SYNTHESIS AND EVALUATION OF MERCAPTO OXADIAZOLE DERIVATIVES OF BIOLOGICAL INTEREST

M. Pharm. Dissertation Protocol Submitted to

Rajiv Gandhi University of Health Sciences, KarnatakaBangalore – 560041

By

Mr. DHADUK KAMLESH GHANSHYAMBHAI B.Pharm

Under the Guidance of

SRI. S. S. PUROHIT M. Pharm, (Ph.D.)

LECTURER,

DEPT. OF PHARMACEUTICAL CHEMISTRY,

Department of Pharmaceutical Chemistry

SET’s College of Pharmacy, S. R. Nagar, Dharwad,

Karnataka -580002

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

ANNEXURE –II

PROFORMA FOR REGISTRATION OF SUBJECT DISSERTATION

1. NAME OF THE CANDIDATE AND ADDRESS

MR. DHADUK KAMLESH GHANSHYAMBHAIDEPT. OF PHARMA CHEMISTRYSET’s COLLEGE OF PHARMACYS.R.NAGAR, DHARWAD- 580002.

2. NAME OF THE INSTITUTION SET’s COLLEGE OF PHARMACYS. R. NAGAR,DHARWAD- 580002.

3. COURSE OF STUDY AND SUBJECT

MASTER OF PHARMACY IN PHARMACEUTICAL CHEMISTRY

4. DATE OF ADMISSION TO THE COURSE

23-06-2011

5. TITLE OF THE TOPIC:

SYNTHESIS AND EVALUATION OF MERCAPTO OXADIAZOLE

DERIVATIVES OF BIOLOGICAL INTEREST.

6.0 BRIEF RESUME OF THE INTENDED WORK: 6.1 Need for the study:

Heterocyclic chemistry is the chemistry branch dealing exclusively with synthesis,

properties and applications of heterocyclics especially vital to drug design.

Incorporation of an oxygen, a nitrogen, a sulfur, or an atom of a related element into

an organic ring structure in place of a carbon atom gives rise to a heterocyclic compound.

Since the heterocyclic atom must form more than one bond in order to be incorporated

into a ring structure, halogens do not form heterocyclic compounds although they may be

substituents on a heterocyclic ring structure. Heterocyclic compounds, like polycyclic

ring compounds, are usually known by non-systematic names1. Azoles are five

membered heterocyclic compounds containing in their rings one or more hetero atoms, at

least one of which is nitrogen. Standard drugs used in some of the medicinally important

derivatives containing pyrazoles (azoles) are Novalgin, Aminopyrine etc. which possess

NSAID properties. Apart from this, imidazoles, triazoles possess different biological

activities like antimalarial, hypertensive and antifungal.2

The presence of Oxygen and Nitrogen in heterocyclic system has attracted the

attention of medicinal chemists because of the diverse biological activities and profound

efficacy.3 Five membered heterocycles with two carbon atoms, two nitrogen atoms and

one oxygen atom are called Oxadiazoles. Depending upon the orientation of the nitrogen

atoms they are described as 1,2,3-, 1,2,4-, 1,2,5-, 1,3,4- Oxadiazoles.

Oxadiazoles possess antitubercular,18 muscle relaxant,4 antiviral,5 analgesic,6 anti-

cancer,19 anticonvulsant,20 anti-inflammatory,21 hypotensive,7 antimicrobial,22

anthelmintic,8 properties. Substituted mercapto oxadiazoles possess antimicrobial9,10,15,16,

antitubercular12, antifungal9,11,13, antitumor14 properties.

So, there is an urge to synthesize more potent derivatives containing these atoms

and this research is an attempt to synthesize better, effective substituted mercapto

oxadiazoles.

6.2 Review of literature:

Extensive literature survey was carried out in libraries of SET’s College of

Pharmacy Dharwad and KLE’s College of Pharmacy Belgaum. Karnataka University,

Dharwad and by visiting various web sites through internet the relevant data has been

collected.

Literature review showed following important Substituted mercapto oxadiazole

derivatives of pharmacological importance.

