ENCLOSURE I
6. BRIEF RESUME OF INTENDED WORK
6.1. Need for study
Research particularly Anti-tubercular drug discovery is a continuous process of many reasons like
- Tuberculosis is a leading cause of infectious disease mortality in the world.
- According to World Health Organization, one third of population is affected with latent Mycobacterium tuberculosis and approximately 3 million people die every year because of tuberculosis10.
- Tuberculosis is an infectious disease caused by mycobacterium tuberculosis bacillus. The disease is transmitted from one person to another by droplets dispersed in the air. When infected person coughs or sneezes these droplets nuclei released in air and inhaled by non infected person. Although tuberculosis primarily affects the lungs, the other organs may also be affected. For example if immunue system is poor, then infection may spread from lungs to other organs of the body. Organs that can be affected include liver, kidneys, spleen and uterus.17
- The unique structure of the cell allows it to be dormant as a latent infection. The slow growth rate of organism and prolonged treatment of the disease make TB difficult to eradicate.
- The treatment is complicated by a long term administration of few anti-tubercular agents. This intensifies drug side effects and by non compliance by patients often results in the development of multidrug resistant strains.
- There is now recognition that new drugs to treat TB are urgently required, specifically for use in shorter treatment regimens than are possible with the current agents and which can be employed to treat multidrug-resistantant and latent diseases.
ENCLOSURE- II
6.2. Review of literature:
One of the main approaches in developing potent drug molecule is based on the structural modification of naturally occurring drugs. Nitrogen containing heterocyclic system is indispensable structural unit for both the chemist and biochemist. Among the various classes of heterocyclic compounds triazoles form an important component of pharmacologically active compounds.
Ram Shankar Upadhayaya et al 1 ., synthesized a novel triazole and performed biological evaluation of synthesized triazole derivatives (1) against Mycobacterium tuberculosis.
Hacer bayrak et al 2., synthesized some new 1,2,4-triazoles and their Mannich and Schiff bases (2) and reported for their antimicrobial activities.
Ashraf M Abdel-Megeed et al 3., carried the synthesis and molecular modeling study of acylated 1,2,4-triazole-3-acetates (3) and reported their anti-inflammatory activity.
Mali RK et al 4., synthesized some antifungal and anti-tubercular 1,2,4-Triazole analogues (4).
Suvarna G Kini et al 5., synthesized, and reported anti-tubercular activity and docking study of novel cyclic azole substituted diphenyl ether derivatives (5).
Iraj Rahavi Ezabadi et al 7., synthesized sulfonamide-1,2,4-triazole derivatives (6) as antifungal and antibacterial agents.
Ilkay Kucukguzel et al 8., synthesized some novel thiourea derivatives obtained from 5-[(4-aminophenoxy)methyl]-4-alkyl/aryl-2,4-dihydro-3H-1,2,4-triazole-3-thiones (7) and evaluated as antiviral/anti-HIV and anti-tuberculosis activity.
Mahendra Shiradkar et al 9., reported the synthesis of clubbed triazoles (8) and evaluated them for anti-tubercular activity.
Elena Banfi et al 11., synthesized and carried antifungal and antibacterial activity of new imidazole and triazole derivatives (9).
Naseer SAM Khalil et al 12., synthesized and reported antimicrobial activity of some novel N- and S-β-D-glucosides of 5-pyridin-3-yl-1,2,4-triazoles (10).
Bahaa G Mohamed et al 13., synthesized 1-acyl-2-alkylthio-1,2,4-triazolobenzimidazoles (11) and reported them for antifungal , anti-inflammatory and analgesic effects.
Jolanta Kutkowaska et al 14., synthesized and reported antimicrobial activity of some new 3,4-disubstituted-1,2,4-triazole derivatives (12).
Neslihan Demibras et al 15., synthesized some 1-(5-phenylamino-[1,3,4] thiadiazol-2-yl) methyl-5-oxo-[1,2,4] triazole and 1-(4-phenyl-5-thioxo-[1,2,4]triazol-3-yl) methyl-5-oxo-[1,2,4]triazole derivatives (13) and reported their antimicrobial activity.
