“MICROWAVE ASSISTED SYNTHESIS OF BIOLOGICALLY ACTIVE 2-SUBSTITUTED 6-FLUORO-7-CHLORO (1,3) BENZOTHIAZOLES”

M. PHARM. DISSERTATION PROTOCOL SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE ,KARNATAKA.

BY Mr. MOHAN KUMAR K.M. B. Pharm.

Under the guidance of, Dr. B.SHIVAKUMAR M. Pharm., Ph. D.

P. G. DEPARTMENT OF PHARMACHEMISTRY S. C. S. COLLEGE OF PHARMACY, HARAPANAHALLI – 583 131. RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE, KARNATAKA.

Annexure – II

MOHAN KUMAR K.M. S/O GURUSWAMY K.M. MANAGER,B.E.O. OFFICE, 01 Name and Address of the Candidate NANDINI NILAYA, 4TH WARD, HIPPITOTA, HARAPANAHALLI (P)-583 131, DAVANAGERE (D), KARNATAKA.

T. M. A. E. SOCIETY’S 02 Name of the Institution S. C. S. College of Pharmacy, Harapanahalli – 583 131.

Course of the Study M. Pharm (Previous) 03 Branch PHARMACEUTICAL CHEMISTRY

04 Date of Admission to course 16 june 2008

“ MICROWAVE ASSISTED SYNTHESIS OF BIOLOGICALLY ACTIVE 2-SUBSTITUTED 05 Title of the Topic 6-FLUORO-7-CHLORO (1,3) BENZOTHIAZOLES ”

06 Brief resume of the intended work :

6.1) Need for the study: Benzothiazoles are bicyclic ring system with multiple applications, 2-amino benzothiazoles were intensively studied as central muscle relaxants. So benzothiazole derivatives have been studied extensively and found to have diverse chemical reactivity and broad spectrum of biological activities like, Antimicrobial 1-3, Antitumor 4-5, Anthelmentic 6-8, antileishmanial 9-10, Anticonvulsant 11-12, Antiinflammatory activity13-15. Therefore we plan to synthesize the chemical entities with chloro fluoro 1,3- Benzothiazole may result the compounds with interesting biological activity. Ranjana Aggarwal et al16. synthesized 1-(6-flurobenzothiazole-2yl)-3-(4-fluro- phenyl)-5-aryl pyrazolines and their iodine mediated oxidation to corresponding pyrazoles and screened for biological activities. Sandhyabawa et al17 synthesized 6-fluro-2-[4-formyl-3(substituted phenyl)pyrazol- 1-yl] benzothiazole and evaluated for their antibacterial activity. Some of the compound showed excellent antibacterial activity comparable to standard.

18 S.N. Sawhney et al . reported and synthesized 2-(3'-5'-dimethyl-4'-substituted-pyrazole-1'- yl) benzothiazoles and evaluated for their anti-inflammatory activity. Some of the compounds showed low to moderate anti-inflammatory activity. B.Shivakumar et al19. reported the synthesis 6-fluro-7-substituted-1,2.,3,4-tetrazolo [5,1-b] benzothiazole and screened for their antimicrobial properties. The tested compounds exhibited weak antimicrobial activity compared to standard. E. Jayachandran et al20. further in continuation of their work reported the synthesis of 2-(3- amino-5-S-methyl-4-corboxamido pyrazol-1-yl)-6-fluro-7-substituted (1,3) benzothiazoles and screened for their antimicrobial and anti-inflammatory activity activity. Some of them have shown moderate to promising anti-inflammatory activity activity and exhibited significantantimicrobial activity. E. Jayachandran et al21. reported the synthesis of 2-[N-P-tolyl sulphon hydrazino ]-6-fluro-7- substituted (1,3) benzothiazoles and screened for their anti-inflammatory activity. Some of these compounds showed promising anti-inflammatory activity. Microwave mediated reactions have been conducted to achieve enhanced reaction rates, cleaner products and manipulate simplicity, shorter reaction time, simple reaction conditions and higher product yields, render the microwave method superior to conventional methods. Besides synthesis of heterocyclic compounds of all types of organic reactions such as oxidation, reduction, alkylation, aromatic substitution, esterification, hydrolysis, decorboxylation and various condensation reactions are conveniently performed under microwave induced organic reactions to develop new chemical entities in a shortest possible time is enormous. Microwave mediated reactions under dry conditions using inorganic are being employed to accomplish greater activity. Every year newer heterocyclic molecules are generated by drug discovery research and development, such activity can be greatly accelerated via microwave-mediated reactions. Due to continued interest in the synthesis of chloro-fluoro (1,3) benzothiazole derivatives22-33. Here we report the practical and efficient method for synthesis of 2-(N2-arylidene hydrazino)-7-chloro-6- fluoro-(1,3) benzothiazoles, starting from appropriate 2-amino and 2-hydrazino-chloro-fluoro benzothiazoles under microwave irradiation and solvent free conditions. The structures of the synthesized compounds will be established on the basis of the spectral properties (IR, NMR and Mass) and elemental analysis. The synthesized compounds will also be screened by for their possible biological and pharmacological activities.

