesis and Antimicrobial Activity of N-(2-alkoxy-2-oxoethyl) Pyridinium

Chloride and Tetrafluoroborates

O.E. Zhuravlev, A.A. Belyaeva, and L.I. Voronchihina

Department of Organic Chemistry, Tver State University, 170002, Russia, Tver, Sadovy alley, 35


Quaternary ammonium compounds containing in the molecule one or two higher radical used for many years in medicine as antimicrobial agents. Clearly, the biological activity of quaternary amine salt affects the nature of the cation and anion, so replacement of traditional halide anion in the structure of salt on the bulk type [PF6]-, [BF4]-, [FeCl4]- and other anions should affect to change their biological activity [1]. Such compounds - quaternary ammonium salts, pyridinium, imidazolium containing bulky hydrophobic or hydrophilic anions and remain in the liquid state in a wide temperature range, known as ionic liquids (IL) have recently attracted the attention of researchers due to their unique properties. Ionic liquids containing as pyridinium cation moiety with a long chain alkyl substituent may be potential antimicrobial agents.

In this work was synthesis of N-(2-alkoxy-2-oxoethyl) pyridinium chlorides and tetrafluoroborates and studied for their antimicrobial activity. In the scheme illustrates the synthesis of chlorides and tetrafluoroborates N-(2-alkoxy-2-oxoethyl) pyridinium. All synthesized compounds were characterized by IR spectroscopy and elemental analysis.

+ R OHtoluen, t

0

CH2 C

O

ORClCH2 C

O

OHCl + N

N+

CH2 C

O

OR Cl- NaBF4, H2O

N+

CH2 C

O

OR BF4-

- NaCl

p-toluenesulfonicacid

R: C2H5, C4H9, C6H13, C10H21, C12H25

Antibacterial activity was studied by the agar diffusion method in Muller—Hinton medium against test cultures of Gram-positive bacteria Staphylococcus aureus ATCC 25923 and Bacillus

subtilis 534 and Gram-negative Escherichia coli ATCC 25922, Shigella sonnei I phase 941, Pseudomonas aeruginosa ATCC 27853, and Salmonella yphimurium 5715. Antifungal activity was studied against yeast-like fungus Candida albicans ATCC 885 – 653.

The study showed that substitution on the Cl- anion BF4- consisting of quaternary pyridinium

salts increases the bacteriostatic activity of the compounds. This study was performed under financial support by the President of the Russian Federation (program for state support of young Russian scientists, candidates of science; project no. 14.Z56.15.4358-MK). References [1] E.V.Kharitonova, O.E. Zhuravlev, V.M. Chervinets, L.I. Voronchikhina, M.A. Demidova, Pharmaceutical Chemistry Journal. 2012. Vol. 46, No. 5, 266-268


321

AUTHOR INDEX

Abdelhamid M.S. ...................................... 131 Abdigalymova B.А. .................................. 205 Abel A.A. .................................................. 120 Abramova T. ..................................... 286, 304 Abulyaisova L.K. ...................................... 268 Abzianidze V..................................... 132, 133 Adekenov S.M. ............. 27, 88, 205, 230, 268 Afanasyeva S............................................. 253 Afiyatullov Sh.Sh. ....................................... 86 Agafontsev A. ........................................... 301 Aksenov A.V. ............................................ 271 Aksinenko A.Yu........................................ 279 Aleksandrova G................................. 134, 135 Aleksanyan D.V. ....................................... 161 Alekseev K.V. ............................................. 81 Alexandrova L.A. ...................... 136, 137, 234 Aliev G. ................................... 28, 56, 64, 129 Alikhanova O. ........................................... 113 Amosova S.V. ........................................... 252 Ananieva I.A. ............................................ 269 Ananikov V.P. ........................................... 172 Anarbaev R.O............................................ 315 Anastyuk S.D. ..................................... 74, 215 Anikina L. ................................................. 253 Anisimova V.A. ........................................ 319 Antokhin A.M. .......................................... 271 Antonenko T.A.......................................... 138 Antonov S.V.............................................. 310 Apanasenko I.E. ........................ 139, 264, 283 Ardashov O. ................................ 63, 127, 251 Arkhipov I.A. .............................................. 76 Artemova N.P............................................ 236 Asatryan N. ......................................... 94, 227 Atazhanova G.A. ....................................... 268 Atazhanova G.А. ............................... 140, 205 Audran G. .................................................... 67 Averina E. ................................. 186, 263, 313 Avtushenko M.S................................ 174, 177 Aztopal N. ................................................. 124 Babina A. .................................................. 141 Babkin V.A. ........................................ 81, 142 Bachurin S. .......................................... 34, 279 Baev D.S. ...................................... 88, 98, 143 Baisarov G.M. ........................................... 230 Bakier S. .................................................... 189 Balaev A.N. ....................................... 196, 243 Balashova T.V. .......................................... 138 Balmagambetova L.T. ............................... 226 Barreto G.E. ................................................ 64 Baryshnikova M.A. ................................... 196 Baskakova Z. ............................................. 213

