FINAL COMPLETION REPORT OF MINOR RESEARCH PROJECT …

FINAL COMPLETION REPORT OF MINOR RESEARCH PROJECT

ENTITLED

“SYNTHESIS OF SOME NEW THIAZINE DERIVATIVES AND THEIR

BIOLOGICAL SCREENING”

FROM 1ST

SEPTEMBER 2017 TO 31ST

AUGUST 2019

SUBMITTED TO

UNIVERSITY GRANTS COMMISSION

WESTERN REGIONAL OFFICE, GANESHKHIND PUNE - 411007

SUBMITTED BY

PRINCIPAL INVESTIGATOR

Mr. UTTAM BANDU CHOUGALE.

M.Sc., NET.

DEPARTMENT OF CHEMISTRY

KARMAVEER HIRE ARTS, SCIENCE, COMMERCE AND EDUCATION

COLLEGE, GARGOTI. 416209

DISTRICT KOLHAPUR (M.S.) INDIA

ACKNOWLEDGEMENT

I express my deep sense of gratitude to the University Grants Commission, WRO

Pune for financial assistance to this minor research project. I wish to express my

wholehearted gratitude to my research supervisor Dr. Savita R. Dhongade (Desai) for her

masterly guidance, valuable suggestions and perceptive criticism while carrying out the

present work. Her untiring enthusiasm, moral support, kind care and help out of the way led

me to overcome all the difficulties I encountered during this endeavor.

I wish to express my wholehearted gratitude to Dr. Hemant V. Chavan whose constant

encouragement make me possible to complete this work.

I wish to express my deep sense of gratitude to Hon. Vice-Chancellor, Pro-Vice-

Chancellor, Director of B. C. U. D. and Head of the Department of Chemistry, Shivaji

University Kolhapur for their valuable guidance, consistent encouragement and inspiration

during the course of the investigation and providing me all the facilities during the research

work.

I am highly obliged to Principal, Karmaveer Hire Mahavidyalaya, Gargoti for providing

the infrastructure and necessary facilities in terms of the well-equipped research laboratory.

My heartily thanks to my research colleagues Mr. Pravin Kharade, Mr.Vitthal Divate, Ms.

Poonam Shetake and Ms. Sonatai Patil for their help in the present work.

This work would not have been completed without the help provided by the NCL

Pune and CFC Shivaji University in recording the IR, H1-NMR, C

13-NMR and Mass spectra

and also by Maratha Mandals Central Research Loboratory, Belgum regarding the

determination of biological activities.

INDEX

Sr.No.

Name of the Chapter

Page No.

1.

Chapter 1: Introduction, Microwave Assisted Organic

Synthesis, Introduction to thiazine

1-11

2.

Chapter 2: Synthesis of Novel Chalcones by

Grinding( Conventional) and Microwave Assisted

(Green) Methods

12-20

3.

Chapter 3: Microwave Assisted Synthesis of 1,3-

thiazine derivatives.

21-39

4.

Chapter 4: Biological Activity- Antimicrobial activity

of chalcones and thiazine derivatives.

40-44

1

CHAPTER 1: INTRODUCTION

Section I

Microwave Assisted Organic Reaction

1.1 Microwave-Assisted Organic Synthesis (MAOS) – A Brief History:

After the invention of burner by Robert Bunsen in 1855, the energy from this heat

source applied to a reaction vessel in a focused manner to synthesize organic compounds in

laboratory. The Bunsen burner was later superseded by the mantle, oil bath or hotplate as

means of applying heat to a chemical reaction. In the last few years, heating and driving

chemical reactions by microwave energy has been an increasingly popular theme in the

scientific community [1, 2]. Microwave energy, originally applied for heating foodstuffs by

Percy Spencer in the 1940s, has found a variety of technical applications in the chemical and

related industries since the 1950s, in particular in the food-processing, drying and polymer

industries. Other applications range from pathology (histoprocessing, tissue fixation) [3] and

medical treatments (diathermy) [4].

In those early days, experiments were typically carried out in sealed Teflon or glass

vessels in a domestic household microwave oven without any temperature or pressure

measurements. The results were often violent explosions due to the rapid uncontrolled

heating of organic solvents under closed vessel conditions. In the 1990s, several groups

started to experiment with solvent-free microwave chemistry (so-called dry-media reactions),

which eliminated the danger of explosions [5]. Here, the reagents were pre-adsorbed onto

either an essentially microwave-transparent (i.e., silica, alumina or clay) or strongly

absorbing (i.e., graphite) inorganic support, that additionally may have been doped with a

catalyst or reagent. Particularly in the early days of MAOS, the solvent-free approach was

very popular since it allowed the safe use of domestic microwave ovens and standard open-

vessel technology. While a large number of interesting transformations using “dry media”

reactions have been published in the literature [5], technical difficulties relating to non-

uniform heating, mixing and the precise determination of the reaction temperature remained

unresolved, in particular when scale-up issues needed to be addressed.In order to nonetheless

achieve high reaction rates, high boiling microwave-absorbing solvents have been frequently

used in open vessel microwave synthesis [6]. However, the use of these solvents presented

serious challenges in relation to product isolation and recycling of the solvent. Because of the

2

recent availability of modern microwave reactors with on-line monitoring of both

temperature and pressure, MAOS in dedicated sealed vessels using standard solvents-a

technique pioneered by Christopher R. Strauss in the mid-1990s [7] has been celebrating a

comeback in recent years. This is clearly evident surveying the recently published (since

2001) literature in the area of controlled microwave-assisted organic synthesis (MAOS).

Since the early days of microwave synthesis, the observed rate accelerations and

sometimes altered product distributions compared to oil bath experiments have led to

speculation on the existence of so-called “specific” or “non-thermal” microwave effects [8].

Historically, such effects were claimed when the outcome of a synthesis performed under

microwave conditions was different from that of the conventionally heated counterpart at the

same apparent temperature. Reviewing the present literature [9], it appears that today most

scientists agree that in the majority of cases the reason for the observed rate enhancements is

a purely thermal/kinetic effect, i.e., a consequence of the high reaction temperatures that can

rapidly be attained when irradiating polar materials in a microwave field, although effects

that are caused by the unique nature of the microwave dielectric heating mechanism

(“specific microwave effects”) clearly also need to be considered.

Since 2001, therefore, the number of publications related to MAOS has increased

dramatically, to such a level that it might be assumed that, in a few years, most chemists will

probably use microwave energy to heat chemical reactions on a laboratory scale [1,2]. Not

only is direct microwave heating able to reduce chemical reaction times significantly, but it is

also known to reduce side reactions, increase yields and improve reproducibility. Therefore,

many academic and industrial research groups are already using MAOS as a technology for

rapid reaction optimization, for the efficient synthesis of new chemical entities or for

discovering and probing new chemical reactivity.

Synthesis of new chemical entities is major bottleneck in drug discovery.

Conventional methods for various chemical syntheses are very well documented and

practiced [10]. Microwave assisted organic synthesis [11] (MAOS) has emerged as a new

“lead” in organic synthesis. Due to its ability to couple directly with the reaction molecule

and by passing thermal conductivity leading to a rapid rise in the temperature, microwave

irradiation has been used to improve many organic syntheses.

3

1.2 Microwave frequency:

Microwaves have wavelengths of 1 mm- 1 m. corresponding to frequencies between

0.3 and 300 GHz. These microwaves radiation lie in the region of the electromagnetic

Spectrum between millimeter wave and radio wave i.e. between IR and radio wave.

1.3 Principle:

The basic principle behind the heating in microwave oven is due to the interaction of

charged particle of the reaction material with electromagnetic wavelength of particular

frequency. The phenomena of producing heat by electromagnetic irradiation are either by

collision or by conduction, sometimes by both [12-18].

1.4 Heating Mechanism:

In microwave oven, material may be heated with use of high frequency

electromagnetic waves. The heating arises from the interaction of electric field component of

the wave with charged particles in the material. Two basic principal mechanisms involve in

the heating of material.

1.5 Dipolar Polarization:

Dipolar polarization is a process by which heat is generated in polar molecules. On

exposure to an oscillating electromagnetic field of appropriate frequency, polar molecules try

to follow the field and align themselves in phase with the field. However, owing to inter-

molecular forces, polar molecules experience inertia and are unable to follow the field. This

results in the random motion and interaction of particles generates heat. Dipolar polarization

can generate heat by either one or both the following mechanisms:

1. Interaction between polar solvent molecules such as water, methanol and ethanol.

2. Interaction between polar solute molecules such as ammonia and formic acid.

The energy in a microwave photon (0.037kcal/mol) is very low, relative to the typical

energy required to break a molecular bond (80-120 kcal/mol). Therefore, microwave

excitation of molecules does not affect the structure of an organic molecule and the

interaction is purely kinetic.

4

Fig. 1.1

1.6 Conduction Mechanism:

The conduction mechanism generates heat through resistance to an electric current.

The oscillating electromagnetic field generates an oscillation of electrons or ions in a

conductor, resulting in an electric current. This current faces internal resistance, which heats

the conductor.

The main limitation of this method is that it is not applicable for materials that have

high conductivity, since such materials reflect most of the energy that falls on them.

1.7 Effects of Solvents:

Every solvent and reagent will absorb microwave energy differently. They each have

a different degree of polarity within the molecule and therefore, will be affected either more

or less by the changing microwave field.

Absorbance

level

Solvents

High

DMSO, EtOH, MeOH, Propanols, Nitobenzene, Formic Acid,

Ethylene Glycol.

Medium Water, DMF, NMP, Butanol, Acetonitrile, HMPA, Methyl Ethyl

Ketone, Acetone, Nitromethane, Dichlorobenzene, 1,2-

Dichloroethane, Acetic Acid, trifluoroacetic Acid,

Low Chloroform, DCM, Carbon tetrachloride,1,4-Dioxane, Ethyl

Acetate, Pyridine, Triethyamine, Toluene, Benzene,

Chlorobenzene, Pentane, Nexane and other hydrocarbons

5

1.8 Conventional Vs Microwave Heating:

Microwave heating is different from conventional heating in many respects. The

mechanism behind microwave Synthesis is quite different from conventional synthesis.

Points enlisted in following table, differ the microwave heating from conventional heating

[19-26].

Sr.

No

CONVENTIONAL MICROWAVE

1 Reaction mixture heating proceeds

from a surface usually inside surface

of reaction vessels.

Reaction mixture heating proceeds

directly inside mixture.

2 The vessel should be in physical

contact with surface source that is at a

higher temperature source. (e.g.

mental, oil bath, steam bath etc.)

No need of physical contact of a

reaction with the higher temperature

source while vessel is kept in

microwave cavities.

3 By thermal or electric source heating

take place.

By electromagnetic wave heating take

place.

4 Heating mechanism involve-

conduction.

Heating mechanism involve-dielectric

polarization and conduction.

5 Transfer of energy occurs from the

wall, surface of vessel to the mixture

and eventually to reacting species.

The core mixture is heated directly

while surface (vessel wall) is source

of loss of heat.

6 In conventional heating, the highest

temperature (for an open vessel) that

can be achieved is limited by boiling

point of particular mixture.

In microwave, the temperature of the

mixture can be raised more than its

boiling point. i.e. superheating take.

7 In the conventional heating the entire

compound in mixture are heated

equally.

In microwave specific component can

be heated specifically.

8 Heating rate is less. Heating rate is several folds high.

The application of microwaves in the organic synthesis community is only now

beginning to receive widespread attention.However, use related to Microwave Assisted

6

Organic Synthesis (MAOS), since the late 1990s has increased dramatically to a point where

it might be assumed that, in the up-coming years, most chemists will probably use quick

bursts of microwave energy [27]. Microwave increases rates of reaction, yields can be

improved, and reaction pathways can be selectively activated or suppressed. Fundamentally,

microwaves heat things differently than conventional means.

1.9 Two Specific Mechanisms of Interaction between Materials and Microwaves:

There are two specific mechanisms of interaction between materials and microwaves:

(1) dipole interactions and (2) ionic conduction. Both mechanisms require effective coupling

between components of the target material and the rapidly oscillating electrical field of the

microwaves.

Microwave reactions under solvent-free conditions are attractive in offering reduced

pollution and offer low cost together with simplicity in processing and handling [28]. The

recent introduction of microwave synthesis has gained acceptance and popularity among the

synthetic chemist community & it includes virtually all types of chemical reactions such as

Vilsmeier [29], phthalimide [30], & quinazolinone [31] derivatives etc. are also enhanced by

the microwave irradiation. It has been reported that the rate of variety of organic reaction

such as Esterification [32] could be enhanced by microwave irradiation.

1.10 Conclusions:

Microwave heating is very convenient to use in organic synthesis. The heating is

instantaneous, very specific and there is no contact required between the energy source and

the reaction vessel. Microwave assisted organic synthesis is a technique which can be used to

rapidly explore „chemistry space‟ and increase the diversity of the compounds produced.

