High throughput screen for small molecule inhibitors of Ran regulated functions

在2011-2012年，我们开发了正交化学发光筛选来鉴定特异性作用抑制剂。我们还设计并测试了基于细胞的测定，以监测抑制剂对RanGTP和importin α 1/importin β调节的核输入和有丝分裂纺锤体组装的影响。我们用20个主屏幕点击测试了这些辅助屏幕。虽然我们确定了至少一个经过验证的初步筛选命中，在二级筛选中也显示了活性，但我们现在正在与NCATS（前身为NCGC）合作，以优化二级筛选，用于使用整套1000种候选抑制剂进行自动二级筛选。我们的目标是确定特异性作用的抑制性化合物，显示预期的生物活性。这些数据是确保R 03项目第二年资金所必需的，我们将专注于最佳识别的抑制剂，并分析其作用机制，并基于结构-活性关系分析优化其性能。

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.