Beloshapkin S.A. ...................................... 165 Belousova P. ............................................. 194 Belov D.S. ................................................. 238 Belyaeva A.A. ........................................... 320 Berberova N.T. ................................. 144, 278 Berestetskii A............................................ 133 Besednova N. .............................................. 73 Bessonov I. ............................................... 209 Bilan M.I. .................................................... 50 Biryukova M.S. ................................... 77, 145 Blanz J. ..................................................... 207 Bobkov D. ................................................. 218 Bobrovsky S.I. .......................................... 146 Bochkarev M.N......................................... 138 Bodrov A.V............................................... 236 Bodyagin V. .............................................. 147 Bogachenkov A. ....................................... 148 Bogdanova E. ............................................ 149 Bogomyakov A. ........................................ 135 Boldyrev S.A. ........................................... 316 Boldyrev V.V. ................................... 114, 149 Boldyreva E.V. ........................... 35, 149, 267 Bolshakova K.................................... 132, 133 Bolshchikov B. ......................... 113, 122, 294 Boltneva N.P. ...................................... 68, 106 Borawska M.H. ......................................... 189 Borisevich S.S. ........................................... 66 Borisov S.A. ....................................... 77, 150 Borodkin G.I. ............................................ 299 Borovikova L. ........................................... 250 Boteva A. .......................................... 179, 214 Boyarskikh U.A. ......................................... 98 Boymirzaev A. .......................................... 134 Brémond P. ................................................. 67 Brimble M. ................................................ 255 Brylev K.A. ......................................... 82, 280 Buczek K. ................................................. 190 Bulgakova E.............................................. 151 Bunev A.S. ................................ 152, 153, 154 Burakova E. .............................................. 155 Burgart Ya.V....................................... 68, 115 Burov O.N................................................. 266 Byankina A.O. ............................................ 74 Bychkov A.L..................... 156, 192, 248, 298 Bykov E.E. ................................................ 157 del Carmen Arias Esparza M. ..................... 56 Cevatemre B. ............................................ 124 Chafeev M.A............................................... 69 Chalova P. ................................................. 286 Charushin V. ....................... 36, 108, 158, 213 Cheblukova V. .......................................... 201


322

Cheplyaeva N. ........................... 284, 285, 311 Cheresiz S. .................................................. 79 Chernonosov A.A........................................ 77 Chernyak E.I. ............................................ 314 Chernysheva N.B. ..................................... 112 Chibunova E. ............................................. 288 Chislov M. ................................................. 288 Chistov A.A. ....................................... 80, 159 Chisty L. .................................................... 133 Chistyachenko J.V. ................................... 150 Chistyachenko Yu.S. ..................... 76, 77, 160 Chistyakov V............................................. 225 Chmutina K.S. ........................................... 210 Chulakov E.N. ........................................... 231 Chulikova N.S. .......................................... 198 Chupakhin O.N. .... 36, 68, 108, 115, 163, 300 Churusova S.G. ......................................... 161 Cooper M.A. ............................................... 37 Dae-Kun Lee ............................................. 193 Danagulyan G. .................................... 70, 162 Danilenko V.N. ........................................... 38 Danilova M. .............................................. 117 Davaa V.V. .................................................. 81 Deev S.L.................................................... 163 Deeva O.A. .......................................... 96, 164 Demchuk D.V. .......................................... 112 Demidova Yu.S. ........................................ 165 Demin A. ................................................... 166 Denisenko V.A. ........................................... 86 Dmitriev A. ................................................. 53 Dogadina A. .............................................. 148 Dolinina E.S. ............................................. 167 Drašar P. .................................................... 206 Drašar P.B. .................................................. 71 Drebushchak T.N. ..................................... 267 Drebushchak V. ......................................... 149 Druzhilovsky D. .......................................... 53 Du L. ......................................................... 169 Dubovskii P.V. .......................................... 168 Dubrovina V. ............................................. 135 Dueva E.V. ................................................ 263 Durandin N.A. ........................................... 157 Dushkin A.V. .................. 72, 76, 77, 104, 150 ....................................... 160, 169, 283, 308 D'yakonov V. ............................ 143, 170, 171 Dzhemilev U.M. ................................ 170, 171 Dzhemileva L.U. ............................... 170, 171 Efremova O. .............................................. 180 Egorova K.S. ............................................. 172 Elinson M.N. ............................. 146, 233, 258 Epishina T.A. ............................................ 279 Erkin A. ..................................................... 173 Ermakova S. ................ 73, 174, 175, 176, 177