6.2) Review of Literature A literature survey done by referring chemical abstracts, journals in internet and also availing facilities at I.I.Sc. Ban galore. Further the references work will be carried out at Kuvempu University Shimoga, Gulbarga University Gulbarga and Indian Institute of Chemical Technology at Hyderabad.

6.3 Objective of the Study It could be noted from the literature that the chloro-fluoro- benzothiazoles associated with substituent at 2nd position are biologically and pharmacologically potent, therefore in continuation of our work on chloro-fluro-benzothiazoles, it has been considered worthwhile to synthesize 2-(N2-arylidene hydrazino)-7-chloro-6-fluoro-(1,3)- benzothiazoles starting from appropriate 2-amino and 2- hydrazino chloro-fluoro benzothiazoles under microwave irradiation and solvent free conditions.The newly synthesized compounds will be characterized by elemental and spectral properties, these synthesized compounds will also be subjected for their possible biological and pharmacological activites. Materials and methods Preparation and screening methods will be followed according to literature procedure. 7.1) Source of data  Journal of Medicinal Chemistry.  Tetrahedron Letter.  Tetrahedron.  Chemical Abstracts.  Indian Journal of Heterocyclic Chemistry.  Indian Journal of Chemistry.  Journal of Chemical Society.  Journal of American Chemical Society.  Standard test books for screening of biological and pharmacological activities. Indian Pharmacopoeia. a) Spectroscopic data: 7.2) Methods of Collection of Data IR of all new compounds will be obtained by (including sampling procedure, if any) using of spectrometers available in our college, NMR and Spectroscopic data will be obtained from I.I.T.Madras and I.I.S.C. Bangalore etc.

b) Pharmacological and Biological Activity:

1) Anti-inflammatory activity34 in-vitro - Selected synthesized compounds will be subjected to screen for inhibit the denaturation of protein as an In-vitro screening model for anti-inflammatory compounds.

2) Antioxidant activity35 – Selected compounds will be evaluated for antioxidant activity.

3) Antimicrobial activity36 –All synthesized compounds will be evaluated for antimicrobial activity by cup plate method. Selected compounds are subjected to MIC.

7.3) Does the study require any investigations or interventions to be -NO- conducted on patients or other humans or animals? If so, please describe briefly.

7.4) Has ethical clearance been obtained from your institution in case of 7.3 -NO- References : 1) Sreenivasa, M.V., Nagappa, A.N. and Nargund, L.V.G., “Various benzothiazolo triazole derivatives possess good antimicrobial activity”. Indian J.Heterocycl. Chem . 8,1998,23.

2) Gopkumar. P., Shivakumar, B., Jayachandran, E., Nagappa, A.N., Nargund, L.V.G., and Gurupadaiah, B.M., “Some 6-fluoro-7-(substituted)-(2-N-p-anilinosulfonamido) benzothiazoles were synthesized and studied for their antimicrobial and antifungal activities”. Indian J.Heterocycl. Chem, 11,2001, 39.

3) .Bhusari, S.R., Pawar, R.P., and Vibute Y.B., “Some new 2-( substituted phenylsulfonamido)-6- substituted benzothiazoles and screened them for antibacterial activity”. Indian J.Heterocycl. Chem., 11,2001,79.

4) Shi, D.F., Bradshaw, T.D., Wrigley, S., Carol J., Mccall, P.L., Malcolm, F. and Stevens, F.G, 2-(4-Aminophenyl) benzothiazoles , comprise a novel mechanistic class of antitumor agents, J. Med.Chem., 39,1996, 3375.

5) Kashiyarna, E.,Hutchinson, I., Chua, M.S., Stinson, S.F., Phillips, L.R.,Kaur, G., Sausville, E.A., Bradshaw, T.D., Westwell, A.D. and Stevens, M.F.G, 2-(4- Aminophenyl) benzothiazoles- antitumor agents, J. Med. Chem,42,1999, 4172.

6) Nargund, L.V.G.,Few novel 8-fluoro-9- substituted(1,3) benzothiazolo (5,1-b)-1,3,4- triazoles studied for their anthelmintic activity., Indian Drugs, 36,1999,137.

7) Bhusari, K.P., Khedekar, P.B., Umathe, S.N., Bahekar, R.H., and Rao R.R.8-bromo-9- substituted(1,3) benzothiazolo [5,1-b]-1,3,4-triazoles screened for their anthelmintic activity. Indian J.Heterocycl. Chem., 9,2000, 275.

