In-silico and In-vitro Studies on Targeting Tumor Apoptosis by Activating Caspase-3

Background: Despite the availability of numerous treatment options, cancer remains the primary cause of death in the current decade. The present study integrates computational and experimental methods to identify and develop a novel promising compound with anticancer properties. The study aims to evaluate the potential anticancer properties of selected molecules based on docking score, compliance with Lipinski’s rule of five, and in-silico safety prediction. Structure drug design by molecular docking methods was used to explore ligand conformations at various target binding sites. The in-silico physicochemical, pharmacokinetic, and toxicology properties of designed molecules were predicted using online software. Further, in-vitro, anticancer properties were studied by MTT Assay and flow cytometry. Results: 2-Butyl-3- (3, 5-diiodo-4- hydroxybenzoyl) benzofuran is one of the selected ligands showed potent anticancer activity against HT- 29 (Human Colon Cancer) cells and A549 (Human lung cancer) at IC50 concentrations. Further apoptosis studies demonstrated the possible mode of action against HT-29 cells. The proposed mechanism of action may be caspase-3 activation. As a consequence of caspase-3 activation, enzymes such as DNase are activated, leading to DNA fragmentation and apoptosis, and DNA fragmentation was 6.5 times greater than in untreated cells. The IC50 concentration of 2-Butyl-3- (3, 5-diiodo-4- hydroxybenzoyl) benzofuran inhibited 50% of HT-29 cells during G0/G1 phase. Conclusion: This research contributes to the burgeoning field of integrated in-vitro and in-silico analysis of biological systems, which can be used to study complicated cancer cell population dynamics. Future studies should investigate the impact on Human lung cancer cells.