Tuberculosis remains a significant global public health challenge. The relentless escalation of antimicrobial resistance necessitates urgent development of novel chemotherapeutic agents with clearly elucidated molecular targets, despite notable advancements in antimicrobial drug development over the past two decades. The present study systematically evaluates a series of three aroylhydrazones for their InhA inhibitory capacity. The compounds demonstrated high antymycobacterial activity against Mycobacterium tuberculosis H37Rv, with MIC values ranging from 0.0730 to 0.4412 µM, favorable cytotoxicity profiles (HEK-293T IC50: 217.5–361 µM), and a high selectivity index (SI: 633.49–3516). Molecular docking analyses revealed stable interactions between the derivatives and two crystallographic structures of InhA, prompting detailed enzymatic characterization. Spectrophotometric analysis of NADH-dependent InhA inhibition demonstrated concentration-dependent activity, with lead compounds achieving 48.6-59.1% enzyme suppression at micromolar concentrations. Comparative analysis with the reference inhibitor triclosan (62.5% inhibition) established the competitive potential of these novel aroylhydrazones. Quantitative structure--activity relationship analysis yielded half-maximal inhibitory concentrations ranging from 5.6 to 32.1 μM, confirming their pharmacological relevance. The current data will facilitate further research to enhance the therapeutic potential of these compounds and clarify their mechanisms of action.