High throughput screening of a general anesthetic binding site

全身麻醉剂结合位点的高通量筛选

基本信息

批准号：
7761812
负责人：
Roderic G Eckenhoff
金额：
$ 2.5万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-20 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7761812
关键词：
Adverse effects Anesthesia procedures Anesthetics Apoferritin Binding Binding Sites Biological Assay Blood - brain barrier anatomy Breathing Calorimetry Cells Chemistry Classification Cognitive Collaborations Collection Data Development Ferritin Fluorescence General Anesthesia General anesthetic drugs Inhibitory Concentration 50 Injectable Laboratories Libraries Ligands Measures Medical Molecular Bank Noise Operative Surgical Procedures Organism Patients Penetration Pharmaceutical Preparations Powder dose form Property Proteins Reagent Retrieval Running Series Signal Transduction Site Testing Thermodynamics Time Titrations Toxic effect Transmembrane Domain Triage Validation Water analog base computerized tools drug candidate flexibility follow-up high throughput screening in vivo novel public health relevance receptor repository small molecule

项目摘要

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.

描述（由申请人提供）：我们建议在 NCGC 进行基于与 GABAA 模拟物脱铁铁蛋白中明确表征的位点结合的全身麻醉剂高通量筛选。该测定基于具有增强 1-氨基蒽荧光的化合物的竞争。该检测采用 1536 孔格式，显示出出色的信号：背景和信号：噪声比，并且当使用阳性对照化合物进行测试时，Z' 因子优于 0.8。 LOPAC 1280 套件经过筛选，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，其中大部分都经过了 ITC 的验证。初步筛选后，将应用一种新颖的计算过滤器，然后与 NCGC 合作进行活性物质的 SAR、聚类和分类。验证屏幕将使用 ITC 来确认和扩展 HTS IC50 值。除了提供几种探针化合物供进一步开发外，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。公共健康相关性：美国每年有超过 4000 万患者使用全身麻醉剂，尽管已知全身麻醉剂具有令人担忧的副作用，但最近发现它具有持久的认知影响。需要新的、更具体的药物来避免这些副作用。我们建议对分子库化合物进行高通量筛选，以结合到有利的麻醉位点，以生成探针化合物，进一步开发为安全的全身麻醉剂。