摘要
瞬时受体电位香草酸型通道第三亚型(transient receptor potential vanilloid 3,TRPV3)与伤害性感受、热感受等重要生理功能密切相关,发现靶向TRPV3通道的选择性小分子拮抗剂对深入研究TRPV3通道的病生理功能和促进TRPV通道的基础研究和新药靶点研究具有重要的意义。本文基于已知的TRPV3小分子拮抗剂,提出基于配体的计算机辅助药物设计的方法进行选择性小分子拮抗剂的骨架跃迁,采用三维形状和静电相似性的虚拟筛选策略,对PKU-CNCL、Specs化合物库进行大规模的虚拟筛选;同时以TRPV1晶体结构为模板,并结合TRPV3的已解析出来的N末端ARD区域的晶体结构,构建了人TRPV3的晶体结构的单体模型,以TRPV3单体模型进行了TRPV3小分子拮抗剂的分子对接分析,将其应用于虚拟筛选和骨架跃迁中。经过两轮的虚拟筛选和生物活性评价,发现了对TRPV3具有选择性抑制作用且抑制活性IC_(50)为18.0±1.1μmol·L-1(n=4)的新型骨架结构化合物V-39,该化合物与TRPV3相互作用的重要氨基酸残基为第310和314位组氨酸以及第577位精氨酸。本文研究结果可以用于进一步的结构改造和优化,有助于指导新型的TRPV3选择性拮抗剂的药物设计。
Transient receptor potential vanilloid member 3 （TRPV3） is a temperature-sensitive cation channel protein, which contributes to nociception, itch, hair growth, emotional control and the pathophysiology of migraine. However, research progress on TRPV3 fundamental molecular biology is rather slow, compared to other TRP channels due to the lack of its selective antagonists. It＇s necessary to identify TRPV3 selective antagonists for the study on TRPV3 physiological functions. In this study, several selective TRPV3 antagonists were identified by ligand-based virtual screening of shape-based similarity and electrostatic matching. The most potent one （V-39） blocked 2-APB-activated currents in a stable human TRPV3 expressed HEK293T cell line with IC50=18.0 ±1.1 μmol·L-1 （n=4）. Besides, the interaction pattern between TRPV3 and its antagonists were studied through docking the antagonists into a homology model （TRPV3HM4） generated from the crystal structure of TPRV1. The docking results show that the binding site of TRPV3 locates between linker domain （of N-terminus and TM1） and TRP Box. There are a π-π stacking interaction and hydrogen bonding interactions between compound V-39 and residues His-310, His-314 and Arg-577 of the pocket. Identification of these antagonists provides new probes for understanding the pharmacological function of TRPV3 channel.
作者
沈燕君
张新然
吴洋
王克威
刘振明
金宏威
张亮仁
张礼和
SHEN Yan-jun;ZHANG Xin-ran;WU Yang;WANG Ke-wei;LIU Zhen-ming;JIN Hong-wei;ZHANG Liang-ren;ZHANG Li-he(State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China)
出处
《药学学报》
CAS
CSCD
北大核心
2018年第6期966-975,共10页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(81373272,81573273,81670637)
