High Throughput Assay:Topoisomerase Enzyme Targets (RMI)

描述（由申请人提供）：本提案旨在进一步开发和利用高通量分子筛选应用，一种新的DNA拓扑异构酶连续光谱分析方法。这些酶是抗癌、抗菌和抗病毒药物治疗的既定靶标，这种强大的检测方法现在可以快速筛选大型化学文库，为这些靶标寻找新的候选药物。由于使用了敏感的分子信标技术，该分析对于高通量筛选应用尤其稳健和经济，并且在反应过程中释放拓扑异构酶以允许单个酶分子周转多个底物分子。通过筛选2000个成员的小分子文库，已经确定了几种新的先导化合物，这些先导化合物专门针对痘病毒家族拓扑异构酶，验证了该分析的实用性。

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.