Estrada M. ................................................. 165 Evseenko V.I............................................. 283 Ezhova N. ................................................. 250 Fang-Fang Wang ...................................... 193 Fedenok L.G. .............................................. 92 Fedorov V. ................................................ 240 Fedorova I.A. ............................................ 269 Fedorova O. .............................................. 158 Fedorova V.A. .................................. 178, 316 Fedotov K.Yu. ............................................ 92 Fedotov V.V. .................................... 297, 305 Fefilova I. .......................................... 179, 214 Feofanov A.V. .......................................... 168 Filimonov D. ............................................... 53 Filippov A.S. ............................................. 252 Fomenko V.V. .......................................... 119 Frantseva V. .............................................. 180 Frolova S.Y. .............................................. 196 Frolova T.S. ...................................... 181, 219 Furkert D. .................................................. 255 Gabrel’yan A.V......................................... 279 Gaidul K.V. ................................................. 72 Galkina I. .................................................. 202 Galyametdinova I...................................... 265 Galyautdinov I.V. ..................................... 182 Ganbaatar J. .............................................. 183 Garkushina I.............................................. 250 Garshinina A.V. ........................................ 316 Gasilova E. ................................................ 135 Gatilov Ju.V. ............................................. 205 Genaev A. ................................................... 63 Gerbst A.G. ................................................. 50 Gibadullina G.G. ....................................... 182 Gil A. ........................................................ 289 Glamazdin I.I. ............................................. 76 Glazunov V. .............................................. 215 Gloriozova T. .............................................. 53 Glotov A. .................................................. 117 Godovalov A.P. ........................................ 184 Gogol E.S. ................................................... 81 Golantsov N.E........................................... 185 Goreva T.V. .............................................. 279 Gorokhovets N.V. ..................................... 129 Goryunov L. .............................................. 318 Gracheva Yu. .................................... 138, 186 Grigorèv I.A. ........................................... 314 Grigoriev V.V. .......................................... 279 Grigoryan A. ............................................... 94 Grischenko L.A........................................... 81 Grishin Yu.K............................................. 263 Gruzdev D. ........................................ 158, 231 Gubanova N.V. ......................................... 181 Gudasheva T.A. ............................ 39, 96, 164


323

Gushchina O.I. .......................................... 187 Guz A.V. ................................................... 159 Hae-Dong Jang .................................. 193, 194 Hardy D. .................................................... 207 Harmatha J. ............................................... 206 Hasan Z. .................................................... 120 Helbert W. ................................................. 215 Ho Young Kang ........................................ 276 Hubina A. .................................................. 126 Hwu J. R. ..................................................... 40 Ibrayeva A.K. ............................................ 226 Icsel C. ...................................................... 124 Ignatova A.A. ............................................ 168 Ihalaynen A.A. .......................................... 271 Il’ina I. ......................................... 63, 127, 188 Imbs T.I. .................................................... 177 Isaev N.P. .................................................. 139 Isakov V. ................................................... 215 Isakova E.B. .............................................. 275 Ishigeev R.S. ............................................. 252 Isidorov V.A.............................. 189, 190, 191 Ivanov A.V. ................................................. 59 Ivanov I.D. ................................................ 123 Ivanov I.V. ................................................ 192 Ivanov V. ................................................... 166 Japertas P................................................... 208 Jasko M.V. ................................................ 281 Jhung S.H. ................................................. 120 Ji-Eun Park ................................................ 194 Jin-Tae Lee .................................................. 47 Kairemo K. .................................................. 41 Kalitnik A.A. ............................................... 74 Kamnev A.A. ............................................ 130 Kandalintseva N. ............................... 201, 309 Kapoor M. ................................................... 40 Karakas D. ................................................. 124 Karapetian R.N............................................ 69 Karelson M. ............................................... 116 Karganova G.G. .................................. 99, 159 Karpenko I.L. ............................ 136, 137, 234 Kataev V.E. ................................................. 75 Katokhin A.V. ........................................... 160 Kazachenko A.S. ......................................... 84 Kazachinskai E.I. ...................................... 293 Kazakova A.N. .......................................... 130 Kazakova O.B. .......................................... 203 Kel A.E........................................................ 93 Kel-Margoulis O.V. .................................... 93 Khabibulina A.G. ...................................... 252 Khachatryan D.S. .............................. 196, 243 Khairullina Z.R. ........................................ 182 Khalikov S.S. ........................ 72, 76, 197, 198 Khalymbadzha I.A. ................................... 163

