Discovering a New <i>N</i> ‐acylhydrazone Tethered Isatin/Benzofuran Scaffold for Colorectal Cancer: Design, Synthesis, Biological Activity, and ADME‐ <i>Tox</i> Modelling

ABSTRACT In this study, we designed and synthesized a novel series of hybrid molecules by linking isatin with the biologically active core of benzofuran through an N ‐acylhydrazone feature. The biological evaluation was conducted on colorectal adenocarcinoma cells (SW480) and nonmalignant colonic epithelial cells (NCM460) to assess cytotoxic and antiproliferative effects. Among the synthesized compounds, a brominated isatin–benzofuran derivative bearing a para‐fluorophenyl group exhibited significant cytotoxic and antiproliferative activity, with a better efficacy profile compared to the reference drug 5‐fluorouracil (5‐FU) and the corresponding parental molecules. Notably, this compound exhibited a slight selectivity toward cancer cells over non‐malignant cells and induced morphological changes, which are probably compatible with any type of programmed cell death. In addition, morphological and fluorescent changes, including nuclear condensation and reduced mitochondrial membrane potential, suggest that the hybrid may trigger mitochondrial‐dependent cell death. In silico ADME analysis predicted favorable biopharmaceutical profiles with no significant safety liabilities, supporting its potential as a lead compound for further development. These findings underscore the value of molecular hybridization as a strategy to enhance biological activity and support the potential of this hybrid as a lead candidate for further studies.