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

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.