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

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 IC₅₀ 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.