High Throughput Assay:Topoisomerase Enzyme Targets (RMI)

高通量检测：拓扑异构酶靶标 (RMI)

• 批准号: 7022489
• 负责人: JAMES T. STIVERS
• 金额: $ 8.15万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7022489
• 关键词: DNA gyrase; DNA topoisomerases; antineoplastics; bacterial proteins; biotechnology; drug discovery /isolation; drug screening /evaluation; enzyme inhibitors; enzyme substrate; high throughput technology; small molecule; smallpox virus; technology /technique development; virus protein

DESCRIPTION (provided by applicant): This proposal seeks to further develop and utilize, in high throughput molecular screening applications, a novel continuous spectroscopic assay for DNA topoisomerases. These enzymes are established targets for anticancer, antibacterial and antiviral drug therapies, and this robust assay now allows for rapid screening of large chemical libraries for new drug candidates for these targets. The assay is especially robust and economical for high-throughput screening applications because sensitive molecular beacon technology is utilized, and the topoisomerase is released during the reaction to allow turnover of multiple substrate molecules by a single molecule of enzyme. The utility of the assay has been validated, by screening a 2000 member small molecule library from which several novel lead compounds have been identified that specifically target a poxvirus family topoisomerase.

描述（由申请人提供）：本提案旨在在高通量分子筛选应用中进一步开发和利用一种新颖的 DNA 拓扑异构酶连续光谱测定法。这些酶是抗癌、抗菌和抗病毒药物治疗的既定靶点，这种强大的检测方法现在可以快速筛选大型化学库，寻找针对这些靶点的新候选药物。该测定对于高通量筛选应用特别稳健且经济，因为利用了敏感的分子信标技术，并且在反应过程中释放了拓扑异构酶，以允许单个酶分子对多个底物分子进行周转。通过筛选 2000 个成员的小分子库，已验证了该测定法的实用性，其中已鉴定出几种专门针对痘病毒家族拓扑异构酶的新型先导化合物。