Synthesis and Evaluation of Novel Bis-Chalcone Derivatives Containing a Thiophene Moiety as Potential Anticancer Agents: In Vitro, In Silico , and Mechanistic Studies

Abstract

Chalcone compounds have demonstrated potent anticancer activities in the past few decades with few adverse consequences. Using Claisen-Schmidt condensation, two new series of bis-chalcone derivatives (<b>5a</b>-<b>c</b> and <b>9a</b>-<b>c</b>) bearing the thiophene moiety have been designed and generated. All compound structures were examined and elucidated by spectroscopic investigations. The MTT assay, gene expression assay, cell cycle analysis, apoptosis assay, Western blotting analysis, zymographic analysis, and molecular docking were used to evaluate the anticancer efficacy of the synthesized compounds against breast, colon, and lung cancer cells. Out of the two synthesized series, four compounds (<b>5a</b>, <b>5b</b>, <b>9a</b>, and <b>9b</b>) showed significant cytotoxic effects against breast (IC₅₀ values of 7.87 ± 2.54 and 4.05 ± 0.96 μM for compounds <b>5a</b> and <b>5b</b>, respectively), colon (IC₅₀ values of 18.10 ± 2.51 and 17.14 ± 0.66 μM for compounds <b>5a</b> and <b>9a</b>, respectively), and lung (IC₅₀ values of 41.99 ± 7.64 and 92.42 ± 30.91 μM for compounds <b>5a</b> and <b>9b</b>, respectively) cancer. These compounds upregulated the proapoptotic genes and caspase-3 and -9 protein, downregulated antiapoptotic and matrix metalloproteinase 2 (MMP-2) gene, and MMP-2 and -9 enzymatic activity. Additionally, these compounds significantly increased early and late apoptosis, necrosis, and induced cell cycle arrest at the subG1 phase with a concomitant decrease in the percentage of cell fractions at G0/G1, S, and G2/M phases. Moreover, the molecular docking was carried out on caspase-3 and -9 and MMP-2 and -9. The results suggest that compounds <b>5a</b>, <b>5b</b>, <b>9a</b>, and <b>9b</b> are potential and effective anticancer drugs.