威氏马兜铃内酯M通过靶向逆转录酶和整合酶的核转位来有效抑制HIV复制。

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

作者信息

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

机构信息

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, Kunming 650223, China; School of Pharmaceutical Science & Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming 650500, China.

Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China.

出版信息

Chin J Nat Med. 2014 Mar;12(3):186-93. doi: 10.1016/S1875-5364(14)60031-5.

DOI:10.1016/S1875-5364(14)60031-5

PMID:24702804

Abstract

AIM

To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder (Thymelaeaceae).

METHODS

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.

摘要

目的

评估瑞香狼毒（瑞香科）中一种瑞香烷二萜类化合物——瑞香狼毒素M的抗HIV活性及作用机制。

方法

采用细胞病变效应试验和ELISA法检测p24定量，评估瑞香狼毒素M对不同HIV毒株的抗HIV活性。通过实时PCR和ELISA分析瑞香狼毒素M对HIV逆转录的抑制作用。采用基于细胞的成像试验观察瑞香狼毒素M对HIV-1整合酶核转位的影响。通过分子对接分析瑞香狼毒素M对LEDGF/p75-IN相互作用的影响。

结果

瑞香狼毒素M能有效抑制不同的HIV-1毒株，包括HIV-1IIIB、HIV-1A17和HIV-19495，诱导细胞病变效应，EC50值在3.81至15.65 ng·mL⁻¹之间。瑞香狼毒素M对HIV-2ROD和HIV-2CBL-20诱导的细胞病变效应也有较高的抑制活性，EC50值分别为18.88和31.90 ng·mL⁻¹。p24定量试验进一步证实了瑞香狼毒素M对三种HIV-1毒株的抑制活性，EC50值在15.16至35.57 ng·mL⁻¹之间。瑞香狼毒素M还能有效抑制HIV-1IIIB诱导的MT-4细胞溶解，EC50值为9.60 ng·mL⁻¹。机制研究表明，瑞香狼毒素M通过破坏整合酶与LEDGF/p75之间的相互作用，靶向HIV-1逆转录酶和整合酶的核转位。