Benzimidazole-Based Derivatives as Apoptotic Antiproliferative Agents: Design, Synthesis, Docking, and Mechanistic Studies

Abstract

A new class of benzimidazole-based derivatives (<b>4a</b>-<b>j</b>, <b>5</b>, and <b>6</b>) with potential dual inhibition of EGFR and BRAF^V600E has been developed. The newly synthesized compounds were submitted for testing for antiproliferative activity against the NCI-60 cell line. All newly synthesized compounds <b>4a</b>-<b>j</b>, <b>5</b>, and <b>6</b> were selected for testing against a panel of sixty cancer cell lines at a single concentration of 10 µM. Some compounds tested demonstrated remarkable antiproliferative activity against the cell lines tested. Compounds <b>4c</b>, <b>4e</b>, and <b>4g</b> were chosen for five-dose testing against 60 human tumor cell lines. Compound <b>4c</b> demonstrated strong selectivity against the leukemia subpanel, with a selectivity ratio of 5.96 at the GI₅₀ level. The most effective in vitro anti-cancer assay derivatives (<b>4c</b>, <b>4d</b>, <b>4e</b>, <b>4g</b>, and <b>4h</b>) were tested for EGFR and BRAF^V600E inhibition as potential targets for antiproliferative action. The results revealed that compounds <b>4c</b> and <b>4e</b> have significant antiproliferative activity as dual EGFR/BRAF^V600E inhibitors. Compounds <b>4c</b> and <b>4e</b> induced apoptosis by increasing caspase-3, caspase-8, and Bax levels while decreasing the anti-apoptotic Bcl2 protein. Moreover, molecular docking studies confirmed the potential of compounds <b>4c</b> and <b>4e</b> to act as dual EGFR/BRAF^V600E inhibitors.