<p>“DESIGN, SYNTHESIS OF SOME NEW PYRAZINE DERIVATIVES AND EVALUATION OF THEIR ANTIMICROBIAL AND POSSIBLE PHARMACOLOGICAL ACTIVITIES”</p><p>M.PHARM DISSERTATION PROTOCOL SUBMITTED TO Rajiv Gandhi University of Health Science Bangalore, Karnataka</p><p>UNDER THE GUIDANCE OF PROF.G. SUDHEENDRA M.Pharm,(PhD) Submitted by ABDUL NAZIR B.Pharm</p><p>DEPARTMENT OF PHARMACEUTICAL CHEMISTRY LUQMAN COLLEGE OF PHARMACY, GULBARGA-585102 2010-2011 RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA,</p><p>BANGALORE</p><p>ANNEXURE II</p><p>PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION</p><p>1. NAME OF THE CANDIDATE ABDUL NAZIR AND ADDRESS LUQMAN COLLEGE OF PHARMACY, OLD JEVERGY ROAD GULBARGA-585102</p><p>2. NAME OF THE INSTITUTION LUQMAN COLLEGE OF PHARMACY, OLD JEVERGY ROAD GULBARGA-585102</p><p>3. COURSE OF STUDY AND SUBJECT M.PHARM (PHARMACEUTICAL CHEMISTRY)</p><p>4. DATE OF ADMISSION OF COURSE 27-06- 2010</p><p>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:</p><p>6.1 Need for the study: Pyrazines are important six-membered heterocyclic compounds1. The 1,4-diazine</p><p> structural moiety can be found in compounds such as food flavours2, low band gap</p><p> conjugated materials3 and modern pyrazine phosphines4. The various condensed</p><p> heterocycles containing pyrazine ring system such as pyrazolo[3,4-b]pyrazine derivatives</p><p> have been reported to possess anticancer activity with low toxicity5-6,antiinflammatory7,</p><p> blood platelet aggregation inhibitors7, bone metabolism improvers8, adenosine</p><p> antagonists9-10 and controlling herbicides11. They also show antifungal and antiparasitic</p><p> activities12-13. In addition, they are used as disperse dyes14 and as fluorescents15.</p><p>Another class of nitrogen heterocycle that is piperazine derivatve have been</p><p> extensively investigated by the organic chemist due to their close association with various</p><p> types of biological activities. They have wide clinical application in the therapy of</p><p> functional diseases and exhibit anthelmintic, antibacterial and insecticidal activities16.</p><p>Combination of biologically active molecule into one molecule and synthesis of totally</p><p> newer moiety has been one of the method of research.</p><p>Based on the above mentioned facts we would like to extend our synthetic effort</p><p> towards the synthesis of new heterocycles containing piperazine and pyrazine moieties</p><p> that are suitabally substituted and form the compounds which are more potent and have</p><p> least undesirable side effects. 6.2 Objective of the study:</p><p>The increasing clinical importance of drug resistant bacterial and fungal pathogens has</p><p> lent additional urgency to antimicrobial research and development of new antibacterial,</p><p> antifungal compounds. Hence the necessary data will be generated by laboratory</p><p> experiment techniques which includes: </p><p> o Synthesis of novel pyrazine derivatives.</p><p> o Elemental analysis of synthesized compounds.</p><p> o To carry out the antimicrobial and biological activity of the synthesized compounds.</p><p>6.3 Review of Literature:</p><p> 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.</p><p> Mostafa H et al (2008)18 have reported the synthesis of some cyclooctane-based pyrazines and quinoxalines using cis,cis -1,5-cyclooctadiene.</p><p> Talaat I et al (2006)19 synthesized new derivatives of pyrazolo[3,4-b]pyrazines</p><p> and related heterocycles were prepared starting from 6-amino-3-methyl-1-phenyl-</p><p>1H-pyrazolo[3,4-b]pyrazine-5-carbonitrile and evaluated for antifungal and</p><p> antibacterial activities. These compounds were showed good anfungal and</p><p> antibacterial activities</p><p> Sadashiva CT et al (2006)20 the cholinergic hypothesis of Alzheimer’s disease (AD) has spurred the development of numerous structural classes of compounds</p><p> with different pharmacological profile aimed at increasing central cholinergic</p><p> neurotransmission. Thus proving a symptomatic treatment for this disease are</p><p> cholinomimetics with the pharmacological profile of acetyl cholinesterase (AchE)</p><p> inhibitors. The novel bioactive 1-[bis(4-fluorophenyl)-methyl]piperazine</p><p> derivatives were synthesized under mild conditions using different aryl/ alkyl</p><p> halides and heterocyclic alkyl halides with 1-[bis(4-fluorophenyl)-</p><p> methyl]piperazine in the presence of powdered potassium carbonate in N,N-</p><p> dimethylformamide. All the synthesized compounds were characterized by</p><p> spectroscopic techniques, elemental analysis and were screened for their efficacy</p><p> as AchE inhibitor. Some derivatives in this class showed good inhibition against</p><p>AchE as compared to neostigmine as standard.