1. Shaharya M. et.al., have reported Oxadiazole mannich bases: Synthesis and

antimycobacterial activity.15

R1=C6H5 , R2=Furfuryl

R1=C6H5, R2=phenyl

2. Islam M. et.al., have reported synthesis and antimicrobial activity of some novel

oxadiazole derivatives.10

R=Phenyl, m-Xyly, P-tolyl, P-Chlorophenyl

3. Pattan SR. et.al., have reported Synthesis and evaluation of some novel

substituted 1,3,4,Oxadiazole and pyrazole derivatives for anti-tuberculer

activity.12

4. Jiao QC. et.al., have reported Synthesis, biological evaluation, and molecular

docking studies of 2-chloropyridine derivatives possessing 1,3,4-oxadiazole

moiety as potential antitumor agents.14

R=2me-C6H4, 4-F-C6H4, 4NH2-C6H4

5. Mayekar AN. et.al., have reported Synthesis and Antimicrobial Studies on New

Substituted 1,3,4-Oxadiazole Derivatives Bearing 6-Bromonaphthalene Moiety.16

6. Parikh PK. et.al., have reported synthesis and biological evaluation of 1,3,4-

oxadiazole derivatives as potential antibacterial and antifungal agents.9

.

7. Kaplancikli ZM. et.al., have reported Synthesis of Some Oxadiazole

Derivatives as New Anticandidal Agents.11

R=P-Cl, m-Cl, H

8. Xu W. et.al. have reported Synthesis and Antifungal Activity of Novel

Sulfone Derivatives Containing 1,3,4-Oxadiazole Moieties.13

6.3 Objectives of Study:

1) To synthesize a new series of substituted mercapto oxadiazole derivatives of

highest purity.

2) To characterize the structure of the newly synthesized compounds by different

analytical techniques such as IR, NMR and Mass spectral data.

3) To evaluate the synthesized compounds for different biological activities.

7.0

MATERIALS AND METHODS:

7.1 Source and Collection of data:

Chemical Abstracts.

Indian Journal of Chemistry.

Indian Journal of Heterocyclic Chemistry.

Journal of Medicinal Chemistry.

Journal of Heterocyclic Chemistry.

European Journal of Medicinal Chemistry.

Bioorganic and Medicinal Chemistry Letters.

World wide web.

J-Gate@ Helinet etc.

7.2 Method of collection of Data:

A) Synthesis of the compounds:

Chemicals and other reagents required for synthesis will be procured from standard

company sources. Compounds will be synthesized by using standard techniques. The

reactions will be monitored by TLC. Purification of the compound will be done by

standard procedures like recrystallization.

B) Characterization of the compounds:

The synthesized compounds will be characterized by preliminary laboratory

techniques such as melting point, boiling point etc. Compounds synthesized will be

confirmed by FTIR, Mass Spectroscopy and NMR spectral data. The Mass and NMR

spectral data of the synthesized compound will be collected by sending the compounds to

research centers like IISc, Bangalore.

C) Antimicrobial evaluation:

C-1) In vitro evaluation of antibacterial activity.17

The MIC determination of the synthesized compounds will be carried out in side-

by-side comparison with Ciprofloxacin and Norfloxacin against Gram-positive

(Staphylococcus aureus, Bacillus subtilis) and Gram-negative bacteria (Klebsiella

pneumoniae, Escherichia coli) by broth microdilution method. Serial dilutions of the test

compounds and reference drugs will be prepared in Mueller Hinton agar. Drugs (10 mg)

will be dissolved in dimethylsulfoxide (DMSO, 1 ml). Further progressive dilutions with

melted Mueller Hinton agar will be performed to obtain the required concentrations of 1,

2, 4, 8, 16, 31.25, 62.5, 125, 250 and 500 mg/ml. The tubes will be inoculated with

105cfu/ml (colony forming unit/ml) and incubated at 37 oC for 18 h. The MIC will be the

lowest concentration of the tested compound that yields no visible growth on the plate.

To ensure that the solvent will have no effect on the bacterial growth, a control will be

performed with the test medium supplemented with DMSO at the same dilutions as used

in the experiments.