8) Amit, B.N., Kamath, R.V., and Khadse, G.B., Some substituted imidazobenzothiazoles were tested for in vivo anthelmintic activity , Indian J.Heterocycl. Chem., 9,2000,309.

9) Delmas, F., Avellaneda, A., Giorgio, C.D., Robin, M., Clercq, E.D., Timon-David, P. and Galy, J.P.,(1,3 benzothiazol-2-yl) amino-9-(10H)-acridinone derivatives were assessed for their in vitro antileishmanial and anti HIV activities., Eur. J. Med. Chem., 39,2004, 685.

10) Delmas, F., Giorgio, C.D., Robin, M., Gasquest, N.A.M., Detang, C., Costa, M., Timon-David, P. and Galy, J.P., Position 2-substitution-bearing 6-nitro and 6-amino benzothiazoles and their corresponding anthranillic acids and assessed the in vitro antiparasitic activity. Antimicrob. Agents Chemother., 46,2002, 2588. 11) Siddiqui, N., Pandeya, S.N., Sen, A.P. and Singh, G.S.,A series of benzothiazolyl guanidines were synthesized for their significant anticonvulsant activity, Pharmak Eftiki, 4,1992,121.

12) Singh, S.P., Misra, R.S., Parmar, S.S. and Brumleve, S.J., Some 2-(4- arylthiosemicarbazidocarbonylthio) benzothiazoles, screened for their anticonvulsant activity. J. Pharm. Sci., 64,1978,1245.

13) Oketani, K., Nagakura, N., Harada, K. and Inoue, T., In vitro pharmacological profiles of 6-hydroxy-5,7-dimethyl-2-(methylamino)-4-(3-pyridylmethyl) benzothiazoles, were assessed for their anti-inflammatory activity. Eur. J. Pharm., 422,2001, 209.

14) Sawhney, S.N., Bhutani, S. and Dharamvir, Some novel 2-(2- benzothiazolyl)-6-aryl-4,5-dihydro-3(2H)-pyridazinone, screened for their anti- inflammatory activity. Indian J. Chem., 26B,1987, 348.

15) Singh, S.P. and Vaid, R.K., Some new 2-(4-butyl-3,5-dimethylpyrazol-1-yl)-6- substituted benzothiazoles, were assessed for their anti-inflammatory activity. Indian J. Chem.,, 25B, 1986, 288.

16) Ranjana Aggarwal, Vinod Kumar, Shiv P Sing. Synthesis of some new 1-(6-flurobenzothiazole-2yl)-3-(4-fluro- phenyl)-5-aryl pyrazolines and their iodine(III) mediated oxidation to corresponding pyrazoles. Ind J Chem., 46B,2007,1332-36.

17) Sandhyabawa, Harish Kumar. Synthesis of 6-fluro-2-[4-formyl-3(substituted phenyl)pyrazol-1-yl] benzothiazoles as potential antibacterial agents. Ind J .Hetero Chem., 14,2005, 249-50.

18) S.N. Sawhney, Tomer RK, OM Prakash, Indra Prakash, Singh SP, Benzothiazole derivatives: Part XI-Synthesis and anti-inflammatory activity of some 2-(3'-5'- dimethyl-4'-substituted-pyrazole-1'-yl) benzothiazoles.Ind J Chem, 20B,1981, 314-16.

19) Shivakumar B, Sojan K Paul, Nagendra Rao R, Jayachandran. E. Synthesis and microbial evaluation of 6-fluro-7-substituted-1,2.,3,4-tetrazolo [5,1-b] benzothiazoles. Ind J. Hetero Chem, 15,2005,71-72.

20) Jayachandran. E, Naragund LVG, Shivakumar B, Kamal Bhatia. Synthesis and Pharmacological screening of 2-(3-amino-5-S-methyl-4-corboxamido pyrazol-1-yl)-6- fluro-7-substituted (1,3) benzothiazoles. Oriental J Chem, 19(1)2002, 139. 21) Sreenivasa Rao D, Jayachandran. E, Sreenivasa GM, Shivakumar. Inhibition of albumin denaturation and anti-inflammatory activity of 2-[N-P-tolyl sulphon hydrazino ]-6-fluro-7-substituted (1,3) benzothiazoles. Oriental J Chem, 21(1),2005,113.

22) Gajaraj Sharma, Sreenivasa, G.M. and Jayachandran,.E., Synthesis of bioactive molecule fluoro benzothiazole comprising quinazolinyl oxadiazoles derivative for biological and pharmacological screening. Ultra chemistry, 4(1), 2008, 103-114.

23) Jitender Kumar, Manjunath Ghate, Palash Pal, Mayukh Baidya and Amith Kumar Das, Synthesis and antimicrobial activity of some new sulphonamidofluorobenzothiazoles. Ind. J.Hetero chem, 17,2007, 199-200.

