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）：该提案旨在在高吞吐量分子筛选应用中进一步发展和利用DNA拓扑异构酶的新型连续光谱测定法。这些酶是针对抗癌，抗菌和抗病毒药物疗法的靶标，而这种强大的测定方法现在可以快速筛选大型化学文库为这些靶标的新药候选物。由于利用了敏感的分子信标技术，因此对于高通量筛选应用而言，该测定法对高通量筛选应用特别强大，并且在反应过程中释放了拓扑异构酶，以允许单个酶分子的多个底物分子的营业率。通过筛选一个2000成员的小分子库来验证该测定的效用，从中已经确定了几种新型的铅化合物，这些化合物专门针对了鼠盆家族拓扑异构酶。