Characterizing Cannabinoid CB2 Receptor Ligands Using DiscoveRx PathHunter™ β-Arrestin Assay

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS SLAS Discovery Pub Date : 2009-01-01 DOI:10.1177/1087057108327329

Debra McGuinness , Asra Malikzay , Richard Visconti , Karen Lin , Marvin Bayne , Frederick Monsma , Charles A. Lunn

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Abstract

The authors have characterized a set of cannabinoid CB₂ receptor ligands, including triaryl bis sulfone inverse agonists, in a cell-based receptor/β-arrestin interaction assay (DiscoveRx PathHunter™). The results were compared with results using a competitive ligand binding assay, and with effects on forskolin-stimulated cAMP levels (PerkinElmer LANCE™). The authors show good correlation between the 3 assay systems tested, with the β-arrestin protein complementation assay exhibiting a more robust signal than the cAMP assay for cannabinoid CB₂ agonists. Further assay validation shows that DiscoveRx PathHunter™ HEK293 CB₂ β-arrestin assay can be carried out from cryopreserved cell suspensions, eliminating variations caused by the need for multiple cell pools during live cell screening campaigns. These results, and the authors’ results evaluating a test set of random library compounds, validate the use of ligand-induced interaction between the human cannabinoid CB₂ receptor and β-arrestin as an appropriate and valuable screening platform for compounds specific for the cannabinoid CB₂ receptor.

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使用DiscoveRx PathHunter™β-阻滞蛋白检测表征大麻素CB2受体配体

作者在基于细胞的受体/β-抑制素相互作用试验（DiscoveRx PathHunter™）中表征了一组大麻素CB2受体配体，包括三芳基双砜逆激动剂。结果与竞争配体结合试验的结果进行了比较，并对福斯克林刺激的cAMP水平（PerkinElmer LANCE™）的影响进行了比较。作者展示了三种检测系统之间良好的相关性，β-抑制蛋白互补检测比大麻素CB2激动剂的cAMP检测显示出更强的信号。进一步的实验验证表明，DiscoveRx PathHunter™HEK293 CB2 β-阻滞蛋白实验可以从冷冻保存的细胞悬液中进行，消除了在活细胞筛选活动中需要多个细胞池引起的变化。这些结果，以及作者对随机文库化合物测试集的评估结果，验证了使用配体诱导的人类大麻素CB2受体和β-抑制素之间的相互作用作为大麻素CB2受体特异性化合物的适当和有价值的筛选平台。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

SLAS Discovery Chemistry-Analytical Chemistry

CiteScore

7.00

自引率

3.20%

发文量

审稿时长

39 days

期刊介绍： Advancing Life Sciences R&D: SLAS Discovery reports how scientists develop and utilize novel technologies and/or approaches to provide and characterize chemical and biological tools to understand and treat human disease. SLAS Discovery is a peer-reviewed journal that publishes scientific reports that enable and improve target validation, evaluate current drug discovery technologies, provide novel research tools, and incorporate research approaches that enhance depth of knowledge and drug discovery success. SLAS Discovery emphasizes scientific and technical advances in target identification/validation (including chemical probes, RNA silencing, gene editing technologies); biomarker discovery; assay development; virtual, medium- or high-throughput screening (biochemical and biological, biophysical, phenotypic, toxicological, ADME); lead generation/optimization; chemical biology; and informatics (data analysis, image analysis, statistics, bio- and chemo-informatics). Review articles on target biology, new paradigms in drug discovery and advances in drug discovery technologies. SLAS Discovery is of particular interest to those involved in analytical chemistry, applied microbiology, automation, biochemistry, bioengineering, biomedical optics, biotechnology, bioinformatics, cell biology, DNA science and technology, genetics, information technology, medicinal chemistry, molecular biology, natural products chemistry, organic chemistry, pharmacology, spectroscopy, and toxicology. SLAS Discovery is a member of the Committee on Publication Ethics (COPE) and was published previously (1996-2016) as the Journal of Biomolecular Screening (JBS).