Nowadays, it could be considered that all of the previously conventionally heated reactions

could be performed using this technique.

7

1.11 References:

1. Leadbeater N.; Chemistry World, 2004, 1, 38.

2. Adam D.; Nature, 2003, 421, 571.

3. Giberson R.T., Demaree R. S. (Eds.); Humana Press, Totowa, New Jersey, 2001.

4. Prentice W. E.; McGraw-Hill, New York, 2002.

5. (a) Kidawi M.; Pure Appl. Chem., 2001, 73, 147.

(b) Varma R.S.; Pure Appl. Chem., 2001, 73, 193.

(c) Varma R.S.; Tetrahedron, 2002, 58, 1235.

(d) Varma R.S.; Kavitha Printers, Bangalore, 2002.

6. Bose A. K., Manhas M. S., Ganguly S. N., Sharma A.H., Banik B.K.; Synthesis, 2002, 15.

7. Strauss C. R., (Ed.: A. Loupy); Wiley-VCH., Weinheim, 2002, 35.

8. (a) Perreux L., Loupy A.; Tetrahedron, 2001, 57, 9199.

(b) Kuhnert N., Angew; Chem. Int. Ed., 2002, 41, 1863.

(c) Strauss C. R.; Angew. Chem. Int. Ed., 2002, 41, 3589.

9. Kappe C. O.; Angew. Chem. Int. Ed., 2004, 43, 6250.

10. (a) William A.S., Aleksel F.B., Ron C.; Royal Society of Chemistry (Great Britain).

(b) Organic Synthesis Coll vol. 5

(c) Comprehensive organic Synthesis book volumes.

11. (a) Kappe C.O.; Angew.Chem. Int. Ed., 2004, 43, 6250.

(b) Kappe C.O., Dallinger D.; Nat. Rev. Drug Disc., 2006, 5, 51.

(c) Nagariya A.K., Meena. A.K., Kiran, Yadav A.K., Niranjan.U.S., Pathak A.K.,

Singh B., Rao M.M.; Journal of pharmacy Research., 2010, 3, 575-580.

(d) Jignasa.K.S., Ketan T.S., Bhumika. S.P., Anuradha K.G.; Der Pharma

Chemica, 2010, 2(1), 1342-353.

12. (a) Adam D.; Nature, 2003, 421, 571-572.

(b) Blackwell H.E.; Org. Biomol. Chem., 2003, 1, 1251-55.

13. Sharma S. V., Rama-Sharma G.V.S., Suresh B.; Indian J. Pham. Sci., 2002, 64, 337-344.

14. Johansson H., Am. Lab.; 2001, 33(10), 28-32.

15. Bradley D.; Mod. Drug Discovery, 2001, 4, 32-36.

16. Larhed M., Hall berg A.; Drug Discovery Today, 2001, 6, 406-416.

17. Wathey B., Tierney J., Lidrom P., Westman; J. Drug Discovery Today, 2002, 7, 373-380.

18. Dzieraba C.D., Combs A.P.; Annual reports in medicinal chemistry, academic Press,

2002, 37, 247-256.

8

19. In http://www.milestonesci.com.

20. In http://www.maos.net.

21. In http://www.cem.com.

22. Kuhnert N.; Angrew Chem. Int. Ed., 2002, 41, 1863-1866.

23. Lidstrom P., Tierney J., Wathey B., Westman J.; Tetrahedron, 2001, 57, 9225-83.

24. Nuchter. M., Ondruschka B., Lautenschlager W.; Chem. Eng. Technol., 2003, 26, 1208-

1216.

25. Nuchter. M., Ondruschka B., Bonrath W., Gum A.; Green Chem., 2004, 6, 128-141.

26. Leadbeater N.E.; Chemistry World, 200, 1, 38-41.

27. Hayes; Microwave synthesis-Chemistry at the speed of light, CEM Publishing

Matthews, NC, 2004.

28. (a) Kuhneut N., Danks T. N.; Green Chem., 2001, 3, 68.

(b) Tanaka K., Toda F.; Chem. Rev., 2000, 100, 1025.

29. K. Dinakaran and P. T. Perumal; Indian J. Chem., 2000, 39B, 135-136.

30. Abdol Reza Hajipoor, Shadpour E. Mallakpour and Gholamhasan Imanzadeh; Indian J.

Chem., 2001, 40B, 250-251.

31. Deepthi K. Santa, Reddy D. Sahadeva, Reddy P. Pratap & Reddy R.S.N.; Indian J.

Chem., 2000, 39B, 220-222.

32. Mitra A. K., De. A. and Karchaundhuri N.; Indian J. Chem., 2000, 43B, 235-239.

33. Usyatinsky A. Ya., Khmelnitsky Y. L.; Tetrahedron Lett., 2000, 41, 5031.

34. Bentiss F., Lagrenée M., Barbry D.; Tetrahedron Lett., 2000, 41, 1539.

35. Alterman M., Hallberg A.; J. Org. Chem., 2000, 65, 7984.

36. Selvi S., Perumal P.T.; J. Heterocycl. Chem., 2002, 39, 1129.

37. Hayes B. L.; CEM Publishing, Matthews, NC, 2002.

38. Ranu B.C., Hajra A., Jana U.; Tetrahedron Lett., 2000, 41, 531.

9

CHAPTER I: Section II

Introduction to Thiazine

N S

1,3-Thiazine

Thiazine is a six membered heterocycle which contains two hetero atoms (N and S)

placed in the six membered heterocyclic rings at 1, 3 or 1, 4 positions. 1, 3 Thiazines are very

useful units in the field of medicinal and pharmaceutical chemistry and have been reported to

exhibit variety of biological activities [1].A large group of dyes has phenothiazine structure

including methylene blue. Thiazines are used in dyes, tranquilizer and insecticide. Thiazine is

fairly basic diuretic supplement which reduces water and increase vascularity, so it also is

used as anabolic agent in medicine [2]. The 1, 3-thiazine nucleus is active core of

cephalosporin which is among the widely used β-lactumantibiotics [3]. The ability of thiazine

to exhibit antitubercular, antibacterial, anti-HIV and cannabinoid receptor agonist has been

reported [4]. Sawantetal synthesized 1,3-thiazines and carried out antimicrobial screening

which reveals that the compounds with methoxy substituent were found better antimicrobial

compounds[5].The potential use of chlorpromazine derivatives of this phenothiazine as an

antimicrobial, increasing activity of antibiotics to which bacteria are susceptible and reverse

resistance of Staphylococcus aureus and Corynebacterium to penicillin strongly supports that

phenothiazine can be exploited for the management of bacterial infections[6].Whereas,

chalcones undergo a variety of chemical reactions and are found useful in synthesis of variety

of heterocyclic compounds like pyrimidine and thiazole derivatives which are synthesized

through the reaction of chalcones with urea and thiourea in the presence of alkaline refluxing

ethanol[7]. F. K. Mohammed etal synthesized new chromene base heterocyclic thiazine from

2-amino-5-hydroxy-4-phenyl-7-methyl-4H [1-chromeno-3-carbonitrile; which showed a

good biological activity[8]. The derivatives of 1, 3 thiazineshaving N-C-S linkage have been

used as antitubercular, antibacterial, antimicrobial, antitumor, insecticidal, fungicidal,

herbicidal agents, tranquilizers and various dyes etc.[9-18] Further, 1, 3-thiazine core

moieties have remarkable potential of anti radiation agents. 1, 3-Thiazines are used in various

10

organic syntheses and transformations as reaction intermediates. In light of these biological

activities it appeared of interest to synthesize 1, 3 thiazine derivatives.

However synthesis of very few pyrazolyl thiazine derivatives like 2-[2-[3-(2-amino-

4-chloro-phenyl)-4-formyl-pyrazol-1-yl]-6-(4-chloro-phenyl)-6H-[1,3]thiazin-4-yl]-N-

phenyl-acetamide, 2-{6-(4-chloro-phenyl)-2-[4-formyl-3-(4-nitro-phenyl)-pyrazol-1-yl]-6H-

[1,3]thiazin-4-yl}-N-phenyl-acetamide, 2-[2-[3-(4-amino-phenyl)-4-formyl-pyrazol-1-yl]-6-

(4-chloro-phenyl)-6H-[1,3]thiazin-4-yl]-N-phenyl-acetamide, 2-{6-(4-chloro-phenyl)-2-[4-

formyl-3-(2-hydroxy-phenyl)-pyrazol-1-yl]-6H-[1,3]thiazin-4-yl}-N-phenyl-acetamide have

been reported[19].

References:

1. Rathod S. P., Charjan A. P., Rajput P. R.; Rasayan. J. Chem., 2010, 3(2), 363-367.

2. Doifode S. K., Wadekar M. P., Ewatkar S. R.; Orient. J. Chem., 2011, 27(3), 1265-1267.

3. Yadav L. D. S., Singh A.; Tetrahedron Lett., 2003, 44, 5637-5640.

4. Dabholkar V. V., Parab S. D.; Heterolett.Org. 2011, 1, 176-188.

5. Sawant R. L., Bhangale L. P., Wadekar J. B.; Int. J. of Drug Design and Discovery, 2,

2011, 637- 641.

6. Kumar S., Shrivastava D. N., Singhal S., Vipin S., Seth A. K., Yadav Y. C.; J. Chem.

Pharm. Res., 2011, 3(1), 563-571.

7. Balaji P. N., Sreevani M. S., Harini P., Rani P. J., Prathusha K., Chandu T. J.; J. Chem.

Pharm. Res., 2010, 2(4), 754-758.

8. Mohammed F. K., Soliman A. Y., Sawy A. S., Badre M.G.; J. Chem. Pharm. Res., 2009,

1(1), 213-224.

9. Rai V. K., Yadav B. S., Yadav L. D. S.; Tetrahedron, 2009, 65, 1306-1315.

10. Fu L. , Li Y., Ye D. and Yin S.; Chem. Nat. Compd., 2010, 46(2), 169-172.

11. Haider F.H.Z.; J. Chem. Pharm. Res., 2012, 4(4), 2263-2267.

12. Dighade A. S., Dighade S. R.; Der. Pharma. Chemica, 2012, 4(5), 1863-1867.

13. Biehl Ed., Sathunuru R.; ARKIVOC, 2004, 14, 51-60.

14. Hossaini Z., Nematpour M., Yavari I.; Monatsh Chem., 2010, 141, 229–232.

15. Fedoseev V. M., Mandrugin A. A., Trofimova T. P., Zefirova O.N.; Moscow Uni. Chem.

B+., 2008, 63(5), 274-277.

11

16. Batra S., Bhowmik S., Mishra A.; R.S.C. Adv., 2011, 1, 1237-1244.

17. Sambhaji P.V., Shivraj B.S.; Organic Chem. Curr. Res., 2012, 1(5), 1-3.

18. Nagaraj A., Reddy C.S., J. Iran. Chem. Soc., 2008, 5(2), 262-267.

19. a. Jupudi Srikanth, Talari Sandeep, Karunakaram Divya, Govindarajan R.; IJRPC, 2013,

3(2), 213-220.

b. Jupudi1Srikanth *, K.1Padmini, Preethi P.1 Jaya, Bharadwaj P.V.P.2 Deepak,

Vengal Rao.; Asian J. Res. Pharm. Sci., 2013, 3(4), 170-177.

12

CHAPTER 2

Synthesis of Novel Chalcones by Grinding (Conventional) and Microwave Assisted

(Green) Methods

2.1 Introduction:

There is growing interest in the synthesis of different heterocyclic compounds with

pharmacological potential [1-5]. Chalcones are the important building blocks in the synthesis

of variety of such heterocyclic compounds [6-8]. Also they constitute an important class of

bioactive natural products [9-11]. Chemically, they consist of open chain flavonoids in which

the two aromatic rings are joined by a three carbon α-β unsaturated carbonyl system. The

presence of reactive α-β unsaturated keto function in chalcones is found to be responsible for

their antimicrobial activity [12-14]. In recent years a variety of chalcones have been reviewed

for their cytotoxic, anticancer, antiviral, insecticidal and enzyme inhibitory properties [13,

14].

A number of chalcones having hydroxyl, alkoxy groups in different position have

been reported to possess antibacterial [15-18], antiulcer [19], antifungal [20], antioxidant

[21], antimalarial [22, 23], anti-diabetic [24] activities. Library of such biological activities of

chalcone derivatives motivated us to synthesize different chalcones derivatives.

2.2 MATERIALS AND METHODS:

Melting points were determined with Melting point apparatus using open capillary

tubes and are uncorrected. The IR spectra were recorded in KBr pellets on a Nicolet 400D

spectrometer and 1H NMR spectra were recorded in CDCl3 with TMS as internal standard on

a Bruker spectrometer at 400 MHz and their chemical shifts are recorded in δ (parts per

million) unit. Mass spectra were recorded on a Shimadzu GCMS-QP 1000 EX mass

spectrometer at 70 eV. Purity of the compounds was checked by TLC on silica-G plates of 2

mm thickness using n-hexane and ethyl acetate as solvent system. The visualization of spot

was carried out in an iodine chamber.

