Integration of virtual screening with high-throughput flow cytometry to identify novel small molecule formylpeptide receptor antagonists
Integration of virtual screening with high-throughput flow cytometry to identify novel small molecule formylpeptide receptor antagonists
复制标题
DOI：
10.1124/mol.105.014068
复制
发表时间：
2005-11-01
影响因子：
3.6
通讯作者：
Oprea, TI
中科院分区：
文献类型：
--
作者：
Edwards, BS;Bologa, C;Oprea, TI
甲酰肽受体(Fpr)家族的G蛋白偶联受体有助于在慢性炎症部位定位和激活组织损伤的白细胞。我们开发了一个FPR同源模型和药效团(分别基于牛视紫质晶体结构和已知的FPR配体)，用于电子筛选类似的48万个类药物小分子。然后用HyperCyt流式细胞仪平台以高通量、直接测量人fpr结合的免洗试验对4,324种与药效团匹配的化合物的子集进行物理筛选，并以40/min的速度分析样品(每个样品类似于2亩L的2,500个细胞)。从52个确认的命中(命中率1.2%)中，我们鉴定了30个潜在的先导化合物(抑制常数，K-I=1-32µM)，代表9个不同的化学家族。一个家族中的四个化合物是弱部分激动剂。其他人都是敌手。这种虚拟筛选方法将物理筛选命中率提高了12倍(与随机化合物收集的0.1%命中率相比)，为识别小分子拮抗剂提供了一种有效的过程。
The formylpeptide receptor (FPR) family of G-protein-coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. We developed a FPR homology model and pharmacophore ( based on the bovine rhodopsin crystal structure and known FPR ligands, respectively) for in silico screening of similar to 480,000 drug-like small molecules. A subset of 4324 compounds that matched the pharmacophore was then physically screened with the HyperCyt flow cytometry platform in high-throughput, no-wash assays that directly measure human FPR binding, with samples (each similar to 2500 cells in 2 mu l) analyzed at 40/min. From 52 confirmed hits (1.2% hit rate), we identified 30 potential lead compounds ( inhibition constant, K-i = 1 - 32 mu M) representing nine distinct chemical families. Four compounds in one family were weak partial agonists. All others were antagonists. This virtual screening approach improved the physical screening hit rate by 12-fold ( versus 0.1% hit-rate in a random compound collection), providing an efficient process for identifying small molecule antagonists.