Development of Hepatitis C Virus entry inhibitors

描述（由申请人提供）：开发丙型肝炎病毒（HCV）感染的新治疗药物是一项主要的公共卫生优先事项。我们已经选择HCV进入作为发现和开发新型抗病毒剂的靶点。这个I期项目的总体目标是开发一种新的基于细胞的膜融合检测，准确地再现了HCV的进入，使用良好的特征，生物相关的细胞reagents.在I期项目期间，我们将开发稳定的细胞系，表达融合形式的天然HCV包膜糖蛋白，并使用这些细胞，以适应基于细胞的膜融合检测HCV的进入。我们将产生稳定的人肝细胞系作为适当的融合靶点，并优化高通量筛选（HTS）的重现性，灵敏度和适用性。我们还将使用单克隆抗体和表达推定的附着受体的细胞系来探测HCV进入的机制，I期项目的成功将使HCV膜融合检测在II期中用于结构多样的小分子文库的HTS，以识别HCV进入的有效抑制剂。先导化合物将在一种新的转基因小鼠模型中筛选体内抑制HCV感染，并探讨其作用机制。因此，有必要开发新的抗HCV药物用于临床。

DESCRIPTION (provided by applicant): The development of new therapeutic agents for hepatitis C virus (HCV) infection is a major public health priority. We have selected HCV entry as a target for the discovery and development of novel antiviral agents. The overall goal of this Phase I project is to develop a novel cell-based membrane fusion assay that accurately recapitulates HCV entry, using well characterized, biologically relevant cellular reagents.During the Phase I project, we will develop stable cell lines that express fusogenic forms of native HCV envelope glycoproteins, and use these cells to adapt a cell-based membrane fusion assay for HCV entry. We will generate stable human hepatocyte cell lines as appropriate targets for fusion, and optimize the assay for reproducibility, sensitivity and suitability for high-throughput screening (HTS). We will also probe the mechanism of HCV entry using monoclonal antibodies and cell lines expressing putative attachment receptors.Success in the Phase I project will enable the HCV membrane fusion assay to be utilized in Phase II for HTS of libraries of structurally diverse small molecules, in order to identify potent inhibitors of HCV entry. Lead compounds will be screened for in vivo inhibition of HCV infection in a novel transgenic mouse model and mechanisms of action will be probed. Promising compunds will be developed for clinical application for HCV infection.