作用于HIV-1gp120小分子HIV进入抑制剂NC-2的虚拟筛选及其作用机制  

Virtual screening of small molecular HIV-1 entry inhibitor NC-2 targeting gp120 and its action mechanism

作　　者：段恒[1] 王玉芹[1] 宋德寿[2] 陈之朋[1] 裘佳寅[1] 陆路[3] 姜世勃[3] 刘叔文[1] 谭穗懿[1] 

机构地区：[1]南方医科大学药学院,广东广州510515 [2]华南农业大学资源环境学院,广东广州510640 [3]复旦大学上海医学院分子病毒学教育部/卫生部重点实验室,上海200032

出　　处：《南方医科大学学报》2013年第6期826-831,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81102482;81202544)~~

摘　　要：目的基于中和抗体VRC01与HIV-1 gp120的结合模式,利用计算机虚拟筛选,从IBS天然产物数据库中筛选靶向HIV-1gp120的小分子HIV-1进入抑制剂,并对其抗病毒活性和机制进行研究。方法运用MM-PBSA方法计算候选化合物与HIV-1gp120结合后自由能的变化;利用HIV-1假病毒、活病毒技术及细胞融合实验,检测化合物抑制HIV-1感染的活性;XTT比色法检测化合物对细胞的毒性;采用酶联免疫吸附测定法(ELISA)研究化合物体外抗病毒活性的机理。结果利用计算机从40 000个化合物中虚拟筛选出19个与gp120结合后自由能降低较大的小分子化合物,其中NC-2具有抑制HIV-1感染和细胞融合的活性,其抑制HIV-1实验株IIIB的IC50是1.95±0.44μmol/L,抑制HIV-1JRFL假病毒的IC50是10.58±0.13μmol/L。酶联免疫吸附法结果表明NC-2体外能抑制HIV-1 gp120与CD4的结合,但不抑制HIV-1 gp41六螺旋的形成。结论该计算机虚拟筛选的方法可为开发作用于HIV-1 gp120的小分子进入抑制剂提供参考。同时,通过计算机辅助设计加病毒活性筛选的方法,得到一个新颖结构的HIV进入抑制剂NC-2。Objective To screen the HW-1 entry inhibitors targeting HIV-1 gp120 from the IBS natural product database by virtual screening based on the binding mode of the neutralizing antibody VRC01 with HIV-1 gp120 and investigate the anti-viral activities of the inhibitors and their action mechanisms. Methods The binding interaction of the candidate molecules binding gp120 and changes of the binding free energy were analyzed by MM-PBSA calculation. The anti-HIV-1 activities of the tested compounds were detected by HIV-1 pseudotyped virus, laboratory-adapted HIV-1 and a cell-cell fusion assay. The cytotoxicity of the studied molecules was examined by XTT colorimetric assay. The mechanisms of the anti-viral activities of the candidate molecules were analyzed using enzyme-linked immunosorbent assay. Results A total of 19 molecules with distinct reduction of the binding free energy after binding with gp120 were screened from 40000 molecules. Among them, NC-2 showed anti-HIV-1 activities against HIV-1 pseudotyped virus and laboratory-adapted HIV-1, and was capable of blocking HIV-1 envelope-mediated cell-cell fusion. The ICs0 of NC-2 for inhibiting HW-lm~ and pseudotyped HW-1j^L infection were 1.95_＋0.44 gmol/L and 10.58_＋0.13 ~raol/L, respectively. The results of ELISA suggested that NC-2 could inhibit the binding of HIV-1 gp120 to CD4 without blocking the formation of gp41 six-helix bundle in vitro. Conclusion This computer-based virtual screening method can be used to screen HIV-1 entry inhibitors targeting gp120. Using this virtual screening approach combined with anti-viral activity screening, we obtained a potent HIV-1 entry inhibitor NC-2 with novel structure.

关 键 词：HIV-1进入抑制剂 GP120 VRC01 计算机辅助设计 药物筛选 

分 类 号：R96[医药卫生—药理学]