Mao Jie, Zhou Lixin, Wu Yuxing, Wang Kaizhen, Ye Xiuquan, Wang Tianyu, Yang Jiamei, Tong Jun, Miao Qi, Jiang Sheng, Xiao Yibei, Zhang Kuojun
Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China.
Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China; Department of Hematology, Tongji Hospital, Frontier Science Center for Stem Cell Research, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
Bioorg Chem. 2025 Mar;156:108158. doi: 10.1016/j.bioorg.2025.108158. Epub 2025 Jan 10.
Hematopoietic progenitor kinase 1 (HPK1), which negatively regulates immune signaling, has emerged as an attractive small-molecule drug target for tumor immunotherapy. Herein, we report the discovery of the 1,2,4-benzotriazine derivatives as new potent HPK1 inhibitors. Notably, compound A29 exhibited improved HPK1 inhibitory activity relative to compound 1 in the ADP-Glo kinase assay (IC = 2.70 and 13.6 nM, respectively). The pronounced inhibitory activity of A29 against downstream p-SLP76 in Jurkat T cells (IC = 8.1 nM) as well as the ability to induce the production of interleukin 2 (IL-2) in human peripheral blood mononuclear cells (PBMCs) confirmed its cellular target engagement and immune stimulatory effect. Consistently, this lead compound significantly enhanced T-cell killing ability against murine colon cancer cells CT26 or MC38 in a co-culture system. Furthermore, A29 was efficacious in a CT26 xenograft mouse model alone, and significantly enhanced the antitumor efficacy of an anti-PD-1 antibody. This work provides a promising lead for the development of effective HPK1 inhibitors for tumor immunotherapy.
造血祖细胞激酶1（HPK1）对免疫信号传导起负调节作用，已成为肿瘤免疫治疗中一个颇具吸引力的小分子药物靶点。在此，我们报告了1,2,4 - 苯并三嗪衍生物作为新型高效HPK1抑制剂的发现。值得注意的是，在ADP - Glo激酶测定中，化合物A29相对于化合物1表现出更高的HPK1抑制活性（IC分别为2.70和13.6 nM）。A29对Jurkat T细胞中下游p - SLP76具有显著的抑制活性（IC = 8.1 nM），以及在人外周血单核细胞（PBMC）中诱导白细胞介素2（IL - 2）产生的能力，证实了其细胞靶点结合和免疫刺激作用。同样，在共培养系统中，这种先导化合物显著增强了T细胞对小鼠结肠癌细胞CT26或MC38的杀伤能力。此外，A29单独在CT26异种移植小鼠模型中有效，并显著增强了抗PD - 1抗体的抗肿瘤疗效。这项工作为开发用于肿瘤免疫治疗的有效HPK1抑制剂提供了一个有前景的先导化合物。
