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目前,临床使用的抗AIDS一线药物主要是HIV逆转录酶抑制剂和蛋白质酶抑制剂。但是,由于这类药物价格昂贵、严重的副作用、毒性、耐药性等问题日益严重,发展新作用机制的抗HIV-1药物已成当务之急。因此,本课题以HIV-1病毒RNA核转运关键蛋白Rev为靶点,建立了新型抗HIV药物筛选模型,以期寻找具有新作用机制的抗病毒药物。在筛选模型中,选择了由HIV-1病毒偏爱性密码子所编码的GFP(GFPHIV)作为报告基因,用于快速简便地分析样品对Rev依赖的RNA核转运的影响。选取抑制剂来普霉素B作为模型的阳性对照对本模型进行了初步评价,计算出Z’因子为0.8220。上述结果初步证明了该模型可用于新型抗HIV药物的筛选。对3000个化合物进行初筛,阳性率为9.3%,复筛阳性率为7.3%。通过抗病毒活性的测定实验以及对阳性化合物进行免疫印迹检测,最后发现IMB7C7这个化合物以病毒RNA核转运为靶点,是具有较好效果的特异性抑制剂。
Currently, clinically used first-line anti-AIDS drugs are mainly HIV reverse transcriptase inhibitors and protease inhibitors. However, the development of new mechanisms of action for anti-HIV-1 drugs has become a top priority because of the high cost of such drugs, serious side effects, toxicity and drug resistance. Therefore, in this project, a novel anti-HIV drug screening model was established with the target protein Rev of RNA core of HIV-1 virus as a target, in order to find antiviral drugs with new mechanism of action. In the screening model, GFP (GFPHIV), encoded by HIV-1 virus-preferred codons, was selected as a reporter for rapid and easy analysis of the effect of samples on RNA-dependent nuclear translocation in Rev. Select the inhibitor leptomycin B as a positive control model of this model was initially evaluated, calculated Z ’factor of 0.8220. The above results preliminary proved that the model can be used for screening of new anti-HIV drugs. The first screening of 3000 compounds, the positive rate was 9.3%, the positive rate of triple screening was 7.3%. Through the determination of antiviral activity and the immunoblotting of positive compounds, IMB7C7 was found to be a specific inhibitor with good effect as a target of viral RNA nuclear translocation.