13

Experimental Procedure:

General procedure for the preparation of Chalcone derivatives:

a) Grinding Method:

Aromatic aldehyde (1 mmol), Substituted acetophenone (1 mmol) and

sodium hydroxide (in catalytic amount) were grinded in mortar using pestle. After

completion of reaction monitored by TLC, Reaction mixture was kept overnight. The

solid mass obtained was isolated by simple work using water. Finally the oven dried

product was recrystallized from ethanol to get desired chalcone in excellent yield.

b) Microwave Irradiation Method:

Aromatic aldehyde (1 mmol), Substituted acetophenone (1 mmol) and sodium

hydroxide (in catalytic ammount) was mixed along with ethyl alcohol:water (1:1) and

irradiated in microwave synthesizer system at 30% power (160W) for 2-3 min. at

65oC . Reaction was monitored by TLC (Ethyl acetate: n-Hexane, 8:2 system).After

completion of the reaction, the reaction mixture was recrystallized from EtOH to

afford the pure product.

CHO

R2

R1

+

sub. aldehyde

O

CH3

R3

sub. acetophenone

NaOH

Grind, RT

O

R3

R2

R1

chalcone derivatives

Scheme 1

Synthesis of (E)-3-(4-ethoxy-phenyl)-1-(4-methoxy-phenyl)-propenone [1a]:

O

H

C2H

5O

O

CH3

MeO

O

H5C

2O

OMe

+NaOH

ChalconeGrind, RT

14

p-Methoxy acetophenone 1.50 g (1 mmol) and p-ethoxy benzeldehyde 2.82 g (1

mmol) were grinded along with sodium hydroxide as in general procedure to give 1a. The

excess of alkali was neuralised by 1:1 HCl and the solid obtained was filtered, washed with

water, dried and recrystallized from ethanol to get Yield 86% of [1a].

Mol. Formula: C18H18O3, Mol. Wt: 282.33, M.P. 116-1180C, IR (KBr): 1040 cm

-1 (C-O),

1209 cm-1

(O-CH3), 1410 cm-1

(CH=CH), 1515 cm-1

(C-C), 1670 cm-1

(C=O), 3019 cm-1

(Ar-

CH). 1H-NMR(CDCl3): 𝛿, 1.42-1.45 (3H, t, CH3), 3.82 (3H, s, OCH3), 4.09-4.12 (2H, q,

OCH2), 8.02-8.04 (2H, d, Ar-2‟,6‟), 7.45–7.47 (1H, d,=CH), 6.97-6.99 (2H, d, Ar-3

‟,5‟), 7.79-

7.81(1H, d,=CH),7.55-7.57(2H, d, Ar-2‟‟,6‟‟), 6.81-6.83 (2H, d, Ar-2‟‟, 6‟‟) ppm.

Synthesis of (E)-3-(2, 4-dichloro-phenyl)-1-(4-methoxy-phenyl) propenone [1b]-:

O

H

Cl Cl

O

CH3

MeO

O

Cl

OMe

Cl

+NaOH

ChalconeGrind, RT

By the same method 4-methoxy acetophenone and 2, 4-di-chlorobenzaldehyde was

grinded with sodium hydroxide to get 1b with 75% yield. Recrystallized was from ethanol.

Mol. Formula: C16H12Cl2O2 Mol. Wt. 307.18, M.P. 155-1580C, IR (KBr):3019 cm

-1 (Ar-CH),

1670 cm-1

(C=O), 1515 cm-1

(C-C), 1410 cm-1

(CH=CH), 1040 cm-1

(-OCH3). 1H-

NMR(CDCl3):𝛿, 3.82 (3H, s, OCH3), 6.97-6.99(2H, d, Ar-3‟,5‟), 8.02-8.04 (2H, d, Ar-2‟,6‟),

7.45-7.47(1H, d, Ar-6‟‟), 7.79-7.81 (1H, d, Ar-5‟‟), 7.48-7.50(1H,d,=CH), 7.20-7.22(1H,

d,=CH), 7.37 (1H,s, Ar3‟‟) ppm.

Synthesis of (E)-3-(4-methoxy-phenyl)-1-(4-methoxy-phenyl) propenone [1c]-:

O

H

MeO

O

CH3

MeO

O

OMe

MeO

+NaOH

ChalconeGrind, RT

15

By using the same method p-methoxy acetophenone and p-methoxy benzaldehyde

were grinded with sodium hydroxide to get 1c with 70% yield. Recrystallized was from

ethanol.

Mol. Formula: C17H16O3 Mol. Wt. 268.304, M.P. 175-1770C, IR (KBr): 3020 cm

-1 (Ar-CH),

1640 cm-1

(C=O), 1515 cm-1

(C-C), 1599, 1528 cm-1

(CH=CH), 1017 cm-1

(-OCH3). 1H-

NMR (CDCl3), 𝛿, 3.87 (3H, s, OCH3), 6.98 (2H, d, Ar-3‟,5‟), 8.02 (2H, d, Ar-2‟,6‟) 7.41(1H,

d,=CH), 7.76 (1H, d,=CH), 7.61 (2H, d, Ar-2‟‟,6‟‟), 6.92 (2H, d, Ar-3‟‟,5‟‟) ppm.

Synthesis of (E)-3-(4-hydroxy-phenyl)-1-(4-methoxy-phenyl) propenone [1d]-:

O

H

OH

O

CH3

MeO

O

OMe

OH

+NaOH

ChalconeGrind, RT

By Using the same method p-methoxy acetophenone and p-hydroxy benzaldehyde

were grinded with sodium hydroxide to get (1d) with 52% yield. Recrystallized was from

ethanol. Mol. Formula: C16H14O3 Mol. Wt. 230.280, M.P. 238-2400C IR (KBr): 1640 cm

-1 (

C=O), 1598, 1528 cm-1

(>C=C<), 1020 cm-1

(OCH3), 3659 cm-1

(OH). 1H-NMR (CDCl3), 𝛿,

3.87 (3H, s, OCH3), 6.89 (2H, d, Ar-3‟,5‟), 7.94 (2H, d, Ar-2‟,6‟), 7.51(1H, d,=CH),

7.71(1H, d,=CH), 7.45 (2H, d, Ar-2‟‟,6‟‟), 6.96 (2H, d, Ar-3‟‟,5‟‟), 8.03(1H, s, OH) ppm.

Synthesis of (E)-3-(4-methoxy-phenyl)-1-(4-ethoxy-phenyl) propenone [1e]-:

O

H

MeO

O

CH3

H5C

2O

O

OC2H

5

MeO

+NaOH

ChalconeGrind, RT

By using the same method p-ethoxy acetophenone and p-methoxy benzaldehyde were

grinded with sodium hydroxide to get (1e) with 65% yield. Recrystallized was from ethanol.

Mol. Formula: C18H18O3 Mol. Wt. 282.330, M.P. 186-1880C. IR (KBr):1654 cm

-1 (C=O),

1599, 1526 cm-1

(>C=C<), 1026 cm-1

(OCH3), 1048 cm-1

(OC2H5). 1H-NMR (CDCl3), 𝛿,

16

1.40-1.42 (3H, t, CH3), 4.02-4.06 (2H, q, CH2), 3.83 (3H, s, OCH3), 6.95 (2H, d, Ar-3‟,5‟),

8.02 (2H, d, Ar-2‟,6‟), 7.50(1H, d, =CH), 7.76 (1H, d, =CH), 6.61 (2H, d, Ar-2‟‟,6‟‟) 6.91

(2H, d, Ar-3‟‟, 5‟‟) ppm.

Synthesis of (E)-3-phenyl-1-(4-ethoxy-phenyl) propenone [1f]-:

O

H

O

CH3

H5C

2O

O

OC2H

5

+NaOH

ChalconeGrind, RT

By Using the same method p-ethoxy acetophenone and benzaldehyde were grinded

with sodium hydroxide to get (1f) with 84% yield. Recrystallization was from ethanol.

Mol. Formula: C17H16O2 Mol. Wt. 252.1151, M.P. 140-1430C. IR (KBr):1648 cm

-1 (C=O),

1576, 1534 cm-1

(>C=C<), 1048 cm-1

(OC2H5). 1H-NMR (CDCl3), 𝛿, 1.39-1.43 (3H, t, CH3),

4.03-4.08 (2H, q, CH2), 6.95 (2H, d, Ar-3‟,5‟), 8.02(2H, d, Ar-2‟,6‟), 7.51 (1H, d, =CH), 7.76

(1H, d, =CH), 7.61 (2H, d, Ar-2‟‟,6‟‟), 7.36-7.39 (3H, m, Ar) ppm.

Synthesis of (E)-3-(4-methoxy-phenyl)-1-(4-bromophenyl) propenone [1g]-:

O

H

MeO

O

CH3

Br

O

Br

MeO

+NaOH

ChalconeGrind, RT

By Using the same method p-bromo acetophenone and p-methoxy benzaldehyde were

grinded with sodium hydroxide to get (1g) with 88% yield. Recrystallized was from ethanol.

Mol. Formula: C16H13BrO2 Mol. Wt. 317.177, M.P. 197-1990C. IR (KBr): 1658 cm

-1 (C=O),

1596, 1540 cm-1

(>C=C<), 1026 cm-1

(OCH3), 722 cm-1

(Br). 1H-NMR (CDCl3), 𝛿, 3.84, (3H,

OCH3), 7.85 (2H, d, Ar-3‟, 5‟), 7.87 (2H, d, Ar-2‟,6‟), 7.58 (1H, d, =CH), 7.61 (1H, d, =CH),

6.92 (2H, d, Ar-2‟‟, 6‟‟), 6.94 (2H, d, Ar-3‟‟, 5‟‟) ppm.

17

Synthesis of (E)-3-(4-methoxy-phenyl)-1-(4-chlorophenyl) propenone [1h]-:

O

H

OH

O

CH3

Cl

O

Cl

OH

+NaOH

ChalconeGrind, RT

By Using the same method p-Chloro acetophenone and p-hydroxy benzaldehyde were

grinded with sodium hydroxide to get (1h) with 30% yield. Recrystallized was from ethanol.

Mol. Formula: C15H11ClO2 Mol. Wt. 301.9942, M.P. 265-2680C, IR (KBr):1650 cm

-1 (C=O),

1590, 1548 cm-1

(>C=C<), 1365 cm-1

(-OH), 654 cm-1

(-Cl). 1H-NMR (CDCl3), 𝛿, 8.92, (1H,

s, OH), 7.55 (2H, d, Ar3‟, 5‟), 7.72 (2H, d, Ar-2‟, 6‟), 7.12 (1H, d, =CH), 7.76 (1H, d, =CH),

7.42 (2H, d, Ar-2‟‟, 6‟‟), 6.56 (2H, d, Ar-3‟‟, 5‟‟) ppm.

Synthesis of (E)-3-(4-methoxy-phenyl)-1-(4-bromophenyl) propenone [1i]-:

O

H

O

CH3

Br

O

Br

+NaOH

ChalconeGrind, RT

By Using the same method p-bromo acetophenone and benzaldehyde were grinded with

sodium hydroxide to get (1i) with 88% yield. Recrystallized was from ethanol.

Mol. Formula: C15H11BrOMol. Wt.285.999, M.P. 155-1570C, IR (KBr): 1652 cm

-1 (C=O),

1568, 1517 cm-1

(>C=C<), 678 cm-1

(Br). 1H-NMR (CDCl3), 𝛿, 7.44 (2H, d, Ar3‟, 5‟), 7.89

(2H, d, Ar-2‟, 6‟), 7.44 (1H, d, =CH), 7.78 (1H, d, =CH), 7.61 (2H, d, Ar-2‟‟, 6‟‟), 7.40 (2H,

d, Ar-3‟‟, 5‟‟), 7.22 (1H, s) ppm.

2.3 RESULTS AND DISCUSSION:

The structures of synthesized compounds were confirmed by IR, 1H-NMR and Mass

spectral analysis. Titled compounds were confirmed by IR spectral data showing sharp bands

in the range between 1030-1660 cm-1

indicated the presence of C=O group. Compounds (1a-

1i) were also confirmed by 1H-NMR spectral analysis. Inspection of the

1H-NMR spectra

suggested that the chalcones were geometrically pure and configured trans.

18

Spectra of some selected compound:

IR Spectrum of (E)-3-(4-Ethoxy-phenyl)-1-(4-methoxy-phenyl)-propenone [1-a]

2.4 CONCLUSION:

Grinding method easily afforded the desired products with higher yields.In comparison with

microwave this method also provides high purity of the product. The synthesized compounds

showed moderate to good antimicrobial activities against Staphylococcus aureus and

Pseudomonas aeruginosa mentioned in detail in chapter biological activity.

