In silico virtual screening for the identification of novel inhibitors against dihydrodipicolinate reductase (DapB) of Mycobacterium tuberculosis , a key enzyme of diaminopimelate pathway.

Tuberculosis (TB) caused by () is one of the most devastating infectious diseases afflicting a large number of human lives. In spite of the availability of several drugs for treating TB, emergence of drug-resistant strains of the pathogen has made treatment and eradication of TB a challenging task. Hence, there is an imperative need for new intervention strategies to target . The enzymes of the diaminopimelate pathway, involved in lysine biosynthesis as well as in cell wall biosynthesis, have been considered as important anti-TB drug targets. One such enzyme is dihydrodipicolinate reductase (DapB), catalyzing the reduction of 2,3-dihydrodipicolinic acid to 2,3,4,5-tetrahydro-dipicolinic acid by utilizing NADH/NADPH as its cofactor. Here, we have generated an antisense knockdown mutant strain of and demonstrated a crucial role of in the and intracellular growth of . Further, virtual screening was performed by employing a library of ~95,000 compounds for the identification of inhibitory molecules against DapB. The molecules with high docking scores were screened against the enzymatic activity of DapB to determine their IC values. Further, hit molecules that inhibited DapB were screened against the growth in broth culture and inside macrophages. The molecules exhibiting inhibition were also evaluated for their cytotoxicity against various mammalian cell lines. In summary, we have identified a lead molecule, B59, with an IC value of 11 µg/mL and MIC value of 20 µg/mL, which can be further optimized to develop potent inhibitory compounds against DapB. IMPORTANCE Non-compliance to lengthy antituberculosis (TB) treatment regimen, associated side effects, and emergence of drug-resistant strains of () emphasize the need to develop more effective anti-TB drugs. Here, we have evaluated the role of dihydrodipicolinate reductase (DapB), a component of the diaminopimelate pathway, which is involved in the biosynthesis of both lysine and mycobacterial cell wall. We showed that DapB is essential for the as well as intracellular growth of . We further utilized DapB, as a target for identification of inhibitors by employing virtual screening, and conducted various screening assays to identify inhibitors with potential to inhibit DapB activity and and intracellular growth of with no significant cytotoxicity against various mammalian cell lines. Altogether, DapB serves as an important drug target and a hit molecule, namely, 4-(3-Phenylazoquinoxalin-2-yl) butanoic acid methyl ester has been identified as an antimycobacterial molecule in our study.