Discovery of novel Cytochrome bd oxidase inhibitors against Mycobacterium tuberculosis.

Drug-resistant Mycobacterium tuberculosis remains a major challenge to effective tuberculosis (TB) control. Cytochrome bd oxidase (Cyt-bd), a critical terminal oxidase within the electron transport chain of M. tuberculosis, maintains bacterial survival under respiratory stress but remains an underexplored therapeutic target. Here, we developed an integrated computational and experimental workflow to identify novel Cyt-bd inhibitors. Among the candidates, TB25, TB25-2, and TB25-14 exhibited substantial antimycobacterial efficacy, each reducing intracellular M. tuberculosis survival in macrophages by over 94 %. Importantly, co-treatment with Q203, a cytochrome bcc oxidase (Cyt-bcc) inhibitor, resulted in pronounced synergistic bactericidal effects. These findings highlight the potential of dual Cyt-bd/Cyt-bcc inhibition as a new strategy for treating drug-resistant and latent TB, and validate the effectiveness of our virtual screening pipeline for discovering new anti-TB agents.