Rajiv Gandhi University of Health Sciences, Karnataka s16

“DESIGN, SYNTHESIS OF SOME NEW PYRAZINE DERIVATIVES AND EVALUATION OF THEIR ANTIMICROBIAL AND POSSIBLE PHARMACOLOGICAL ACTIVITIES”

M.PHARM DISSERTATION PROTOCOL SUBMITTED TO Rajiv Gandhi University of Health Science Bangalore, Karnataka

UNDER THE GUIDANCE OF PROF.G. SUDHEENDRA M.Pharm,(PhD) Submitted by ABDUL NAZIR B.Pharm

DEPARTMENT OF PHARMACEUTICAL CHEMISTRY LUQMAN COLLEGE OF PHARMACY, GULBARGA-585102 2010-2011 RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA,

BANGALORE

ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. NAME OF THE CANDIDATE ABDUL NAZIR AND ADDRESS LUQMAN COLLEGE OF PHARMACY, OLD JEVERGY ROAD GULBARGA-585102

2. NAME OF THE INSTITUTION LUQMAN COLLEGE OF PHARMACY, OLD JEVERGY ROAD GULBARGA-585102

3. COURSE OF STUDY AND SUBJECT M.PHARM (PHARMACEUTICAL CHEMISTRY)

4. DATE OF ADMISSION OF COURSE 27-06- 2010

5. TITLE OF THE TOPIC: “DESIGN,SYNTHESIS OF SOME NEW PYRAZINE DERIVATIVES AND EVALUATION OF THEIR ANTIMICROBIAL AND POSSIBLE PHARMACOLOGICAL ACTIVITIES” 6. Brief Resume of the intended work:

6.1 Need for the study: Pyrazines are important six-membered heterocyclic compounds1. The 1,4-diazine

structural moiety can be found in compounds such as food flavours2, low band gap

conjugated materials3 and modern pyrazine phosphines4. The various condensed

heterocycles containing pyrazine ring system such as pyrazolo[3,4-b]pyrazine derivatives

have been reported to possess anticancer activity with low toxicity5-6,antiinflammatory7,

blood platelet aggregation inhibitors7, bone metabolism improvers8, adenosine

antagonists9-10 and controlling herbicides11. They also show antifungal and antiparasitic

activities12-13. In addition, they are used as disperse dyes14 and as fluorescents15.

Another class of nitrogen heterocycle that is piperazine derivatve have been

extensively investigated by the organic chemist due to their close association with various

types of biological activities. They have wide clinical application in the therapy of

functional diseases and exhibit anthelmintic, antibacterial and insecticidal activities16.

Combination of biologically active molecule into one molecule and synthesis of totally

newer moiety has been one of the method of research.

Based on the above mentioned facts we would like to extend our synthetic effort

towards the synthesis of new heterocycles containing piperazine and pyrazine moieties

that are suitabally substituted and form the compounds which are more potent and have

least undesirable side effects. 6.2 Objective of the study:

The increasing clinical importance of drug resistant bacterial and fungal pathogens has

lent additional urgency to antimicrobial research and development of new antibacterial,

antifungal compounds. Hence the necessary data will be generated by laboratory

experiment techniques which includes:

o Synthesis of novel pyrazine derivatives.

o Elemental analysis of synthesized compounds.

o To carry out the antimicrobial and biological activity of the synthesized compounds.

6.3 Review of Literature:

 Jiri K et al (2010)17 have reported solvent-free condensation reaction were carried out by heating terpene-monooximes with an excess of amines.Employing camphor- and nopinone-derived monooximes,two classes of terpene-fused pyrazines derivatives were synthesized.

 Mostafa H et al (2008)18 have reported the synthesis of some cyclooctane-based pyrazines and quinoxalines using cis,cis -1,5-cyclooctadiene.

 Talaat I et al (2006)19 synthesized new derivatives of pyrazolo[3,4-b]pyrazines

and related heterocycles were prepared starting from 6-amino-3-methyl-1-phenyl-

1H-pyrazolo[3,4-b]pyrazine-5-carbonitrile and evaluated for antifungal and

antibacterial activities. These compounds were showed good anfungal and

antibacterial activities

 Sadashiva CT et al (2006)20 the cholinergic hypothesis of Alzheimer’s disease (AD) has spurred the development of numerous structural classes of compounds

with different pharmacological profile aimed at increasing central cholinergic

neurotransmission. Thus proving a symptomatic treatment for this disease are

cholinomimetics with the pharmacological profile of acetyl cholinesterase (AchE)

inhibitors. The novel bioactive 1-[bis(4-fluorophenyl)-methyl]piperazine

derivatives were synthesized under mild conditions using different aryl/ alkyl

halides and heterocyclic alkyl halides with 1-[bis(4-fluorophenyl)-

methyl]piperazine in the presence of powdered potassium carbonate in N,N-

dimethylformamide. All the synthesized compounds were characterized by

spectroscopic techniques, elemental analysis and were screened for their efficacy

as AchE inhibitor. Some derivatives in this class showed good inhibition against

AchE as compared to neostigmine as standard.

