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Discovery of Novel HPK1 Inhibitors via High-Throughput Virtual Screening, Biological Evaluation, and Molecular
Xinglong Chi1,2, Yiqun Chu1,3, Roufen Chen4
1Affiliated Yongkang First People's Hospital and School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, China.
Abstract:
Hematopoietic progenitor kinase 1 (HPK1) acts as a negative regulator of T-cell receptor signaling, making it a high-priority target for cancer immunotherapy. Addressing the scarcity of structurally novel HPK1 inhibitors, we constructed and validated a comprehensive high-throughput virtual screening (HTVS) workflow to screen the ChemDiv database. This hierarchical protocol, characterized by a hybrid docking strategy and MM/GBSA rescoring, identified 10 candidates for biological testing. Experimental evaluation confirmed compound 8 as a potent hit, exhibiting an IC50 of 1585 nM and 91.65% inhibition at 25 μM. Subsequent 500 ns molecular dynamics simulations validated the stability of the compound 8-HPK1 complex (ΔGbinding = -41.59 kcal/mol), driven by critical interactions with residues GLU92 and CYS94. These findings establish compound 8 as a promising scaffold for structural optimization and demonstrate the robustness of our computational framework for HPK1 inhibitor discovery.
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