LUCIFERASE REPORTER MYCOBACTERIOPHAGES FOR EVALUATING NORBORNENE-BASED ANTITUBERCULOSIS DRUG SUSCEPTIBILITY TESTING ON MYCOBACTERIUM TUBERCULOSIS
Â Objective: In 2015, 9.6 million people around the world became sick with tuberculosis (TB) disease and 1.5 million TB-related deaths worldwide. Recent increasing incidence of multidrug-resistant (MDR; resistance to at least rifampicin (RIF) and isoniazid [INH]) and extensively drug-resistant (MDR resistance plus resistance to a fluoroquinolone and an aminoglycoside) makes TB a serious concern. Lot of research is needed to deal with this infectious disease for a better alternative in treatment or modification of these older TB drugs. The present study aimed at evaluating antimycobacterial activity of norbornene (NOR) derived INH copolymer with poly ethylene glycol (NOR- polyethylene glycol [PEG]-INH) a novel nanocarrier along with the anti-TB drug using luciferase reporter phages (LRPs).
Methods: NOR derived INH accounts for 74% of INH, 24% of NOR, and 2% of PEG. H37Rv control strain, a sensitive, and a resistant strain of Mycobacterium TB (MTB) used in this study. The in vitro activity of the drug was evaluated using absolute concentration method. The resistant strain was evaluated using LRP assay to observe the minimum inhibitory concentration of the drug.
Results: Reduction in light units was observed for the resistant strain exposed to plain INH and NOR-PEG-INH separately. 24% of reduction was observed in strains exposed to plain INH whereas 37% of reduction was observed in strains exposed to NOR-PEG-INH.
Conclusion: NOR-based INH had better antimycobacterial activity compared to plain INH and RIF. Antimycobacterial activity of INH and RIF increases even with very low dosage with NOR conjugate.
2. Dutta PK, Das JK. An overview of information, education and communication (IEC) activities in revised national tuberculosis control programme (RNTCP) and role of NGOs. Indian J Prev Soc Med 2012;43(4):366-71.
3. Amit D, Dharmeshkumar P. A profile of adverse effects of anti-tubercular drugs. GCSMC J Med Sci 2016;1:37-41.
4. Schaberg T, Rebhan K, Lode H. Risk factors for side-effects of isoniazid, rifampin and pyrazinamide in patients hospitalized for pulmonary tuberculosis. Eur Respir J 1996;9(10):2026-30.
5. Orme IM Development of new vaccines and drugs for TB: Limitations and potential strategic errors. Future Microbiol 2011;6(2):161-77.
6. Dye C, Espinal MA. Will tuberculosis become resistant to all antibiotics? Proc Biol Sci 2001;268(1462):45-52.
7. Sosnik A, Carcaboso AM, Glisoni RJ, Moretton MA, Chiappetta DA. New old challenges in tuberculosis: Potentially effective nanotechnologies in drug delivery. Adv Drug Deliv Rev 2010;62(4-5):547-59.
8. Ruckmani K, Kumaresan C. Sustained release aerosol for pulmonary drug delivery system: A review. Int J Pharm Pharm Sci 2013;5:126-30.
9. Pandey R, Khuller GK. Subcutaneous nanoparticle-based antitubercular chemotherapy in an experimental model. J Antimicrob Chemother 2004;54(1):266-8.
10. Mane SR, Dinda H, Sathyan A, Das Sarma J, Shunmugam R. Increased bioavailability of rifampicin from stimuli-responsive smart nano carrier. ACS Appl Mater Interfaces 2014;6(19):16895-902.
11. Vijayakameshwara RN, Shivshankar RM, Abhinoy K, Jayasri DS, Raja S. Norbornene derived doxorubicin copolymers as drug carrires with pH responsive hydrazone linker. Biomacromolecules 2012;13:221-30.
12. Shivshankar RM, Vijayakameswara RN, Raja S. Reversible pH and lipid sensitive vesicles from amphiphilic norbornene-derived thiobarbiturate homopolymers. ACS Macrolett 2011;1:482-8.
13. Shivshankar RM, Koushik C, Himari D, Jayasri DS, Raja S. Stimuli responsive nanocarrier for an effective delivery of multifrontline tuberculosis drugs. Polym Chem 2014;5:2725-35.
14. NIRT Manual. Standard Protocols for Mycobacteriology Laboratory, Version 1.1, November; 2010.
15. Azger D, Vanaja K, Narayana PR. Constructiona and evaluation of luciferase reporter phages for detection of active and non-replicating tubercle bacilli. J Microbiol Methods 2009;7(31):18-25.
16. Sarvamangala D, Nagasejitha P, Phrabu S, Srinivas L, Murthy US. Preparation and evaluation of isoniazid nano-conjugates for improving the therapeutic efficiency. Int J Pharm Sci Res 2015;6:739-45.
17. Jacob WR, Bartella RG, Udani R, Kalkut T, Sarkis GJ, Hatfull GF, et al. Rapid assessment of drug susceptibility testing of Mycobacterium tuberculosis by means of luciferase reporter phage. Science 1993;260:819-22.
18. Sarkis GJ, Jacobs WR Jr, Hatfull GF. L5 luciferase reporter mycobacteriophages: A sensitive tool for the detection and assay of live mycobacteria. Mol Microbiol 1995;15(6):1055-67.
19. Schlossberg D. Tuberculosis: A Sourcebook for Nursing Practice. New York: Springer Publications; 1995.
20. Banaiee N, Bobadilla V, Osornio JS, Luciferase reporter mycobacteriophages for detection, identification and antibiotic susceptibility testing of MTB in Mexico. J Clin Microbiol 2001;39:3883-8.
21. Suarez S, Oâ€™Hara P, Kazantseva M, Newcomer CE, Hopfer R, McMurray DN, et al. Airways delivery of rifampicin microparticles for the treatment of tuberculosis. J Antimicrob Chemother 2001;48(3):431-4.
22. Sharma A, Sharma S, Khuller GK. Lectin-functionalized poly (lactide-co-glycolide) nanoparticles as oral/aerosolized antitubercular drug carriers for treatment of tuberculosis. J Antimicrob Chemother 2004;54(4):761-6.