High throughput screening of a general anesthetic binding site

描述（由申请人提供）：我们在NCGC提出了一种高通量的全麻药物筛选方法，该筛选方法基于与GABAA模拟物apoferittin中一个特征良好的位点的结合。该分析是基于与增强的1-氨基蒽荧光化合物的竞争。该分析已采用1536孔格式，并显示出良好的信噪比和信噪比，当与阳性对照化合物进行测试时，Z′因子优于0.8。对LOPAC 1280进行了筛选，检索到文库中唯一已知的全麻药物，以及其他几种化合物，其中大多数已通过ITC验证。在主筛选之后，将应用一种新的计算过滤器，然后与NCGC合作进行SAR，聚类和活动分类。验证屏幕将使用ITC来确认和扩展HTS IC50值。除了为进一步开发提供几种探针化合物外，该项目还可能提供有用的实验室试剂并为蛋白质配体化学提供信息。

DESCRIPTION (provided by applicant): We propose a high throughput screen at NCGC for general anesthetics based on binding to a well characterized site in a GABAA mimic, apoferritin. The assay is based on competition of compounds with enhanced 1-aminoanthracene fluorescence. The assay has been taken to a 1536-well format, and shows excellent signal: background and signal: noise ratios, and when tested with positive control compounds, achieves Z' factors better than 0.8. The LOPAC 1280 set has been screened which resulted in retrieval of the only know general anesthetic in the library, and several other compounds most of which were validated by ITC. After primary screen, a novel computational filter will be applied, followed by SAR, clustering and triage of actives in collaboration with NCGC. Validation screens will use ITC to confirm and extend the HTS IC50 values. In addition to providing several probe compounds for further development, this project is likely to provide useful laboratory reagents and inform protein-ligand chemistry. PUBLIC HEALTH RELEVANCE: General anesthetics are used in over 40 million patients in the US each year, and, although known to have worrisome side effects, have recently been discovered to have durable cognitive effects. New and more specific drugs are required to avoid these side effects. We propose a high throughput screen of the molecular library compounds for binding to a favorable anesthetic site in order to generate probe compounds for further development as safe general anesthetics.