Vera Klimesova et al 16., synthesized 1,2,4-triazole 3-benzylsulfanyl derivatives (14) and reported their antimycobacterial activity.
In view of this we are planning to synthesize a series of Triazole derivatives of biological interest under the following stages.
1) Synthesis of the titled compounds by adopting different chemical synthetic methodologies.
2) Chemical characterization of synthesized novel substituted Triazoles by different physical and chemical analytical methods like Chomatography, Spectroscopic methods.etc.
3) Evaluation of all the synthesized compounds for their anti-tubercular activity.
ENCLOSURE- III
6.3. Objective of Study:
Many of the drugs available in the market for the treatment of microbial infections and many nitrogen-containing heterocyclic compounds have been reported to posse’s relatively high toxicity compared to other heterocyclic derivatives.19
Based on the literature survey we planned to synthesize some novel Triazoles, which are less toxic and more potent anti-tubercular agents.
Hence we planned to synthesize some novel Triazoles and carryout their anti-tubercular activity.
ENCLOSURE - IV
7. MATERIALS AND METHODS
7.1. Source of data:
The survey of literature on the area of present investigation will be done by referring to chemical abstract, all the national and international journals pertaining to synthetic, medicinal and pharmaceutical chemistry.
The information about Pharmacological and biological activities will be collected by referring to Indian Journal of Pharmacology, Indian Journal of Physiology and Pharmacology, European Journal of Pharmacology, Annals of surgery, Gut, Journals of Pharmacological experiment and therapeutics etc. for this purpose, the library facilities available in our College, Gulbarga University, Gulbarga, Indian Institute of Science, Bangalore and Indian Institute of Chemical technology, Hyderabad will be made use.
The day-to-day development in the area will be update by literature survey through e- publishing, Internet and current periodicals in our library and elsewhere.
All the basic facilities required for the synthesis of proposed compounds are available in our College laboratories. For purification of the products TLC, and HPLC and other facilities such as vaccum pump, rotary evaporators are also available in our College laboratory.
ENCLOSURE-V
7.2 . Method of Collection of data:
The chemical structures of the synthesized compounds are established on the basis of physical, chemical, analytical data, and physical constant like melting point, boiling point etc.
UV, IR, NMR and MASS spectral data is used for the characterization of the synthesized compounds by sending the sample to other advanced research center like IICT-Hyderabad, IIT-Chennai and IISc, Bangalore.
The main objective of the present investigation is to explore newer molecules with potent anti-tubercular activity. Anti-tubercular activity can be measured by Agar micro-dilution method.6,18.
Anti-tubercular activity by micro-dilution method.
The susceptibility and determination of MIC of the synthesized compound can be done against M. tuberculosis H37Rv organism, were performed by agar micro-dilution method where serial two fold dilutions of each test compound were added into 7H11 agar and M. tuberculosis H37Rv used as a test organism. MIC is the concentration of the compound that completely inhibits the growth and colony forming ability of M. tuberculosis.
Procedure
In 24 well plates, 3ml middle brook 7H11 agar medium with OADC supplement is dispensed in each well. The test compound added to the middle brook medium agar before in duplicate. So that the final concentration in each well is 25, 12.5, 6.25, 3.125, 1.56 µg/mL respectively. The known CFU of H37Rv culture was dispensed on the top of agar in each well in negative pressure biosafety hood. The plates are then incubated at 37 0 C / 5 % CO2 incubator. The concentration at which complete inhibition of colonies was observed was taken as MIC of test drug.
ENCLOSURE-VI
8. REFERENCES
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2. Bayrak H, Demirbas A, Karaoglu SA, Neslihan D. Synthesis of some new 1,2,4-triazoles their Mannich and Schiff bases and evaluation of their antimicrobial activities. Eur J Med Chem 2009; 44: 1057-1066.
3. Abdel-Megeed AM, Abdel-Rahman HM, Gamal-Eldien SA, El-Gendy MA. Design synthesis and molecular modeling study of acylated 1,2,4-triazole-3-acetates with potential anti-inflammatory activity. Eur J Med Chem 2009; 44: 117-123.