 Narendra SC et al (2006)21 a series of novel substituted 1-[bis(4-fluorophenyl)-

methyl]piperazine derivatives (4a–g) and (5h–m) have been synthesized. The

synthesized compounds were characterized by elimental analysis. All the

synthesized compounds were evaluated in vitro for their efficacy as antimicrobial

agents against representative strains of Gram-positive by paper disc diffusion and

micro dilution methods. Among the newly synthesized compounds.

 Preeti C et al (2006)22 a series of substituted piperazine derivatives have been

synthesized and tested for antimicrobial activity. The antibacterial activity was

tested against Staphylococcus aureus, Pseudomonas aeruginosa, Streptomyces

epidermidis and Escherichia coli, and antifungal activity against Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger. All synthesized compounds

showed significant activity against bacterial strains but were found to be less

active against tested fungi. In vitro toxicity tests demonstrated that compounds 4d

and 6a showed very less toxicity against human erythrocytes.

 Patel HS et al (2005)23 Novel N-substituted piperazine derivatives containing

sulfonyloxy aniline moiety have been prepared. The various 4-sulfonyloxy aniline

(SA) derivatives have been prepared by the condensation reaction of N-Acetyl

Sulfanilyl chloride (ASC) and sodium phenates followed by hydrolysis. The SA

derivatives are then reacted with chloro acetyl chloride to give corresponding (N-

Chloroacetyl) derivatives. These derivatives are then reacted with N-phenyl

piperazine to yield the corresponding piperazine derivatives.

 Jean F et al (2000)24 synthesized the 2,6-Diamino-3,5-diaryl-1,4-pyrazine

derivatives as novel antioxidant.These compounds showed good antioxidant

activity.

7 Materials and Method

7.1 Method of collection of data:

The research work will be carried out in the laboratory by various experimental

technique includes:

1. The reactions will be monitored by TLC technique and Rf values data will be

generated.

2. Percentage of yield, melting point for each compound will be determined and recorded.

3. Synthesis of target molecules and their physical constant, solubility, elemental

analysis data will be done.

4. Spectroscopic data of new compounds i.e. I.R., NMR, Mass will be done for

structural confirmation of few synthesized compounds.

7.2 Source of data:

 From available literature.  From library based books.  Web sites - www.pubmed.com

- www.google.com

- www.scirus.com

www.herbmed.com

7.3 Assessment of toxic effect25

As per OECD-425 SYNTHETIC STRATEGY:

N R R N NH + Cl N

N R N N N

R2-CHO

N R N N N N=CH-R2

SCHEME 7.4 Screening of Biological activity

The synthesized compounds are subjected to carry out antimicrobial and other

biological activities.

A) Antimicrobial activity26 and other biological activities. The antimicrobial activity of test compounds is carried out by cup-plate method

using bacterial (Stephalococcus aureus, Basillus Subtillus, Escherachia coil) as well

as fungal (Candida albicans, Aspragillus niger) organisms.

B) Analgesic activity27 In present study the analgesia is assessed by employing tail-flick method using albino

rats of either sex.

7.5 Does the study require any investigations or interventions to be conducted on

Patients or humans or animals? If so, please describe briefly

Yes Male and female rats will be used for the pharmacological investigation

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

The present study is approved from institutional animal ethic committee (IAPC copy enclosed). 8. List of references:

1. Brown DJ. The pyrazines: Supplement 1: Wiley: New York, 2002. 2. Maga JA, Sizer. C.E.J. Agr. Food Chem. 1973;21:22. 3. Wen L, Nietfeld JP, Amb CM, Rasmussen SC. Journal of org. chem. 2008;78:8529. 4. (a) Imamoto T, Sugita K, Yoshida K. journal of Am. Chem. 2005;127:11934. (b) Das AK, Bulak E, Sarkar B, Lissner F, Schleid T, Niemeyer M, Feilder J, Kaim W. Organometallics. 2008;27:218. 5. Suzuki, S.; Inoue, A. Jpn. Kokai Tokkyo Koho JP 02 172 988 [90 172 988] (Cl. C07D487/04) (4 Jul.1990); Chem. Abstr. 1990, 113, 218276t. 6. Suzuki, S. S.; Inoue, A. Jpn. Kokai Tokkyo Koho JP 02 240 084 [90 240 084] (Cl. C07D487/04) (25 Sep.1990); Chem. Abstr. 1991, 114, 178382m. 7. Sado, T.; Inoue, A. Jpn Kokai Tokkyo Koho JP 02 101 078[90 101 078] (Cl.C07D487/04) (12 Apr. 1990), Chem. Abstr.1990, 113, 78422k. 8. Imaizumi, K.; Sado, T. Jpn Kokai Tokkyo Koho JP 06 80 570 [94 80 570] (Cl. A61K31/495) (22 Mar. 1994), Chem. Abstr. 1994, 121, 91797w. 9. Akahane, A.; Kuroda, S.; Itani, I.; Tabuchi, S.; Sato, Y.;Matsuoka, N.; Tada, M.; Matsuoka, H.; Oku, T.; Tanaka, A.(Fujisawa Pharmaceutical Co., Ltd Japan) A. PCT Int. Appl.(2001), 69 pp. CODEN: PIXXD2 WO 2001040230 Al20010607, Chem. Abstr. 2001, 135, 33489a. 10. Akahane, A.; Tanaka, A. (Fujisawa Pharmaceutical Co., Ltd Japan) A. PCT W: JP, Chem. Abstr. 2003, 138, 24732a. 11. Int. Appl. (2002), 34 pp. CODEN: PIXXD2 WO 2002100864 Al 20021219 Designated States; Takabe, F.; Shibayama, A.;Yamaguchi, M.; Yamaji, M.; Hanai, R.; Sadohara, H. Jpn Kokai Tokkyo Koho JP 09 59 276 [97 59 276] (Cl.C07D471/04) (4 Mar. 1997); Chem. Abstr. 1997, 126.277477a. 12. El-Emary, T. I.; El-Dean, A. M. Kamal; El-Kashef, H. S. Farmaco. 1998, 53, 383. 13. El-Kashef, H. S.; El-Emary, T. I.; Gasquet, M.; Timon-David, P.; Maldonado, J.; Vanelle, P. Pharmazie. 2000,55(8), 572.

