Pharmacology of GPR55 in Yeast and Identification of GSK494581A as a Mixed-Activity Glycine Transporter Subtype 1 Inhibitor and GPR55 Agonist
Pharmacology of GPR55 in Yeast and Identification of GSK494581A as a Mixed-Activity Glycine Transporter Subtype 1 Inhibitor and GPR55 Agonist
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DOI：
10.1124/jpet.110.172650
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发表时间：
2011-04-01
影响因子：
3.5
通讯作者：
Dowell, Simon J.
中科院分区：
文献类型：
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作者：
Brown, Andrew J.;Daniels, Dion A.;Dowell, Simon J.
GPR 55是由L-α-溶血磷脂酰肌醇激活的G蛋白偶联受体，并被认为在疼痛信号传导、骨形态发生以及可能在血管内皮细胞中发挥作用。它对某些大麻素（与大麻素CB 1和CB 2受体相互作用的分子）具有亲和力，但由于缺乏选择性药理学工具，对其在细胞系统和组织中的功能作用的研究受到限制。在这里，我们提出了我们的特性GPR 55在酵母酿酒酵母和人胚肾（HEK 293）细胞。我们描述了GSK 494581 A（1-{2-氟-4-[1-（甲基氧基）乙基]苯基}-4-{[4 '-氟-4-（甲基磺酰基）-2-联苯基]羰基}哌嗪），其是通过多样性筛选鉴定的GPR 55的选择性小分子配体。GSK 494581 A是一系列苯甲酰哌嗪类药物之一，最初被鉴定为甘氨酸转运蛋白亚型1（GlyT 1）的抑制剂并获得专利。GPR 55和GlyT 1之间的结构-活性关系在该系列中是不同的。最具GPR 55选择性的实例是GSK 575594 A（3-氟-4-（4-{[4 '-氟-4-（甲基磺酰基）-2-联苯基]羰基}-1-哌嗪基）苯胺），其对GPR 55（pEC（50）= 6.8）的选择性是GlyT 1（pIC（50）= 5.0）的约60倍。在GPR 55和GlyT 1上具有活性的几个范例已经在广泛的其他分子靶点上进行了分析，并且在大麻素受体和所有其他测试的靶点上没有活性。苯甲酰基哌嗪激动剂激活人GPR 55，但不激活啮齿动物GPR 55，这表明这些直系同源物之间相对低水平的序列同一性（75%）转化为配体结合位点的重要功能差异。
GPR55 is a G protein-coupled receptor activated by L-alpha-lysophosphatidylinositol and suggested to have roles in pain signaling, bone morphogenesis, and possibly in vascular endothelial cells. It has affinity for certain cannabinoids (molecules that interact with the cannabinoid CB1 and CB2 receptors), but investigation of its functional role in cell-based systems and in tissue has been limited by a lack of selective pharmacological tools. Here, we present our characterization of GPR55 in the yeast Saccharomyces cerevisiae and in human embryonic kidney (HEK293) cells. We describe GSK494581A (1-{2-fluoro-4-[1-(methyloxy)ethyl]phenyl}-4-{[4'-fluoro-4-(methylsulfonyl)-2-biphenylyl]carbonyl}piperazine), a selective small-molecule ligand of GPR55 identified through diversity screening. GSK494581A is one of a series of benzoylpiperazines originally identified and patented as inhibitors of the glycine transporter subtype 1 (GlyT1). The structure-activity relationship between GPR55 and GlyT1 is divergent across this series. The most GPR55-selective example is GSK575594A (3-fluoro-4-(4-{[4'-fluoro-4-(methylsulfonyl)-2-biphenylyl] carbonyl}-1-piperazinyl) aniline), which is approximately 60-fold selective for GPR55 (pEC(50) = 6.8) over GlyT1 (pIC(50) = 5.0). Several exemplars with activity at GPR55 and GlyT1 have been profiled at a broad range of other molecular targets and are inactive at cannabinoid receptors and all other targets tested. The benzoylpiperazine agonists activate human GPR55 but not rodent GPR55, suggesting that the relatively low level of sequence identity between these orthologs (75%) translates to important functional differences in the ligand-binding site.