McGuinness Debra, Malikzay Asra, Visconti Richard, Lin Karen, Bayne Marvin, Monsma Frederick, Lunn Charles A
New Lead Discovery, Schering-Plough Research Institute, Kenilworth, New Jersey, USA.
J Biomol Screen. 2009 Jan;14(1):49-58. doi: 10.1177/1087057108327329.
The authors have characterized a set of cannabinoid CB(2) receptor ligands, including triaryl bis sulfone inverse agonists, in a cell-based receptor/beta-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 beta-arrestin protein complementation assay exhibiting a more robust signal than the cAMP assay for cannabinoid CB(2) agonists. Further assay validation shows that DiscoveRx PathHunter HEK293 CB(2) beta-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 CB(2) receptor and beta-arrestin as an appropriate and valuable screening platform for compounds specific for the cannabinoid CB(2) receptor.
作者在基于细胞的受体/β-抑制蛋白相互作用检测（DiscoveRx PathHunter）中对一组大麻素CB(2)受体配体进行了表征，其中包括三芳基双砜反向激动剂。将结果与使用竞争性配体结合检测的结果以及对福司可林刺激的环磷酸腺苷（cAMP）水平的影响（珀金埃尔默公司的LANCE检测）进行了比较。作者表明，所测试的三种检测系统之间具有良好的相关性，对于大麻素CB(2)激动剂，β-抑制蛋白蛋白质互补检测比cAMP检测表现出更强的信号。进一步的检测验证表明，DiscoveRx PathHunter HEK293 CB(2)β-抑制蛋白检测可以从冷冻保存的细胞悬液中进行，消除了活细胞筛选过程中因需要多个细胞库而导致的差异。这些结果，以及作者评估一组随机文库化合物测试集的结果，验证了人源大麻素CB(2)受体与β-抑制蛋白之间的配体诱导相互作用作为大麻素CB(2)受体特异性化合物合适且有价值的筛选平台的用途。
