Department of Organic Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel.
Department of Immunology, The Weizmann Institute of Science, Rehovot 76100, Israel.
Cell Chem Biol. 2019 Jan 17;26(1):98-108.e5. doi: 10.1016/j.chembiol.2018.10.011. Epub 2018 Nov 15.
The c-Jun NH2-terminal kinase (JNK) signaling pathway is central to the cell response to stress, inflammatory signals, and toxins. While selective inhibitors are known for JNKs and for various upstream MAP3Ks, no selective inhibitor is reported for MKK7--one of two direct MAP2Ks that activate JNK. Here, using covalent virtual screening, we identify selective MKK7 covalent inhibitors. We optimized these compounds to low-micromolar inhibitors of JNK phosphorylation in cells. The crystal structure of a lead compound bound to MKK7 demonstrated that the binding mode was correctly predicted by docking. We asserted the selectivity of our inhibitors on a proteomic level and against a panel of 76 kinases, and validated an on-target effect using knockout cell lines. Lastly, we show that the inhibitors block activation of primary mouse B cells by lipopolysaccharide. These MKK7 tool compounds will enable better investigation of JNK signaling and may serve as starting points for therapeutics.
c-Jun NH2-末端激酶（JNK）信号通路是细胞对应激、炎症信号和毒素反应的核心。虽然已经有针对 JNK 和各种上游 MAP3K 的选择性抑制剂，但尚未有报道针对 MKK7（激活 JNK 的两个直接 MAP2K 之一）的选择性抑制剂。在这里，我们使用共价虚拟筛选，鉴定出了选择性的 MKK7 共价抑制剂。我们对这些化合物进行了优化，使其成为细胞中 JNK 磷酸化的低微摩尔抑制剂。与 MKK7 结合的一个先导化合物的晶体结构表明，对接正确地预测了结合模式。我们在蛋白质组水平上和针对 76 种激酶的面板上证明了我们抑制剂的选择性，并使用敲除细胞系验证了靶标效应。最后，我们表明这些抑制剂可阻断脂多糖激活原代小鼠 B 细胞。这些 MKK7 工具化合物将使 JNK 信号的研究更加深入，并可能成为治疗的起点。
