Design, synthesis, biological and computational evaluation of novel S/N-glycerolyl and peptide-conjugated [1,2,4]triazolo[1,5-a]pyrimidine derivatives as potent CDK2 inhibitors for anticancer therapy.

Targeting Cyclin-Dependent Kinase 2 (CDK2) remains a critical strategy in anticancer drug discovery. This study unveils a highly promising series of novel [1,2,4]triazolo[1,5-a]pyrimidine (TP) derivatives, achieved through innovative S/N-glycerolylation and peptide conjugation strategies. We report the rational design, efficient multi-step synthesis (yields up to 85 %), and comprehensive biological and computational evaluation. Notably, peptide conjugate 14c emerged as an exceptionally potent and selective lead, exhibiting outstanding anticancer activity against MCF-7 (IC = 0.7 μM), HCT-116 (IC = 1.1 μM), and MGC-803 (IC = 1.5 μM) cell lines, coupled with remarkable selectivity over non-cancerous HEK293 cells (SI = 22.4). Mechanistic validation confirmed 14c as a superior CDK2 inhibitor (IC = 0.21 μM, ∼2-fold more potent than roscovitine) with excellent kinase selectivity (S = 0.13). This potent inhibition effectively induced G1 cell cycle arrest (68.7 % at 0.7 μM), Rb dephosphorylation (75 % reduction), and significant apoptosis (38.5 %). Compound 14c demonstrated high metabolic stability in human liver microsomes (t½ = 68.5 min, CLint = 8.5 mL/min/kg), supporting its potential for favorable pharmacokinetics. Molecular docking and extensive 100 ns MD simulations revealed a stable, high-affinity binding mode (Kd ∼175-207 nM), elucidating the structural basis for its efficacy. These findings highlight peptide conjugate 14c as a breakthrough lead compound, demonstrating significant promise for development into a novel, selective CDK2- targeted anticancer therapeutic.