Kharenko A.V. .......................................... 310 Kharitonov Yu.V. ............................. 199, 290 Khazanov V. ............................................... 42 Khlebnikova T. ........................................... 78 Kholshin S. ....................................... 201, 309 Khomenko T. .................................... 239, 315 Khramtsova E.A. ........................................ 87 Khripach V.................................................. 43 Khursan S.L. ............................................... 66 Khusainova N. .......................................... 202 Khusnutdinova E.F. .................................. 203 Khvostov M.I. ............................................. 72 Khvostov M.V. ............... 77, 88, 98, 150, 160 Kilmetiev A.S. .......................................... 204 Kim A.V. .................................................. 139 Kiselev O.I. ......................................... 44, 108 Kiselev S.V. .............................................. 236 Kishkentaeva A.S. .................................... 205 Klochkov S. .............................................. 253 Klochkov V.V. .......................................... 236 Kmoníčková E. ......................................... 206 Kobzeva T.V. ............................................ 291 Kochegarova K.V. .................................... 289 Kochetkov S.N. ........................... 45, 234, 312 Kolchanov N.A. .......................................... 49 Kolesov B. ................................................ 149 Kolotaev A.V. ................................... 196, 243 Kolovanov E. .................................... 207, 208 Kolyada M. ............................................... 144 Komarova N.I. .................................... 98, 315 Kondratenko E.I. ....................................... 131 Konev V. ..................................................... 78 Kononova A. ............................................... 79 Kontoghiorghes G.J. ................................ 291 Kopitsyna M. ............................................ 209 Kopra J. ..................................................... 116 Korchagina D. ....... 63, 95, 188, 239, 251, 315 Korchagina T. ........................................... 180 Kornakova T.A. ........................................ 210 Korobeynikov V. ........................................ 79 Korolev Yu.M. .......................................... 310 Korotkikh M.O. ................................ 212, 244 Korovina A.N. .......................................... 312 Korshun V.A. ...................................... 80, 159 Kostyleva S.A. .................................. 261, 262 Kostyro V.V. ............................................... 81 Kostyro Ya.A .............................................. 81 Kotovskaya S. ........................................... 213 Koval O.A. ................................................ 315 Kovalenko Yu. .......................................... 149 Kovaleva K. ................................................ 79 Kovaleva N.V. ............................................ 68 Kovalskaya A.V. ....................................... 178


324

Kozlov V.A. .............................................. 161 Kozlovskaya L.I. ................................. 99, 159 Krasilnikova A.A. ............................... 82, 280 Krasnov V. ................................ 158, 166, 231 Krasnova A. ...................................... 221, 223 Krasnykh O. .............................. 115, 179, 214 Krasotkina J....................................... 147, 220 Kravchenko A.O. ................................ 74, 215 Kravchenko I. ............................................ 235 Krivorotov D. ............ 132, 216, 217, 218, 270 Krivtsov I. ................................................. 158 Krogsgaard-Larsenn N. ............................. 255 Krutikov V. ............................................... 173 Kubilius R. ................................................ 207 Kucheryavaya D. ....................................... 300 Kudryavtsev K.V. ....................................... 83 Kukhanova M.K. ....................................... 281 Kukina T.P. ....................................... 181, 219 Kumpins V. ............................................... 200 Kurkin A.V........................................ 232, 238 Kurkutov E.O. ........................................... 252 Kusaykin M. ................................................ 73 Kustov P. ................................................... 220 Kutyakov S.V. ........................................... 159 Kuzmitsky B.B. ......................................... 200 Kuznetsov A. ............................................. 221 Kuznetsov B.N. ........................................... 84 Kuznetsov V. ............. 132, 133, 217, 218, 270 Kuznetsova S.A. .................................. 84, 114 Kuznetsova T.S. ................................ 263, 313 Kuznetsova V.A. ............................... 180, 284 Kverka M. ................................................. 206 Kyu Lee ..................................................... 276 Lagunin A. .................................................. 53 Lakatosh S.A. .............................. 85, 245, 275 Lanevskij K. .............................................. 208 Larkina E.A. .............................................. 187 Lashin V. ................................................... 207 Latypova R.M. .......................................... 262 Lavrenov S.N. ................................... 157, 222 Lavrik O.I. ........................................... 46, 315 Lavrinenko V. ........................................... 141 Lebedeva N.A. .......................................... 315 Lepehova S.A. ............................................. 81 Leshchenko E.V. ......................................... 86 Leshchinskaya A. ...................................... 250 Leshina T.V. ................................................ 87 Lesnichaya M. ........................................... 134 Leung E. .................................................... 255 Levdansky A.V. .......................................... 84 Levdansky V.A. .......................................... 84 Levit G.L. .................................................. 231 Lipatnikov K. ............................................ 223

