ZAP-70选择性共价抑制剂及降解剂的设计合成和抗炎活性研究

ZAP-70激酶在T细胞的发展和免疫信号通路中发挥重要作用，是治疗自身免疫性疾病的潜在靶点。目前报道的ZAP-70小分子抑制剂存在选择性低和活性弱等问题。本项目以ZAP-70为靶标，采用基于结构的药物设计方法，通过共价抑制策略和蛋白水解靶向嵌合体技术，设计和合成ZAP-70激酶域的首个靶向共价抑制剂和蛋白降解剂，研究它们的体内外抗炎活性。前期工作中我们设计了一类咪唑并嘧啶类化合物，初步优化得到了苗头化合物Z13，它对ZAP-70表现出了较强的激酶抑制活性（IC50 = 55 nM），对同家族Syk和其它激酶显示出了高选择性，并能够抑制T细胞的增殖、分化和活化。质谱实验确证了Z13与ZAP-70的C346形成共价键。本项目将基于苗头化合物进行系统的构效关系研究和开发蛋白降解剂，为设计选择性和高活性的ZAP-70抑制剂提供新的思路，为进一步开发治疗自身免疫性疾病药物提供重要的先导化合物。

ZAP-70 kinase plays critical roles in the development and immune signaling of T cell. It has been reported as a potential target for autoimmune diseases. The ZAP-70 inhibitors described to date are either nonselective or very weak. We report herein the structure-guided development of the first-in-class potent targeted covalent inhibitor and PROTAC protein degrader of ZAP-70, and evaluation of their anti-inflammatory activity in vitro and in vivo. In previous work, we have developed a series of imidazo[1,2-c]pyrimidine analogues targeting ZAP-70. Primary optimization of these compounds produced a hit compound Z13, displaying potent inhibitory activity to ZAP-70 (IC50 = 55 nM), and good selectivity for ZAP-70 over structurally related Syk and other kinases. In addition, Z13 also showed potent inhibitory effects on T cell proliferation, division and activiation. A mass spectrometry analysis further confirmed the covalent linkage between the inhibitor and ZAP-70 protein at C346. We will systematically study the structure-activity relationship and develop the protein degrader of ZAP-70 based on our hit compound. This project will give a new perspective for developing selective and highly potent ZAP-70 inhibitors, and provided us with an important lead compound for further development for treatment of autoimmune diseases.