| The invasion for HIV breaking into the target cells requires fusion,reverse transcription,gene integration,expression of packaging proteins and release of virus particles and other steps.Based on the entire life cycle of HIV,the drug is divided into inhibitors of fusion,reverse transcription inhibitors integrase inhibitors and protease inhibitors.Integrase inhibitors can prevent the integration of viral DNA and target cell nuclear DNA;and the integrase is foreign enzymes for cells,so the inhibitors do not act on the host cell itself.In cocktail therapy,inhibitors do not adversely affect the compatibility of other types of drugs.At present,the world’s main method of treating AIDS is drug control.Natural medicine structure is diverse,rich in sources,with great potential.By reviewing the literatures and the research done in our laboratory,it was found that natural drug A might have a good anti-HIV effect.And by the means of surface plasmon resonance Biacore SPR analysis,there is a good combination between A and integrase.So this article attempts to find the effective part or effective molecules attached to integrase from the natural drug A.The experimental process is as follows:First,with 60%ethanol leaching drugs,to obtain A crude extract.Second,with Biacore SPR technology,rapid high-throughput screening of drugs can be combined with the integration of enzymes,select the RU value of more than 100 drugs,the crude drug concentration of 500μg/mL obtained when the RU change value of 144,to determine the effective.Thirdly,the efficacy of the selected drugs was verified by CCK8 and CCK-8 in vitro.The results of toxicity test were as follows:TC50 was 547μg/mL,the maximum non-toxic measure was250.7μg/m L,the IC50 was101.77μg/m L.Fourthly,the specific effective molecular solution was obtained by specific enrichment with amine coupling method,and its RU change was 447.Fifthly,the chemical molecular structure and the binding mechanism of the molecule were analyzed by HPLC-MS and computer molecular simulation.The effective molecules were P and Q,and the computerized docking score of the isomer was 104 and 94,while the positive control of Remgway’s score of 112,indicating that the effective molecules have a good effect of inhibition of integrase function. |