24) Gangadhar Shaw, Sreenivasa, G.M. and Jayachandran,.E, Synthesis of fluoro substituted benzothiazoles incorporated with 1,3,4-thiadiazoles for biological and pharmacological screening. Oriental. J. chemistry, 24(2), 2008, 475-484.

25) Snehal Patel, Sreenivasa, G.M. and Jayachandran,.E, Niladry Ghose and Jayaraj Kumar, Synthesis of fluoro substituted benzothiazoles comprising sulphonamido quinazolinyl imidazole for biological and pharmacological screening.Ultra Science, 20(2), 2008, 281.

26) Kore Srinivas, Sreenivasa, G.M. and Jayachandran,.E, and D. Sreenivasa Rao, Synthesis and pharmacological screening of bioactive molecule fluoro benzothiazole comprising sulfonamido imidazolinone derivatives. Asian J. Chem, 19, 2007, 4193-4202.

27) Jayachandran,.E, Nargund, L.V.G., Akhiles Roy, Sreenivasa, G.M, Krupanidhi,.K, Synhesis of 1-[6'- fluoro-7' substituted- (1,3') benzothiazol-2-yl] amino-4-substituted-3 chloro azetidin-2 -one for antimicrobial screening.Oriental. J. chemistry, 23(3), 2007, 829-835.

28) Jayachandran,.E, L.V.G. Nargund, Akhiles Roy, Sreenivasa, G.M, Krupanidhi,.K, Synhesis of 3-[6'- fluoro-7' substituted- (1,3') benzothiazol-2'-yl] amino-2-substituted- (1,3)thiazolidin-4 -one for biological screening.Oriental. J. chemistry, 23(3), 2007,943-950.

29) Ranjana Aggarwal, Vinod Kumar, Shiv P Sing., Ind. J. Chem. 46B, 2007,1332-1136.

30) Jayachandran,.E, Sreenivasa, G.M., Shivakumar,. B, and Rao S.D, Anthelmintic activity of 8-fluoro-9-substituted (1,3)-benzothiazolo(5,1-B)-1,2,4-triazole on Perituma Posthuma. Indian drugs., 43(4), 2007, 343-347. 31) Jayachandran,.E, Sreenivasa, G.M., Nargund, L.V.G., Shanmukha,.I, Gaikwad,.P.M., Synthesis of fluoro substituted benzothiazole for pharmacological and biological screening. Oriental. J. chemistry, 22(3), 2006, 537-544.

32) Sreenivasa Rao D, Jayachandran,.,E, Sreenivasa G.M, Shivakumar,. B, Oriental .J. Chemisry, 21(1), 2005, 113-116.

33) Shivakumar,. B, Sojan K Paul, Nagendra Rao R, Jayachandran. E, Ind.J. Hetero Chem .15, 2005, 71-72.

34) Matior Rachman A.K.M., “Anti-inflammatory activity of Sida cordifolia Linn” .Indian J. pharmacol, 38,2006, 207-208 pp.

35) Annie Shirwaikar, K Rajendran & C Dinesh Kumar, “In-vitro antioxidant studies of Annona Squamosa Linn. Leaves”. .Indian J. Experimental Biology. 42, 2004, 803.

36) Neeta S., K. Avani . “A study of the antimicrobial activity of Elephscaber” Indian J. pharmacol., 37, 2005, 126-128 pp. 9 Signature of the Candidate (Mr. MOHAN KUMAR K.M.) Remarks of the Guide The Mentioned Title Compound will be Synthesized and Screened for the activity. They are confirmed by 10 the Spectroscopical Studies under the guidance of me in the P.G.Department of Pharmaceutical Chemistry of our College.

11.I. Name and Designation of GUIDE: Dr. B. SHIVAKUMAR (in Block Letters) Professor, P.G. Dept. of Pharmaceutical Chemistry, Guide ship Reference No. of RGUHS S.C.S. College of Pharmacy, ACA/PGT-M.Ph/SCS/02/99- Harapanahalli-583131. 2000/5821 Date: 05.06.1999 Davangere-dist.

11.2.Signature

11.3.Co-Guide (if any) Dr. B.H.M. JAYAKUMAR SWAMY Professor, P.G. Dept. of Pharmaceutical Chemistry, S.C.S. College of Pharmacy, 11 Harapanahalli-583131. Davangere-dist.

11.4.Signature

11.5. Head of the Department Dr. E. JAYACHANDRAN Professor and Head, Guide ship Reference No. of RGUHS P.G. Dept. of Pharmaceutical Chemistry, ACA-2/RP-TEA/01/97-98/3-6-98 S.C.S. College of Pharmacy, Harapanahalli-583131. Davangere-dist.

11.6. Signature

12.1 Remarks of the Principal

12 12.2. Signature (Dr. S. RAMACHANDRA SETTY)