19

2.5 REFERENCES:

1. Dua R., Shrivastava S., Sonwane S. K. and Shrivastava S. K.; Adv. Biol. Res., 2011,

5(3), 120-144.

2. Batovska D. I., Todorova I. T.; Curr. Clin. Pharmacol., 2010, 5(1), 1-29.

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4. Prashar H., Chawla A., Sharma A.K. and Kharb R.; Int. J. Pharm. Sci. Res., 2012,

3(7), 1913-1927.

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7. Ooi T., Ohara D., Fukumoto K. and Maruoka K.; Org. Lett. 2005, 7(15), 3195-3197.

8. Zhang Y., Li X., Li J., Chen J., Meng X., Zhao M. and Chen B.; Org. Lett. 2012,

14(1), 26-29.

9. Rozmer Z. and Perjesi P.; Phytochem. Rev., 2016, 15, 87-120.

10. Zhang E.H., Wang R.F., Guo S.Z. and Liu B.; J. Evidence-Based Complementary

Altern. Med., 2013, 1-22.

11. Orlikova B., Tasdemir D., Golais F., Dicato M. and Diederich M.; Genes. Nutr., 2011,

6, 125–147.

12. Prasad Y.R., Rao A.L. and Rambabu R.; Eur. J. Chem., 2008, 5(3), 461-466.

13. Choudhary A.N., Juyal V.; Int. J. Pharm. Pharm. Sci., 2011, 3(3), 125-128.

14. Bhuiyan M.M.H, Hossain M.I., Mahmud M.M and Al-Amin M.; Chemistry Journal,

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2005, 40, 103-112.

16. Yu D.C., Panfilova L.V., Boreko E.I.; Pharm. Chem., 1982, 16, 103-105.

17. Liu X.L., Xu Y.J., Go M.L.; Euro. J. Med. Chem., 2008, 43, 681-1687.

18. Bozic D.D., Milenkovic M., Ivkovic B. and Cirkovic I.; Indian J. Med. Res., 2014,

140(1), 130-137.

19. Avila H.P., Smania E.F.A., Monache F.D. and Junior S. A.; Bioorg. Med. Chem.

2008, 16, 9790-9794.

20. Zhang M. et. al. ACS Omega., 2018, 3, 18343-18360.

21. Jeffery J.A., Pamela E.O., Jared L.R., Jeffery N.J., Peter D.M., Linda M.O., Pamela

S.W. and Beth L.E.; Bioorg. Med. Chem. Lett., 1996, 6(8), 995-998.

https://www.ncbi.nlm.nih.gov/pubmed/?term=Batovska%20DI%5BAuthor%5D&cauthor=true&cauthor_uid=19891604

https://www.ncbi.nlm.nih.gov/pubmed/?term=Todorova%20IT%5BAuthor%5D&cauthor=true&cauthor_uid=19891604

https://www.ncbi.nlm.nih.gov/pubmed/19891604

https://www.ncbi.nlm.nih.gov/pubmed/?term=Bo%26%23x0017e%3Bi%26%23x00107%3B%20DD%5BAuthor%5D&cauthor=true&cauthor_uid=25222788

https://www.ncbi.nlm.nih.gov/pubmed/?term=Milenkovi%26%23x00107%3B%20M%5BAuthor%5D&cauthor=true&cauthor_uid=25222788

https://www.ncbi.nlm.nih.gov/pubmed/?term=Ivkovi%26%23x00107%3B%20B%5BAuthor%5D&cauthor=true&cauthor_uid=25222788

https://www.ncbi.nlm.nih.gov/pubmed/?term=Cirkovi%26%23x00107%3B%20I%5BAuthor%5D&cauthor=true&cauthor_uid=25222788

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22. Lahtchev K.L., Batovska D.I., Parushev S.P., Ubiyvovk V.M., Sibirny A.A.; Euro. J.

Med. Chem., 2008, 43, 2220-2228.

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26. Rao Y. K., Fang S.H., Tzeng Y.M.; Bioorg. Med. Chem., 2009, 17, 7909-7914.

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21

CHAPTER 3

Microwave Assisted Synthesis of 1, 3-thiazine derivatives.

3.1 Introduction:

1,3-thiazine is two heteroatom ( N & S at 1,3 position ) containing six membered

heterocyclic compound. Because of its diverse biological activities [1] found useful in the

field of medicinal and pharmaceutical chemistry. Thiazine is used as anabolic agent in

medicine [2], also in dyes, tranquilizers, and insecticides. Antituberculer, antibacterial and

anti-HIV activities have been reported [3]. Cephalosporin is among widely used beta lactum

antibiotics whose active core is made up of 1,3-thiazine nucleus. Sawantet.al. reported that

compounds with methoxy substituent shows better antimicrobial activity [4]. The potential

use of chlorpromazine derivatives of this phenothiazine as an antimicrobial, increasing

activity of antibiotics to which bacteria are susceptible and reverse resistance of

staphylococcus aureus and corynebacteriatopenicillin strongly supported that phenothiazine

can be exploited for the management of bacterial infections [5].

On the other hand, Chalcones being building blocks of variety of heterocyclic

compounds like pyrimidine and thiazole derivatives which are synthesized from chalcone

with urea and thiourea in basic medium [6]. F.K.Mohammedet. al. synthesized new chromene

base heterocyclic thiazine from 2-amino-5-hydroxy-4-phenyl-7-methyl-4H[1-chromeno-3

carbonitrile; which showed a good biological activity[7]. The derivatives of 1,3thiazines

having N-C-S linkage have been used as antitubercular, antibacterial, antimicrobial,

antitumor, insecticidal, fungicidal, herbicidal agents, in tranquilizers and various dyes etc.8-

16. Further, 1,3-thiazine core moieties have remarkable potential as anti radiation agents. 1,3-

Thiazines are used in various organic synthesis and transformations as reaction

intermediates[8-16]. In light of these biological activities it appeared of interest to synthesize

1,3 thiazine derivatives.

However, synthesis of very few pyrazolyl thiazine derivatives like 2-[2-[3-(2-amino–

4-chloro-phenyl)-4-formyl–pyrazol-1-yl]–6-(4-chloro-phenyl)-6H-[1,3]thiazin–4-yl]-N-

phenyl-acetamide, 2-{6-(4-chloro-phenyl)–2-[4formyl-3-(4-nitro-phenyl)pyrazol-1-yl]-6H-

[1,3]thiazin-4-yl}-N-phenyl-acetamide, 2[2-[3-(4-amino-phenyl)-4-formyl-pyrazol-1-yl]-6-

(4-chloro-phenyl)-6H-[1,3]thiazin-4-yl]-N-phenyl-acetamide, 2-{6-(4-chloro-phenyl)-2-[4-

22

O

R2R1

R3

R1

N

R3

S

N

NH

R4 R5

R2

R4NHNH2

R5N=C=S 20% 140 W

4(a-g)Chalcone

formyl-3-(2hydroxy-phenyl)-pyrazol-1-yl]-6H-[1,3]thiazin-4-yl}-N-phenyl-acetamide have

been reported [17].

3.2 Experimental:

Materials and Method:

Melting points were determined with Melting point apparatus using open capillary

tubes and are uncorrected. The IR spectra were recorded in KBr pellets on a Nicolet 400D

spectrometer and 1H NMR spectra were recorded in CDCl3 with TMS as internal standard on

a Bruker spectrometer at 400MHz. Mass spectra were recorded on a Shimadzu GCMS-QP

1000 EX mass spectrometer at 70 eV. Purity of the compounds was checked by TLC on

silica-G plates of 2 mm thickness using n-hexane and ethyl acetate as solvent system. The

visualization of spot was carried out in an iodine chamber. Biological activities are predicted

by using using cup-plate agar diffusion method

General outline for the synthesis of thiazine:

In scheme 2, Chalcones prepared in scheme 1 were subjected for further sequencial

reaction to obtain the targeted comound thiazine derivatives in good to excellent yield.

Initially the chalcones, phenyl hydrazine and phenyl isothiocyanate were subjected for one

pot three component microwave irradiation to offered the intermediate compounds 4(a-g).

These intermediate compounds on further irradiation with substituted chalcones to generate

the desired products thiazine derivatives. The advantage of this method includes the short

reaction time, easy work-up and excellent yields.

Synthesis of 2-phenyl-2, 5-dihydro-pyrazole-1-carbothioicacid phenyl amide derivatives

4(a-g).

23

N N

NH

S

R2R1

R3

R5

R4

Table-1: General Characteristics and Elemental Analysis data of the Compounds [4(a-g)]

General Synthetic Procedure for the 2-phenyl-2, 5-dihydro-pyrazole-1-carbothioic acid

phenyl amide derivatives [4(a-g)]:

In 100 mL round bottom flask 0.01 mole chalcone [1(a-b)], 0.01 mole substituted

hydrazine, 0.01 mole substituted isothiocynate were mixed in ethanol (30 mL) and the

reaction mixture was irradiated with microwaves at 20% power (140 W) for 5-7 min. The

completion of the reaction was confirmed by TLC (Ethyl acetate: Hexane). The reaction

mixture was allowed to cool at room temperature, poured into crushed ice. The solid

separated was filtered, washed with little 1:1 ethanol and purified by recrystallization from

ethanol to give pure products. The products thus obtained [4(a-g)] were directly used for next

step.

Comp.

R

1

R2

R

3

R4

R

5 Yield

%

M.P.

0C

Mol. Formula

4-a OCH3 H OC2H5 C6H5 C6H5 81 213 C31H29N3O2S

4-b OCH3 H OC2H5 H C6H5 75 201 C25H25N3O2S

4-c OCH3 H OC2H5 C6H5 C3H5

78 210 C28H29N3O2S

4-d OCH3 H OC2H5 H C3H5

84 205 C22H25N3O2S

4-e OH Cl Cl C6H5 C6H5 79 235 C28H21Cl2N3OS

4-f OH Cl Cl C6H5 C3H5

74 225 C25H21Cl2N3OS

4-g OH Cl Cl H C6H5 85 207 C22H17Cl2N3OS

24

R1

NH

R3

S

N

NH

R4R5

R2O

R6

R7

R8

N S

R6 R7R8

N

R5

NR4 R3

R1

R2

20% 140 W

carbothioic acid phenyl amide

1,3-thiazine

N S

N

N

R8R7R6

R5

R4

R2

R3

R1

Synthesis of 1, 3-thiazine derivatives [6(a-j)]:

Table-2: General Characteristics and Elemental data of the Compounds [6(a-j)]

25

Table-2: General Characteristics and Elemental data of the Compounds [6(a-j)]

General Synthetic Procedure for the thiazine derivatives [6(a-j)]:

In a 100 mL round bottom flask 0.001 mole 2-phenyl-2, 5-dihydro-pyrazole-1-

carbothioic acid phenyl amide derivatives [4(a-g)], 0.001 mole chalcone [5(a-b)] were

mixed in ethanol (30 mL). The reaction mixture was irradiated with microwaves at 20%

power (140 W) for 5-7 mins. The completion of the reaction was confirmed by TLC (Ethyl

acetate: Hexane). The reaction mixture was allowed to cool at room temperature, poured

into crushed ice. The solid separated was filtered, washed with little 1:1 ethanol and

purified by recrystallization from ethanol to furnish pure products [6(a-j)].

Co

mp.

R1

R2

R3

R4

R5

R6

R7

R8

Yield

%

M.P

. oC

Mol. Formula

6-a OCH3 H OC2H5 C6H5 C6H5 OCH3 H OC2H5 86 245 C49H47N3O4S

6-b OCH3 H OC2H5 H C6H5 OCH3 H OC2H5 75 238 C43H43N3O4S

6-c OCH3 H OC2H5 H C6H5 OH Cl Cl 78 230

C46H39Cl2N3O3S

6-d OCH3 H OC2H5 C6H5 C3H5 OCH3 H OC2H5 84 242

C46H47N3O4S

6-e OCH3 H OC2H5 H C3H5 OCH3 H OC2H5 79 233

C40H43N3O4S

6-f OH Cl Cl C6H5 C6H5 OH Cl Cl 85 255

C43H31Cl4N3O2S

6-g OH Cl Cl C6H5 C6H5 OCH3 H OC2H5 77 245

C46H39Cl2N3O3S

6-h OH Cl Cl C6H5 C3H5 OCH3 H OC2H5 79 249

C43H39Cl2N3O3S

6-i OH Cl Cl H C6H5 OCH3 H OC2H5 81 267

C40H35Cl2N3O3S

6-j OH Cl Cl H C6H5 OH Cl Cl 69 252

C37H27Cl4N3O2S

26

3.3 Spectral Analysis of some representative compounds:

a) 5-(4-Ethoxy-phenyl)-3-(4-methoxy-phenyl)-2- phenyl-2,5- dihydro- pyrazole-1-

carbothioic acid phenyl amide [4-a]:

Molecular Formula: C31H29N3O2S,

Melting Point: 2130C,

IR (KBr)νmax:. 1020(C-N), 1040(C-O), 1160(OCH3) , 1200(C=S), 1230(N-N), 1609(C-C),

3018(Ar-CH), 3500(NH) cm-1

.

