USP7多重变构调控机制和小分子抑制剂选择性识别机制的结构生物学研究

去泛素化蛋白酶7（USP7）在靶向p53的药物研发和肿瘤免疫治疗中发挥着关键作用。基于USP7的变构调控机制，开发高选择性的变构抑制剂一直是肿瘤靶向药物研发的热点，但仍面临以下问题：1）USP7变构调控机制精确而复杂。2）高通量筛选方案的局限性。3）高分辨率USP7全长-变构抑制剂复合体三维空间结构的缺乏。基于以上问题，本项目将首先解析不同激活状态下USP7全长及其复合体的晶体结构，揭示USP7多重变构调控机制的结构生物学基础。然后以USP7的变构调控机制为导向，优化建立精准的高通量筛选方案，结合计算机辅助的虚拟筛选和实体库筛选，寻找结构新颖的苗头化合物。通过解析USP7-小分子抑制剂的共晶结构，结合生物化学、生物物理学和细胞水平检测等手段，初步阐明小分子化合物的选择性识别机制。本项目将为进一步理性设计优化高选择性的USP7变构抑制剂提供思路和方向，具有重要的创新性和研究价值。

Ubiquitin specific protease 7 (USP7) plays a critical role in drug development targeting p53 and cancer immunotherapy. Based on the allosteric regulation mechanism of USP7, the development of highly selective allosteric inhibitors has been highly pursued in anticancer drug discovery, but it still encounters the following problems: 1) The multiple-layer allosteric regulation of USP7 is precise and complicated. 2) Limitation of high throughput screening method. 3) Lacking of high-resolution co-crystal structure of full-length USP7 with allosteric inhibitor complex. To solve these problems, we will first determine the crystal structures of USP7 under different activation states, and reveal the structural basis of USP7 multiple allosteric regulation mechanisms. Then, guided by the allosteric regulation mechanism of USP7, an accurate high-throughput screening method is established to identify new allosteric USP7 inhibitors combined with virtual screening and compound library screening. By solving the co-crystal structures of USP7 with small-molecule inhibitors, together with biochemistry, biophysics and cell-based assays, the selective recognition mechanism of inhibitors is preliminarily elucidated. This project will provide directions for further rational design and optimization of USP7 allosteric inhibitors with highly selective, and has important innovation and potential research value.