Antiviral Screening Assays Based on HCV Replicons

描述（由申请人提供）：明确定义和 丙型肝炎病毒（HCV）的强大细胞培养系统对于 促进对HCV的抗病毒药物的鉴定。Apath，LLC建议 开发一系列基于细胞的检测方法，用于检测和筛选化合物 具有抗丙型肝炎病毒（HCV）的抗病毒活性。这些试验将 基于含有组成型复制亚基因组HCV的细胞系， 复制子。已发表的HCV细胞培养复制系统中没有一个是 足够稳健以有效地用于抗病毒筛选测定。 最近，Lohmann等人描述了由以下构建的亚基因组HCV复制子： 基因型Ib RNA，并且他们分离了含有该基因型的Huh7细胞系。 自主复制的复制子含有类似HCV复制子的细胞系 是在查尔斯·M.大米和授权专为 阿帕斯这些细胞已被广泛分析，并已被证明， 含有表现出自主HCV RNA复制的HCV复制子。我们 我建议最大限度地利用这些细胞系作为药物发现工具 通过格式化用于化合物筛选的测定和通过修饰复制子以 使系统更适合于高通量筛选。该系统 代表了用于识别和测试抑制剂的最佳可用系统， 因此，它可能成为发现新药的重要工具。 拟定商业应用： Apath，LLC建议开发一系列基于细胞的检测方法，用于检测和筛选 具有抗丙型肝炎病毒（HCV）的抗病毒活性的化合物。 这些测定是关键的 用于评估具有生物化学鉴定的抗HCV活性的先导化合物，还 协助寻找治疗HCV感染的新抗病毒药物。

DESCRIPTION (provided by applicant): Identification of a well-defined and robust cell-culture system for hepatitis C virus (HCV) is critical for facilitating efforts to identify antivirals against HCV. Apath, LLC proposes to develop a series of cell-based assays for testing and screening of compounds with antiviral activity against hepatitis C virus (HCV). These assays will be based on cell lines which contain a constituitively replicating subgenomic HCV replicon. None of the published HCV cell-culture replication systems are sufficiently robust to be used effectively in an antiviral screening assay. Recently, Lohmann et al. described a subgenomic HCV replicon constructed from genotype lb RNA and they isolated Huh7 cell lines which contain this autonomously replicating replicon. Cell lines containing a similar HCV replicon have been made in the laboratory of Charles M. Rice and licensed exclusively to Apath. The cells have been analyzed extensively and have been demonstrated to contain an HCV replicon which exhibits autonomous HCV RNA replication. We propose to maximize the usefulness of these cell lines as drug-discovery tools by formatting assays for compound screening and by modifying the replicon to make the system more amenable to high throughput screening. This system represents the best available system for identifying and testing inhibitors of HCV and thus could be an essential tool for the discovery of new drugs. PROPOSED COMMERCIAL APPLICATION: Apath, LLC proposes to develop a series of cell-based assays for testing and screening of compounds with antiviral activity against hepatitis C virus (HCV). Such are assays critical for evaluating lead compounds with biochemically identified activity against HCV and also to assist in efforts to identify new antivirals for treatment of HCV infections.