Anti-HIV-1 Screening And Mechanism Of PM-19 Derivative

AIDS,a human acquired immunodeficiency syndrome,is a major infectious disease caused by HIV-1 virus that poses a serious threat to social public health.At the end of 2015,the global HIV-1 virus carriers have 36.7 million people,looking for effective treatment is still important.K₇[PTi₂W₁₀O₄₀]·6H₂O?PM-19?has strong anti-HIV-1 ability and low toxicity,and it has been shown that Heteropolyacid compounds have good antiviral and antitumor activity.With PM-19 as the lead compound,the development of safe and effective anti-HIV-1 agents will provide a new way for the treatment of AIDS.Firstly,the quantitative analysis method of fake virus infectivity was optimized.The sensitivity,reproducibility and the correlation between the method and the luciferase method were investigated.The results showed that AlphaLISA was used to detect p24 protein and the minimum detection limit was 2.1 pg/mL,with good sensitivity.The reproducibility of p24protein in four different pseudovirus subtypes was tested.The coefficient of variation of the repeat test was less than 10%,and the coefficient of variation was about 10%,which showed good reproducibility.In addition,four pseudoviruses were titrated using p24 antigen and luciferase method,and the results were consistent with the results of 50%histological cell infection(TCID₅₀).There was a good linear correlation between the two methods.Secondly,the cytotoxicity and anti-HIV-1 virus activity of 28 heteropoly acid PM-19derivatives were analyzed by CCK-8 cell proliferation assay and fake virus single life cycle experiment.Half of the toxicity of the compound TC₅₀ and anti-HIV-1 virus half inhibitory concentration IC₅₀.The results showed that the cytotoxicity of PM-19 was 25.96±0.89?g/mL,and the phosphonium salt derivative had no significant difference from PM-19.However,the TC₅₀ of pyridine salt,imidazole salt,ammonium salt and aryl ammonium salt derivative was significantly higher than that of PM-19,which indicated that the modified derivative could effectively reduce the cytotoxicity.In addition,PM-19 had a dose-dependent inhibition of viral proliferative activity with an IC₅₀0 of 0.013±0.004?g/m L.Screened 28 PM-19 derivatives have different inhibitory effects on HIV-1 virus.Wherein the anti-HIV-1 virus activitives of the pyridiniums,adamantane ammonium salt and serine derivatives is better than that of PM-19.Finally,six kinds?X-4,X-5,X-7,X-11,X-15,X-19?with highly effective and low toxicity derivatives were detected by surface plasmon resonance?SPR?to carry out anti-HIV-1 mechanism of action.It were found that the X-4?X-5and X-7 has a good affinity and inhibit integrase activity;X-4 and X-5 dose dependent reverse transcriptase inhibitory activity.The results showed that X-4 had good in vitro antiviral activity and had multiple targets.Further experiments showed that X-4 pairs of B and A/D Type of viral infection has a selective inhibitory effect on the A and C subtype virus infection no effect.In this study,the antiviral activity and mechanism of twenty-eight PM-19 derivatives were screened on the basis of optimizing the antiviral drug screening method.It was found that a highly effective and low toxicity pyridinium X-4,which has good inhibitory activity against B and A/D subtype fake virus,and its mechanism may be related to the inhibition of reverse transcriptase and integrase 3'processing activity.The study will provide experimental basis for the development of new agents of HIV-1 inhibitors.