Discovery of pyrimidine-based dual CDK2/TrkA inhibitors: Synthesis, antiproliferative activity, and mechanistic insights.

A series of newly synthesized pyrimidine analogues 12a-o was developed as dual CDK2/TrkA inhibitors. Synthesis of the target compounds was achieved via a condensation reaction of methyl trifluoroacetoacetate with thiourea, followed by alkylation with various benzyl chlorides. Subsequent chlorination with POCl3 and final nucleophilic substitution with the appropriate amines. The compounds were preliminarily screened for antiproliferative activity at a single concentration (10 μM) against National Cancer Institute (NCI) 60 cancer cell lines. The most active compounds 12b, 12g, 12k, and 12l were further investigated using MTT assay against MCF-7 (breast cancer), HCT-116 (colon cancer), and A-549 (lung cancer) cell lines, using Imatinib as a reference drug. Compound 12k was the most potent derivative (IC50 = 0.05, 0.02, and 0.04 μM, respectively) compared to imatinib (IC50 = 0.09, 0.06, and 0.12 μM, respectively). The therapeutic safety of compound 12k was assessed in WI-38 normal lung fibroblast cells demonstrating high safety margins. In the kinase selectivity profile, it showed moderate inhibition activity against TrkA and CDK2 kinases, with IC50 values of 6.81 and 10.13 μM, respectively. Moreover, cell cycle arrest was noticed at G2 and sub-G1 phases and induced apoptosis by 79.39%. Molecular docking studies of compound 12k demonstrated binding patterns compatible with inhibition for both targets, CDK2 and TrkA. Furthermore, its physicochemical properties and ADMET profile showed a good drug-like profile. Lastly, this study provides a significant contribution in developing novel effective dual-acting anticancer agents.