Development of Hepatitis C Virus entry inhibitors

描述（由申请人提供）：开发新的治疗丙型肝炎病毒（HCV）感染的药物是一个主要的公共卫生优先事项。我们选择HCV作为发现和开发新型抗病毒药物的靶点。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试验，使用具有良好特征的生物相关细胞试剂，准确地概括HCV的进入。在I期项目中，我们将开发表达天然HCV包膜糖蛋白融合形式的稳定细胞系，并使用这些细胞适应HCV进入的基于细胞的膜融合试验。我们将生成稳定的人肝细胞系作为合适的融合靶点，并优化实验的重现性、灵敏度和高通量筛选（HTS）的适用性。我们还将利用单克隆抗体和表达推测的附着受体的细胞系来探索HCV进入的机制。I期项目的成功将使HCV膜融合试验能够在II期用于结构多样化小分子文库的HTS，以确定HCV进入的有效抑制剂。我们将在一种新型转基因小鼠模型中筛选先导化合物来抑制HCV感染，并探讨其作用机制。有希望的化合物将被开发用于HCV感染的临床应用。

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.