Lipeeva A.V. ............................................... 88 Ljushina G. ............................................... 214 Lkhagvamaa E. ......................................... 183 Lobasenko V. ............................ 180, 285, 311 Lobov A.N. ............................................... 178 Logashenko E.B. ........................... 89, 93, 109 Lomovskiy I.O. ........................................... 90 Lomovsky O.I. ............ 90, 156, 192, 241, 248 ...................................................... 293, 298 Lomteva N.A. ........................................... 131 Lukyanenko E.R. .............................. 232, 238 Lushchekina S.V. ................................ 68, 106 Luzina O.A. ........................................ 91, 273 Makara N.S. ................................................ 66 Makarov A.A. ................................... 170, 171 Makarov V.A. ........................................... 129 Makhaeva G.F..................................... 68, 106 Malyar Yu.N. ............................................ 114 Malyarenko (Vishchuk) O. ....................... 175 Malysheva L. .............................................. 78 Malyuga A.A. ........................................... 198 Manakov A. .............................................. 149 Markov A.V. ................................. 89, 93, 109 Markova I.D. ............................................... 92 Markova L.N............................................. 115 Marque S.R.A. ............................................ 67 Maslova V. ................................................ 214 Matevosyan K.R. ...................................... 196 Matuszek A. .............................................. 255 Mayorov A. ............................................... 179 Medvedeva N.I. ........................................ 203 Medvedeva N.R. ....................................... 259 Melnikov V. .............................................. 225 Merkhatuly N. ........................................... 226 Meteleva E.S. .................................... 160, 283 Mikaelyan A. ...................................... 94, 227 Mikhailenko M.A. .................................... 114 Mikhalchenko O.S. ........................... 127, 289 Mikheev A. ............................................... 244 Milaeva E. ............. 48, 97, 138, 144, 186, 242 Milekhin D. ............................................... 147 Milyukov V. .............................................. 228 Minin A. .................................................... 166 Miniyarova L.R......................................... 262 Min-Ji Seo ................................................. 194 Minzanova S. .................................... 228, 307 Miranda M.A. ............................................. 87 Mirchink E.P. ............................................ 275 Mironov A.F. ............................................ 187 Mironov V. ....................................... 228, 307 Mironov М.Е. ........................................... 229 Mironova L. ...................................... 228, 307 Mitrofanov D.A. ....................................... 271


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Mochulskaya N. ........................................ 213 Mokhov V. ........................................ 285, 311 Mokrov G.V. ............................................... 96 Mordvinov V.A. ........................................ 160 Morozov A. ............................................... 209 Morozov P.G. ............................................ 266 Morozov S.V. ............................................ 314 Morozov S.Yu. .......................................... 129 Moshkin M.P. .............................................. 49 Moskwa J. ................................................. 189 Movchan N. ............................................... 242 Mukhatova E. ............................................ 144 Mukusheva G.K. ....................................... 230 Mulyukova A.R. ................................ 170, 171 Murashkevich A. ....................................... 158 Murzin D.Yu. ............................................ 165 Musalov M.V. ........................................... 252 Musalova M.V. ......................................... 252 Musikhina A. ............................................. 300 Musiyak V.V. ............................................ 231 Myasnyanko I.N. ....................................... 232 Myung-Hee Kang .............................. 193, 194 Myz S.A. ........................................... 149, 267 Narkevitch I............................................... 216 Nasybullin R.F. ......................................... 233 Naushabekova D.D. .................................. 226 Nazarov A.A. .............................................. 97 Nazarova О.А............................................ 205 Nechepurenko I.V. ...................................... 98 Nefedov A. ................................................ 251 Neganova M. ..................................... 186, 279 Negria S.D. ................................................ 234 Nesterkina M. ............................................ 235 Nevinsky G. .............................. 121, 249, 318 Nevskaya K. .............................................. 166 Nifantiev N.E. ............................................. 50 Nikitina L.E. .............................................. 236 Nikolaeva E.A. .......................................... 178 Nikolskaya T.A. ........................................ 187 Nosova E. .................................................. 213 Nosova Y.N. ................................................ 97 Novikova G.V. .......................................... 237 Novoselov A.M. ........................................ 238 Nuzhdina A. .............................................. 209 Odarchenko T. ........................................... 239 Odinokov V.N. .................................. 182, 261 Ogienko A.A. .................................... 149, 267 Ogienko A.G. .................................... 149, 267 Okhlopkova Zh.M. .................................... 276 Okmanovich K.A. ..................................... 243 Omarova A.T. ........................................... 226 Omelkov A. ............................................... 240 Orlov A. .................................................... 220

Orlov A.A. ................................................ 159 Orlov D.V. .......................................... 90, 241 Orshanskaya Y.R. ..................................... 316 Osipenko A. .............................................. 250 Osipov V.N. ...................................... 196, 243 Osipova V. ........................................ 144, 242 Oslovsky V. .............................................. 295 Osolodkin D.I. ............................ 99, 159, 263 Ostapenko G.I. .......................... 152, 153, 154 Ostroukhova L.A. ............................... 81, 142 Ovcharenko V. .......................................... 135 Ovchinnikova L. ....................................... 121 Pai Z. ........................................................... 78 Pakharukova M.Y. .................................... 160 Palyulin V.A. ................ 61, 99, 100, 106, 263 Panchenko V.N. ................................ 120, 289 Pankrushina N. .................................. 212, 244 Panov A.A................................................. 245 Pantileev A.S. ............................................. 96 Parfenyuk E.V........................................... 167 Patchev V. ................................................... 51 Patrushev S. .............................................. 101 Pavlova A............................ 63, 127, 188, 251 Pavlova A.P. ............................................. 153 Peltek S.E. ................................................... 49 Perlovich G.L. ................................... 102, 246 Pershina A. ................................................ 166 Petrenko N.I. ............................................. 128 Petrov K. ................................................... 265 Petrova E.N. ................................................ 81 Petunov S. ................................................. 218 Piepponen P. ............................................. 116 Pisarev O. .......................................... 247, 250 Poddel’sky A.I. ......................................... 278 Podgorbunskikh E.M. ....................... 156, 248 Pogodin P. ................................................... 53 Pokrovsky A.G. ...... 52, 79, 88, 128, 229, 254 Pokrovsky M.A. .......................... 88, 128, 229 Pokushalov E.A. ....................................... 119 Politanskaya L. ................................. 121, 249 Polonik S. .................................................. 103 Polovinkina M. ......................................... 242 Polyakov N.E. ............... 72, 92, 104, 139, 160 .............................................. 264, 283, 291 Polyakova G.G. ......................................... 219 Polyakova I. .............................................. 250 Ponomarev K. ........................................... 251 Popadyuk I.I. ..................................... 105, 109 Popov A.V. ............................................... 123 Popov Yu. ................................. 180, 285, 311 Poroikov V. ................................................. 53 Posvyatenko A.V. ..................................... 172 Potapov V.A. ............................................ 252


