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

开发 HTS 测定法来筛选 HCV 核心 NS3h 相互作用的抑制剂

基本信息

批准号：
8099275
负责人：
ARTHUR Donny STROSBERG
金额：
$ 19.8万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8099275
关键词：
Adopted Adverse effects Affect Affinity Antibodies Award Biological Assay Capsid Proteins Complex Core Protein Development Dimerization Dimethyl Sulfoxide Dose Drug Combinations Drug Industry Energy Transfer Enzymes Europium FDA approved Fluorescence Resonance Energy Transfer Future Glutathione S-Transferase Glycoproteins Goals Grant Hepatitis C Hepatitis C virus Individual Inhibitory Concentration 50 Interferons Investigation Label Laboratories Lead Libraries Life Life Cycle Stages Lipids Liver diseases Miniaturization Modeling Molecular Bank Molecular Probes Mutation Network-based Patients Pegylated Interferon Alfa Peptide Hydrolases Peptide antibodies Peptides Performance Pharmaceutical Preparations Phase III Clinical Trials Phosphoproteins Plague Polymerase Production Proteins RNA Binding Regimen Ribavirin Screening procedure Serine Protease Signal Transduction Solutions Structural Protein Tertiary Protein Structure Testing Time United States National Institutes of Health Vaccines Validation Variant Viral Viral Proteins Virion Virus Virus Inhibitors Work abstracting allophycocyanin base cytotoxicity fluorophore helicase hepatitis C virus NS3 protein hepatoma cell high throughput screening inhibitor/antagonist interest novel particle programs protein protein interaction replicase response small molecule virus core

项目摘要

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.

描述（由申请人提供）：摘要丙型肝炎病毒（HCV）是肝病的主要原因，全球约有1.3亿人感染。尚未开发疫苗，唯一可用的治疗方法是卵甲状干扰素α和利巴韦林的组合。新型靶蛋白的缺乏，开发新型直接作用治疗的进展受到了极大的阻碍。我们已经根据抑制HCV核心的二聚化（病毒式衣壳蛋白的二聚化，这对于病毒颗粒的组装至关重要）开发了高通量筛选测定法。这些现在由制药行业采用的测定方法使我们能够鉴定出新型HCV生产抑制剂。基于我们的核心核测定法的筛选效果被验证的命中被用作分子探针，以研究核心与HCV的NS3解旋酶的相互作用。我们将开发新的能量测定的转移，以鉴定Core-NS3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，以了解HCV非结构蛋白如何参与病毒颗粒的组装。这些抑制剂还可以作为与其他抗病毒药物相结合的药物的基础，以开发比当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：叙事性丙型肝炎会影响全球1.3亿个人，其中300万人生活在美国，而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。我们将基于抑制核心之间的相互作用，CapsID蛋白（用于组装病毒颗粒的组合蛋白和NS3解旋酶，这是病毒复制必不可少的HCV蛋白，因此，我们将开发针对病毒有效分子探针的特定转移转移测定法。这些化合物可能导致开发口服的药物，用于联合治疗乙型肝炎。