EGCG乙酸酯的HIV融合抑制活性及其作用机制研究  

HIV fusion-inhibiting activity of peracetate ester of (-)-epigallocatechin gallate and the mechanism

作　　者：杨洁[1] 李润明[1] 毛芹超[1] 陈之朋[1] 姜世勃[1] 姜志宏[1,2] 刘叔文[1] 

机构地区：[1]南方医科大学药学院,广州510515 [2]香港浸会大学中医药学院

出　　处：《中国新药杂志》2010年第21期1967-1972,共6页Chinese Journal of New Drugs

基　　金：国家自然科学基金(30729001;U0832001);广州市科技计划项目(2007J1-C0251)

摘　　要：目的:研究EGCG乙酸酯的HIV融合抑制活性,并探讨其作用靶点。方法:采用HPLC分析pH对于EGCG乙酸酯稳定性的影响以及EGCG乙酸酯在细胞中的水解产物;采用酶联免疫测定法、细胞-细胞融合方法以及假病毒实验检测EGCG乙酸酯抑制HIV与靶细胞融合的活性;应用分子对接软件研究EGCG与gp41的结合位点。结果:EGCG通过其苯环结构和羟基分别与gp41 NHR区域的疏水残基和正电荷残基结合从而抑制病毒的进入。用乙酰基保护EGCG活性酚羟基形成EGCG乙酸酯,稳定性明显增强。EGCG乙酸酯在酶联免疫反应中无活性,但在细胞实验中能有效抑制HIV介导的细胞融合及HIV假病毒感染,其半数抑制浓度IC50分别为(4.93±0.57)和(1.45±0.32)μg.mL-1。EGCG乙酸酯在细胞中的水解产物被证明为EGCG。结论:EGCG乙酸酯在细胞中能转化为母体药物,从而具有HIV融合抑制活性,且靶向作用于HIV gp41。该前药有望研发为稳定、高效的抗HIV药物,用于HIV的治疗。Objective：To investigate the HIV fusion-inhibiting activity of the peracetate ester of（-）-epigallocatechin gallate [（-）-EGCG] and explore the mechanisms.Methods： The effect of different pH values on the stability of the peracetate ester of（-）-EGCG solution and the hydrolysis of the peracetate ester of（-）-EGCG were measured by HPLC.The inhibiting activity of the peracetate ester of（-）-EGCG on HIV fusion was determined by ELISA,HIV env-mediated cell-cell fusion assay and pseudovirus assay.The binding sites of the peracetate ester of（-）-EGCG on gp41 was analyzed with a computer-aided molecular docking technique.Results： EGCG inhibited the HIV entry through its benzene ring and hydroxyl bound to the hydrophobic charged resides,and the positively charged resides located in the viral gp41 NHR region.The stability of EGCG significantly was enhanced after we introduced peracetate protection groups on the reactive hydroxyls of（-）-EGCG.ELISA showed that the peracetate ester of（-）-EGCG had no inhibiting activity against HIV in vitro,but exhibited the potential in inhibiting HIV-mediated cell fusion and HIV env pseudovirus with IC50 of（4.93±0.57） and（1.45±0.32） μg·mL-1,respectively.EGCG as hydrolysis of the peracetate ester of（-）-EGCG inside the cells was identified with HPLC.Conclusion： The peracetate ester of（-）-EGCG can inhibit HIV fusion by targeting gp41 when it is converted into its parent compound inside the cells.These results suggest that the peracetate ester of（-）-EGCG is a promising prodrug for further development as an effective and stable anti-HIV agent for HIV treatment.

关 键 词：EGCG乙酸酯 前药 HIVgp41 HIV融合抑制剂 

分 类 号：R965.1[医药卫生—药理学] R978.7[医药卫生—药学]