“SYNTHESIS OF SOME NEW DERIVATIVES OF NITROGEN HETEROCYCLES AND EVALUATION OF THEIR BIOLOGICAL ACTIVITIES”
M.PHARM DISSERTATION PROTOCOL
SUBMITTED TO
Rajiv Gandhi University of Health Sciences, Bangalore, Karnataka
By
MOHAMMED HAMZA MOHD AHMED
UNDER THE GUIDANCE OF
Dr. G. K. KAPSE
M.Pharm, Ph.D
Professor
POST GRADUATE DEPARTMENT OF PHARMACEUTICAL CHEMISTRY
LUQMAN COLLEGE OF PHARMACY, GULBARGA-585102.
2010-11
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA BANGALORE
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the candidate Andaddress / MOHAMMED HAMZA MOHD AHMED
Luqman college of pharmacy,
old jewargi road,
gulbarga-585102.
2. / Name of the institution / Luqman college of pharmacy,
old jewargi road,
gulbarga-585102.
3. / Course of study and subject / m.pharm
(Pharmaceutical Chemistry)
4. / Date of Admission of course / SEPTEMBER 2010
5. / TITLE OF THE TOPIC / “SYNTHESIS OF SOME NEW DERIVATIVES OF NITROGEN HETEROCYCLES AND EVALUATION OF THEIR BIOLOGICAL ACTIVITIES”
6. / Brief Resume of the intended work:
6.1 Need for the study:
The chemistry of N-bridged heterocycles derived from 1,2,4-triazole has received considerable attention in recent year due to their usefulness in different areas of biological activities and as industrial intermediates.
1, 2, 4-triazole moiety appears frequently in the structure of various natural products1 and the synthesis of compounds incorporating this moiety has attracted widespread attention of chemists as well as biologists, mainly due to their diverse biological activities in pharmaceutical and agrochemical fields2. A large variety of 1,2,4-triazole derivatives possess antibacterial3 , antifungal4, antimycobacterial5, antiviral6, anti-inflammatory7, anticonvulsant8, antidepressant9, antitubercular10, antitumoral11, antihypertensive12, analgesic13, enzyme inhibitor14, hypoglycemic15, sedative, hypnotic16, antiparasitic, herbicidal, insecticidal and plant growth activities17. Thus several potent drugs possessing triazole nucleus have been applied in medicine like, alprazolam (anxiolytic agent, tranquilizer), anastrozole, letrozole, vorozole (antineoplastics, non-steroidal competitive aromatase inhibitors), estazolam (hypnotic, sedative, tranquilizer), etoperidone (antidepressant), fluconazole, itraconazole, terconazole (antifungl agents), ribavirine (antiviral agent), benatradin (diuretic), rilmazafon (hypnotic, anxiolytic, used in the case of neurotic insomnia), nefazodone (antidepressant, 5-HT2 A-antagonist), rizatriptan (antimigrain agent), trapidil (hypotensive), prazodone (antidepressant, anxiolytic, selectively inhibits central serotonine uptake) and triazolam (sedative and hypnotic)18.
Schiff bases are considered as a very important class of organic compounds which have wide applications in many biological aspects19. Many Schiff bases containing 1, 2, 4-triazole moiety exhibit antibacterial, antifungal20 and antitumoral activities21.
The substituted nicotinic acid is among the various heterocycles that have received most attention during last three decades as potential biomolecules. Nicotinic acid derivatives exhibit antibacterial, antioxidant, anti-inflammatory and anticarcinogenic activities. It is seen from the current literature that pyridine congeners are associated with different biological properties like pesticidal22-23, insecticidal24 and antifungal25 activities.
A large number of antibiotics contain amide linkage. Several derivatives of amides were prepared and found to possess antimicrobial activities. Literature survey reveals that various drugs e.g. penicillin26 (antibacterial), pyrazinamide27 (antitubercular), indinavir28, ritonavir29. (Protease inhibitors as anti-AIDS) etc contain their particular activities due to the amide linkage present in their structure.
