INHIBITION OF THE HIV 1 REV TRANSACTIVATOR

该项目的目标是确定HIV-1Rev的新抑制剂 用于治疗艾滋病的蛋白质。Rev函数是绝对必需的 用于艾滋病毒复制。在第一阶段，我们构建了一个报告质粒 含HIV-1外显子6和外显子7拼接之间的荧光素酶基因 供体和受体部位。在这个结构中，荧光素酶只表达。 在Rev.存在的情况下，进行了瞬时共转染实验 适用于自动化高通量筛选。在第二阶段，一个完整的 将进行10万份样本的自动化高通量筛选， 包括50,000种真菌提取物和等量的来自 一个药物化学图书馆。潜在的REV抑制剂将导致 荧光素酶活性下降。几个后续化验将是 用于鉴定特定作用于REV和TO的先导化合物 排除细胞毒性化合物。铅生物将被参照，铅 部分纯化和结构鉴定。其作用机制 这些实体表现出特定的REV抑制特性，例如 抑制REV-RRE相互作用或与细胞因子相互作用， 将在体外进行研究。特定的REV抑制剂铅萃取物和 化合物将在艾滋病毒复制检测系统中进行测试，代表 病毒感染的急性和慢性阶段。在第三阶段，选定的销售线索 将接受合成孔径雷达化学反应，并进入临床前和 临床发展。 拟议的商业应用：目前还没有有效的治疗方法 治疗艾滋病的药物。HIV-1 Rev的小分子量抑制剂 反式激活剂，作为这个项目的结果，具有重要的意义 在艾滋病治疗中作为抗病毒药物的潜力。

The goal of this project is to identify novel inhibitors of the HIV-1 Rev protein for the treatment of AIDS. Rev function is absolutely required for HIV replication. In Phase I, we constructed a reporter plasmid containing the luciferase gene between the HIV-1 exon 6 and exon 7 splice donor and acceptor sites. In this construct, luciferase is expressed only in the presence of Rev. A transient co-transfection assay has been adapted for automated high throughput screening. In Phase II, a fully automated high-throughput screen of 100,000 samples will be carried out, including 50,000 fungal extracts and an equal number of compounds from a medicinal chemistry library. Potential Rev inhibitors will cause a decrease in luciferase activity. Several follow-up assays will be utilized to qualify lead compounds which act specifically on Rev and to exclude cytotoxic compounds. Lead organisms will be refermented, the lead moiety purified and its structure elucidated. The mechanism of action of those entities exhibiting specific Rev-inhibitory properties, e.g. inhibition of Rev-RRE interactions or interaction with cellular factors, will be investigated in vitro. Specific Rev inhibitor lead extracts and compounds will be tested in HIV replication assay systems, representing acute and chronic stages of viral infection. In Phase III, selected leads will be subjected to SAR chemistry and entered into pre-clinical and clinical development. PROPOSED COMMERCIAL APPLICATION: Currently there is no effective therapy for AIDS. Small molecular weight inhibitors of the HIV-1 Rev transactivator, identified as a result of this project, have significant potential as anti-viral agents in the treatment of AIDS.