Margaréta VOJTIČKOVÁ under the direction of: Assoc. Prof. Andrej BOHÁČ Dr. Gilles HANQUET 27 th...
50
Development of VEGFR-2 Inhibitors by Ynamide Based Click Chemistry Margaréta VOJTIČKOVÁ under the direction of: Assoc. Prof. Andrej BOHÁČ Dr. Gilles HANQUET 27 th September 2013, Strasbourg
Margaréta VOJTIČKOVÁ under the direction of: Assoc. Prof. Andrej BOHÁČ Dr. Gilles HANQUET 27 th September 2013, Strasbourg
Margarta VOJTIKOV under the direction of: Assoc. Prof. Andrej
BOH Dr. Gilles HANQUET 27 th September 2013, Strasbourg
Slide 2
Summary 1) Introduction 2) Angiogenesis 3) Aim of the project
4) Study towards target ynamides 5) Azides 6) Click Chemistry 7)
Biological assays 8) General conclusion 2 IntroductionAngiogenesis
Aim of the project YnamidesAzides Click Chemistry Biological assays
General conclusion
Slide 3
Introduction WHO: Cancer is uncontrolled growth and spread of
cells Cancer leading cause of mortality 7.6 million dead people in
2008 (13 % of all deaths) neccessity to develop new active
compounds for cancer treatment 3
Slide 4
Angiogenesis 4
Slide 5
Aim of the project PDB: 1Y6A tyrosine kinase complex prepared
in low ca 10 % yield over 5 steps sensitive N-aryloxazole-2-amine
group 5 Harris, P. A.; Cheung, M.; Hunter, R. N.; Brown, M. L.;
Veal, J. M.; Nolte, R. T.; Wang, L.; Liu, W.; Crosby, R. M.;
Johnson, J. H.; Epperly, A. H.; Kumar, R.; Luttrell, D. K.;
Stafford, J. A. J. Med. Chem. 2005, 48, 1610.
Slide 6
1,3-oxazole / 1,2,3-triazole replacement (me-too or me-better)
exchange of heterocyclic core: activity and/or selectivity higher
stability synthetic feasibility better physical and chemical
properties for bioavailability lower toxicity inhibitor novelty
6
Slide 7
Predicted 1,2,3-triazoles interaction analysis, molecular
modelling, docking 7 triazolic analogues of PDB : 1Y6A Discovery
Studio Visualizer 3.5 software Drug like properties prediction
toolkit Molinspiration 7 http://www.rcsb.org/pdb/home/home.do
http://accelrys.com/products/discovery-studio/visualization-download.php
Molinspiration Property Calculation Service
http://www.molinspiration.com/cgi-bin/properties
1,4-regioisomers
Slide 8
docking scores and ligand possess in VEGFR2 are less
interesting significantly higher price of ruthenium catalyst 8
1,5-regioisomers
Slide 9
Retrosynthetic approach 9
Slide 10
Ynamides Literature background 10 chemistry of ynamides
exploded in last decade heteroatom-substituted alkynes ynamines
unstable, the firts report in 1892 ynamides right balance between
stability and reactivity Bode, J. Liebigs Ann. Chem. 1892, 267,
268. DeKorver, K. A.; Li, H. ; Lohse, A. G. ; Hayashi, R. ; Lu, Z.
; Zhang, Y. ; Hsung, R. P. Chem. Rev. 2010, 110, 5064. Evano, G. ;
Jouvin, K.; Coste, A. Synthesis 2013, 45, 17.
Slide 11
Ynamides Literature background - preparation first synthesis of
electron deficient ynamines Viehe, 1972 Zhdankin & Stang, 1994
synthesis of ynamines using alkynyl iodonium triflate / tosylate
salts 11 Janousek, Z. ; Collard, J. ; Viehe, H. G. Angew. Chem.
Int. Ed. 1972, 11, 917. Murch, P.; Williamson, B. L.; Stang, P. J.
Synthesis 1994,1255. Kitamura, T.; Tashi, N.; Tsuda, K.; Fujiwara,
Y. Tetrahedron Lett. 1998, 39, 3787.
Slide 12
expansion of chemistry of ynamides Witulski, Rainer 12
Witulski, B.; Stengel, T. Angew. Chem. Int. Ed. 1998, 37, 489.