C-2) In vitro evaluation of antifungal activity.23,24

The MIC determination of the synthesized compounds will be carried out in side-

by-side comparison with Fluconazole and Griseofulvin against Candida Albicans,

Candida neoformans, Aspergillus niger and Aspergillus flavus by broth microdilution

method. Serial dilutions of the test compounds and reference drugs will be prepared in

sabouraud dextrose agar broth. Drugs (10 mg) will be dissolved in dimethylsulfoxide

(DMSO, 1 ml). Further progressive dilutions with melted sabouraud dextrose agar broth

will be performed to obtain the required concentrations of 1, 2, 4, 8, 16, 31.25, 62.5, 125,

250 and 500 mg/ml. MIC values were read after 1 day for Candida species and Candida

neoformans, and 2 days for Aspergillus niger, Aspergillus flavus in 37 oC. The inoculum

sizes contained approximately 1105 cells/ml. The MIC will be the lowest concentration

of the tested compound that yields no visible growth on the plate. To ensure that the

solvent will have no effect on the fungal growth, a control will be performed with the test

medium supplemented with DMSO at the same dilutions as used in the experiments.

7.3 Does the study require any investigation or interventions to be conducted on

patients or other humans/animals? If so please describe briefly.

No.

7.4 Has ethical clearance been obtained from your institution in case of 7.3?

Not applicable

8.0

LIST OF REFERENCES:

1. http://www.3rd1000.com/chem301/chem302a.htm

2. http://prr.hec.gov.pk/Chapters/206-1.pdf

3. Alagwadi KR, Suresh S, Pattan SR, Pujar GV, Javali MC. Synthesis and

antimicrobial evaluation of some 2-substituted oxadiazoles. Indian J heterocycl

Chem 2007;17:93-94.

4. Almasirad A, Vousooghi N, Tabatabai SA, Kebriaeezadeh A, Shafiee A.

Synthesis anticonvulsant and muscle relaxant activities of substituted 1,3,4-

oxadiazole, 1,3,4-thiadiazole and 1,2,4-triazole. Acta Chim slov 2007;54:317-24.

5. Kim RM, Rouse EA, Chapman KT, Schleif WA, Olsen DB, Stahlhut M. et al.

P1’ oxadiazole protease inhibitors with excellent activity against native and

protease inhibitors–resistant HIV-1. Bioorg Med Chem Lett 2004;14:4651-54.

6. Vagadevi HM, Vaidya VP. Studies in naphthofurans: Part III- Synthesis of 2-

substituted naphtho [2,1-b] furans, 2-(2’-aryl-3’-acetyl-1’,3’,4’-oxadiazolyl)

aminonaphtho [2,1-b] furans and their biological activities. Indian J Heterocycl

Chem 2001;10:253-60.

7. Tyrkov AG, Tyurenkov IN, Timchenko MV, Perfilova VN. Hypertensive

activity of 3-aryl-5-nitromethyl-1,2,4-oxadiazoles and their alkyl substituted

derivatives. Pharm Chem J 2006;40:240-42.

8. Manjunath SY, Biradar JS, Raga B. Synthesis and anthelmintic activity of

triheterocycles: [5’-(5”-substituted-3”-phenylindol-2”-yl)-1’,3’,4’,-oxadiazol-2’-

yl-thiomethyl] benzimidazoles. Indian J Heterocycl Chem 2009;18:321-24.

9. Parikh KP, Marvaniya HM, Sen DJ, synthesis and biological evaluation of

1,3,4-oxadiazole derivatives as potential antibacterial and antifungal agents.

International J of Drug Development & Research 2011;3(2):248-255.

10. Islam M, siddiqui AA, Ramadoss R, Bhakt A, Goyal S, synthesis and

antimicrobial activity of some novel oxadiazole derivatives. Acta Poloniae

Pharmaceutica n Drug Research 2008;65(4):441-447.

11. Kaplancikli ZA, Synthesis of Some Oxadiazole Derivatives as New Anticandidal Agents. Molecules 2011;16:7662-7671.

12. Pattan SR, Rabara PA, Pattan JS, Bukitagar AA, Wakale VS, Musmade DS, Synthesis and evaluation of some novel substituted

1,3,4,Oxadiazole and pyrazole derivatives for anti-tuberculer activity. Indian J of

Chem 2009;48B:1453-1456.

