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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.
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. 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 wikst roelide M on the three HIV-1 was was 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.