Witulski, B.; Gmann, M. Synlett 2000, 1793. Witulski, B.; Stengel,
T. Angew. Chem. Int. Ed. 1998, 38, 2426. Witulski, B.; Stengel, T.;
Fernandez-Hernandez, J.M. Chem. Commun. 2000, 1965. Witulski, B.;
Alayrac, C. Angew. Chem. Int. Ed. 2002, 41, 3281. Rainier, J. D.;
Imbriglio, J. E. J. Org. Chem. 2000, 65, 7272. Rainier, J. D.;
Imbriglio, J. E. Org. Lett. 1999, 1, 2037.
Slide 13
Elimination protocols Zemlicka, 1994 preparation of ynamides
via lithium-halogen exchange Brckner, 2000 preparation of ynamides
from formamides via lithium- halogen exchange 13 Brckner, D.
Synlett 2000, 1402. Brckner,D. Tetrahedron 2006, 62, 3809. Yang, B.
H.; Buchwald, S. L. J. Organomet. Chem. 1999, 576, 125.
Slide 14
Direct N-alkynylation using bromoalkynes Buchwald copper
catalyzed (CuSO 4. 5H 2 O, CuI, Cu 2 O, Cu(OAc) 2 N-arylations of
amides Tam, 1996 improvement of the methodology using KHMDS
Skrydstrup, 2008 second generation of Hsungs protocol mild base - K
3 PO 4 or K 2 CO 3 higher yields 52 91 % 14 J. Am. Chem. Soc. 2002,
124, 7421. Tetrahedron 2006, 62, 3823 J. Org. Chem. 2008, 73,
9447
Slide 15
Ynamides Literature background - utilization 2013 Evano modular
indole synthesis 2006 Cintrant - -addition of ynamides
hydrostannylation 2006 Hsung et al. Lindlar type hydrogenation of
ynamides 15 Org. Lett. 2013, 15, 3122. Tetrahedron Lett. 2006, 47,
3139 J. Org. Chem. 2006, 71, 4170.
Slide 16
Tam 2006 ruthenium catalyzed [2+2]-cycloaddition of alkenes
(bi- and tricyclic) with ynamides Saito 2012 total synthesis of
(-)-Herbindoles A-C - [2+2+2]-cycloaddition Click chemistry K. B.
Sharpless 2001 16 Riddell, N.; Villeneuve, K.; Tam, W. Tetrahedron
2005, 7, 3681. Villeneuve, K.; Riddell, N.; Tam, W. Tetrahedron
2006, 62, 3823. Saito, N.; Ichimaru, T.; Sato, Y. Org. Lett. 2012,
14, 1914.
Slide 17
Ynamides - Proposed synthesis 17
Slide 18
Preparation of model ynamide A) Corey-Fuchs approach published
by Brckner crucial step - selection of EWG (Boc, Piv, Ts) B)
Bestmann-Ohira reaction one step synthesis, all the attempts failed
18 Brckner, D. Synlett 2000, 1402. Brckner,D. Tetrahedron 2006, 62,
3809.
Slide 19
Preparation of target ynamide Pathway A: via Corey-Fuchs
approach problematic preparation of Corey-Fuchs precursor 19
Slide 20
Pathway A: formylation in good yield tosylation - p-TsCl,
without or with (Et 3 N, pyridine, NaH, n-BuLi) base Pathway B:
tosylation in good yield formylation HCOOH + DCC, CDI; BtCHO;
HCOOEt + base; Vilsmeier-Haack formylation; Eschenmoser salt;
acetic-formic anhydride + base 20
Slide 21
21 Application of Corey-Fuchs approach on tosylated
N-formamide.
Slide 22
Pathway C: Transformation of trichloroacetamides to ynamides
Speziale, Smith - 1962; Himbert, Regitz 1972 key step conversion of
trichloroenamine to lithiated ynamine 22 Speziale, A. J.; Smith, L.
R. J. Am. Chem. Soc. 1962, 84, 1868. Himbert, G.; Regitz, M.
Chemische Berichte 1972, 105, 2963.
Slide 23
Pathway D: Direct N-alkynylation of arylamines using iodonium
triflate salts Preparation of triflate salt 23 Tanaka, K.;
Takeishi, K. Synthesis 2007, 18, 2920. Kerwin, S.; Nadipuram, A.
