High throughput screen for small molecule inhibitors of Ran regulated functions

2011-2012年，我们开发了正交化学发光筛选来鉴定特异性抑制剂。我们还设计并测试了基于细胞的检测方法，以监测抑制剂对RanGTP和输入蛋白α 1/输入蛋白β调节的核输入和有丝分裂纺锤体组装的影响。我们用20个主屏幕来测试这些辅助屏幕。虽然我们已经确定了至少一个经过验证的主筛命中，并且在次级筛中也显示出活性，但我们现在正在与NCATS（以前的NCGC）合作，优化次级筛，以实现包含全部1000种候选抑制剂的自动化次级筛。我们的目标是确定特异性抑制化合物，显示预期的生物活性。该数据是为该项目第二年获得R03资助所必需的，我们将重点关注已确定的最佳抑制剂，并根据结构-活性关系分析分析其作用机制并优化其性能。

In 2011-2012 we developed orthogonal chemiluminescence screen to identify aspecifically acting inhibitors. We also designed and tested cell-based assays to monitor the effects of the inhibitors on the RanGTP and importin alpha1/importin beta-regulated nuclear import and mitotic spindle assembly. We tested those secondary screens with a group of 20 primary screen hits. Although we identified at least one validated primary screen hit that displayed activity also in the secondary screens, we are now collaborating with NCATS (formerly NCGC) to optimize the secondary screens for automated secondary screens with the entire set of 1000 candidate inhibitors. Our objective is to identify specifically acting inhibitory compounds which display the expected biological activity. This data is required to secure R03 funding for the second year of the project where we will focus on the best identified inhibitor(s) and analyze their mechanism of action and optimize their performance based on structure-activity relationship analysis.