13. Xu M, He J, He M, Han F, Chen X, Pan Z, Wang J, Synthesis and

Antifungal Activity of Novel Sulfone Derivatives Containing 1,3,4-Oxadiazole

Moieties. Molecules 2011;16: 9129-9141.14. Jiao QC, Zhu HL, Cheng K, Zheng QZ, Zhang XM, Synthesis, biological

evaluation, and molecular docking studies of 2-chloropyridine derivatives

possessing 1,3,4-oxadiazole moiety as potential antitumor agents. Bioorganic &

Medicinal Chemistry 2010;18:7836–7841.

15. Shaharyar M, Ali MA, Oxadiazole mannich bases: Synthesis and

Antimycobacterial activity. Bioorg. Med. Chem. Lett 2007.

16. Mayeker AN, Yathirajan HS, Narayana B, Sarojini BK, Suchetha N,

Synthesis and Antimicrobial Studies on New Substituted 1,3,4-Oxadiazole

Derivatives Bearing 6-Bromonaphthalene Moiety. Int. J of Chem 2010;2(1):38-

54.

17. Talath S, Gadad AK. Synthesis, antibacterial and antitubercular activities of

some 7-[  4-(5-amino-[1,3,4]thiadiazole-2-sulfonyl)-piperazin-1-yl] fluoroquino-

lonic derivatives. Eur J Med Chem 2006;41:918-24.

18. Chaudhari BR, Shinde DB, Shingare MS. Synthesis of some 1,4-benzothiazinyl

thiosemicarbazides, triazoles, oxadiazoles, thiadiazoles and their antitubercular

activity. Indian J Heterocycl Chem 1995;4:187-90.

19. Holla BS, Poorjary KN, Bhat KS, Mithun A, Poojary B. Synthesis and anticancer

activity studies on some 2-chloro-1,4-bis-(5-substituted-1,3,4-oxadiazol-2-

ylmethyleneoxy) phenylene derivatives. Indian J chem 2005;44B:1669-73.

20. Lankau HJ, Unverferth K, Grunwald C, Hartenhauer H, Heinecke K, Bernoster K

et al. New GABA-modulating 1,2,4-oxadiazole derivatives and their

anticonvulsant activity. Eur J Med Chem 2007;42:873-79.

21. Husain A, Alam MM, Zaman MS, Ahuja P. Synthesis and biological evaluation

of 2-[3-(4-methoxy phenyl)propan-3-one]-5-(substituted phenyl)-1,3,4-

oxadiazoles. Indian J Hetrocycl Chem 2008;17:265-66.

22. Rajak H, Gupta AK, Kharya MD, Mishra P. Synthesis and antimicrobial activity

of new 2,5-disubstituted 1,3,4-oxadiazoles. Indian J Hetrocycl Chem 2009;19:25-

23. Ryu CK, Park RE, Ma MY, Nho JH. Synthesis and antifungal activity of 6-

arylamino-phthalazine-5,8-diones and 6,7-bis(arylthio)-phthalazine-5,8-diones.

Bioorg Med Chem Lett 2007;17:2577–80.

24. Mcginnis MR, Rindali MG, Lorian EV, editor. Antibiotics in Laboratory

Medicine. 4th ed. Baltimore (USA): Williams and Wilkins; 1996. p. 176.

9. SIGNATURE OF THE STUDENT

10. REMARK OF THE GUIDEThe above mentioned information and literature has been extensively investigated,

verified and was found to be correct. The present study will be carried out under my

supervision and guidance.

11. 11.1 NAME AND DESIGNATION OF THE GUIDE

11.2 SIGNATURE

SRI. S. S. PUROHIT M. Pharm. , (Ph.D.), LecturerDEPT. OF PHARMA - CHEMISTRY,SET’s COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD- 580002.

11.3 NAME AND DESIGNATION OF CO-GUIDE

11.4 SIGNATURE

---------------------

11.5 HEAD OF THE DEPARTMENT

11.6 SIGNATURE

Dr. S. D. JOSHI M. Pharm, Ph.D.PROFESSOR AND HEAD DEPT. OF PHARMA - CHEMISTRY,SET’s COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD- 580002.

12. 12.1 REMARK OF THE PRINCIPAL

12.2 SIGNATURE

The above mentioned information is correct and I

recommend the same for approval.

Dr. V.H. KULKARNI M. Pharm, Ph.D. PROFESSOR & PRINCIPAL, SET’s COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD- 580002.