Synlett 2004, 1404. potentialy explosive
Slide 24
using bromoalkynes 2 possible reagents: Tams protocol -COOMe
EWG small yield of alkynylation step 28 %; overall yield 26 % (3
steps) 24 Pathway D: Direct N-alkynylation of arylamines low
boiling point
Slide 25
Skrydstrup protocol 2 key ynamides prepared using COOMe -
Overall yield 88 % over 3 steps alkynylation step 97 % using Boc-
EWG overall yield 52 % over 3 steps 25 Skrydstrup., T. ;
Dooleweerdt, K. ; Birkedal, H. ; Ruhland, T. J. Org. Chem. 2008,
73, 9447.
Slide 26
Azides synthesis of several aromatic azides as partners for
Click Chemistry discovered 140 years ago by Grie polar mesomeric
structures Preparation: from diazonium salts 26 Grie, P. Philos.
Trans. R. Soc. London 1864, 13, 377. Grie, P. Justus Liebigs Ann.
Chem. 1865, 135, 131. Capitosti, S. M.; Hansen, T. P.; Brown, M. L.
Org. Lett. 2003, 5, 2865.
Slide 27
SN aromatic SN aromatic from non-activated aromatic halides Ma,
Zhu 2004 copper-catalyzed reaction, EtOH / water Liang 2005 mild
conditions 27 Zhu, W.; Ma, D. Chem. Comm. 2004, 888. Andersen, J.;
Madsen, U.; Bjrkling, F.; Liang, X. Synlett 2005, 14, 2209.
Slide 28
synthesis of appropriate azides azidation and Suzuki-Miyaura
cross-coupling key steps 28 Preparation of target azides
Slide 29
azide with ortho-substituted pyridyl cycle 57 % overall yield
over 2 steps azide with meta-substituted pyridyl cycle 45 % overall
yield (2 steps) 29 Trokowski, R.; Akine, S.; Nabeshima, T. Dalton
Trans. 2009, 46, 10359. Li, W.; Nelson, D. P.; Jensen, M. S.;
Hoerrner, R. S.; Cai, D; Larsen, R. D. Org. Synth. 2005, 11,
393.
Slide 30
C-H palladium activated acetoxylation recently developed method
for heteroatom-directed functionalization Ph(IOAc) 2, Pd(OAc) 2 in
acetanhydride and benzene 30 Daugulis, O.; Zaitsev, V. G. Angew.
Chem., Int. Ed. 2005, 44, 4046.
Slide 31
Preparation of MOM- protected azidobiaryl V.47d and
unsuccessful preparation of hydroxylated azidobiarym V.40d 31
Doyagez, E. S. Synlett 2005, 10, 1636. Soni, A.; Dutt, A.;
Sattigeri, V.; Cliffe, I. A. Synth. Commun. 2011, 41, 1852.
Slide 32
Pyrrole azide V.43 traces after Suzuki coupling azidation not
performed 32 Gu, Z. ; Zakarian, A. Org. Lett. 2010, 12, 4224.
Morrison, M. D. ; Hanthorn, J. J. ; Pratt, D. A. Org. Lett. 2009,
11, 1051.
Slide 33
Proposed new retrosynthetical approach Prepared 2 new azides
V.78a and V.78b 33 14% over 3 steps 19% over 4 steps
Slide 34
Preparation of urea azide V.42 Overall yield 37 % over 7 steps
34 Kotha, S.; Shah, V. R. Eur. J. Org. Chem. 2008, 1054. Ganesh,
T.; Thepchatri, P.; Du, L. L. Y.; Fu, H; Snyder, J. P.; Sun, A.
Bio. Med. Chem. Lett. 2008, 4982. Chandrappa, S.; Vinaya, T.;
Ramakrishnappa, T.; Rangappa, K. S. Synlett 2010, 3019. Deng, Q.
H.; Wang, J. C.; Xu, Z. J.; Zhou, C. Y.; Che, C. M. Synthesis 2011,
18, 2959.
Slide 35
Preparation of pyrimidine azide V. 42 2 synthetical pathways
performed 35 Yaziji, V,; Rodriguez, D.; Guierrez-de-Terran, H.;
Coehlo, A. ; Caamano, O. ; Garcia-Mera, X. ; Brea, J. ; Loza, M. I.
; Cadavid, M. I. ; Sotelo, E. J. Med. Chem. 2011, 54, 457. Chang,
L. C. W.; Ijzerman, A. P.; Brussee, J. Oct. 1, 2004, United States
Patent US 2007/0032510. Ye, C.; Gao, H.; Boatz,Drake, G. W.;
Twamley, B.; Shreeve, J. M. Angew. Chem. Int. Ed. 2006, p.
7262.
