ANTIVIRAL STRATEGY DIRECTED TOWARD HCV RNA TRANSLATION

丙型肝炎病毒(HCV)感染了世界人口的0.5%-1.5%。它 会导致急性和慢性肝炎和肝硬变，并与 与肝细胞癌有关。我们已经提出了一种抗丙型肝炎病毒的战略 依赖于丙型肝炎病毒RNA翻译的独特属性，该属性由 位于丙型肝炎病毒5‘端的内部核糖体进入位点(IRES) 未翻译的区域。这一策略的目标是识别抗病毒药物 专门抑制丙型肝炎病毒翻译而不影响帽的化合物- 细胞信使核糖核酸的依赖翻译。在第一阶段，我们成功地 建立了稳定表达双报告结构的细胞系，CAT- IRES-FLuci，并在我们的自动化设施中筛选出73,000种真菌 提取物和化合物对该细胞株的抑制活性 翻译控制下的萤火虫荧光素酶(FLuci)信号 艾丽斯。该屏幕将显示21个点击数 弗卢西抑制和四甲基偶氮唑盐(一种细胞毒性)抑制的差异 指标)，目前正在进行后续调查。在第二阶段， 我们将完成对75,000种真菌提取物和55,000种真菌提取物的筛选 化合物。命中得分，并在高吞吐量中重新确认 筛选将根据效力、选择性、 细胞毒性和抗病毒活性。真菌提取物中的活性物质 将被提纯并确定它们的化学结构。抗丙型肝炎病毒药物 将与我们的合作伙伴Biochem Pharma同步开发 三. 拟议的商业应用：丙型肝炎病毒感染全球0.5%-1.5% 并构成一个主要的健康问题，因为没有有效的 可用的预防或治疗。化合物和天然产品 抑制丙型肝炎病毒RNA翻译可开发成新型抗病毒药物 具有巨大的商业潜力。

Hepatitis C virus (HCV) infects 0,5-1.5% of the world population. It causes acute and chronic hepatitis and cirrhosis and is highly associated with hepatocellular carcinoma. We have proposed an anti-HCV strategy that relies on the unique property of HCV RNA translation, which is directed by the internal ribosomal entry site (IRES) located in the HCV 5' untranslated region. The goal of this strategy is to identify antiviral compounds that specifically inhibit HCV translation without affecting cap- dependent translation of cellular mRNA. In phase I, we have successfully established a stable cell line expressing a dual-reporter construct, CAT- IRES-Fluci, and have screened in our automated facilities 73,000 fungal extracts and compounds against this cell line for activities that inhibit the firefly luciferase (Fluci) signal under the translational control of IRES. This screen results in identification of 21 hits that show difference between Fluci inhibition and MTT inhibition (a cytotoxic indicator), which are now being investigated in follow-up. In phase II, we will complete screening of a total of 75,000 fungal extracts and 55,000 compounds. Hits that score and are reconfirmed in high throughput screening will be prioritized with respect to potency, selectivity, cytotoxicity, and antiviral activity. Active compounds in fungal extracts will be purified and their chemical structure determined. Anti-HCV drugs will be developed with our collaborative partner, Biochem Pharma, in phase III. PROPOSED COMMERCIAL APPLICATION: HCV infects 0.5-1.5% of the world population and poses a major health problem since there is no effective prevention or treatment available. Compounds and natural products that inhibit HCV RNA translation can be developed into novel antiviral drugs with enormous commercial potential.