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Poveshchenko A.F. ................................... 280 Pozmogova G. ................................... 295, 296 Pozmogova T.N. ......................................... 82 Prazdnova E.V. ......................................... 266 Preobrazhenskaya M.N. ............................ 157 Priporova A.A. .......................................... 269 Prosenko A. ....................................... 201, 309 Proshin A.N. ...................................... 106, 288 Proskurin G.V. ............................................ 80 Pukhov S. .................................................. 253 Pulina N............................................. 221, 223 Pyshnii D. .................................................... 60 Qiuhong D. ................................................ 176 Radchenko E.V. ................................ 100, 106 Radilov A. ......................................... 216, 218 Radnaeva L. .............................................. 287 Rakhmanova M.E........................................ 91 Rakhmatullina А.А. .................................. 236 Ramsh S. ................................................... 218 Raskildina G.Z. ......................................... 130 Ratushnyak A.S. ........................................ 303 Razumova Yu. ........................................... 254 Redkozubova O. ........................................ 186 Renko J.-M. ............................................... 116 Reynisson J. ...................... 107, 255, 256, 315 Reznik V. .................................................. 265 Richardson A............................................. 255 Richardson R.J. ........................................... 68 Robinson E. ............................................... 255 Rodin I.A. .................................................. 271 Romanov V.E. ........................................... 257 Rudakova Ya.I. ......................... 152, 153, 154 Rudik A. ...................................................... 53 Rusinov G. ................................................ 158 Rusinov V.L. ..... 108, 163, 259, 260, 297, 305 Ryabchikova E.I. ....................................... 156 Rybalkina E.Yu. ........................................ 161 Ryzhakov A.M. ......................................... 306 Ryzhkov F. ................................................ 258 Saarma M. ................................................. 116 Sabutskii Yu. ............................................. 103 Saifina L. ................................................... 265 Sakki J. ...................................................... 116 Sal’nikova O.I. .......................................... 219 Salakhutdinov N.F. ........ 63, 79, 89, 91, 93,95 ................................. 98, 105, 109, 118, 119 ............................... 127, 141, 145, 165, 188 ............................... 224, 239, 251, 254, 273 ............................................... 289, 315, 316 Salimova E.V. ........................................... 262 Salnikov G................................................... 63 Salnikova O.I. ........................................... 212 Salomatina O.V. ............ 89, 93, 105, 109, 224

Saloutin V.I. ........................................ 68, 115 Sametov V.P. ............................................ 182 Sang-Hyun Lee ................................. 193, 194 Sapozhnikova I.M. .................................... 259 Saranina I. ................................................. 155 Savateev K.V. ........................... 260, 297, 305 Savchenko R.G. ........................................ 261 Savinova A.O. ............................................. 81 Savvateeva L.V. ........................................ 129 Sazonov A. ................................................ 166 Sazonovas A. ............................................ 208 Sedenkova K.N. ........................................ 263 Seelmann. I ............................................... 110 Seitkalieva M.M. ...................................... 172 Selyutina O.Yu. .................. 92, 104, 139, 264 Semenov V................................................ 265 Semenov V.V. ........................................... 112 Semenova E. ............................................... 79 Semenova M. ............................................ 118 Semenova M.N. ................................ 111, 112 Semenyuk Yu.P. ....................................... 266 Serbin A. ................................... 113, 122, 294 Serebryakova M.V. ................................... 129 Seredenin S.B. .................................... 96, 164 Sergeevichev D.S. ..................................... 119 Serkov I.V. ................................................ 106 Shakhtshneider T.P. .......................... 114, 267 Sham Nikam ............................................... 54 Shamilova S.T........................................... 268 Shapovalova E.N. ..................................... 269 Shashkov A.S. ............................................. 50 Shchegol’kov E.V. .............................. 68, 115 Shchegoleva N. ......................................... 166 Shchetinina I.A. ........................................ 271 Shchipalkin A. .......................................... 270 Shchur I.V. ................................................ 115 Shestakova T.S. ........................................ 163 Shestopalov M.A. ............................... 82, 280 Shestopalova L.V. ............................... 82, 280 Shevchenko N. .......................................... 175 Shevelev O. ............................................... 166 Shevlyakova O.A. ..................................... 271 Shevtsov P.N. ........................................... 279 Shevtsova E. ..................................... 186, 279 Shilov A.G. ............................................... 264 Shipitsyn A.V. .......................................... 272 Shishkin E. ........................................ 285, 311 Shpakovsky D. .......................... 138, 144, 242 Shpigun O.A. .................................... 269, 271 Shteingarts V. ........................... 121, 249, 318 Shtil А.А. .................................................... 55 Shtro A.A. ......................................... 273, 316 Shubenkova E.G. ...................................... 274


