蛋白别构位点识别方法发展及在药物设计中的应用

别构调节是一种通过调节分子结合到蛋白活性位点以外的别构位点进行蛋白功能调节的方式。由于别构位点相对活性位点多样性强，开发结合蛋白别构位点的小分子药物选择性高，毒副作用小，已成为国际各大医药公司新发展方向。然而，当前发现的蛋白别构位点主要来源于实验中随机发现，缺乏合理识别方法，严重影响了蛋白别构利用和相关小分子的药物开发。在前期的工作中，我们构建了包含当前已知别构蛋白数据系统，初步分析了别构位点特征，发展了针对部分蛋白的别构位点识别方法，在相应蛋白发现新的别构位点及别构调节小分子。本课题拟在此基础上, 系统分析别构位点特征标识，设计构象循环遍历和残基偶联策略,发展普适性的蛋白别构位点识别方法，并在多巴胺受体3及Sirtuin3为代表的体系上(通过生物化学检测)验证该识别方法的有效性，从而为各种蛋白的别构位点识别提供实用工具，也为重要疾病靶标发展新型别构小分子药物提供先导化合物候选。

Allostery is the most direct, rapid and efficient way of regulating protein function induced by the binding of a ligand at a site topographically distinct from the orthosteric site, also termed as the allosteric site. From the perspective of drug discovery, because allosteric sites evolved under lower sequence-conservation pressure compared with the evolutionarily conserved active sites, targeting allosteric sites can provide unprecedented advantages in terms of higher specificity and lower toxicity. Thus, the discovery and development of small-molecule allosteric drugs has drawn increasing attention as potential therapeutic agents in the international pharmaceutical industry. Currently, most of the known allosteric sites are serendipitously discovered by experiments due to devoid of feasible methods to identify the allosteric sites, which severely hinder the exploitation of allosteric regulation and the development of small-molecule allosteric drugs. In the previous study, we comprehensively constructed the Allosteric Database (ASD), preliminarily analyzed the structural characteristics of allosteric sites, developed a method (Allosite) to detect the allosteric sites of specific proteins, and found novel allosteric sites and their allosteric modulators in proteins of interest. Based on these results, in this project we systematically analyze the unique features of allosetric sites, design the strategy of conformational sampling and residue coupling, and develop a universal method to identify the allosetric sites. Subsequently, the dopamine receptor 3 and Sirtuin 3 will be chosen as models to validate the efficiency of our method through biochemical tests. This project will provide a useful tool to identify allosteric sites in various proteins and the lead compounds to the development of novel small-molecule allosteric drugs aimed at the diseases-assocaited therapy.