Antiviral Screening Assays Based on HCV Replicons

描述(申请人提供)：丙型肝炎病毒(丙型肝炎病毒)最多 在美国流行的慢性血液传播感染是一个全球健康问题。 美国约有400万人，1.7亿人 全世界的人都是慢性感染者。高达40%的慢性肝病 这种疾病与丙型肝炎病毒有关，每年导致多达10,000人死亡。 尽管有所改善，但治疗仍然不太理想，而且是新的治疗方法 选择是非常必要的。APath，LLC已经开发出基于细胞的检测方法 抗丙型肝炎病毒活性化合物的检测与筛选 病毒(丙型肝炎病毒)。这些检测是基于Huh7细胞系，这些细胞系含有 建设性地复制亚基因组丙型肝炎病毒复制子。这些细胞已经被 经过广泛分析，并已被证明含有一个丙型肝炎病毒复制子， 展示自主的丙型肝炎病毒RNA复制。我们已经将这些细胞系作为 通过格式化用于化合物筛选的分析来实现药物发现工具 适用于高通量筛选。在此阶段II提案中，我们描述了 我们计划使用这些基于细胞的分析方法来鉴定具有抗丙型肝炎病毒抗体的化合物 活动。为了实现这一目标，我们将计划利用独特的天然资源 化合物，并筛选40,000多口井，包含多达200,000个离散 化合物，以识别抑制复制丙型肝炎病毒复制子的“命中”。我们 将确认二次丙型肝炎病毒复制子中所有“命中”的抗病毒活性 定量和细胞毒性检测。我们将鉴定出最具代表性的化合物 有望成为潜在的抗丙型肝炎病毒的化学和生物学特征 毒品。我们还计划通过生化和生物化学来确定铅的目标 遗传方法。

DESCRIPTION (Provided by applicant): Hepatitis C virus (HCV) is the most prevalent chronic blood-borne infection in the U.S and a global health problem. Approximately 4 million individuals in the U.S. and 170 million individuals worldwide are chronically infected. As much as 40 percent of chronic liver disease is HCV related and this results in up to 10,000 deaths each year. Despite improvements, treatment of remains suboptimal, and new therapeutic options are critically needed. Apath, LLC has developed cell-based assays for testing and screening of compounds with antiviral activity against hepatitis C virus (HCV). These assays are based on Huh7 cell lines that contain a constructively replicating subgenomic HCV replicon. The cells have been analyzed extensively and have been demonstrated to contain an HCV replicon that exhibits autonomous HCV RNA replication. We have promoted these cell lines as drug-discovery tools by formatting assays for compound screening that are amenable to high throughput screening. In this Phase II proposal we describe our plans to use these cell-based assays to identify compounds with anti-HCV activity. Toward this goal we will plan to utilize a unique source of natural compounds and to screen over 40,000 wells containing up to 200,000 discrete compounds to identify "hits" that inhibit replication of the HCV replicon. We will confirm antiviral activity of all "hits" in secondary HCV replicon quantitation and cytotoxicity assays. We will identify compounds with the most promising chemical and biological profiles to pursue as potential anti-HCV drugs. We also plan to identify targets of leads through biochemical and genetic approaches.