Tuberculosis (TB) remains a major global health threat, causing significant morbidity and mortality. The emergence of drug-resistant strains of Mycobacterium tuberculosis has further highlighted the urgent need for novel therapeutic targets and treatments. Targeting aminoacyl-tRNA synthetases, essential enzymes in protein synthesis, has emerged as a promising strategy for developing new anti-TB drugs. This project aims to investigate the feasibility of using collision-induced affinity selection mass spectrometry (CIAS-MS) for the screening of tRNA synthetases as drug targets. To date, I have applied this approach to leucyl-tRNA synthetase (LeuRS). Among the 1500 compounds tested, 24 compounds were identified from CIAS-MS screening, and 10 compounds showed antimycobacterial activity, divided into 6 series. One of these compounds, when combined with Bedaquiline (BDQ), an ATP synthase inhibitor, demonstrated enhanced antibacterial activity, indicating a synergistic effect.