Coumarin-1,2,3-Triazole Conjugates as Molecular Scaffolds for the Selective Induction of ROS-Driven Apoptosis in Cancer Cells in the Development of Pyruvate Kinase M2 Inhibitors

The increasing incidence of cancer and the emergence of drug resistance underscore the urgent need for new therapeutic options. This study aimed to synthesize and evaluate new coumarin-triazole hybrids for their cytotoxic selectivity and safety profile, providing insights into their potential as anticancer scaffolds. Twelve novel coumarin-based compounds were obtained in good yields (40-80%) and evaluated through <i>in vitro</i>, <i>in silico</i>, and <i>in vivo</i> assays. Among them, compound <b>7f</b> displayed the highest antiproliferative potency and selectivity, with selectivity indices (SI) of 4.61 for B16F-10, 3.38 for HCT116, and 2.98 for 4T1, and an average SI of 2.18 across SCC-4, SCC-9, and SCC-25 cell lines. Mechanistic assays indicated that the cytotoxic effect of <b>7f</b> is associated with oxidative stress-induced apoptosis, as evidenced by elevated ROS levels and activation of caspases 3/7. <i>In vivo</i> toxicity assessment in C57BL/6 mice revealed no significant changes in body weight, food intake, or macroscopic organ alterations at doses of up to 400 mg/kg, indicating a favorable safety profile. Additionally, <i>in vitro</i> biochemical evaluation and molecular docking suggested that <b>7f</b> interacts with Pyruvate Kinase M2 (PKM2) and inhibits its glycolytic activity in a dose-dependent manner. Overall, the findings identify compound <b>7f</b> as a selective and low-toxicity coumarin hybrid that exerts cytotoxic effects through ROS-mediated apoptosis, representing a promising lead structure for future optimization in anticancer drug development.