arobusthomogeneousbindingassayfor42nicotinicacetylcholinereceptor
文献类型：期刊论文
| 作者 | Hui Xin1; Gao Jie1; Xie Xin1 ; Naoki Suto2; Tsuyshi Ogiku2; Wang Mingwei1
|
| 刊名 | actapharmacologicasinica
 |
| 出版日期 | 2005 |
| 卷号 | 026期号:010页码:1175 |
| 关键词 | α4β2 烟碱酸 乙酰胆碱受体 微生物凝集素 实验室检查 |
| ISSN号 | 1671-4083 |
| 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 α4β2 nicotinic acetylcholine receptor （nAChR） modulators. Methods： Membrane preparation of HEK293 cells expressing α4β2 nAChR, ^3Hcytisine 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 R JR 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 ^3Hcytisine binding to α4β2 nAChR, were identified initially following an HTS campaign. Secondary screening confirmed that 17 compounds with novel chemical structures possessed relatively high binding affinity to α4β2 nAChR （Ki〈2μmol/L）. Eight compounds displayed antagonistic effects with 〉50% inhibi- tion 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 α4β2 nAChR modulators in an HTS setting. Its application may be expanded to other membrane receptors and ion channels. |
| 语种 | 英语 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/283024]  |
| 专题 | 中国科学院上海药物研究所 |
| 作者单位 | 1.中国科学院上海药物研究所 2.Discovery and Pharmacology Research Laboratories, Tanabe Seiyaku Co, 2-50, Kawagishi 2-Chome, Toda, Saitama 335-8505 |
| 推荐引用方式 GB/T 7714 | Hui Xin,Gao Jie,Xie Xin,et al. arobusthomogeneousbindingassayfor42nicotinicacetylcholinereceptor[J]. actapharmacologicasinica,2005,026(010):1175. |
| APA | Hui Xin,Gao Jie,Xie Xin,Naoki Suto,Tsuyshi Ogiku,&Wang Mingwei.(2005).arobusthomogeneousbindingassayfor42nicotinicacetylcholinereceptor.actapharmacologicasinica,026(010),1175. |
| MLA | Hui Xin,et al."arobusthomogeneousbindingassayfor42nicotinicacetylcholinereceptor".actapharmacologicasinica 026.010(2005):1175. |
入库方式： OAI收割
来源：上海药物研究所
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