Shuai Wen, Yang Panpan, Xiao Huan, Zhu Yumeng, Bu Faqian, Wang Aoxue, Sun Qiu, Wang Guan, Ouyang Liang
State Key Laboratory of Biotherapy and Cancer Center, Innovation Center of Nursing Research, West China Hospital, Sichuan University, No. 17, Section 3, Renmin South Road, 610041, Chengdu, China.
Angew Chem Int Ed Engl. 2024 Dec 9;63(50):e202411037. doi: 10.1002/anie.202411037. Epub 2024 Oct 31.
c-Jun N-terminal kinases (JNKs) including JNK1/2/3 are key members of mitogen-activated protein kinase family. Wherein JNK3 is specifically expressed in brain and emerges as therapeutic target, especially for neurodegenerative diseases. However, developing JNK3 selective inhibitors as chemical probes to investigate its therapeutic potential in diseases remains challenging. Here, we adopted the covalent strategy for identifying JNK3-selective covalent inhibitor JC16I, with high inhibitory activity against JNK3. Despite targeting a conserved cysteine in the vicinity of ATP pocket in JNK family, JC16I exerted a greater than 160-fold selectivity for JNK3 over JNK1/2. Importantly, even at low concentration, JC16I showed enhanced and long-lasting inhibition against cellular JNK3. In addition, its alkyne-containing probe JC-P1 could label JNK3 in SH-SY5Y cell lysate and living cells, with good proteome-wide selectivity. JC16I selectively suppressed the abnormal activation of JNK3 signaling and sufficiently exhibited neuroprotective effect in Parkinson's diseases (PD) models. Overall, our findings highlight the potential of developing isoform-selective and cell-active JNK3 inhibitors by covalent drug design strategy targeting a conserved cysteine. This work not only provides a valuable chemical probe for JNK3-targeted investigations in vitro and in vivo but also opens new avenues for the treatment of PD.
包括JNK1/2/3在内的c-Jun氨基末端激酶（JNKs）是丝裂原活化蛋白激酶家族的关键成员。其中JNK3在大脑中特异性表达，并成为治疗靶点，尤其是针对神经退行性疾病。然而，开发JNK3选择性抑制剂作为化学探针来研究其在疾病中的治疗潜力仍然具有挑战性。在此，我们采用共价策略鉴定出JNK3选择性共价抑制剂JC16I，它对JNK3具有高抑制活性。尽管靶向JNK家族ATP口袋附近的一个保守半胱氨酸，但JC16I对JNK3的选择性比对JNK1/2高160倍以上。重要的是，即使在低浓度下，JC16I对细胞内JNK3也表现出增强且持久的抑制作用。此外，其含炔基的探针JC-P1可在SH-SY5Y细胞裂解物和活细胞中标记JNK3，具有良好的全蛋白质组选择性。JC16I在帕金森病（PD）模型中选择性抑制JNK3信号的异常激活，并充分显示出神经保护作用。总体而言，我们的研究结果突出了通过靶向保守半胱氨酸的共价药物设计策略开发亚型选择性和细胞活性JNK3抑制剂的潜力。这项工作不仅为JNK3靶向的体内外研究提供了有价值的化学探针，也为PD的治疗开辟了新途径。
