INHIBITION OF HEPATITIS C VIRUS HELICASE

丙型肝炎病毒（HCV）感染了世界人口的1.5%以上。它 引起肝炎和肝硬化，并与肝- 细胞癌我们建议鉴定抑制HCV的化合物 解旋酶被认为对HCV RNA和蛋白质很重要 合成.我们的策略包括筛选化合物和真菌 提取物，以确定解旋酶抑制剂和验证 在各种HCV复制模型中的抗病毒活性。到目前为止， 证明了重组HCV NS3蛋白在人血清中的解旋酶活性。 闪烁邻近测定（SPA），并格式化和验证 用于高通量筛选的测定。在第一阶段，我们将完成 筛选100，000种实体（50，000种化合物和50，000种真菌 提取物）。命中，定义为抑制SPA的化合物和提取物 信号在高通量筛选中，将通过再筛选来确认。在 此外，作为SPA信号的内在抑制剂的命中物将被 识别和排除。此外，选择性命中，抑制 将鉴定HCV解旋酶的活性，但不鉴定哺乳动物解旋酶的活性 作为选择性打击。在第二阶段，我们将优先确定 无毒的选择性解旋酶抑制剂， 黑猩猩肝细胞模型和小鼠模型。 拟定商业应用： HCV感染超过1.5%的世界和美国人口，并构成了主要的 健康问题，因为没有有效的治疗或预防 available.抑制HCV解旋酶的化合物和天然产物可 开发成为具有巨大商业价值的新型抗HCV药物 潜力

Hepatitis C virus (HCV) infects over 1.5% of the world population. It causes hepatitis and cirrhosis and is highly associated with hepato- cellular carcinoma. We propose to identify compounds that inhibit HCV helicase, which is thought to be important for HCV RNA and protein synthesis. Our strategy includes screening of compounds and fungal extracts to identify helicase inhibitors and verification of the antiviral activity in various HCV replication models. To date, we have demonstrated the helicase activity of a recombinant HCV NS3 protein in a scintillation proximity assay (SPA) and formatted and validated the assay for high throughput screening. In phase I, we will complete screening of 100,000 entities (50,000 compounds and 50,000 fungal extracts). Hits, defined as compounds and extracts that inhibit the SPA signal in high throughput screen, will be confirmed by re-screening. In addition, hits that are intrinsic inhibitors of the SPA signal will be identified and excluded. Furthermore, selective hits that inhibit the activity of HCV helicase but not a mammalian helicase will be identified as selective hits. In phase II, we will prioitize the hits to identify nontoxic, selective helicase inhibitors that inhibit HCV replication in a chimpanzee hepatocyte model and mouse model. PROPOSED COMMERCIAL APPLICATION: HCV infects over 1.5% of the world and US population and poses a major health problem since there is no effective treatment or prevention available. Compounds and natural products that inhibit HCV helicase can be developed into novel anti-HCV drugs with enormous commercial potential.