Discovery, synthesis, and anti-cancer activity of N-CF₃-benzimidazoles

Benzimidazole is a privileged heterocyclic pharmacophore widely used in medicinal chemistry, while the N-trifluoromethyl (N-CF₃) moiety represents a valuable modification strategy to optimize the druggability of small molecules. However, systematic studies on the antitumor activity of N-CF₃-benzimidazoles remain scarce to date. In this study, the N-CF₃-substituted benzimidazole scaffold was identified as a core active antitumor motif through phenotypic screening of our in-house compound library. Leveraging our established synthetic methodology, a library of 39 structurally diverse N-CF₃-benzimidazole derivatives was prepared and systematically evaluated for their in vitro antiproliferative activity against three human cancer cell lines (786-O, A498, HepG2) using the CCK-8 assay. The results showed that most compounds exhibited varying degrees of antiproliferative activity at 10 µM. Notably, the hit compound 15 displayed micromolar GI₅₀ values and showed the most potent activity against 786-O cells. Furthermore, in silico target prediction of representative derivatives suggested potential interactions with a panel of kinases and epigenetic modulators, providing preliminary insights into their antitumor mechanism. This work characterizes the antitumor potential of N-CF₃-benzimidazoles, offering a critical underpinning for the development of novel antitumor drug candidates.