H1

NMR (CDCl3, 200.13MHz): δ 1.30-1.32 (t, 3H, CH3), 3.86(s, 3H, OCH3), 3.99-4.03(q,

2H, CH2), 5.94-5.95(d, 1H, =CH), 5.97-5.99 (d, 1H, CH), 6.87-6.97(m, 8H, Ar-H), 7.44-

7.72(m, 10H, Ar-H), 8.65(d, 1H, NH) ppm.

13C NMR(CDCl3, 50.32 MHz): δ14.68(OCH2CH3), 55.42(OCH3), 61.00(O-CH2- CH3),

63.58(>CH-), 102.00(2×=CH), 104.00(2×=CH), 119.36(2×=CH), 125.00(2×=CH),

125.38(=C<), 126.37(=C<), 127.63(2×=CH), 128.30(2×=CH), 129.00(2×=CH),

129.70(2×=CH), 132.64(=C<), 133.64(=C<), 136.34(=C<), 138.00(=C<), 139.00(=C<),

148.00(=C<), 160.89(2×=C<), 178.62 (C=S).

Elemental Analysis:

Calcd.:C(73.39%), H(5.78%), N(8.34%).

Found: C(73.35%), H(5.76%), N(8.28%).

b) 6-(4-ethoxyphenyl)-2-[5-(4-ethoxyphenyl)-3-(4-methoxyphenyl)-2-phenyl-2, 5-

dihydro-1H-pyrazol-1-yl]-4-(4-methoxyphenyl)-3-phenyl-3, 6-dihydro-2H-1,

3-thiazine [6-a]:



IR (KBr)νmax:. 1025 (C-N), 1060(C-O), 1165(O-CH3), 1235(N-N), 1595 (C=C),

3018 (Ar-CH)cm-1

.

27

H1

NMR (CDCl3, 200.13MHz): δ 1.39-1.46(t, 6H, 2×CH3), 3.88 (s, 6H, 2×CH3), 4.02-4.12

(q, 4H, 2×CH2), 4.80-4.86(d, 2H, 2×CH), 5.14(s, 1H, CH), 5.94-5.99(d,2H, 2× CH), 6.89-

6.99(m, 8H, 2×C6H4), 7.38- 7.45 (m, 8H, 2×C6H4), 7.69-7.97(m,10H,2×C6H5) ppm.

13C-NMR(CDCl3, 50.32 MHz):δ 13.47(CH2-CH3), 14.81(CH2-CH3), 42.22(CH),

55.17(OCH3), 55.72(OCH3), 61.10(CH2), 63.64(CH2), 69.62(CH), 72.90(CH),

112.00(2×=CH), 113.31(2×=CH), 113.87(2×=CH), 114.93(2×=CH), 119.36(2×=CH),

122.40(2×=CH), 123.46(=C<), 125.37(=CH), 125.98(=C<), 127.00(2×=CH),

127.74(2×=CH), 128.00(2×=CH), 129.00(2×=CH), 129.33(2×=CH), 130.02(2×=CH),

130.82(=CH), 131.48(=C<), 133.99(=C<), 138.58(=C<), 139.08(=C<), 143.42(=C<),

143.99(=C<), 146.00(=C<), 146.63(=C<), 158.21(=C<), 160.00(=C<), 160.39(C-O),

161.00(C-O).

GCMS m/z (%): 774 [M+, 1%], 282(100%), 655(20%), 502(20%), 378(18%), 253(80%),

225(32%), 165(23%), 147(20%), 135(55%),92 (30%), 77(32%).

Elemental Analysis:

Calcd.:C(76.04%), H(6.12%), N(5.48%).

Found: C(76.01%), H(6.12%), N(5.48%).

c) 3-Allyl-6-(4-ethoxyphenyl)-2-[5-(4-ethoxyphenyl)-3-(4-methoxyphenyl)-2, 5-

dihydro- 1H- pyrazol-1-yl]-4-(4-methoxyphenyl)-3-(prop-2-en-1-yl)-3, 6-dihydro- 2H-1,

3- thiazine [6-e]:



IR (KBr) νmax:. 1051.01(OCH3), 1076.35 (C-N), 1240.00(C-O), 1572.72 (C=C), 2927.36

(CH2), 3006.48 (Ar-CH), 3369.03 (NH) cm-1

.

H1

NMR (CDCl3, 200.13MHz): δ 1.39-1.48(t, 6H, 2×CH3), 3.55-3.56 (d, 2H,CH2), 3.88(s,

6H, 2×CH3), 4.02-4.15 (q, 4H, 2×CH2), 4.84-4.90(d, 2H, 2×CH), 5.05-5.06(d, 2H, =CH2),

5.31(s, 1H, CH), 5.76-5.82(q, 1H, =CH), 6.89-7.01(m,8H, 2×C6H4), 7.50- 7.57 (m, 8H,

2×C6H4), 11.00 (s, 1H, NH) ppm.

28

13C NMR(CDCl3, 50.32 MHz):δ 13.47(CH2-CH3), 14.68(CH2-CH3), 42.28(CH),

55.00(OCH3), 55.42(OCH3), 56.53(CH2), 61.10(OCH2), 63.58(OCH2), 69.62(CH),

71.76(CH), 112.00(2×=CH), 113.73(2×=CH), 114.07(2×=CH), 114.81(2×=CH),

118.76(=CH2), 126.23(=CH), 127.00(2×=CH), 127.58(2×=CH), 128.00(2×=CH),

130.03(2×=CH), 132.89(=CH), 133.21(=CH), 133.97(=C<), 137.69(=C<), 138.58(=C<),

139.08(=C<), 145.02(=C<), 146.71(=C<), 158.22(=C<), 160.00(=C<), 160.52(=C<),

161.02(=C<).

GCMS m/z (%): 661 [M+, 1%], 282(100%), 253(80%), 225(32%), 165(23%),

147(20%), 135(55%),92 (30%), 77(32%).

Elemental Analysis:

Calcd.:C(72.65%), H(6.58%), N(6.39%).

Found: C(72.59%), H(6.55%), N(6.35%).

d) 4-{5-(2, 4-dichlorophenyl)-1-[6-(4-ethoxyphenyl)-4-(4-methoxyphenyl)-3-

phenyl-3, 6-dihydro-2H-1, 3-thiazin-2-yl]-2, 5-dihydro-1H-pyrazol-3- yl} phenol [6-i]:

Molecular Formula: C40H35Cl2N3O3S,


IR (KBr)νmax: 696.18(C-Cl), 1051.01(OCH3), 1076.35 (C-N), 1240.00(C-O),

1572.72 (C=C), 3006.48(Ar-CH), 3369.03(NH) cm-1

.

H1

NMR (CDCl3, 200.13MHz): δ 1.35-1.44(t, 3H, CH3), 3.88(s, 3H, CH3), 4.02-4.15 (q,

2H, CH2), 4.84-4.90(d, 2H, 2×CH), 5.05-5.06(d, 2H, =CH), 5.31(s, 1H, CH), 6.05-6.11(m,

8H, 2×C6H4), 6.55-6.64(m, 5H, C6H5), 7.04- 7.16 (m, 4H, C6H4), 7.57-7.59(d, 2H, 2×=CH),

7.81(s, 1H, =CH), 9.40(s, 1H, OH) ppm.

13C NMR(CDCl3, 50.32 MHz):δ14.89(CH2-CH3), 42.27(CH), 55.19(OCH3), 63.24(OCH2),

69.62(CH), 113.87(=CH), 114.47(2×=CH), 116.12(2×=CH), 119.28(2×=CH),

29

123.13(2×=CH), 126.23(=CH), 126.76(=CH), 127.50(=CH), 128.00(2×=CH),

128.63(2×=CH), 129.00(=CH), 129.43(=CH), 130.07(2×=CH), 131.41(2×=CH),

132.39(=C<), 133.00(C-Cl), 133.37(C-Cl), 134.00(=CH), 134.48(=C<), 138.79(=C<),

143.34(=C<), 145.02(=C<), 146.77(=C<), 158.21(=C<), 159.44(OH), 160.45(=C<).

GCMS m/z (%): 707 [M+, 1%], 282(100%), 267(18%), 251(19%), 225(32%),

211(18%), 165(28%), 147 (20%), 135(35%), 107(18%), 92(30%), 77(32%).

Elemental Analysis:

Calcd.:C(67.83%), H(5.05%), N(5.98%).

Found: C(67.79%), H(4.98%), N(5.93%).

e) 3-chloro-4-{2-[5-(2, 4-dichlorophenyl)-3-(4-hydroxyphenyl)-2, 5-dihydro-1H-

pyrazol-1-yl]-4-(4-hydroxyphenyl)-3-phenyl-3, 6-dihydro-2H-1, 3-thiazin-6-yl}

phenol [6-j]:

Molecular Formula: C37H27Cl4N3O2S


IR (KBr)νmax: 696.18(C-Cl), 1076.37 (C-N), 1589.08 (C=C), 3006.48 (Ar-CH),

3326.61 (NH), 3412.72 (OH) cm-1

.

H1

NMR (CDCl3, 200.13MHz):δ 4.40-4.46(d, 2H, 2×CH), 4.84=4.90(d, 2H, 2×CH), 5.05-

5.06(d, 1H, =CH), 5.25(s, 1H, OH), 5.43(s, 1H, CH), 5.55(s, 1H, =CH), 6.14-6.20(m,4H,

C6H4), 6.55-6.64(m, 5H, C6H5), 7.04- 7.16 (m, 4H, C6H4), 7.57-7.59(d, 3H, 3×=CH),

7.81(s, 1H, =CH), 8.91(s, 1H, =CH), 9.40(s, 1H, OH) ppm.

13C NMR(CDCl3, 50.32 MHz):δ 42.25(CH), 69.63(CH), 116.00(4×=CH), 119.29

(2×=CH),123.46(=CH), 125.48(=CH), 126.00(=CH), 126.48(=CH), 127.02(4×=CH),

127.79(=CH),128.00(=CH), 128.58(=CH), 129.00(=CH), 129.47(2×=CH), 131.30(=C<),

132.27(C-Cl),132.99(C-Cl), 133.27(C-Cl), 134.00(C-Cl), 134.67(=CH), 135.02(=C<),

135.81(=C<),138.58(=C<), 143.39(=C<), 145.02(=C<), 146.65(=C<), 159.60(2×OH).

30

GCMS m/z (%):717 [M+, 1%], 282(100%), 704(20%), 267(18%), 253(80%),

251(19%),225(32%), 211(18%), 165(28%),147 (20%), 135(35%), 107(18%),

92(30%), 77(32%), 65(18%).

Elemental Analysis:

Calcd.:C(61.79%), H(3.83%), N(5.87%).

Found: C(61.76%), H(3.78%), N(5.84%).

3.4 Conclusion:

In summary, we have developed efficient method for the selective synthesis of novel

thiazine derivatives, which attains completion in 5-7 min in an aqueous medium and may

provide a useful route for the rapid drug discovery. Method affords excellent yields in very

short time with high purity.

31

Spectral Evidences of Synthesized Compounds:

Spectrum: IR Spectrum of 5-(4-ethoxy-phenyl)-3-(4-methoxy-phenyl)-2-phenyl-2, 5-

dihydro-pyrazole-1-carbothioic acid phenyl amide [4-a]

32

Spectrum: H1 NMR Spectrum of 5-(4-ethoxy-phenyl)-3-(4-methoxy-phenyl)-2-phenyl-2, 5-

dihydro-pyrazole-1-carbothioic acid phenyl amide [4-a]

33

Spectrum: 13

C NMR Spectrum of 5-(4-ethoxy-phenyl)-3-(4-methoxy-phenyl)-2-phenyl-2,

5-dihydro-pyrazole-1-carbothioic acid phenyl amide [4-a]

34

Spectrum: IR Spectrum of 6-(4-ethoxyphenyl)-2-[5-(4-ethoxyphenyl)-3-(4-methoxyphenyl)-

2-phenyl-2, 5-dihydro-1H-pyrazol-1-yl]-4-(4-methoxyphenyl)-3-phenyl-3, 6-dihydro-2H-1,

3-thiazine [6-a]

35

Spectrum: H1 NMR Spectrum of 6-(4-ethoxyphenyl)-2-[5-(4-ethoxyphenyl)-3-(4-

methoxyphenyl)-2-phenyl-2, 5-dihydro-1H-pyrazol-1- yl]-4-(4-methoxyphenyl)-3-phenyl-3,

6-dihydro-2H-1, 3-thiazine [6-a]

36

Spectrum: 13

CNMR Spectrum of 6-(4-ethoxyphenyl)-2-[5-(4-ethoxyphenyl)-3-(4-



37

Spectrum: Mass Spectrum of 6-(4-ethoxyphenyl)-2-[5-(4-ethoxyphenyl)-3-(4-



38

3.5 References:

1. S. P. Rathod, A. P. Charjan, P. R. Rajput; Rasayan. J. Chem., 2010, 3(2), 363-367.

2. S. K. Doifode, M. P. Wadekar, S. R. Ewatkar; Orient. J. Chem., 2011, 27(3), 1265-1267.

3. V. V. Dabholkar, S. D. Parab; Hetero. lett. org., 2011, 1, 176-188.

4. R. L. Sawant, L. P. Bhangale, J. B. Wadekar; Int. J. of drug design and discovery, 2011, 2,

637-641.

