Development of a complex scintillation proximity assay for high-throughput screening of PPAR gamma modulators
文献类型：期刊论文
| 作者 | Wu, B; Gao, J; Wang, MW
|
| 刊名 | ACTA PHARMACOLOGICA SINICA
 |
| 出版日期 | 2005-03 |
| 卷号 | 26期号:3页码:339-344 |
| 关键词 | peroxisome proliferator-activated receptor gamma retinoid X receptor alpha scintillation proximity assay high-throughput screening |
| ISSN号 | 1671-4083 |
| DOI | 10.1111/j.1745-7254.2005.00040.x |
| 文献子类 | Article |
| 英文摘要 | Aim: To develop a complex high-throughput screening (HTS) assay based on scintillation proximity assay (SPA) technology for identification of novel peroxisome proliferator-activated receptor gamma (PPAR gamma) modulators. Methods: Full-length PPAR gamma and retinoid X receptor alpha (RXR alpha), biotinylated PPAR response element (PPRE), [H-3]BRL49653 and streptavidin-coated FlashPlate or microbead were used to develop an HTS assay based on SPA technology. This 'ABCDE' method was validated against conventional hydroxyapatite (HA) assay and applied to large-scale screening of 16 000 synthetic compounds and natural product extracts. Results: (1) IC50 values of positive control compounds (BRL49653 and troglitazone) obtained from the 'ABCDE' method and HA assay were comparable and consistent with those reported elsewhere; (2) Approximately 178 showing more than 70% competitive inhibition on BRL49653 binding to PPARy, were identified initially by the 'ABCDE' method (microbead); (3) Secondary screening using FlashPlate and cross-reactivity studies with RAR alpha, beta, gamma and RXR alpha, beta, gamma confirmed that 12 compounds possessed specific PPARy binding properties including 2 with IC50 values less than 0.5 mu mol/L and novel chemical structures. Conclusions: The 'ABCDE' method using either FlashPlate or microbead, is a highly efficient, automatable, and robust tool to screen potential PPARy modulators in HTS setting. Its application may be expanded to other nuclear receptors that form heterodimers upon activation. |
| WOS关键词 | PROLIFERATOR-ACTIVATED RECEPTOR ; RETINOID-X-RECEPTOR ; PEROXISOME-PROLIFERATOR ; BINDING ; SUPERFAMILY ; DNA |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| CSCD记录号 | CSCD:1998382 |
| WOS记录号 | WOS:000227647800013 |
| 出版者 | ACTA PHARMACOLOGICA SINICA |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/273896]  |
| 专题 | 国家新药筛选中心 |
| 通讯作者 | Wang, MW |
| 作者单位 | 1.Chinese Acad Sci, Grad Sch, Shanghai, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Biol Sci, Natl Ctr Drug Screening, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wu, B,Gao, J,Wang, MW. Development of a complex scintillation proximity assay for high-throughput screening of PPAR gamma modulators[J]. ACTA PHARMACOLOGICA SINICA,2005,26(3):339-344. |
| APA | Wu, B,Gao, J,&Wang, MW.(2005).Development of a complex scintillation proximity assay for high-throughput screening of PPAR gamma modulators.ACTA PHARMACOLOGICA SINICA,26(3),339-344. |
| MLA | Wu, B,et al."Development of a complex scintillation proximity assay for high-throughput screening of PPAR gamma modulators".ACTA PHARMACOLOGICA SINICA 26.3(2005):339-344. |
入库方式： OAI收割
来源：上海药物研究所
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