特异性RORγ拮抗剂的设计、合成和抗炎作用机制研究

The retinoic acid receptor-related orphan nuclear receptors α and γ are considered to be the master regulators of development of TH17 cells, a subset of T cells that have been implicated in the pathology of several autoimmune diseases, including multiple sclerosis (MS) and rheumatoid arthritis (RA). Given the important role of RORα in the regulation of circadian rhythm and metabolism, there was some concern that a dual RORγ-selective antagonist could cause undesirable side effects, suggesting that a RORγ-selective compound would be more appropriate. Some pharmaceutical such as Pfizer put a lot of effort in developing RORγ antagonist as drug for autoimmune diseases. Here, we have screened and then optimized a series of small molecule antagonists. Among them, compound S4-3 show an IC50 less than 10 nM agonist RORγ. Based on our previous experimental data, we plan to deeply investigate the structure-activity relationship. Anti-inflammation effect will also be evaluated on cell-based assy. In this project, rational drug design, synthesis, activity evaluation and structure study will be performed at the same time. In such way, the selectivity and anti-inflammation mechanism will be elucidated. This study will provide important probe for functional study. The study will provide RORγ ligand being considered more attractive candidates for therapeutics aimed at treating TH17-mediated autoimmune diseases such as multiple sclerosis, rheumatoid arthritis.

孤儿核受体RORα和RORγ两种亚型直接调控TH17细胞分化，是多发性硬化症等自身免疫性疾病潜在的药物作用靶标。因RORα在生理节奏和代谢过程中起重要的调节作用，RORα/RORγ双效拮抗剂将引起不良副作用，所以RORγ特异性拮抗剂的发现就显得格外迫切。Pfizer等国际制药公司已投入大量精力开发RORγ小分子拮抗剂作为自身免疫类疾病药物。申请人利用合理药物设计成功筛选了一批小分子化合物，通过初步优化获得化合物S4-3对RORγ的体外抑制活性优于现有RORγ拮抗剂，IC50值小于10nM。本申请拟在上述工作基础上，采用设计、合成、活性测试相结合的方法，对S4-3化合物进行深入构效关系研究，同时开展拮抗剂与RORγ复合物的结构生物学研究，阐明化合物选择性的结构机制和抗炎作用机制。本课题将为进一步研究RORγ在免疫系统中的功能提供探针分子，并可能为新型自身免疫疾病药物开发提供重要的先导化合物。

本项目完成RORγ靶标抑制剂虚拟筛选平台的搭建，同时已完成对多个数据库虚拟筛选工作。根据筛选结果购买两批化合物，并对其进行活性测试，获得多个有活性化合物。相关成果发表在European Journal of Medicinal Chemistry和中国药物化学杂志上，同时申请了专利。此外，根据组合库设计方法得到活性好的化合物XY11和XY018，并对其开展靶标新应用方面的研究。我们首次发现并证明了RORγ可以驱动AR从而间接促进前列腺癌症的发展。XY018可以有效抑制AR蛋白的表达，并有效抑制前列腺癌细胞的生长，移植瘤动物模型实验结果显示XY018可以抑制肿瘤的生长。该部分成果发表一篇文章在Nature Medicine杂志上。我们通过对新骨架化合物进行的分子、细胞及动物水平的生物活性测试，探索了新的药物作用机制。