M1胆碱受体选择性别构激动剂的虚拟筛选和体外活性研究

目的:M1毒蕈碱型乙酰胆碱受体(M1受体)在改善学习和记忆等高级认知功能障碍中起重要作用,本文利用计算机辅助药物设计和高表达各M受体亚型的CHO细胞(Chinese hamster ovary cell,中国仓鼠卵巢细胞),以期筛选获得新型M1受体选择性别构激动剂。方法:通过计算机辅助药物设计方法,对已知具有M1受体选择性作用别构激动剂与M1受体的晶体结构进行对接,确定活性对接口袋,据此进行化合物库虚拟筛选;利用高表达各M受体亚型的CHO细胞,对化合物进行体外活性检测。结果:虚拟筛选得到184个化合物,其中,体外实验显示化合物AJ-292和AG-205-6对M1受体有明显的激动效果,而对M3、M5受体则无影响。结论:综合利用虚拟筛选、结构分析以及特异性活性分析,筛选出具有M1受体高选择性激动作用的化合物AJ-292和AG-205-6,为设计开发新型的M1选择性别构激动剂奠定了基础。

Virtual Screening and in Vitro Activity Study on M1 Muscarinic Acetylcholine Receptor Selective Allosteric Agonists

ABSTRACT Objective: M1 muscarinic acetylcholine receptors (M1R) play important roles in learning and memory. Based on the active pocket obtained from the docking of M1R allosteric agonists and M1R, we screened out the selective allosteric M1R agonists. Methods: Virtual screening was established via computer-aided drug design based on the allosteric site in M1R crystal structure. The se- lective interaction modes between the compounds and M1R were comprehensively analyzed and used for virtual screening of the com- pound candidates from several compound libraries. The in vitro activity of compounds was studied using human recombinant muscarinic acetylcholine receptor cell lines. Results: AJ-292 and AG-205-6 showed significant agonizing effects on CHO-M1 cells, but they showed no influence on CHO-M3 and CHO-M5 cells among the 184 compounds obtained by virtual screening. Conclusion: Based on the com- bination of virtual screening, structural analysis and specific activity analysis, two compounds were screened out, which show high selec- tive activities on CHO-M1 cells. This work lays the foundation for the design and development of novel M1R allosteric agonists.