INHIBITION OF THE HIV 1 REV TRANSACTIVATOR

该项目的目标是识别HIV-1 Rev的新型抑制剂， 蛋白质治疗艾滋病。Rev函数是绝对必需的 for HIV HIV HIV复制.在第一阶段，我们构建了一个报告质粒， 在HIV-1外显子6和外显子7剪接之间含有荧光素酶基因 供体和受体位点。在该构建体中，荧光素酶仅表达 在Rev. A存在的情况下，已进行瞬时共转染试验 适用于自动化高通量筛选。在第二阶段，全面 将对10万个样品进行自动化高通量筛选， 包括50，000种真菌提取物和相同数量的化合物， 药物化学图书馆潜在的Rev抑制剂将导致 荧光素酶活性降低。将进行几项后续试验， 用于鉴定特异性作用于Rev的先导化合物， 排除细胞毒性化合物。铅生物将被重新发酵， 部分纯化并阐明其结构。的作用机制 表现出特定Rev抑制特性的那些实体，例如， 抑制Rev-RRE相互作用或与细胞因子的相互作用， 将在体外进行研究。特定Rev抑制剂电极导线浸提液和 化合物将在HIV复制测定系统中进行测试，代表 病毒感染的急性和慢性阶段。在第三阶段， 将接受SAR化学检查，并进入临床前和 临床发展。 拟定商业应用：目前尚无有效治疗方法 治疗艾滋病 HIV-1 Rev的小分子量抑制剂 反式激活因子，确定作为该项目的结果，具有显着的 作为抗病毒药物治疗艾滋病的潜力。

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.