Development of Hepatitis C Virus entry inhibitors

描述(由申请人提供)：丙型肝炎病毒(丙型肝炎病毒)感染的新治疗药物的开发是一项主要的公共卫生优先事项。我们选择进入丙型肝炎病毒作为发现和开发新型抗病毒药物的目标。这个第一阶段项目的总体目标是开发一种新的基于细胞的膜融合分析方法，使用特征良好的、生物相关的细胞试剂准确地概括丙型肝炎病毒进入的情况。在第一阶段项目中，我们将开发稳定的表达融合形式的天然丙型肝炎病毒包膜糖蛋白的细胞系，并使用这些细胞来适应基于细胞的膜融合分析以适应丙型肝炎病毒进入。我们将建立稳定的人肝细胞系作为合适的融合目标，并优化检测方法的重复性、灵敏度和高通量筛选(HTS)的适用性。我们还将使用单抗和表达可能的附着受体的细胞系来探索丙型肝炎病毒进入的机制。第一阶段项目的成功将使丙型肝炎病毒膜融合试验能够在第二阶段用于结构多样化的小分子文库的HTS，以确定有效的丙型肝炎病毒进入的抑制剂。将在一种新的转基因小鼠模型中筛选先导化合物对体内丙型肝炎病毒感染的抑制作用，并探讨其作用机制。有希望开发出治疗丙型肝炎病毒感染的药物。

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.