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: 6695860
• 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 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6695860
• 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鉴定和开发丙型肝炎病毒NS 5 B聚合酶的选择性无毒抑制剂，并最终测试它们作为慢性丙型肝炎的治疗药物。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。进展为慢性感染（70-80%）、肝病（>50%）的高比率以及慢性丙型肝炎的全球分布使其成为发病率和死亡率的主要原因。HCV疫苗的生产是一个困难的挑战，应该追求基于分子的HCV治疗剂的密集开发。HCV研究中的一个主要问题是研究HCV复制和鉴定有效的基于分子的治疗方法的模型系统的开发。我们已经开发了在E. coli中，并纯化mg量的纯化的酶活性NS 5 B聚合酶蛋白。此外，NS 5 B聚合酶的晶体结构已被解析，并且基于细胞的稳健复制子系统可用于测试HCV RNA复制的抑制。这些发展允许虚拟筛选作为候选NS 5 B聚合酶抑制剂的小分子。本项目的具体目标是：1）表达和纯化足够量的重组HCV NS 5 B聚合酶用于体外测定和晶体生长研究; 2）使用体外测定测试已经通过基于结构的虚拟筛选鉴定的其他化合物直接抑制HCV NS 5 B聚合酶的能力。这些结果和现有的NS 5 B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）进行NS 5 B、模板和抑制剂的共晶体生长研究，目的是使用这种三元复合物结构来设计和优化NS 5 B聚合酶抑制剂。