A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor
文献类型：期刊论文
| 作者 | Hui, X; Gao, J; Xie, X ; Suto, N; Ogiku, T; Wang, MW
|
| 刊名 | ACTA PHARMACOLOGICA SINICA
 |
| 出版日期 | 2005-10 |
| 卷号 | 26期号:10页码:1175-1180 |
| 关键词 | nicotinic acetylcholine receptor scintillation proximity assay high-throughput screening |
| ISSN号 | 1671-4083 |
| DOI | 10.1111/j.1745-7254.2005.00202.x |
| 文献子类 | Article |
| 英文摘要 | Aim: To develop a homogeneous high-throughput screening (HTS) assay based on scintillation proximity assay (SPA) technology for identification of novel alpha 4 beta 2 nicotinic acetylcholine receptor (nAChR) modulators. Methods: Membrane preparation of HEK293 cells expressing alpha 4 beta 2 nAChR, [H-3]cytisine and wheat germ agglutinin (WGA)-coupled microbeads were used to develop an HTS assay based on SPA technology. This method was validated against a conventional filter binding approach and applied to large-scale screening of a library containing 32 000 synthetic compounds. Intracellular calcium measurement was carried out to verify the bioactivities of the hits found by the SPA assay. Results: IC50 values of 2 reference compounds (epibatidine and RJR 2403) determined by SPA and filter binding methods were comparable and consistent with those reported elsewhere. A total of 54 compounds, showing more than 60% competitive inhibition on [H-3]cytisine binding to alpha 4 beta 2 nAChR, were identified initially following an HTS campaign. Secondary screening confirmed that 17 compounds with novel chemical structures possessed relatively high binding affinity to alpha 4 beta 2 nAChR (K-i < 2 mu mol/L). Eight compounds displayed antagonistic effects with > 50% inhibition on ABT-594-induced calcium mobilization while none showed any agonist activity. Conclusions: This homogeneous binding assay is a highly efficient, amenable to automation and robust tool to screen potential alpha 4 beta 2 nAChR modulators in an HTS setting. Its application may be expanded to other membrane receptors and ion channels. |
| WOS关键词 | SCINTILLATION-PROXIMITY ASSAYS ; ABT-594 ; PHARMACOLOGY ; CELLS |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| CSCD记录号 | CSCD:2208486 |
| WOS记录号 | WOS:000232346000004 |
| 出版者 | BLACKWELL PUBLISHING |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/273790]  |
| 专题 | 国家新药筛选中心 |
| 通讯作者 | Wang, MW |
| 作者单位 | 1.Tanabe Seiyaku Co Ltd, Discovery & Pharmacol Res Labs, Toda, Saitama 3358505, Japan 2.Chinese Acad Sci, Grad Sch, Chinese Acad Sci,Shanghai Inst Mat Med, Shanghai Inst Biol Sci,Nat Ctr Drug Screening, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Hui, X,Gao, J,Xie, X,et al. A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor[J]. ACTA PHARMACOLOGICA SINICA,2005,26(10):1175-1180. |
| APA | Hui, X,Gao, J,Xie, X,Suto, N,Ogiku, T,&Wang, MW.(2005).A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor.ACTA PHARMACOLOGICA SINICA,26(10),1175-1180. |
| MLA | Hui, X,et al."A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor".ACTA PHARMACOLOGICA SINICA 26.10(2005):1175-1180. |
入库方式： OAI收割
来源：上海药物研究所
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