5. S. Kumar, D. N. Shrivastava, S. Singhal, S. Vipin, A. K. Seth, Y. C. Yadav; J. Chem.

Pharm. Res., 2011, 3(1), 563-571.

6. P. N. Balaji, M. S. Sreevani, P. Harini, P. J. Rani, K. Prathusha, T. J. Chandu; J. Chem.

Pharm. Res., 2010, 2(4), 754-758.

7. F. K. Mohammed, A. Y. Soliman, A. S. Sawy, M. G. Badre; J. Chem. Pharm. Res., 2009,

1(1), 213-224.

8. V. K. Rai, B. S. Yadav, L. D. S. Yadav; Tetrahedron, 2009, 65, 1306-1315.

9. L. Fu, Y. Li, D. Ye, S. Yin; Chem. Nat. Compd., 2010, 46(2), 169-172.

10. F. H. Z. Haider; J. Chem. Pharm. Res., 2012, 4(4), 2263-2267.

11. A. S. Dighade, S. R. Dighade; Der. Pharma. Chemica, 2012, 4(5), 1863-1867.

12. Z. Hossaini, M. Nematpour, I. Yavari; Monatsh Chem., 2010, 141, 229–232.

13. V.M. Fedoseev, A. A. Mandrugin, T. P. Trofimova, O. N. Zefirova; Moscow Uni. Chem.

B+, 2008, 63(5), 274-277.

14. S. Batra, S. Bhowmik, A. Mishra; RSC Adv., 2011, 1, 1237-1244.

15. P. V. Sambhaji, B. S. Shivraj; Organic Chem. Curr. Res., 2012, 1(5), 1-3.

16. A. Nagaraj, C. S. Reddy; J. Iran. Chem. Soc., 2008, 5(2), 262-267.

17. a. Srikanth Jupudi, Sandeep Talari, Divya Karunakaram, R. Govindarajan; IJRPC, 2013,

3(2), 213-220.

b. Srikanth Jupudi1, K. Padmini, Preethi P. Jaya, Bharadwaj P.V.P.Deepak, Vengal Rao;

Asian J. Res. Pharm. Sci., 2013, 3(4), 170-177.

18. D. A. Filimonov, V. V. Poroikov; In Chemoinformatics Approaches to Virtual

Screening. Eds Alexander Varnek and Alexander Tropsha.Cambridge (UK): RSC

Publishing, 2008, 182-216

19.S. A. Kryzhanovskii, R. M. Salimov, A. A. Lagunin, D. A. Filimonov, T. A. Gloriozova,

V. V. Poroikov; Pharmaceut. Chem. J., 2012, 45(10), 605-611.

39

20. O. A. Filz, A. A. Lagunin, D. A. Filimonov, V. V. Poroikov; SAR & QSAR Environ. Res.,

2012, 23(3-4), 279-296.

21. P. Eleftheriou, A. Geronikaki, D. Hadjipavlou-Litina, P. Vicini, O. Filz, D. V. Filimonov,

V. V. Poroikov, S. S. Chaudhaery, K. K. Roy, A. Saxena; Eur. J. Med. Chem., 2012,

47(1), 111-124.

40

CHAPTER 4

Biological Activity

Antimicrobial activity of chalcone and thiazine derivatives

4.1 Introduction:

Antimicrobials are the agents which are used to kills (Cidal) or stop the growth (static)

of microorganisms. Depending on the microorganism against which they act, they are grouped as

antibiotics- acts against bacteria, antifungals- act against fungi etc. Again depending upon their

function they are classified as follows.

1) Disinfectants- kill microbes on non-living surfaces.

2) Antiseptics- used on living surfaces to reduce infections during the surgery.

3) Antibiotics- kill microorganisms within the body. In 1928 Alexander Fleming became the

first to discover a natural antibiotic- Penicillin and was successfully used to treat a

Streptococcus infection in 1942.

4) Antifungal- kills or prevents the further growth of fungi. They kill the fungal organisms

without harming the host.

5) Antiviral- used to treat viral infections. For specific viral, a specific antiviral agent is

required.

6) Antibacterial- kills the bacteria (bactericidal) or prevents the growth of bacteria

(bacteriostatic)

Most of the chalcone and thiazine derivatives from literature shows antimicrobial

activity, which inspired us to carry out the same activity results for our synthesized chalcone and

thiazine derivatives.

41

4.2 Antibacterial activity of chalcone derivatives:

Antimicrobial activity of all synthesized compounds was determined by disc diffusion

method [1,2,3]. All human pathogenic bacteria viz. Staphylococcus aurenus (737), Pseudomonas

aeruginosa (1688) were used for activity determination. Preparation of nutrient broth, Subculture,

base layer medium, agar medium and peptone water was done as per the standard procedure. Disc

measuring 6.25 mm in diameter were punched from Whatman no.1 filter paper. Stock solution of

synthesized compounds diluted in dimethyl sulfoxide (1% DMSO) to give final concentration of

500 µg/ml and 1000 µg/ml. A reference standard for both gram positive and gram negative bacteria

was made by dissolving accurately weighed quantity of chloramphenicol (500 and 1000 µg/ml,

respectively) in sterile distilled water separately. The incubation was carried out at 37 0C for 24 hrs.

All the experiments were carried out in triplicate. Simultaneously, controls were maintained by

employing 0.1 mL of dimethyl sulfoxide which did not reveal any inhibition. Zones of inhibition

produced by each compounds was measured in mm. The results of antibacterial studies are given in

Table 1.

Table 1: Antimicrobial activity of the synthesized compounds

Compound

Antimicrobial activity (%inhibition)

Staphylococcus aureus (737) Pseudomonas aeruginosa (1688)

500 µg/ml 1000 µg/ml 500 µg/ml 1000 µg/ml

1a 21.4 32.6 ------ ------

1b 24.3 33.6 ------ ------

1c ------ ------ ------ ------

1d ------ ------ ------ ------

1e 23.8 33.6 ------ ------

1f -------- -------- ------ ------

1g 25.0 34.8 ------ ------

1h 22.6 32.8 ------ ------

1i 23.0 33.3 ------ ------

Chloramphenicol 42.3 55.2 63.7 78.9

DMSO 1.4 ------ 1.2 --------

42

4.3 Conclusion:

The results revealed that majority of the synthesized compounds showed varying degrees of

inhibition against Gram positive bacteria shown in Table 2. The 1g showed excellent activity

against Staphylococcus aureus at both concentration i.e. 500 µg/ml and 1000 µg/ml. The

compounds 1g, 1b, 1e, 1i and 1h, 1a have shown good to moderate activity against Staphylococcus

aureus at both concentration i.e. 500 µg/ ml and 1000 µg/ ml. Three of the chalcones with anti-

staphylocaccal activity (1c, 1d and 1f) gave no inhibitory zones probably due to their low diffusion

potential into agar media.

Finally, no activity was observed for compounds against Pseudomonas aeruginosa, a Gram

negative organism. It is widely known that Gram positive and negative organism have significantly

different membrane composition and architecture which would explain the selectivity of the present

compounds against Gram positive Staphylococcus aureus.

4.4 MICROBIAL TESTING OF THIAZINE DERIVATIVES:

The new compounds synthesized in the present investigation were screened for their

antibacterial activities.

The antimicrobial activity of these compounds was tested using cup-plate agar diffusion

[4,5,6] method at 40 PPM concentration using 7mm size filter paper disc. At similar condition

standard drug streptomycin sulfate was used.

Medium for growth

For antibacterial activities against E. coli MacConkey agar was used with following composition

[8].

a) Peptone 20gms.

b) Lactose 10gms.

c) Bile Salt 5gms.

d) Sodium chloride 5gms.

e) Neutral red 0.075gms.

f) Agar 12gms.

g) Distilled water 1000ml.

43

For antibacterial activities against S. albus nutrient agar with following composition was used

a) Peptone 5gms.

b) Beef extract 3gms

c) Sodium chloride 8gms

d) Agar 15gms

e) Distilled water 1000ml

Activities were determined by using the cultivated plates of Whatmann filter paper (7mm).

All the compounds were dissolved in DMF (40ppm) Plates were soaked in the solution. These

plates were placed on the medium previously seeded with organisms in Petri dishes and stored in an

incubator at 270 for 48 hours for E. coli and 24 hours for S. albus respectively. An inhibition zone

was measure in mm and recorded in the following table.

Table-:3 Zone of inhibition in mm

Comp.No E.coli S.albus

6a 12 8

6b 9 16

6c 11 14

6d 9 16

6e 12 14

6f 12 8

6g 15 10

6h 9 17

6i 11 10

6j 13 10

44

4.5 Results and Discussion:

In Thiazine derivatives tested compounds 6-a, 6-g and 6-j showed greater degree of antibacterial

activity against E.coli. However, the compounds 6-b, 6-d and 6-h showed greater degree of

antibacterial activity against S.albus.

1) We have reported efficient synthesis of 1,3-thiazine derivatives. Clean reaction, low

reaction times was the main advantages of this method. Satisfactory yield of products

and easy workup make this a useful protocol for green synthesis of this class of

compounds.

2) Also the synthesized thiazine derivatives were screened for their antimicrobial activities.

4.5 References:

1. Rao Y. K., Fang S. H., Tzeng Y. M.; Bioorg. Med. Chem., 2009, 17: 7909-7914.

2. Papo N., Shai Y.; Peptides, 2003, 24, 1693-1703.

3. Te SC.; Int. J. Mol. Sci., 2009, 10, 2440-2475.

4. F. Kavangh; Analytical Microbiology, 1963, 125, (Academic press, New York).

5. H. W. Seeley and P. J. Van Demark; Microbes in Action, A Laboratory Manual of

Microbiology, 1975, 55, (DBT Traporevala Sons and Pvt. Ltd, Bombay).

6. Y. L. Nene and P. N. Thapliyal; Fungicides in Plant Disease Control (Oxford and IBH

Publication, New Delhi), 1982, 192.

7. L. Oramay et.al. The Significance of Bacillus cereus food poisoning in Hungary, in the

Microbiology of dried foods, 1969, 279.

8. P. Chakraborty; A Text Book of Microbiology, New Central Book Agency (P) Ltd , Culcutta

, India , 2000, 211.

9. C. V. Subramanian, Hyphomycetes; Indian Council of Agri. Research, New Delhi, 1971,

802 and 810.

10. P. B. Godkar ; Text Book of Medical Laboratory Technology, Bhalani Publishing House,

Bombay, India, 1996, 326 , 332 , 382.

45

Published

Papers

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

Executive

Summary

64

Annexure – VI


BAHADUR SHAH ZAFAR MARG

NEW DELHI – 110 002

Annual Report of the work done on the Minor Research Project

(Reports to be submitted within 6 weeks after completion of each year)

1. Project report No. 1st/Final : 1

st year report (Annual)

2. UGC Reference No.

F. 47-1161/14(WRO) dated 28th

July 2017 / 8 September 2017

3. Period of report from 01st September 2017 to 31

st August 2018

4. Title of the Research Project:

“Synthesis of Some New Thiazine Derivatives and Their Biological Screening.”

5. (a) Name of the Principal Investigator : Mr.Uttam Bandu Chougale

(b) Department: Chemistry

(c) College where work has progressed: Karmaveer Hire Arts, Science, Commerce and

Education College, Hu. Muralidharnagar, Gargoti 416209.

6. Effective date of starting the project: 01st September 2017

7. Grant approved and expenditure incurred during the period of the report:

a. Total amount approved Rs.2, 50,000/-(First Year) (Amount transferred by UGC to

Principal, Karmaveer Hire Arts, Science, Commerce and Education College, Gargoti.

Bank of India Ac / No. 092810110018163 On 8th

September 2017).

65

b. Total expenditure Rs. 2,53,094/-

c. Report of the work done: separate sheet attached

Brief objective of the project

1. To study the synthesis of novel heterocyclic molecules by multicomponent reactions

(MCR)

2. To check purity of all synthesized compounds using thin layer chromatography.

3. To synthesize variety of chalcones and its use as intermediate for further synthesis of

heterocycles.

4. To use microwave strategy for the synthesis of heterocyclic compounds a green

technique.