</p><p> Narendra SC et al (2006)21 a series of novel substituted 1-[bis(4-fluorophenyl)-</p><p> methyl]piperazine derivatives (4a–g) and (5h–m) have been synthesized. The</p><p> synthesized compounds were characterized by elimental analysis. All the</p><p> synthesized compounds were evaluated in vitro for their efficacy as antimicrobial</p><p> agents against representative strains of Gram-positive by paper disc diffusion and</p><p> micro dilution methods. Among the newly synthesized compounds.</p><p> Preeti C et al (2006)22 a series of substituted piperazine derivatives have been</p><p> synthesized and tested for antimicrobial activity. The antibacterial activity was</p><p> tested against Staphylococcus aureus, Pseudomonas aeruginosa, Streptomyces</p><p> epidermidis and Escherichia coli, and antifungal activity against Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger. All synthesized compounds</p><p> showed significant activity against bacterial strains but were found to be less</p><p> active against tested fungi. In vitro toxicity tests demonstrated that compounds 4d</p><p> and 6a showed very less toxicity against human erythrocytes.</p><p> Patel HS et al (2005)23 Novel N-substituted piperazine derivatives containing</p><p> sulfonyloxy aniline moiety have been prepared. The various 4-sulfonyloxy aniline</p><p>(SA) derivatives have been prepared by the condensation reaction of N-Acetyl</p><p>Sulfanilyl chloride (ASC) and sodium phenates followed by hydrolysis. The SA</p><p> derivatives are then reacted with chloro acetyl chloride to give corresponding (N-</p><p>Chloroacetyl) derivatives. These derivatives are then reacted with N-phenyl</p><p> piperazine to yield the corresponding piperazine derivatives.</p><p> Jean F et al (2000)24 synthesized the 2,6-Diamino-3,5-diaryl-1,4-pyrazine</p><p> derivatives as novel antioxidant.These compounds showed good antioxidant</p><p> activity.</p><p>7 Materials and Method</p><p>7.1 Method of collection of data: </p><p>The research work will be carried out in the laboratory by various experimental</p><p> technique includes:</p><p>1. The reactions will be monitored by TLC technique and Rf values data will be</p><p> generated.</p><p>2. Percentage of yield, melting point for each compound will be determined and recorded.</p><p>3. Synthesis of target molecules and their physical constant, solubility, elemental</p><p> analysis data will be done.</p><p>4. Spectroscopic data of new compounds i.e. I.R., NMR, Mass will be done for</p><p> structural confirmation of few synthesized compounds.</p><p>7.2 Source of data:</p><p> From available literature.  From library based books.  Web sites - www.pubmed.com</p><p>- www.google.com</p><p>- www.scirus.com</p><p> www.herbmed.com</p><p>7.3 Assessment of toxic effect25</p><p>As per OECD-425 SYNTHETIC STRATEGY:</p><p>N R R N NH + Cl N</p><p>N R N N N</p><p>R2-CHO</p><p>N R N N N N=CH-R2</p><p>SCHEME 7.4 Screening of Biological activity</p><p>The synthesized compounds are subjected to carry out antimicrobial and other </p><p> biological activities. </p><p>A) Antimicrobial activity26 and other biological activities. The antimicrobial activity of test compounds is carried out by cup-plate method</p><p> using bacterial (Stephalococcus aureus, Basillus Subtillus, Escherachia coil) as well</p><p> as fungal (Candida albicans, Aspragillus niger) organisms.</p><p>B) Analgesic activity27 In present study the analgesia is assessed by employing tail-flick method using albino</p><p> rats of either sex.</p><p>7.5 Does the study require any investigations or interventions to be conducted on </p><p>Patients or humans or animals? If so, please describe briefly</p><p>Yes Male and female rats will be used for the pharmacological investigation</p><p>7.6 Has ethical clearance been obtained from your institution in case of 7.3?</p><p>The present study is approved from institutional animal ethic committee (IAPC copy enclosed). 8. List of references:</p><p>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.</p><p>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.</p><p>25. New OECD 425 guideline for testing animals; 2001: 1/26, 1-6.</p><p>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.</p><p>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</p><p>11. NAME AND DESIGNATION OF PROF.G. SUDHEENDRA M.Pharm,(PhD) 11.1 GUIDE </p><p>11.2 SIGNATURE 11.3 CO-GUIDE (IF ANY) -</p><p>11.4 SIGNATURE - 11.5 HEAD OF DEPARTMENT PROF.G. SUDHEENDRA M.Pharm,(PhD)</p><p>11.6 SIGNATURE</p><p>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.</p><p>12.1 SIGNATURE OF THE </p><p>PRINCIPAL</p><p>PROF. SYED SANAULLAH M. Pharm, (PhD)</p>