High Throughput Assay:Topoisomerase Enzyme Targets (RMI)

描述(由申请人提供)：本提案寻求在高通量分子筛选应用中进一步开发和利用一种新的DNA拓扑异构酶的连续光谱分析方法。这些酶是抗癌、抗菌和抗病毒药物治疗的既定靶点，现在这种强大的检测方法允许快速筛选大型化学库，以寻找这些靶点的新药候选。由于采用了灵敏的分子信标技术，在反应过程中释放了拓扑异构酶，允许单个酶分子对多个底物分子进行周转，因此该方法对于高通量筛选应用尤其可靠和经济。通过筛选2000个成员的小分子文库，已经鉴定出几种新的先导化合物，它们专门针对痘病毒家族的拓扑异构酶，从而验证了该方法的实用性。

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.