5. To carried out synthesis of some bioactive 1,3-thiazine derivatives.

6. Further synthesized compounds characterized by IR, NMR and Mass spectroscopy

techniques

i. Work done so far and results achieved and publications, if any, resulting from the work (Give

details of the papers and names of the journals in which it has been published or accepted for

publicationp)

Conference Attended-:

1. Attended National Conference on “Recent Trends in Chemical Science and Its

Interdisciplinary Applications” Organized by Department of Chemistry, Yashwantrao Patil

Science College, Solankur on 6-January2018.

2. Attended National Seminar on “Innovative Trends in Physics and Chemistry” Organized by

Department of Physics and Chemistry, Baba Naik Science Mahavidyalaya, Shirala on 7-

February 2018.

66

67

O

H

R2R1

O

CH3

R3

O

R2R1

R3

+NaOH

Aldehyde AcetophenoneChalcone

Report of the work done in First Year

1) Literature surveys were done on the history of heterocyclic chemistry which was began in the

1800's, in step with the development of organic chemistry. Survey revealed that After World

War II, there was an enormous explosion research in the field of heterocycles. About one half of

over six million compounds recorded in Chemical Abstracts are heterocyclic. Heterocyclic

chemistry is one of the most complex and intriguing branch of organic chemistry and

heterocyclic compounds constitute the largest and most varied family of organic compounds.

Many broader aspects of heterocyclic chemistry are recognized as disciplines of general

significance that impinge on almost all aspects of modern organic chemistry, medicinal

chemistry and biochemistry. Heterocyclic compounds offer a high degree of structural diversity

and have proven to be broadly and economically useful as therapeutic agents.

2) Extensive literature survey were done on the synthesis of different chalcone derivatives as an

precursors for the next step of thiazine synthesis. To carry out experimental work, purchased

chemicals, glass wares and equipments.

3) This study concern with the synthesis of chalcones by either Microwave assisted or by Grinding

technique. Also taken the precaution in choosing the starting compounds whose combination

not only gives the better yield but also the diverse biological activities. Microwave and Grinding

techniques provides a powerful way to do synthetic chemistry in green approach.

4) In the research work, different chalcone derivatives have been synthesized from propely

substituted aromatic aldehydes and acetophenones under basic reaction medium by both

Grinding as well as microwave irradiation method.

Scheme 1. Synthesis of Chalcone derivatives.

68


d) Grinding Method: Aromatic aldehyde (1 mmol), Substituted acetophenone (1 mmol) and

sodium hydroxide (in catalytic ammount) was grinded in mortar using pestale. After

completion of reaction monitored by TLC, Reaction mixture was kept overnight. The solid

mass obtained was isolated by simple work using water. Finally the oven dried product was

recrystallized from ethanol to get desired chalcone in excellent yield.

e) Microwave Irradiation Method: Aromatic aldehyde (1 mmol), Substituted acetophenone

(1 mmol) and sodium hydroxide (in catalytic ammount) were mixed along with ethyl

alcohol: water (1:1) and irradiated in microwave synthesizer system at 30% power (160W)

for 2-3 min.at 65oC. Reaction was monitored by TLC (Ethyl acetate: n-Hexane, 8:2 systm).

After completion of the reaction, the reaction mixture was recrystallized from EtOH to

afford the pure product.

Table 1-: Physical constants of the synthesized compounds

Compound

Code

R1

R2

R3

Molecular

Formula

Molecular

Weight

Yield

%

MP (0C)

1a -OC2H5 H -OCH3 C18H18O3 282.33 86 116-118

1b -Cl -Cl -OCH3 C16H12 Cl2O2 307.18 75 155-158

1c -OCH3 H -OCH3 C17H16O3

268.30 70 175-177

1d -OH H -OCH3 C16H14O3

254.28 52 238-240

1e -OCH3 H -OC2H5 C18H18O3

282.33 65 186-188

1f H H -OC2H5 C17H16O2

252.11 84 140-142

1g -OCH3 H -Br C16H13Br O2

317.17 88 197-199

1h -OH H -Br C15H11Br O2

301.99 30 265-268

1i H H -Br C15H11Br O

285.99 88 155-157

69

70

Annexure – VI




Final Report of the work done on the Minor Research Project

(Reports to be submitted within 6 weeks after completion of each year)

1 Project report No. 1st/Final : Final Year Report

2 UGC Reference No.

F. 47-1161/14(WRO) dated 28th

July 2017 / 8th

September 2017.

3. Period of report from 01st September 2017 to 31

st August 2019.

4. Title of the Research Project:

“Synthesis of Some New Thiazine Derivatives and Their Biological Screening.”

5. (a) Name of the Principal Investigator : Mr.Uttam Bandu Chougale

(b) Department: Chemistry

(c) College where work has progressed: Karmaveer Hire Arts, Science, Commerce and

Education College, Hu. Muralidharnagar, Gargoti 416209.

6. Effective date of starting the project: 01st September 2017

7. Grant approved and expenditure incurred during the period of the report:

a. Total amount approved Rs. 3, 60,000/- (Two Years)

b. Total expenditure Rs. 3,61,652/-

c. Report of the work done: separate sheet attached

71

8. Brief objective of the project-:

1. To study the synthesis of novel heterocyclic molecules by multicomponent reactions (MCR)



heterocycles.

4. To use microwave strategy for the synthesis of heterocyclic compounds a green technique.


6. To characterize synthesized compounds by IR, NMR and Mass spectroscopy techniques.

In research work :

a) Carried out Synthesis of variety of chalcones from properly substituted aromatic

aldehydes and acetophenones.

b) Synthesis was carried out by using both Grinding as well as Microwave irradiation

techniques.

c) Synthesized chalcones were screened for their anti-microbial activity. Library of such

bioactive chalcones has been obtained.

d) Microwave assisted efficient synthesis of 1,3-thiazine has been carried out by using

prepared chalcones and phenyl hydrazine, phenyl isothiocynate, aromatic aldehydes ,

acetophenones in two sequential steps. The synthesized thiazine derivatives also

screened for their biological activity by using PASS.

e) Further synthesized compounds characterized by IR, NMR and Mass Spectroscopic

Techniques.

72

9. Work done so far and results achieved and publications, if any, resulting from the work

(Give details of the papers and names of the journals in which it has been published or

accepted for publication)



Interdisciplinary Applications” Organized by Department of Chemistry, Yashwantrao

Patil Science College, Solankur on 6-January2018.

2. Attended National Seminar on “Innovative Trends in Physics and Chemistry” Organized

by Department of Physics and Chemistry, Baba Naik Science Mahavidyalaya, Shirala on

7-February 2018.

3. Presented poster entitled “Microwave Assisted Synthesis of Some Novel Thiazine

Derivatives and Prediction of Their Bioactivity” at International Conference on

“Materials and Environmental Science” Organized by Yashwantrao Patil Science

College, Solankur and The New College, Kolhapur on 7-8 December 2018.

Published Papers-:

1. Savita Dhongade*, Poonam Shetake, Vitthal Divate and Uttam Chougale;

Environmentally Benign Protocol For The Synthesis Of Biologically Significant

Pyrano[3,2-c]Chromine-3-Carbonitriles. European Journal of Biomedical And

Pharmaceutical Sciences. 2018, 5(1), 407-410.

2. U.B.Chougale1*

, P.R.Kharade1, S.R.Dhongade

2; Synthesis of Some Chalcone

Derivatives and Screening of Their Antimicrobial Activity. Research Journal of Life

Sciences, Bioinformatics, Pharmaceutical and Chemical Science. 2019, 5(3), 343-349.

3. U.B.Chougalea*

, P.R.Kharadea, H.V.Chavan

b, S.R.Dhongade

c; Microwave Assisted

Synthesis of Some Novel Thiazine Derivatives and Prediction of Their Bioactivity.

Materials Today: Proceedings 2018, (ICMES 2018). (This paper is under review process

and will be published in Materials Today: Proceedings in upcoming issue.

73

74

N N

NH

S

R1

R2

R3

R5

R4

O

R2R1

R3

R1

N R2

R3

S

N

NR4

R5

R4NHNH2

R5N=C=S 20% 140 W

4(a-g)

Final Report of the work done on the project

1) In scheme 2, Chalcones prepared in scheme 1 were subjected for further sequencial reaction

to obtain the targeted comound thiazine derivatives in good to excellent yield. Initially the

chalcones, phenyl hydrazine and phenyl isothiocyanate were subjected for one pot three

component microwave irradiation to offered the intermediate compounds 4(a-g). These

intermediate compounds on further irradiation with substituted chalcones to generate the

desired products thiazine derivatives. The advantage of this method includes the short

reaction time, easy work-up and excellent yields.

Synthesis of 2-phenyl-2,5-dihydro-pyrazole-1-carbothioic acid phenyl amide derivatives 4(a-g).

Table-: General Characteristics and Elemental Analysis data of the Compounds [4(a-g)]

75

R1

N R2

R3

S

N

NR4

R5

O

R6

R7

R8

N S

R6 R7R8

N

R5

NR4

R2

R3

R1

20% 140 W

General Synthetic Procedure for the 2-phenyl-2, 5-dihydro-pyrazole-1-carbothioic acid

phenyl amide derivatives [4(a-g)].

In 100 mL round bottom flask 0.01 mole chalcone [1(a-b)], 0.01 mole substituted hydrazine, 0.01

mole substituted isothiocynate were mixed in ethanol (30 mL) and the reaction mixture was

irradiated with microwaves at 20% power (140 W) for 5-7 min. The completion of the reaction was

confirmed by TLC (Ethyl acetate: Hexane). The reaction mixture was allowed to cool at room

temperature, poured into crushed ice. The solid separated was filtered, washed with little 1:1

ethanol and purified by recrystallization from ethanol to give pure products. The products thus

obtained [4(a-g)] were directly used for next step.

Synthesis of thiazine derivatives [6(a-j)]-:

Comp.

R

1

R2

R

3

R4

R

5 Yield

%

M.P.

0C

Mol. Formula




78 210 C28H29N3O2S


84 205 C22H25N3O2S



74 225 C25H21Cl2N3OS


76

N S

N

N

R8R7R6

R5

R4

R2

R3

R1

Table-: General Characteristics and Elemental Analysis data of the Compounds [6(a-j)]

Comp.

R1

R2

R3

R4

R5

R6

R7

R8

Yield

%

M.P

. oC

Mol. Formula




C46H39Cl2N3O3S


C46H47N3O4S


C40H43N3O4S


C43H31Cl4N3O2S


C46H39Cl2N3O3S


C43H39Cl2N3O3S


C40H35Cl2N3O3S


C37H27Cl4N3O2S

77

General Synthetic Procedure for the thiazine derivatives [6(a-j)]-:


carbothioic acid phenyl amide derivatives [4(a-g)], 0.001 mole chalcone [5(a-b)] were mixed

in ethanol (30 mL). The reaction mixture was irradiated with microwaves at 20% power (140

W) for 5-7 mins. The completion of the reaction was confirmed by TLC (Ethyl acetate:

Hexane). The reaction mixture was allowed to cool at room temperature, poured into crushed

ice. The solid separated was filtered, washed with little 1:1 ethanol and purified by

recrystallization from ethanol to furnish pure products [6(a-j)].

MICROBIAL TESTING-:




method at 40 PPM concentration using 7mm size filter paper disc. At similar condition standard

drug streptomycin sulfate was used.

Medium for growth

For antibacterial activities against E. coli MacConkey agar was used with following composition.

a) Peptone 20gms.

b) Lactose 10gms.

c) Bile Salt 5gms.



f) Agar 12gms.


For antibacterial activities against S. albus nutrient agar with following composition was used.

a) Peptone 5gms.



d) Agar 15gms


78

79

Annexure - VII




PROFORMA FOR SUBMISSION OF INFORMATION AT THE TIME OF SENDING THE

FINAL REPORT OF THE WORK DONE ON THE PROJECT

1. Title of Project: “Synthesis of Some New Thiazine Derivatives and Their Biological

Screening.”

2. NAME AND ADDRESS OF THE PRINCIPAL INVESTIGATOR: Mr.Uttam Bandu

Chougale, Department of Chemistry, Karmaveer Hire Arts, Science, Commerce and

Education College, Gargoti.

3. NAME AND ADDRESS OF THE INSTITUTE: Karmaveer Hire Arts, Science, Commerce

and Education College, Gargoti, Dist-Kolhapur, Pin-416209.

4. UGC APPROVAL LETTER NO. AND DATE:

F. 47-1161/14(WRO) dated 28th

July 2017

F.47-1161/14(WRO) dated 8th

September 2017

5. DATE OF IMPLEMENTATION: 01/09/2017

6. TENURE OF THE PROJECT: Two years (01/09/2017 to 31/08/2019)

7. TOTAL GRANT ALLOCATED: 3,60,000/-

8. TOTAL GRANT RECEIVED: 2,50,000/- ) (Amount transferred by UGC to Principal, Karmaveer

Hire Arts, Science, Commerce and Education College, Gargoti, Bank of India Ac/No.

