A novel series of 2-mercaptobenzimidazole-based 3-cyanopyridin-2-one hybrids (7a–l) was designed and synthesized
as potential dual inhibitors of EGFR and BRAFV600E kinases. The antiproliferative activity was evaluated
against four human cancer cell lines (MCF-7, A-549, HT-29, and Panc-1) and a normal epithelial cell line (MCF-
10A). Compound 7c exhibited the highest potency (GI₅₀ = 6 μM) and was comparably effective to the reference
doxorubicin (GI50 = 5 μM) against the four cancer cell lines analyzed. Compound 7c also showed potent EGFR
(IC₅₀ = 0.12 μM) and BRAFV600E (IC₅₀ = 0.05 μM) inhibition, induced apoptosis via caspase-3/8/9 activation,
modulated Bax/Bcl-2 expression, and demonstrated strong antioxidant activity. Molecular docking, molecular
dynamics (MD) simulations, and DFT analyses revealed key pharmacophoric interactions within the ATP-binding
sites of EGFR and BRAFV600E kinases, while in silico ADME predictions supported drug-likeness and showed no
PAINS/Brenk structural alerts; however, these predictions do not establish safety, and experimental toxicology
will be required. These findings identify compound 7c as a promising dual-target lead candidate for further
optimization in anticancer drug development.