Mapping Therapeutic Networks and Molecular Interactions: Isatin Mannich Bases as Antitubercular leads with Integrated In vitro Screening

Abstract Tuberculosis (TB), caused primarily by Mycobacterium tuberculosis, remains a major global health concern and continues to necessitate the discovery of novel therapeutic agents with improved efficacy. In the present study, isatin was selected as a promising heterocyclic scaffold for the design and development of novel Mannich base derivatives targeting tuberculosis. An integrated computational and experimental approach was employed to investigate the therapeutic potential of the designed compounds. Network pharmacology analysis was initially performed to identify potential molecular targets, protein–protein interactions, and disease-associated biological pathways of the isatin derivatives. The predicted targets and enriched pathways provided insights into the possible molecular mechanisms underlying their antitubercular activity. Subsequently, molecular docking studies were carried out against selected tuberculosis-associated protein targets to evaluate the binding affinity and interaction patterns of the designed derivatives within the active sites. Based on the computational findings, the selected isatin mannich bases were synthesized and structurally characterized using UV-Visible, IR, 1H NMR, 13C NMR, and mass spectroscopic techniques. The synthesized compounds were further subjected to in vitro antitubercular evaluation to determine their biological activity. The computational investigations indicated favourable interactions of the designed derivatives with the selected therapeutic targets, while spectral characterization confirmed the proposed molecular structures. The in vitro findings further demonstrated the biological potential of the synthesized compounds. Overall, the integration of network pharmacology, molecular docking, synthetic chemistry, spectral characterization, and in vitro evaluation provides a rational strategy for the identification of novel isatin-based mannich derivatives as potential antitubercular agents. Keywords: Isatin, mannich bases, Tuberculosis, Network pharmacology, Spectral characterization.