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Shubin V.G................................................ 299 Shults E.E. ................... 88, 101, 128, 183, 199 ....................................... 204, 229, 257, 290 Sidorova Y.A. ........................................... 116 Silchenko A.S............................................ 177 Silman V.V.................................................. 96 Silnikov V. ........................................ 155, 304 Silyukov O. ............................................... 288 Simakov A.V. ............................................ 165 Simakova I.L. ............................................ 165 Simonov A.Yu. ......................................... 275 Sinitsyna E. ............................................... 117 Sinitsyna O.I.............................................. 181 Sivtceva S.V. ............................................. 276 Sizova N.V. ............................................... 277 Smirnova I.E. ............................................ 203 Smolyaninov I.V. ...................................... 278 Sobin F. ..................................................... 223 Sokolov D.N................................ 91, 145, 273 Sokolov V.B. ............................................. 279 Sokolova A. ............................... 118, 254, 316 Sokolova O.O. ........................................... 233 Sokov S.A. ........................................ 152, 154 Solis Arias P.E ............................................ 56 Solís Herrera H............................................ 56 Solodnikov S.Yu. .............................. 115, 214 Solomatin Ya.A. ........................................ 222 Solovieva A.O. .................................... 82, 280 Solovieva O. .............................................. 180 Solovyev A.G. ........................................... 129 Solyev P.N. ............................................... 281 Sorokina I. ................................................. 282 Sorokoumova G.M. ................................... 137 Spasov A. .......................... 180, 284, 285, 311 Staloverova N.A. ....................................... 237 Stankevich V.K. .......................................... 81 Startseva V.A. ........................................... 236 Stetsenko D. ................................................ 60 Stolpovskaya E.V. ....................................... 81 Strelnikov A.G. ......................................... 119 Streshnev P.P. ........................................... 159 Su In Cho .................................................. 276 Su W. ......................................... 169, 308, 317 Sukhov B. .......................................... 134, 135 Suleimenova Zh.E. .................................... 226 Suleymanova I........................................... 116 Suntsova L.P. ............................................ 283 Surov A.O. ................................................ 246 Suslov E.V. ....................................... 165, 251 Suzdalev K.F. ............................................ 284 Swiecicka I. ............................................... 190 Talanova I.O.............................................. 306 Taran A. .................................................... 285

Taranchenko V.F. ..................................... 271 Tarasenko Yu. ................................... 286, 304 Taraskin V. ............................................... 287 Tarasov D.................................................. 292 Tennikova T. ..................................... 117, 126 Teplyakova O.I. ........................................ 197 Teplyakova T.V. ....................................... 293 Terekhova I. .............................................. 288 Tets V........................................................ 173 Tikunova N. .............................................. 155 Timofeeva M.N................................. 120, 289 Timoshenko M.A. ..................................... 290 Timoshnikov V.A. .................................... 291 Titova Yu. ................................................. 158 Tkachev A. ................................................ 301 Tolstikova T.G. ................. 63, 72, 77, 88, 127 ...................................... 143, 145, 150, 160 ...................................... 188, 251, 282, 303 Tovbin D. .................................................. 292 Trefilova A................................ 115, 179, 214 Trestsova M. ............................................. 300 Tretyakov E. ............................................. 121 Tretyakova E.V. ........................................ 262 Trofimov B.A. ...................... 59, 81, 134, 135 Trofimov N. .............................................. 149 Trofimova E.G. ......................................... 293 Trofimova N.N. .......................................... 81 Trunzer M. ................................................ 207 Tsarkova A.I. ............................................ 306 Tsay S.-C. ................................................... 40 Tsipisheva I.P. .......................................... 178 Tsirulnik S.A. ........................................... 284 Tsvetkov V................ 113, 122, 294, 295, 296 Tsyganov D.V. .......................................... 112 Tsypysheva I.P. ........................................... 66 Tugarova A.V. .......................................... 130 Tumanyan A. ............................................ 162 Tuominen R.K. ......................................... 116 Turks M. ................................................... 200 Tykheev G. ............................................... 287 Tyurenkov I.N............................................. 69 Ufimtseva E.G. ......................................... 123 Uimin M. .................................................. 166 Ulomsky E.N. ................... 259, 260, 297, 305 Ulukaya E. ................................................ 124 Urazova T.S. ............................................. 298 Urbagarova B. ........................................... 287 Usanov N.N. ............................................. 312 Usoltsev A.N............................................. 299 Usov A.I. ..................................................... 50 Ustinov A.V. ....................................... 80, 159 Ustyuzhanina N.E. ...................................... 50 Utepova I. ................................................. 300


