Development of a HTS assay to screen for inhibitors of HCV core NS3h interactions

描述（由申请人提供）：摘要丙型肝炎病毒（HCV）是肝脏疾病的主要原因，全世界约有1.3亿人感染。目前还没有开发出疫苗，唯一可用的治疗方法是聚乙二醇干扰素α和利巴韦林的组合。由于缺乏新的靶蛋白，开发新的直接作用治疗方法的进展受到很大阻碍。我们已经开发了基于抑制HCV核心（病毒衣壳蛋白，其对于病毒颗粒的组装是必不可少的）的二聚化的高通量筛选测定。这些检测方法现在被制药业采用，使我们能够识别HCV生产的新型抑制剂。基于我们的核心-核心测定的筛选的验证命中被用作分子探针来研究核心与HCV的NS 3解旋酶的相互作用。我们将开发新的能量转移试验，以确定核心NS 3解旋酶相互作用的高亲和力抑制剂。所得化合物将构成新的分子探针，用于了解HCV非结构蛋白如何参与病毒颗粒的组装。这些抑制剂也可以作为药物的基础，与其他抗病毒药物联合使用，以开发比目前基于干扰素的不令人满意的方案更普遍适用的治疗方法。 公共卫生相关性：丙型肝炎影响全球1.3亿人，其中300万人生活在美国，唯一的治疗方法是干扰素和利巴韦林的组合，对不到一半的患者有效。我们将开发一种基于能量转移的特异性检测方法，用于筛选病毒的有效分子探针，该方法基于抑制核心（病毒颗粒组装所必需的衣壳蛋白）和NS 3解旋酶（病毒复制所必需的HCV蛋白）之间的相互作用。这些化合物可能导致开发用于联合治疗丙型肝炎的口服药物。

DESCRIPTION (provided by applicant): Abstract Hepatitis C virus (HCV) is a major cause of liver disease, with about 130 million people infected worldwide. No vaccine has been developed, and the only treatment available is a combination of pegylated interferon alpha and ribavirin. Progress in developing novel direct-acting treatments has been greatly hampered by the dearth of novel target proteins. We have developed high-throughput screening assays based on the inhibition of dimerization of HCV core, the viral capsid protein, which is essential for the assembly of the viral particle. These assays, now adopted by the pharmaceutical industry, have allowed us to identify novel inhibitors of HCV production. A validated hit of a screen based on our core-core assays was used as a molecular probe to study the interaction of core with the NS3 helicase of HCV. We will develop new transfer of energy assays to identify high-affinity inhibitors of the core-NS3 helicase interaction. The resulting compounds will constitute novel molecular probes for understanding how HCV non structural proteins participate in the assembly of the viral particle. These inhibitors may also serve as the basis of drugs to be combined with other anti-viral agents to develop treatments that will be more generally applicable than the current interferon-based unsatisfactory regimen. PUBLIC HEALTH RELEVANCE: Narrative Hepatitis C affects 130 million individuals worldwide, 3 million of which live in the US, and the only treatment, a combination of interferon and ribavirin, is effective for less than half of the patients. We will develop a specific transfer-of energy-based assay screening assay for potent molecular probes of the virus, based on inhibition of interaction between core, the capsid protein essential for assembly of the viral particle, and NS3 helicase, a HCV protein essential for viral replication. These compounds may lead to the development of orally available drugs for combination treatment of Hepatitis C.