Artesunate induces tumor cell cycle arrest in the G0/G1 phase by targeting cyclin-dependent kinase 4 (CDK4)

Non-small cell lung cancer (NSCLC) remains the leading cause of cancer-related mortality worldwide, highlighting the urgent need for novel therapeutic targets. Cyclin-dependent kinase 4 (CDK4) plays a critical oncogenic role in NSCLC; however, current CDK4 inhibitors face limitations, including structural homogeneity and acquired resistance. Through HuProt™ human proteome microarray screening, we identified CDK4 as a direct cellular target of artesunate, which was validated by molecular docking, with a binding energy of -7.069 kcal/mol. Artesunate induced profound G0/G1 cell cycle arrest in both p53-wild (A549) and p53-deficient (H1299) NSCLC models via inhibition of the CDK4–Rb–E2F signaling axis, as demonstrated by the dose-dependent suppression of Rb phosphorylation at Ser780/Ser795. Collectively, these results establish artesunate as a structurally distinct CDK4 inhibitor scaffold, providing a strategic template for overcoming the limitations of aminopyrimidine-based kinase inhibitors.