Dichlorophthalic Anhydride Derivatives as Anti‐Apoptotic and Antiproliferative Agents by Multi‐Targeted Mechanism: Biological Evaluation and Molecular Docking Studies

ABSTRACT Twelve derivatives of 4,5‐dichlorophthalic anhydrides were subjected to in vitro evaluation to assess their potential as antiproliferative agents. Compounds 1, 7, 9 , and 11 exhibited noteworthy activity against the breast MCF‐7 cancer cell line as indicated by their respective IC 50 values. Flow cytometry techniques were utilized to analyze the distribution of cancer cells throughout the cell cycle, assess apoptosis, measure the activation of caspase‐3/7, and identify DNA fragmentation using fluorescence microscopy. The four compounds that were evaluated predominantly arrested MCF‐7 cells during the G0/G1 and/or S phases. Interestingly, compounds 7 and 9 exhibited the highest rates of apoptosis among the investigated substances, with percentages of 38.81% and 33.35%, respectively. The caspase assay results indicated that compounds 7 and 9 were the most effective inducers, with percentages of 26.08% and 23.17% respectively. The active compounds exhibit tubulin destabilization. Significant proportions of fragmented DNA (25.67%, 28.81%, and 32.56%) were identified by 7, 9 and 11 , respectively. Compound 7 showed mild inhibition against VEGFR‐2 compared to sorafenib, the standard inhibitor.