In view of the above mentioned facts and in continuation of our interest in the synthesis of heterocycles containing 1,2,4-triazole moiety to identify new candidate that may be value in designing new, potent, selective and less toxic antimicrobial agents, we plan herein for the synthesis and antimicrobial evaluation of some novel structure hybrids incorporating the 1,2,4-triazole moiety with isonicotinic acid and other aryl moieties. This combination is thought in an attempt to investigate the influence of such hybridization and structure variation on the anticipated biological activities, hoping to add some synergistic biological significance to the target molecules.
6.2 Review of Literature:
Abundant references are available with regard to the study of 1,2,4-triazoles. The literature survey reveals that the compound containing 1,2,4-triazole are reported to possess diverse biological activities. Few important are below:
1. Nasser SA et al (2010)30 synthesized some novel 1,2,4-triazole fused acyclic and 21-28 membered macro cyclic compounds with potential antimicrobial activity.
2. Manikrao A.M et al (2009)31 accelerated synthesis of 1,2,4-triazoles derivative with antibacterial activity against Staphylococcus aureus, Klebsiella pneumonia, Escherichia coli and Pseudomonas aeurogenosa using cup-plate method.
3. Somani R.R et al (2009)32 synthesised some 1,2,4-triazole analogues for evaluation of their antifungal activity against C.albicans A. niger.
4. Neslihan D et al (2009)33 synthesized some new 1,2,4-triazoles and their Mannich and Schiff bases and screened for their antimicrobial activity against E. coli ATCC 25922, Y. pseudotuberculosis ATCC 921, Pseudomonas aeriginosa ATCC 27853, Enterococcus faecalis ATCC29212, S. aureus ATCC 25923, B. cereus 709 ROMA, C. tropicalis ATCC 13803 and C. albicans ATCC 60193.
5. Neslihan D et al (2010)34 synthesized some new 1,2,4-triazole derivatives and have shown the activity against E. Coli ATCC 35218, klebsiella pneumonia ATCC 13883, Yersinia pseudotuberculosis ATCC 911, Enterobacter aerogenes ATCC 13048, Pseudomonas aeurogenosa ATCC 10145, staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 29212, Bacillus cereus 709 ROMA, Candida tropicalis ATCC 13803, Candida glabrata 66032.
6. Jianxin FA et al (2006)35 synthesized some new 1,2,4-triazole derivatives and screened for their biological activity in vitro against Gibberella zeae, Alternatia solani.
7. Erhan P et al (2001)36 synthesized 1-acylthiosemicarbazides, 1,3,4-oxadiazoles, 1,3,4-thiadiazoles and 1,2,4-triazole-3-thiones has described activity as anti-inflammatory by carrageenan Paw Edema Test (CPE) method.
8. Neslihan D et al (2009)37 synthesized some 1,2,4-triazoles starting from isonicotinic acid hydrazide and evaluation of their antimicrobial activities against E. coli ATCC 25922, Y. pseudotuberculosis ATCC 911, P. aeruginosa ATCC 27853, Enterococcus aureus ATCC 25923, Bacillus cereus 709 ROMA, C.tropicalis ATCC 13803 and C. albicans 60193, by Agar-well diffusion meyhod.
9. Birsen T et al (2007)38 prepared 5-aryl-3-alkylthio-1,2,4-triazoles and sulfones with anti-inflammatory activity by Carrageenan-indused edema test in mice and also analgesic activity.
10. Nesrin GK et al (2007)39 synthesized 1-acylthiosemicarbazides, 1,2,4-triazole-5(4H)-thiones, 1,3,4-thiadiazoles and hydrazones containing 5-methyl-2-benzoxazolinones and evaluation for their analgesic activity by acetic acid-induced writhing test and hot plate test in mice and anti-inflammatory activity by carrageenan-induced hind paw oedema model in mice and also antimicrobial activity against Candida krusei, Candida albicans and Candida parapsilosis.
11. Goyal PK et al (2010)40 synthesized and characterized 4H-1,2,4-triazole derivatives which have shown analgesic activity with Eddy’s hot plate method.