Slide 36
Click Chemistry K. B. Sharpless, 2001 Requirements: modular and
wide in scope higly efficient and give high yields no or
inoffensive by-poducts stereospecific readily available reagents no
solvents or benign solvents simple purification non-chromatographic
techniques 36 K.B. Sharpless Kolb, H. C.; Finn, M. G.; Sharpless,
K. B. Angew. Chem. Int. Ed. 2001, 40, 2004. Becer, C. R.;
Hoogenboom, R.; Schubert, U. S. Angew. Chem. Int. Ed. 2009, 48,
4900.
Slide 37
Huisgen thermal 1,3-dipolar cycloaddition of alkynes and azides
Click Chemistry CuAAC, RuAAC MW accelerated Click Chemistry Cintrat
and Ijsselstijn, 2006 Click Chemistry with ynamides 2004 Eycken
3-component MW accelerated Click chemistry reaction 37 Huisgen, R.
Angew. Chem., Int. Ed. 1963, 2, 633. Lovell, T.; Hilgraf, R.;
Rostovtsev, V. V.; Noodleman, L.; Sharpless, K. B.; Fokin, V. V. J.
Am. Chem. Soc., 2005, 127, 210. Cintrat, J.C.; IJsselstijn, M.
Tetrahedron 2006, 62, 3837. Appakkuttan, P.; Dehaen, W.; Fokin, V.
V.; Eycken, E. V. Org. Lett. 2004, 6, 4223.
Slide 38
Preparation of In Silico predicted triazoles via CuAAC in mild
conditions using the prepared azides and ynamides Classical
retrosynthetical approach 38
Slide 39
39
Slide 40
Preparation of triazole III.26 with MeCOO- protecting group 40
Problematic deprotection of EWG EntryConditionsResults 11 M KOH in
MeOH, rt, overnight Starting material VI.12 + products of
decomposition VI.14 + VI.15 21 M KOH in MeOH, rt, 20 minStarting
material VI.12 31 M KOH in MeOH, reflux, 20 minProducts of
decomposition VI.14 + VI.15 4 0.5 M KOH in ethylene glycol + water,
rt, 20 min Starting material VI.12 50.5 M KOH in ethylene glycol +
water, reflux, 20 min Starting material VI.12 + products of
decomposition VI.14 + VI.15
Slide 41
with Boc- protecting group 41 EntryConditionsResults 15 equiv
TBAF, THF, rt, overnightStarting material VI.16 25 equiv TBAF, THF,
reflux, 30 minProducts of decomposition 312 M HCl / EtOAc = 1 /
2.3, rt, 1 hour Expected product III.26 + Starting material VI.16 +
Products of decomposition 4TFA, rt, 1 hExpected product III.26 +
products of decomposition [.[. Coleman, C. M.; OShea; D. F. J. Am.
Chem. Soc. 2003, 4054. Englund, E. A.; Gopi, H. N.; Appella; D. H.
Org. Lett. 2004, 213
Slide 42
Alternative retrosynthetical approach preparation of triazole
III.24 42
Slide 43
preparation of triazole III.23 problem with solubility and
purification of the resulting triazole 43 not prepared
Slide 44
44 Prepared triazoles
Slide 45
Biological assays IC 50 biological activity radiometric protein
kinase assay 3 active 2 not active 45
Slide 46
redocking biological activities not corresponded to docking
score newer version Dock 3.6 with PDB : 1Y6A triazolic compounds
much more worse binding energies than oxazolic predicted energies
of triazoles ca 21 % less favorable compare to oxazolic isosters
46
Slide 47
influence of isosteric oxazole / triazole replacement oxazolic
core in PDB : 1Y6A surronded by liphophilic amino acid residues
less favourable to bind triazole dipole moment 47 Discovery Studio
http://accelrys.com/products/discovery-studio/visualization-download.php
(visited 2 nd August 2013)
Slide 48
General conclusion study towards preparation of target ynamides
prepared 7 azides suitable for Click Chemistry approach 48
Slide 49
prepared 5 desired 1,2,3-triazoles 3 active VEGFR-2 inhibitors
2 not active inhibitors 49
Slide 50
Acknowledgement To my supervisors Assoc. Prof. Andrej BOH Dr.
Gilles. HANQUET To members of jury my co-workers from ECPM and
Faculty of Natural Sciences in Bratislava Financial support
Comenius University COST STSM 0602 French Embassy in Bratislava my
parents and all my friends 50