High throughput screening of a general anesthetic binding site

描述（由申请人提供）：我们提出在NCGC基于与GABAA模拟物脱铁铁蛋白中充分表征的位点的结合进行全身麻醉剂的高通量筛选。该测定基于具有增强的1-氨基蒽荧光的化合物的竞争。该测定已采用1536孔格式，并显示出优异的信号：背景和信噪比，并且当用阳性对照化合物测试时，实现了优于0.8的Z'因子。筛选了LOPAC 1280套件，检索到了库中唯一已知的全身麻醉剂，以及其他几种化合物，其中大部分已由ITC验证。在初步筛选后，将应用一种新的计算过滤器，然后与NCGC合作对活性物质进行SAR、聚类和分类。验证屏幕将使用ITC确认和扩展HTS IC 50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。 公共卫生相关性：在美国，每年有超过4000万患者使用全身麻醉剂，尽管已知有令人担忧的副作用，但最近发现具有持久的认知影响。需要新的和更具体的药物来避免这些副作用。我们提出了一个高通量筛选的分子库化合物结合到一个有利的麻醉位点，以产生探针化合物，进一步发展为安全的全身麻醉药。

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.