Molecular hybridization-driven FAK inhibitors: N-2,4-diarylaminopyrimidine-3-sulfamoyl-benzamide derivatives with improved antitumor potency

Abstract

A series of novel 2,4-diarylaminopyrimidine derivatives incorporating a 3-sulfamoylbenzamide moiety were designed and synthesized as potent focal adhesion kinase (FAK) inhibitors. Three compounds (5f, 10b, and 10c) demonstrated FAK inhibitory activities comparable to the reference inhibitor TAE226. In cellular assays, most analogues exhibited significant antiproliferative effects against four FAK-overexpressing tumor cell lines (HCT116, HeLa, MDA-MB-231, and A375), with IC₅₀ values below 1 μM. Notably, compound 10c displayed superior potency, showing IC₅₀ values ranging from 0.08 to 0.31 μM across the tested cell lines, and exhibited approximately 2-fold enhanced activity over TAE226 in inhibiting HCT116, MDA-MB-231, and A375 cell proliferation. Further mechanistic studies revealed that 10c effectively suppressed colony formation, migration, and adhesion of HCT116 cells, while inducing apoptosis via ROS elevation. Molecular docking analysis suggested that the enhanced activity of 10c may arise from an additional hydrogen bond interaction with the Arg426 residue of FAK, leading to stronger binding affinity compared to TAE226. In vivo, 10c showed good oral bioavailability and significantly inhibited HCT116 xenograft tumor growth (TGI = 62.22 % at 50 mg/kg), outperforming TAE226 (50.98 %) without inducing weight loss or hepatorenal toxicity. These results highlight 10c as a promising FAK-targeted candidate worthy of further preclinical investigation.

Keywords: 2,4-Diarylaminopyrimidine; 3-Sulfamoylbenzamides; Anti-tumor activity; FAK inhibitors; Molecular hybridization.