092810110018163 on 08 September 2017)

9. FINAL EXPENDITURE: 3,61,652/- (1,11,652/- balance amount to be obtained)

10. TITLE OF THE PROJECT: “Synthesis of Some New Thiazine Derivatives and Their

Biological Screening.”

11. OBJECTIVES OF THE PROJECT:

1. To study the synthesis of novel heterocyclic molecules by multicomponent reactions (MCR)



heterocycles.

4. To use microwave strategy for the synthesis of heterocyclic compounds a green technique.


6. To characterize synthesized compounds by IR, NMR and Mass spectroscopy techniques.

80

In research work:

a) Carried out Synthesis of variety of chalcones from properly substituted aromatic

aldehydes and acetophenones.

b) Synthesis was carried out by using both Grinding as well as Microwave irradiation

techniques.

c) Synthesized chalcones were screened for their anti-microbial activity. Library of such

bioactive chalcones has been obtained.

d) Microwave assisted efficient synthesis of 1,3-thiazine has been carried out by using

prepared chalcones and phenyl hydrazine, phenyl isothiocynate, aromatic aldehydes ,

acetophenones in two sequential steps. The synthesized thiazine derivatives also

screened for their biological activity.

e) Further synthesized compounds characterized by IR, NMR and Mass Spectroscopic

Techniques.

12. ACHIEVEMENTS FROM THE PROJECT



Interdisciplinary Applications” Organized by Department of Chemistry, Yashwantrao Patil

Science College, Solankur on 6-January2018.

2. Attended National Seminar on “Innovative Trends in Physics and Chemistry” Organized by

Department of Physics and Chemistry, Baba Naik Science Mahavidyalaya, Shirala on 7-

February 2018.

3. Presented poster entitled “Microwave Assisted Synthesis of Some Novel Thiazine

Drivatives and Prediction of Their Bioactivity” at International Conference on “Materials

and Environmental Science” Organized by Yashwantrao Patil Science College, Solankur

and The New College, Kolhapur on 7-8 December 2018.

81

Published Papers-:

1. Savita Dhongade*, Poonam Shetake, Vitthal Divate and Uttam Chougale; Environmentally

Benign Protocol For The Synthesis Of Biologically Significant Pyrano[3,2-c]Chromine-3-

Carbonitriles. European Journal of Biomedical And Pharmaceutical Sciences. 2018, 5(1),

407-410.

2. U.B.Chougale1*

, P.R.Kharade1, S.R.Dhongade

2; Synthesis of Some Chalcone Derivatives

and Screening of Their Antimicrobial Activity. Research Journal of Life Sciences,

Bioinformatics, Pharmaceutical and Chemical Science. 2019, 5(3), 343-349.

3. U.B.Chougalea*

, P.R.Kharadea, H.V.Chavan

b, S.R.Dhongade

c; Microwave Assisted

Synthesis of Some Novel Thiazine Derivatives and Prediction of Their Bioactivity.

Materials Today: Proceedings 2018, (ICMES 2018). (This paper is under review process

and will be published in Materials Today: Proceedings in upcoming issue.

4. During the period of project Six students from B.Sc. III (Chemistry) has done their project

work in this field.

13. SUMMARY OF THE FINDINGS-:

1) Literature survey was done on the history of heterocyclic chemistry which was began in the

1800's, in step with the development of organic chemistry. Survey revealed that After World

War II, there was an enormous explosion research in the field of heterocycles. About one

half of over six million compounds recorded in Chemical Abstracts are heterocyclic.

Heterocyclic chemistry is one of the most complex and intriguing branch of organic

chemistry and heterocyclic compounds constitute the largest and most varied family of

organic compounds. Many broader aspects of heterocyclic chemistry are recognized as

disciplines of general significance that impinge on almost all aspects of modern organic

chemistry, medicinal chemistry and biochemistry. Heterocyclic compounds offer a high

degree of structural diversity and have proven to be broadly and economically useful as

therapeutic agents.

2) Extensive literature survey was done on the synthesis of different chalcone derivatives as an

precursors for the next step of thiazine synthesis. To carry out experimental work, purchased

chemicals, glass wares and equipments.

82

3) This study concern with the synthesis of chalcones by either Microwave assisted or by

grinding technique. Also taken the precaution in choosing the starting compounds whose

combination not only gives the better yield but also the diverse biological activities.

Microwave and grinding techniques provides a powerful way to do synthetic chemistry in

green approach.

4) In the research work, synthesis of different chalcone derivatives has been synthesized from

propely substituted aromatic aldehydes and acetophenones under basic reaction medium by both

Grinding as well as microwave irradiation method.

O

H

R2R1

O

CH3

R3

O

R2R1

R3

+NaOH

Aldehyde AcetophenoneChalcone

Scheme1. Synthesis of Chalcone derivatives


a) Grinding Method: Aromatic aldehyde (1 mmol), Substituted acetophenone (1 mmol)

and sodium hydroxide (in catalytic ammount) was grinded in mortar using pestale. After

completion of reaction monitored by TLC, Reaction mixture was kept overnight. The

solid mass obtained was isolated by simple work using water. Finally the oven dried

product was recrystallized from ethanol to get desired chalcone in excellent yield.

b) Microwave Irradiation Method: Aromatic aldehyde (1 mmol), Substituted

acetophenone (1 mmol) and sodium hydroxide (in catalytic ammount) was mixed along

with ethyl alcohol: water (1:1) and irradiated in microwave synthesizer system at 30%

power (160W) for 2-3 min.at 65oC . Reaction was monitored by TLC (Ethyl acetate : n-

Hexane, 8:2 system ). After completion of the reaction, the reaction mixture was

recrystallized from EtOH to afford the pure product.

83

Table 1-: Physical constants of the synthesized compounds

Compound

Code

R1

R2

R3

Molecular

Formula

Molecular

Weight

Yield

%

MP (0C)

1a -OC2H5 H -OCH3 C18H18O3 282.33 86 116-118

1b -Cl -Cl -OCH3 C16H12 Cl2O2 307.18 75 155-158

1c -OCH3 H -OCH3 C17H16O3

268.30 70 175-177

1d -OH H -OCH3 C16H14O3

254.28 52 238-240

1e -OCH3 H -OC2H5 C18H18O3

282.33 65 186-188

1f H H -OC2H5 C17H16O2

252.11 84 140-142

1g -OCH3 H -Br C16H13Br O2

317.17 88 197-199

1h -OH H -Br C15H11Br O2

301.99 30 265-268

1i H H -Br C15H11Br O

285.99 88 155-157

Table 2-: Antimicrobial activity of the synthesized compounds

Compound Antebacterial activity (%inhibition)

Staphylococcus aureus (737) Pseudomonas aeruginosa (1688)

500 µg/ ml 1000 µg/ ml 500 µg/ ml 1000 µg/ ml

1a 21.4 32.6 ------ ------

1b 24.3 33.6 ------ ------

1c ------ ------ ------ ------

1d ------ ------ ------ ------

1e 23.8 33.6 ------ ------

1f -------- -------- ------ ------

1g 25.0 34.8 ------ ------

1h 22.6 32.8 ------ ------

1i 23.0 33.3 ------ ------

Chloramphenicol 42.3 55.2 63.7 78.9

DMSO 1.4 ------ 1.2 --------

84

O

R2R1

R3

R1

N R2

R3

S

N

NR4

R5

R4NHNH2

R5N=C=S 20% 140 W

4(a-g)

5) Synthesized Chalcones were characterized by using IR, 1H & Mass spectral techniques.

6) Synthesized Chalcones were screened for their antimicrobial activity.

7) Further synthesized compounds were characterized by IR, NMR and Mass spectroscopy

techniques.

5) In scheme 2, Chalcones prepared in scheme 1 were subjected for further sequencial reaction to

obtain the targeted comound thiazine derivatives in good to excellent yield. Initially the

chalcones, phenyl hydrazine and phenyl isothiocyanate were subjected for one pot three

component microwave irradiation to offered the intermediate compounds 4(a-g). These

intermediate compounds on further irradiation with substituted chalcones to generate the desired

products thiazine derivatives. The advantage of this method includes the short reaction time,

easy work-up and excellent yields.

Synthesis of 2-phenyl-2,5-dihydro-pyrazole-1-carbothioic acid phenyl amide derivatives

4(a-g).

85

N N

NH

S

R1

R2

R3

R5

R4

Table-: General Characteristics and Elemental Analysis data of the Compounds [4(a-g)]

General Synthetic Procedure for the 2-phenyl-2,5-dihydro-pyrazole-1-carbothioic acid

phenyl amide derivatives [4(a-g)].

In 100 mL round bottom flask 0.01 mole chalcone [1(a-b)], 0.01 mole substituted hydrazine, 0.01

mole substituted isothiocynate were mixed in ethanol (30 mL) and the reaction mixture was

irradiated with microwaves at 20% power (140 W) for 5-7 min. The completion of the reaction was

confirmed by TLC (Ethyl acetate: Hexane). The reaction mixture was allowed to cool at room

temperature, poured into crushed ice. The solid separated was filtered, washed with little 1:1

ethanol and purified by recrystallization from ethanol to give pure products. The products thus

obtained [4(a-g)] were directly used for next step.

Comp.

R

1

R2

R

3

R4

R

5 Yield

%

M.P.

0C

Mol. Formula




78 210 C28H29N3O2S


84 205 C22H25N3O2S



74 225 C25H21Cl2N3OS


86

R1

N R2

R3

S

N

NR4

R5

O

R6

R7

R8

N S

R6 R7R8

N

R5

NR4

R2

R3

R1

20% 140 W

N S

N

N

R8R7R6

R5

R4

R2

R3

R1

Synthesis of thiazine derivatives [6(a-j)]-:

Table-: General Characteristics and Elemental Analysis data of the Compounds [6(a-j)]

87

General Synthetic Procedure for the thiazine derivatives [6(a-j)]


carbothioic acid phenyl amide derivatives [4(a-g)], 0.001 mole chalcone [5(a-b)] were mixed

in ethanol (30 mL). The reaction mixture was irradiated with microwaves at 20% power (140

W) for 5-7 mins. The completion of the reaction was confirmed by TLC (Ethyl acetate:

Hexane). The reaction mixture was allowed to cool at room temperature, poured into crushed

ice. The solid separated was filtered, washed with little 1:1 ethanol and purified by

recrystallization from ethanol to furnish pure products [6(a-j)].

Comp.

R1

R2

R3

R4

R5

R6

R7

R8

Yield

%

M.P

. oC

Mol. Formula




C46H39Cl2N3O3S


C46H47N3O4S


C40H43N3O4S


C43H31Cl4N3O2S


C46H39Cl2N3O3S


C43H39Cl2N3O3S


C40H35Cl2N3O3S


C37H27Cl4N3O2S

88

MICROBIAL TESTING-:




method at 40 PPM concentration using 7mm size filter paper disc. At similar condition standard

drug streptomycin sulfate was used.

Medium for growth

For antibacterial activities against E. coli MacConkey agar was used with following composition.

a) Peptone 20gms.

b) Lactose 10gms.

c) Bile Salt 5gms.



f) Agar 12gms.


For antibacterial activities against S. albus nutrient agar with following composition was used.

a) Peptone 5gms.



d) Agar 15gms


Activities were determined by using the cultivated plates of Whatmann filter paper (7mm).

All the compounds were dissolved in DMF (40ppm) Plates were soaked in the solution. These

plates were placed on the medium previously seeded with organisms in Petri dishes and stored in an

incubator at 270 for 48 hours for E. coli and 24 hours for S. albus respectively. An inhibition zone

was measure in mm and recorded in the following table.

89

Results and Discussion-:

In Thiazine derivatives tested compounds 6-a, 6-g and 6-j showed greater degree of antibacterial

activity against E.coli. However, the compounds 6-b, 6-d and 6-h showed greater degree of

antibacterial activity against S.albus.

1) We have reported efficient synthesis of 1,3-thiazine derivatives. Clean reaction, low

reaction times was the main advantages of this method. Satisfactory yield of products

and easy workup make this a useful protocol for green synthesis of this class of

compounds.

2) Also the synthesized thiazine derivatives were screened for their antimicrobial activities.

9. CONTRIBUTION TO THE SOCIETY-:

Two research papers published. Six students from B.Sc. III (Chemistry) have done their project

work in this field. The proposed work was utilized for training the B.Sc. students in handling the

various chemicals at the milimole level and developing the skills of the students in monitoring

the reactions at the various levels using the TLC techniques. The students were also trained in

the isolation and purification techniques such as extraction, column chromatography and

crystallization. This opportunity was also utilized in the developing the skills of the students in

characterization technique by studying the IR & NMR spectra of the various compounds those

synthesized.

Comp.No E.coli S.albus

6a 12 8

6b 9 16

6c 11 14

6d 9 16

6e 12 14

6f 12 8

6g 15 10

6h 9 17

6i 11 10

6j 13 10

90

91

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FINAL COMPLETION REPORT OF MINOR RESEARCH PROJECT …