Optimizing Hepatitis C virus NS5B Polymerase Inhibitors

优化丙型肝炎病毒 NS5B 聚合酶抑制剂

• 批准号: 6772443
• 负责人: Y SUDHAKARA BABU
• 金额: $ 30万
• 依托单位: BIOCRYST PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-05 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6772443
• 关键词: drug design /synthesis /production; drug screening /evaluation; enzyme inhibitors; hepatitis C; hepatitis C virus; microorganism disease chemotherapy; protein purification; recombinant virus; replicase; replicon; virus genetics; virus replication

DESCRIPTION (provided by applicant): The over-all objective of this project is identifying and developing selective nontoxic inhibitors of the hepatitis C virus NS5B polymerase and eventually testing them as therapeutics for chronic hepatitis C. At least four million people in the United States have chronic hepatitis C and the recent NIH Consensus Conference suggested that this number is actually two-three fold greater. The high rate of progression to chronic infection (70-80%), liver disease (>50%), and the worldwide distribution of chronic hepatitis C, makes it a major cause of morbidity and mortality. The production of HCV vaccines is a difficult challenge and an intensive development of molecular based HCV therapeutics should be pursued. A major problem in HCV research has been the development of model systems to study HCV replication and identifying effective molecular based therapeutics. We have developed methods for expressing in E. coli and purifying mg quantities of purified enzymatically active NS5B polymerase protein. In addition, crystal structures of NS5B polymerase have been solved and robust cell based replicon systems can be used to test for inhibition of HCV RNA replication. These developments permit virtual screening of small molecules as candidate NS5B polymerase inhibitors. The specific objectives of this project are: 1) to express and purify sufficient quantities of recombinant HCV NS5B polymerase for in vitro assays and crystal growth studies; 2) To test additional compounds, already identified by structure-based virtual screening, for their ability to directly inhibit the HCV NS5B polymerase using an in vitro assay. These results and current NS5B structural information will be used to further design, synthesize and test modifications of lead compounds; 3) To conduct co-crystal growth studies of NS5B, a template, and an inhibitor with the goal of using this ternary complex structure to design and optimize NS5B polymerase inhibitors.

描述（由申请人提供）：该项目的总体目标是确定和开发丙型肝炎病毒NS5B聚合酶的选择性无毒抑制剂，并最终将其作为慢性丙型肝炎的治疗方法进行测试。在美国至少有400万人患有慢性丙型肝炎，最近的NIH共识会议表明，这个数字实际上是2 - 3倍。慢性丙型肝炎发展为慢性感染（70-80%）、肝脏疾病（bbb50 %）的高进展率，以及慢性丙型肝炎在世界范围内的分布，使其成为发病率和死亡率的主要原因。丙型肝炎病毒疫苗的生产是一项艰巨的挑战，应大力开发基于分子的丙型肝炎病毒疗法。HCV研究的一个主要问题是开发模型系统来研究HCV复制和确定有效的分子治疗方法。我们已经开发了在大肠杆菌中表达和纯化纯化酶活性NS5B聚合酶蛋白的方法。此外，NS5B聚合酶的晶体结构已经得到解决，基于细胞的复制子系统可以用于测试HCV RNA复制的抑制。这些进展允许虚拟筛选小分子作为候选NS5B聚合酶抑制剂。该项目的具体目标是：1)表达和纯化足够数量的重组HCV NS5B聚合酶，用于体外检测和晶体生长研究；2)通过体外实验测试已经通过基于结构的虚拟筛选确定的其他化合物直接抑制HCV NS5B聚合酶的能力。这些结果和现有的NS5B结构信息将用于进一步设计、合成和测试先导化合物的修饰；3)开展NS5B、模板、抑制剂共晶生长研究，利用该三元配合物结构设计优化NS5B聚合酶抑制剂。