14. Rangnekar, D. W.; Dhamnaskar, S. V. J. Chem. Technol.Biotechnol. 1990, 49, 311. 15. Hofmann, J.; Sicker, D.; Mann, G. Ger. (East) DD 276688 (Cl. C07D487/04) (7 Mar. 1990); Chem. Abstr. 1990, 113, 231409h. 16. Coyne WE. Medicinal chemistry, edited by Alfred burger (Wiley-Interscience, New York), 1970. 17. Jiri K, Miroslav L, Filip B. One-step and solvent free synthesis of terpine-fused pyrazines. ARKIVOC. 2010;ii:315-322. 18. Mostafa HA, Madeleine H, Mehdi MB, John AJ. Synthesis of some cyclooctane based pyrazines and quinoxalines. ARKIVOC. 2008;166-179. 19. Talaat I. Synthesis and biological activity of some new pyrazolo(3,4-b)pyrazines. Journal of Chinese Chem. Soc. 2006;53:391-401. 20. Sadashiva C, Narendra S, Ponnappa K, Veerabasappa G, Kanchugarakoppal S. Synthesis and efficacy of 1-[bis(4-fluorophenyl)-methyl]piperazine derivatives for acetylcholinesterase inhibition, as a stimulant of central cholinergic neurotransmission in Alzheimer’s disease. Bioorganic & Medicinal Chemistry 2006; 16: 3932-3936. 21. Narendra SC, Sadashiva CT, Kavitha CV, Rangappa KS. Synthesis and in vitro antimicrobial studies of medicinally important novel N-alkyl and N-sulfonyl derivatives of 1-[bis(4-fluorophenyl)-methyl]piperazine. Bioorganic & Medicinal Chemistry 2006; 14: 6621-6627. 22. Preeti C, Rupesh K, Akhilesh K, Devender S, Vibha Y, Anil K, Sharma G, Ramesh C. Synthesis and antimicrobial activity of N-alkyl and N-aryl piperazine derivatives. Bioorganic & Medicinal Chemistry 2006; 14: 1819-1826. 23. Patel HS, Desai HD and Mistry HJ. Synthesis and Antimicrobial Activity of Some New Piperazine Derivaties Containing Aryl Sulfonyloxy Group. E-Journal of Chemistry 2004; 1: 93-98. 24. Jean FC, Maggi B, Catherine DT, Frederique D, Cecile M, Jean FR, Jacqueline MB. 2,6-diamino-3,5-diaryl-1,4-pyrazine derivatives as novel antioxidant. George Thieme verlag Stuttgart New York. 2001;0039-7881.

25. New OECD 425 guideline for testing animals; 2001: 1/26, 1-6.

26. Shahrukh T, Nikul P, Keshav C. Novel 3-[4-(diethylamino)phenyl]-4-substituted- 1-ylsulfonyl) sydnones: synthesis, characterization and antimicrobial studies. Org. Commun 2010 ;( 3) 2: 30-38.

27. Kulkarni SK. Handbook of experimental pharmacology. 1st ed. Dehli Vallabha Prakashen; 1987:950,959-966. 9. SIGNATURE OF CANDIDATE (MR. ABDUL NAZIR) 10. REMARKS OF THE GUIDE The candidate is working under my direct supervision in laboratories of L.C.P Gulbarga-585102

11. NAME AND DESIGNATION OF PROF.G. SUDHEENDRA M.Pharm,(PhD) 11.1 GUIDE

11.2 SIGNATURE 11.3 CO-GUIDE (IF ANY) -

11.4 SIGNATURE - 11.5 HEAD OF DEPARTMENT PROF.G. SUDHEENDRA M.Pharm,(PhD)

11.6 SIGNATURE

12. REMARKS OF THE CHAIRMAN & We will provide all the necessary facilities required for the proposed research work. PRINCIPAL Hence, the plan of work is recommended for registration.

12.1 SIGNATURE OF THE

PRINCIPAL

PROF. SYED SANAULLAH M. Pharm, (PhD)