HCV NS5B POLYMERASE MUTATIONS: BIOLOGY/PHARMACOLOGY

HCV NS5B 聚合酶突变：生物学/药理学

• 批准号: 7381286
• 负责人: CURT H. HAGEDORN
• 金额: $ 7.43万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-27 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381286
• 关键词: RNA; bioinformatics; gene mutation; hepatitis C; infection; model; mutant; oligonucleotides; pharmacology; proteins; public health

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. An estimated four to ten million people in the United States have chronic hepatitis C. Although treatment for HCV has improved only an estimated 20% of patients are able to take current therapies because of contraindications. Developing new therapeutics or vaccines for hepatitis C remains a major unsolved public health challenge. The hypothesis that this study will test is that variants of the HCV NS5B polymerase, arising due to the high mutation rate of HCV, can result in resistance to NS5B polymerase inhibitors and that identifying these mutants will be essential for developing effective new antivirals. Identifying variants of NS5B which are resistant to model polymerase inhibitors should aid in developing inhibitors that target motifs in the NS5B polymerase that are essential for HCV replication. In addition, some variants of NS5B may have more favorable properties for determining the structure of a ternary complex of NS5B, an RNA template, and an inhibitor. Progress has been made on the bioinformatics analysis of HCV sequence data obtained by colleagues at the CDC (J. Med. Virol.). In addition, over 20 mg of one NS5B variant was purified and is currently being used in co-crystal growth studies. The Specific Objectives for the next year are: To identify additional NS5B mutations occurring during chronic HCV infection and select several variants to study; To further improve methods of expressing and purifying sufficient quantities of recombinant HCV NS5B polymerase proteins for crystal growth studies; and To initiate crystal growth studies of at least two other NS5B variants, an oligonucleotide template, and an inhibitor. This will permit structure based approaches to be used in developing therapeutic NS5B inhibitors with the goal of linking strengths regarding HCV proteins (Hagedorn Lab) with expertise in Medicinal Chemistry for an NIH grant applications.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。据估计，美国有 4 至 1000 万人患有慢性丙型肝炎。尽管 HCV 治疗有所改善，但由于禁忌症，估计只有 20% 的患者能够接受当前的治疗。开发丙型肝炎新疗法或疫苗仍然是一个尚未解决的重大公共卫生挑战。本研究将测试的假设是，由于 HCV 高突变率而产生的 HCV NS5B 聚合酶变体可能导致对 NS5B 聚合酶抑制剂的耐药性，并且识别这些突变体对于开发有效的新型抗病毒药物至关重要。鉴定对模型聚合酶抑制剂具有抗性的 NS5B 变体应该有助于开发针对 NS5B 聚合酶中 HCV 复制所必需的基序的抑制剂。此外，NS5B的一些变体可能具有更有利的特性来确定NS5B、RNA模板和抑制剂的三元复合物的结构。 CDC 同事获得的 HCV 序列数据的生物信息学分析已取得进展 (J. Med. Virol.)。此外，还纯化了超过 20 毫克的一种 NS5B 变体，目前正用于共晶生长研究。明年的具体目标是： 识别慢性 HCV 感染期间发生的其他 NS5B 突变，并选择几种变体进行研究；进一步改进表达和纯化足量重组 HCV NS5B 聚合酶蛋白用于晶体生长研究的方法；启动至少两种其他 NS5B 变体、寡核苷酸模板和抑制剂的晶体生长研究。这将允许基于结构的方法用于开发治疗性 NS5B 抑制剂，其目标是将 HCV 蛋白（哈格多恩实验室）的优势与药物化学专业知识联系起来，以申请 NIH 拨款。