4. Mali RK, Somani RR, Toraskar MP, Mali KK, Naik PP, Shirodkar PY. Synthesis of some Antifungal and Anti-tubercular 1,2,4-Triazole Analogues. International J Chem Tech Research 2009; Vol 1(2): 168-173.
5. Kini SG, Bhat AR, Bryant B, Williamson JS, Dayan FE. Synthesis antitubercular activity and docking study of novel cyclic azole substituted diphenyl ether derivatives. Eur J Med Chem 2009; 44: 492-500.
6. Panda G, Parai MK, Srivastav AK, Chaturvedi V, Manju YK, Sinha S. Design synthesis and antitubercular activity of compounds containing aryl and heteroaryl groups with alkylaminoethyl chains. Ind J Chem 2009; 48: 1121-1127.
7. Ezabadi IR, Camoutsis C, Zoumpoulakis P, Geronikaki A, Sokovic M, Glamocilija J, Ciric A. Sulfonamide-1,2,4-triazole derivatives as antifungal and antibacterial agents: Synthesis biological evaluation lipophilicity and conformational studies. Bioorg and Med Chem 2008; 16: 1150-1161.
8. Kucukguzel I, Tatar E, Kucukguzel G, Rollas S, Clercq ED. Synthesis of some novel thiourea derivatives obtained from 5-[(4-aminophenoxy)methyl]-4-alkyl/aryl-2,4-dihydro-3h-1,2,4-triazole-3-thiones and evaluation as antiviral/anti-HIV and anti-tuberculosis agents. Eur J Med Chem 2008; 43: 381-392.
9. Shiradkar M, Suresh Kumar GV, Dasari V, Tatikonda S, Akula KC, Shah R. Clubbed triazole: A novel approach to antitubercular drugs. Eur J Med Chem 2007; 42: 807-816.
10. Satoskar RS, Bhandarkar SD, Rege NN. Pharmacology and Pharmacotherapeutics. 20th edtn. Popular Prakashan. Bombay. 2007: 734-750.
11. Banfi E, Scialino G, Zampieri D, Mamolo MG, Vio L, Ferrone M, Fermeglia M, Paneni MS, Pricl S. Antifungal and antimycobacterial activity of new imidazole and triazole derivatives a combined experimental and computational approach. J Antimicro Chemotherapy 2006; 58: 76-84.
12. Nasser SAM Khalil. Efficient synthesis structure and antimicrobial activity of some novel N- and S-β-D-glucosides of 5-pyridin-3-yl-1,2,4-triazoles. Carbohydrate Res 2006; 341: 2187-2199.
13. Mohamed BH, Abdel-alim M Abdel-Alim, Hussein MA. Synthesis of 1-acyl-2-alkylthio-1,2,4-triazolobenzimidazoles with antifungal, anti-inflammatory and analgesic effects. Acta pharm 2006; 56: 31-48.
14. Kutkowska J, Modzelewska-Banachiewicz B, Ziolkowska G, Rzeskt W, Urbanik-Sypniewska T, Zwolska Z, Prus M. Antimicrobial activity of 3,4-disubstituted-1,2,4-triazole derivatives. Acta Poloniae Pharmaceutica-Drug Research 2005; 62: 303-306.
15. Demibras N, Karaoglu SA, Demibras A, Sancak K. Synthesis and antimicrobial activities of some new 1-(4-phenylamino-[1,3,4]thiadiazol-2-yl)methyl-5-oxo-[1,2,4]triazole and 1-(4-phenyl-5-thioxo-[1,2,4]triazol-3-yl)methyl-5-oxo-[1,2,4]triazole derivatives. Eur J Med Chem 2004; 39: 793-804.
16. Klimesova V, Zahajska L, Waisser K, Kaustova J, Mollmann U. Synthesis and antimycobacterial activity of 1,2,4-triazole 3-benzylsulfanyl derivatives. IL Farmaco 2004; 59: 279-288.
17. Harsh Mohan. Text book of Pathology. 4th ed. Jaypee Brothers. 2003: 134-144.
18. Siddiqi S. Clinical Microbiology Handbook, ASM Press: Washigton D.C, 1992.I
19. Nolan CM, Chalhub EG, Nash DG and Yamauchi T. Antimicrob Agents Chemother. 1979: 171-175.