Screening of diphenylterephthalamide derivatives against Mycobacterium tuberculosis: an in silico and in vitro study.

The β‑carbonic anhydrase (CanB) enzyme has been validated as a potential target for the development of novel antitubercular compounds against Mycobacterium tuberculosis. This enzyme plays a crucial role in maintaining intracellular pH homeostasis, bacterial survival, and pathogenesis, making it a highly promising and druggable target for novel antitubercular therapies. Various inhibitors have been identified, including sulfonamides, dithiocarbamates, carboxylic acids, phenolic compounds, and diphenylterephthalamide derivatives. In this study, we employed a combination of in silico techniques and in vitro biological evaluation to further develop CanB inhibitors belonging to the N,N'-diphenylterephthalamide class. To this end, the structural similarity approach was applied to mine the National Cancer Institute database and prioritize available compounds. Biological evaluation demonstrated that twelve of these compounds exhibited potent antimycobacterial activity with MIC values ranging from 0.125 to 2 μg/mL. Furthermore, selected compounds were efficient inhibitors of CanB, reducing its esterase activity to less than 35%, and showed high affinity for the enzyme (Kd between 0.12 and 3.10 μM), as demonstrated by a thermal shift assay. Overall, this work clearly suggests that the diphenylterephthalamide scaffold represents an effective tool for developing CanB inhibitors with antimycobacterial properties.