6.3 Objective of Study:
It is well established that various derivatives of 1,2,4-triazole exhibit broad spectrum of pharmacological properties such as antibacterial and antifungal activities etc. Many of the available therapeutically important medicines such as ketoconazole, itraconazole, voriconazole and fluconazole contain this heterocyclic nucleus.
Prompted by these observations, we have planned to prepare some new 1,2,4-triazoles and evaluate for their biological activities.
The present investigation includes:
· Synthesis of various derivatives of substituted 1,2,4-triazoles following literature method.
· The chemical structure of the compounds synthesized could be established on the basis of elemental analysis and IR, NMR and Mass spectral studies.
· The compounds of the above type containing different heterocyclic moiety would be tested for antimicrobial activity against different strains of pathogenic organisms by following literature methods. Similarly few of the compounds would also be screened for anti-inflammatory and analgesic activities.
7. / Materials and Methods:
7.1 Method of collection of data:
1. From available literature.
2. From library based books
3. Web sites
- www.sciencedirect.com.
- http://jgate-helinet.informindia.co.in
- www.pubmed.com.
- www.scirus.com.
- www.herbmed.com.
7.2 Synthetic strategy:
Scheme
The various new compounds will be synthesized by conventional synthetic strategy and their yield, physical constant and analytical profile will be determined. All the reactions will be monitored by TLC techniques and chemical tests as applicable.
7.3 Biological screening:
1. Anti-microbial activity:-
All the compounds will be screened for antibacterial activity and antifungal activity using Cup & Plate method in our laboratory and the activities will be compared with the known standard anti-microbial agents 41-43.
2. Anti-inflammatory activity:-
Carrageenan-induced edema in Rats:-
The method of Kamal M. Dawood et al will be followed44
In the dose response experiment, albino rats are randomly assigned into various groups of 6 animal each depending upon number of test compounds as below:
a. Group-I Animals (Control).
b. Group-II Animals (standard drug) Ibuprofen 10mg/Kg
c. Each selected derivatives (test compounds) will be evaluated for anti-inflammatory activity at lower dose and higher dose, each dose of selected derivatives is treated as one group.
3. Analgesic activity:-
In present study the analgesia is assessed by employing tail-flick method using albino rats of either sex.46
a. Group-I Animals (Control).
b. Group-II Animals (standard drug) Ibuprofen 10mg/Kg
c. Each selected derivatives (test compounds) will be evaluated for anti-inflammatory activity at lower dose and higher dose, each dose of selected derivatives is treated as one group
7.4 Source of data:
The present investigation includes the synthesis of target molecules and the newly synthesized molecules will be characterized to generate the data as follows:
· The reactions will be monitored by TLC technique and Rf values will be recorded.
· Percentage of yield , reaction will be determined.
· The physical constant, solubility and elemental analysis will be carried out for newly synthesized compounds.
· IR, NMR and Mass spectral data will be obtained for newly synthesized compounds to elucidate their structures.
Biological Activity:
· Antimicrobial (antibacterial and antifungal) screening of compounds can be done by Cup and Plate method and diameter zone of inhibition in mm is recorded.
· Each compound is evaluated at 50µg/ml and 100µg/ml and the activity will be compared with that of the standard.
Pharmacological activities:
· Pharmacological screening for anti-inflammatory activity by rat paw edema method and the percent reduction of edema in mm will be recorded and the activity is compared with that of the standard drug.
· Pharmacological screening for analgesic activity using Eddy’s Hot-Plate method will be carried out and the activity is compared with that of the standard drug.
7.4-1 Assessment of toxic effect:
As per OECD-425 guidelines.
7.4-2 Screening of Statistical analysis:
Data are expressed as mean ± S.E.M. Results of in vivo tests were compared by
ANOVA followed by Dunnett’s ‘t’ test.
7.5 Does the study require any investigations or interventions to be conducted on patients or humans or animals? If so, please describe briefly.
Albino rats and mice of Wistar strain will be used for the evaluation of anti-inflammatory and analgesic activities.
7.6 Has ethical clearance been obtained from your institution in case of 7.3?
The present study is approved from Institutional Animal Ethics Committee (IAEC copy enclosed).
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