靶向Dyskerin-NOP10相互作用的特异性小分子：优化设计合成及抑制端粒酶活化

端粒酶在肿瘤细胞中特异性高表达，但该酶结构复杂，至今没有筛选出高选择性抑制剂。Dyskerin-NOP10-NHP2三聚体蛋白是端粒酶的核心组份，它在端粒的活化和组装中起关键作用。区别于靶标端粒酶全酶或某个片段蛋白的研究新思路，本申请拟靶标Dyskerin-NOP10蛋白-蛋白相互作用，进而影响Dyskerin复合体的形成，抑制端粒酶的活化。申请拟构建蛋白复合物的结构模型；揭示蛋白结合界面的热点狭缝区域及关键残基，结合NOP10识别底物Dyskerin的结构特点和预研的活性分子，优化设计合成多样性的化合物库。表达、纯化Dyskerin、NOP10蛋白；分析活性化合物与抑制Dyskerin-NOP10相互作用间的构效关系；用Pull down、ITC、MST等实验确认化合物调控Dyskerin-NOP10之间的相互作用；验证先导化合物靶向调控上述Dyskerin-NOP10复合物的形成。

Telomerase is highly expressed in tumor cells, but the structure of the enzyme is complex, so high selective inhibitors have not been screened until now. Dyskerin-NOP10-NHP2 trimer protein is the core component of telomerase, which plays a key role in the stability, activation and assembly of telomere. Targeted interaction of Dyskerin-NOP10 protein-protein is a idea of this study, which is different from target telomerase holoenzyme or a fragment protein, thereby affecting the formation of Dyskerin complex and inhibiting the activation of telomerase. With the help of recent advances in biophysics, we have constructed a structural model of protein complexes, revealing the slit region of hot spots and potential key residues for the interface of protein-protein binding. According to the structural characteristics of substrate Dyskerin identified by NOP10, combined with the active tool molecule preselected and screened, we will optimize the design and synthesis of title compounds. The target proteins of Dyskerin and NOP10 will be expressed and purified in vitro, and the structure-activity relationship between active compounds and Dyskerin-NOP10 inhibition was analyzed in a timely manner. Pull down, ITC fusion protein and MST experiments will be used to confirm the interaction between active compounds regulating Dyskerin-NOP10 protein-protein; Western blot will be confirmed that the active compounds targeting the Dyskerin-NOP10 complexes; finally lead compound with inhibitor telomerase activity and regulation of Dyskerin-NOP10 protein interaction will be discovered.