IDH作为三羧酸循环限速酶可通过突变促进肿瘤发生发展，正成为抗癌新靶点。申请人前期采用理论计算揭示了mIDH1/2抑制剂的作用模式和选择性机制，并应用于虚拟筛选，发现了迄今虚筛来源的活性最高的mIDH1抑制剂V-10。此外，我们首次发现Asp312是影响mIDH2抑制剂选择性的关键残基，据此设计并获得了高选择性的A-61，它能有效逆转mIDH2介导的致癌代谢产物2-HG的生成及组蛋白高甲基化，诱导癌细胞分化并抑制其增殖。而且我们还基于R140Q变构口袋附近的Cys308设计并得到了mIDH2共价抑制剂，这一全新的作用方式未见文献报道。本项目拟以V-10和A-61为先导化合物开展结构优化及逆转组蛋白高甲基化与抗癌作用研究，以期发现活性强、选择性高、且成药性好的mIDH1/2抑制剂供临床前研究。此外，本项目还将设计并合成高活性和选择性的mIDH2共价抑制剂，以探索其用于肿瘤代谢治疗的可行性。

IDH, a rate-limiting enzyme in tricarboxylic acid cycle, could result in tumorgenesis through mutation and it has been recognized as a new target for developing antitumor agents. In previous studies, we had investigated the mechanism of action and the selectivity of mIDH1/2 inhibitors by theoretical calculations and then applied to subsequent virtual screening, leading to identify the hit V-10 (IC50:0.69μM), which showed the best activity derived from virtual screening. For the first time, we had discovered Asp312 is an important residue which decides the selectivity of mIDH2 inhibitors. And according to this, we designed and discovered A-61, a compound with high selectivity (SI>283), which could efficiently reverse the production of 2-HG (IC50: 9.1nM) and the hypermethylation of histone resulting from mIDH2 and induce the differentiation of cancer cells. In addition, based on the residue Cys308 in the allosteric pocket of IDH2 R140Q, we had also found the covalent inhibitors of mIDH2 that had never been reported before. In this proposal, we will use the previously identified V-10 and A-61 as our lead compounds to discover novel mIDH2 inhibitors with high inhibitory activity and high selectivity as well as good druggability for preclinical research, which can effectively reverse mIDH2-induced histone hypermethylation. What is more, we hope to develop covalent agents of mIDH2 with high activity and investigate their application in cancer metabolism.