Wikstroelide M potently inhibits HIV replication by targeting reverse transcriptase and integrase nuclear translocation  

作　　者：ZHANG Xuan HUANG Sheng-Zhuo GU Wan-Gang YANG Liu-Meng CHEN Huan ZHENG Chang-Bo ZHAO You-Xing WAN David Chi-Cheong ZHENG Yong-Tang 

机构地区：[1]Key Laboratory of Animal Models and Human Disease Mechanisms of Chinese Academy of Sciences&Yunnan Province,KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research of Common Diseases,Kunming Institute of Zoology,Chinese Academy of Sciences [2]School of Pharmaceutical Science&Yunnan Key Laboratory of Pharmacology for Natural Products,Kunming Medical University [3]Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences [4]School of Biomedical Sciences,KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research of Common Diseases,Faculty of Medicine,The Chinese University of Hong Kong [5]School of Pharmacy,China Pharmaceutical University

出　　处：《Chinese Journal of Natural Medicines》2014年第3期186-193,共8页中国天然药物（英文版）

基　　金：supported,in part,by grants from the National Natural Science Foundation of China(Nos.81102483,81001462);the 973 Program(No.2009CB522306);the Key Scientific and Technological Program of China(Nos.2009-ZX09501-029,2012ZX10001-006,2012ZX10001-007,2012ZX-09103-101-022),and Yunnan(No.2010GA001)

摘　　要：AIM： To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder （Thymelaeaceae）. METHOD： The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA. The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA. The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay. The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking. RESULTS： Wikstroelide M potently inhibited different HIV-1 strains, including HIV-lmn, HIV-1AI7, and HIV-19495, induced a cytopathic effect, with ECs0 values ranging from 3.81 to 15.65 ng.mL-I. Wikstroelide M also had high inhibitory activities against HIV-2noD and HIV-2cBL_20-induced cytopathic effects with ECs0 values of 18.88 and 31.90 ng.mL 1. The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay, with ECs0 values ranging from 15.16 to 35.57 ng.mL-1. Wikstroelide M also potently inhibited HIV-lnm induced cytolysis in MT-4 cells, with an ECs0 value of 9.60 ng.mL ~. The mechanistic assay showed that wikstroelide M targeted HIV-I reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75. CONCLUSION： Wikslroelide M may be a potent HIV-1 and HIV-2 inhibitor, the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear Iranslocation through dismpting the interaction between integrase and LEDGF/p75.AIM:To evaluate the anti-HIV activity and mechanism of action of wikstroelide M,a daphnane diterpene from Daphne acutiloba Rehder(Thymelaeaceae).METHOD:The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA.The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA.The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay.The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking.RESULTS:Wikstroelide M potently inhibited different HIV-1 strains,including HIV-1IIIB,HIV-1A17,and HIV-19495,induced a cytopathic effect,with EC50 values ranging from 3.81 to 15.65 ng·mL-1.Wikstroelide M also had high inhibitory activities against HIV-2ROD and HIV-2CBL-20-induced cytopathic effects with EC50 values of 18.88 and 31.90 ng·mL-1.The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay,with EC50 values ranging from 15.16 to 35.57 ng·mL-1.Wikstroelide M also potently inhibited HIV-1IIIB induced cytolysis in MT-4 cells,with an EC50 value of 9.60 ng·mL-1.The mechanistic assay showed that wikstroelide M targeted HIV-1 reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75.CONCLUSION:Wikstroelide M may be a potent HIV-1 and HIV-2 inhibitor,the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear translocation through disrupting the interaction between integrase and LEDGF/p75.

关 键 词：Wikstroelide M Daphnane diterpene Daphne acutiloba Rehder HIV Reverse trascriptase INTEGRASE Nucleartranslocation Lens epithelium-derived growth factor （LEDGF/p75） Molecular docking 

分 类 号：R965[医药卫生—药理学]