Nitrite-reductase (NIRB) as potential anti-tubercular target and a method to detect the severity of tuberculosis disease

摘要

The present invention discloses functional nitrite reductase as a potential drug target for anti-tubercular drug development. The present invention also relates to the development of an easy method for identification of nitrite in clinical samples as well as its correlation with the severity of the disease. Presence of active as well as dormant/latent stages of Mycobacterium tuberculosis (MTB) could be identified from nitrite in clinical samples like sputum of potential TB patients.

主权项

1. A method for screening inhibitors against nitrite reductase useful as anti-tubercular drug comprising:
i. infecting Thp1 macrophage culture in wells of a plate with Mycobacterium tuberculosis; ii. washing the infected macrophage culture with 1×PBS solution followed by adding fresh medium containing 50 mM of sodium nitrate; iii. adding 2.5 μl of a test inhibitor(s) against nitrite reductase in DMSO to the infected macrophage culture in a well of the plate on 0 d; iv. adding 2.5 μl of one or more standard inhibitors of narG, nirB and glnA1 at their respective IC90 to the infected macrophage culture in separate wells of the plate on 5 d and incubating the plate for nearly 8 d; v. estimating the level of ammonia/nitrite in the culture medium for determining the viable cell count; and vi. determining reduction in the viable cell count by the test inhibitor(s) as compared to the standard inhibitor(s) to obtain an anti-tubercular drug target.