酶学及热力学研究：天然产物抗HIV机制及作用特点

本项目用酶学和热力学方法研究一类有效的天然物质抗HIV的作用机理和特征。为了克服HIV对逆转录酶抑制剂的耐药性，本项目发现天然活性成分及其改构物芳基四氢萘化合物RM-1不仅抑制野生型HIV-1逆转录的活性与奈韦拉平相当，且对耐药病毒也有高抑制活性（RM-1可抑制8种耐药毒株中的6株；而奈韦拉平全部耐药）。.在此基础上，本项目以RM-1为模型化合物，研究这类化合物的药理作用机制和特点，包括1.解析对逆转录酶多重催化的抑制环节；2.用等温滴定量热法（ITC）研究有代表性化合物热力学特征，分析抑制剂和酶结合的自由能(ΔG)，焓(ΔH)，熵(-TΔS)和Ki（EC50）相关性，从微观和深层次上揭示抑制剂与酶的结合内涵；3.结合分子模拟方法，从结构生物学和构效关系的角度解析化合物与野生和耐药毒株逆转录酶结合模式的异同，为设计和研究新型NNRTIs提供理论和结构依据。

Human immunodeficiency virus infects vital cells in the human immune system leading to acquired immunodeficiency syndrome (AIDS) which could not be cured. The clinical therapeutics obstacle was HIV drug resistance, which was caused by HIV mutating and reproducing itself in the presence of antiretroviral drugs, leading to treatment failure. Therefore, keeping development of novel anti-HIV drugs is highly required. .Reverse transcription is a unique characteristic of HIV which can be blocked by non-nucleoside reverse transcriptase inhibitors. NNRTIs are important components for the combination of antiretroviral (ARV) drugs for the standard antiretroviral therapy..We have found abstracts from Machilus exhibited inhibitory effects on HIV replication. Structural modification and SAR analysis study showed that aryl tetrahydronaphthalene derivatives inhibited HIV reverse transcription process (compound RM-1 has be most potent activity which was relative to Nevirapine. Furthermore, this compound had good bioactivities on NNRTIs-resistant HIV replication as well..This study will focus on the mechanisms and pharmacological characteristics of compound RM-1, which represents aryl tetrahydronaphthalene derivatives. The research will include the effects of the compound on RT's multi-catalytic activities; the effects on drug-resistant reverse transcriptase activities; effects on RT catalytic kinetics; and the thermodynamic signatures with RT. The compound-RT interaction sites will be analyzed by work station calculating based on our experimental results. This research will theoretically and substantially contribute to novel NNRTIs development.