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Utkin Y.N. ................................................. 168 Varizhuk A. ....................................... 295, 296 Varlamov A.V. .......................................... 185 Varlamova A.I. ............................................ 76 Vasil’eva N.Yu............................................ 84 Vasil’yev K.J. ............................................ 271 Vasilenko D............................................... 186 Vasiliev V.N.............................................. 315 Vasilyev A................................................. 148 Vasilyev E. ................................................ 301 Vassiliev P......................... 180, 285, 302, 311 Vavilov N.V. ............................................. 184 Vechkapova S.O................................ 125, 303 Vicente M. A. ............................................ 289 Vigorov A. ........................................ 158, 231 Vinogradova D.V. ..................................... 279 Vishchuk O. .............................................. 176 Vityazeva S. .............................................. 135 Vlakh E. ............................................ 117, 126 Vlasenko N.G. ........................................... 197 Vlassov V. ................................................... 60 Vohtancev I. .............................................. 304 Voinkov E.K. .................................... 297, 305 Volcho K.P. ................... 63, 95, 120, 127, 165 ............................... 188, 239, 251, 289, 315 Volkova A.N. ............................................ 128 Volkova T.G...................................... 288, 306 Volotova E.V. ............................................. 69 Vorobiev E. ............................................... 311 Voronchihina L.I. ...................................... 320 Vorontsova E.V. ........................................ 280 Voskressensky L.G. .................................. 185 Voutilainen M. .......................................... 116 Vyshtakalyuk A................................. 228, 307 Weike Su ................................................... 195 Weiner L.M. ................................................ 92 Xu W. ........................................................ 308 Yagunov S. ........................................ 201, 309 Yakovlev I. ................................................ 216 Yakubovskaya G.A. .................................. 236 Yakutkina O.V. ......................................... 310 Yanalieva L. .............................................. 311 Yanvarev D.V. .......................................... 312 Yarkov S.A.......................................... 96, 164 Yarkova M.A. ..................................... 96, 164 Yarovaya O.I. ...... 79, 118, 141, 145, 254, 316 Yashin N. .................................................. 313 Yermak I.M. ........................................ 74, 215

Yong-Beom Kwon .................................... 193 Young-A Jang ............................................. 47 Young-Ho Kim ......................................... 193 Young-In Kwon ................................ 193, 194 Yu J. .......................................................... 317 Yun Jeong Choe ........................................ 276 Yunusov M.S. ........................................... 178 Yurchenko E. ............................................ 103 Yushkova T. .............................................. 223 Yushkova Yu.V. ....................................... 314 Yuskovets V.............................................. 216 Zaikin P.A. ................................................ 299 Zakharenko A.L. ....................................... 315 Zakharenko A.M. ...................................... 174 Zakharov A. ................................................ 53 Zakharova O. .................... 121, 249, 315, 318 Zakirova N.F. ............................................ 272 Zamay A.S. ............................................... 114 Zambrzycka M. ......................................... 190 Zamyatnin Jr. A.A. ................................... 129 Zanimkhanova P.Z. ..................................... 88 Zapara T.A. ............................................... 303 Zaporozhets T. ............................................ 73 Zaretskaya M.A ........................................ 137 Zarubaev V.V. .................................. 178, 273 Zashikhina N. ............................................ 126 Zefirov N.S. .......... 61, 99, 100, 106, 263, 313 Zefirova O.N. .............................................. 61 Zenkova M.A. ......................... 60, 89, 93, 109 Zernii E.Yu. .............................................. 129 Zhabinskii V. .............................................. 43 Zhang Q. ................................................... 317 Zhenhua Li ................................................ 195 Zhijiang Jiang ........................................... 195 Zhivetyeva S. ............................................ 318 Zhu F. ........................................................ 176 Zhukova N. ....................................... 145, 282 Zhukova S.V. ............................................ 315 Zhukovskaya O.N. .................................... 319 Zhuravlev O.E........................................... 320 Zhuravleva O.I. ........................................... 86 Zídek Z...................................................... 206 Zlotskii S.S. .............................................. 130 Zobov V. ........................................... 265, 307 Zubareva K.E. ..................................... 82, 280 Zueva I. ..................................................... 265 Zvyagintseva T. .......... 73, 174, 175, 176, 177


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MedChem 2015 - Новосибирский Институт Органической ...web.nioch.nsc.ru/medchem2015/images/MedChem_2015_-_book... · 2015. 7. 28. · 2nd Russian Conference