Integrated In Silico and In Vitro Study of Copper Nanocatalyzed Carbonyl-Functionalized Triazoles-Inducing S Phase Cell Cycle Arrest and Apoptosis in MCF-7.

The demand for novel, selective anticancer agents, driven by drug resistance and systemic toxicity of current treatments, underscores the importance of targeted drug discovery. Present research involved cytotoxic screening of a series of synthesized copper nanocatalyzed carbonyl-functionalized triazoles (3a-p), where 3i and 3j have shown highest selectivity index (SI) scores of 2.30 and 4.44, respectively. Computational validation of the lead compounds demonstrated specific interaction with BCL2-associated X protein (BAX) and BCL2, characterized by strong binding affinities ranging between -6.73 and -7.70 kcal/mol. Corresponding protein-ligand complexes demonstrated robust conformational stability throughout their 100 ns of molecular dynamics simulation. Subsequent in vitro validation using MCF-7 cells firmly corroborated the in silico findings, by revealing significant upregulation of BAX (p < 0.001) and downregulation of BCL2 (p < 0.001). Compound induced cellular stress, elevated the ROS-producing cell population up to 40%. Resulting cellular oxidative stress, rapidly depleted the glutathione reserves up to 50% (p < 0.001), consequently compromising the mitochondrial membrane potential leading to mitochondrial dysfunction. Furthermore, the compound induced S-phase cell cycle arrest (upto 51.5%), played a pivotal role in promoting apoptosis by activating DNA damage response pathways. In conclusion, this study has successfully identified two lead compounds (3i & 3j) that modulate multiple converging oncogenic pathways, providing compelling preclinical candidates for targeted management of breast cancer.