Sphingosine and its analog, the immunosuppressant 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol, interact with the CB1 cannabinoid receptor
Sphingosine and its analog, the immunosuppressant 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol, interact with the CB1 cannabinoid receptor
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DOI：
10.1124/mol.105.020552
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发表时间：
2006-07-01
影响因子：
3.6
通讯作者：
Selley, Dana E.
中科院分区：
文献类型：
--
作者：
Paugh, Steven W.;Cassidy, Michael P.;Selley, Dana E.
鞘氨醇-1-磷酸（S1 P）和大麻素受体是G蛋白偶联受体，分别介导S1 P和内源性大麻素的作用。大麻素受体还介导δ（9）-四氢大麻酚（大麻中的主要精神活性成分）的作用，而S1 P受体有助于2-氨基-2-（2-[4-辛基苯基]乙基）-1，3-丙二醇（FTY 720）的免疫抑制作用。FTY 720是一种鞘氨醇类似物，在动物模型和临床试验中可以预防肾移植排斥反应并抑制多种自身免疫性疾病。我们现在报告FTY 720和鞘氨醇都与CB 1而不是CB 2大麻素受体相互作用。FTY 720和鞘氨醇抑制CB 1选择性拮抗剂[H-3] N-（哌啶基）-5-（4-氯苯基）-1- 2，4-二氯苯基-4-甲基-1H-吡唑-3-甲酰胺（[H-3] SR 141716 A）和大麻素激动剂[H-3]（-）-顺式-3-[2-羟基-4-（1，1-二甲基庚基）苯基]-反式4-（3-羟丙基）环己醇（[H-3] CP 55，940）在CB 1表达细胞系和小鼠小脑中以浓度依赖性方式。然而，这些化合物没有显著改变[3 H] CP 55，940与CB 2受体的结合。在G蛋白激活试验中，FTY 720和鞘氨醇抑制鸟苷5 '-O-通过大麻素激动剂R-（+）-[2，3-二氢-5-甲基-3-[（吗啉基）甲基]吡咯并[1，2，3-de] 1，4-苯并恶嗪基]-（1-萘基）甲酮甲磺酸盐（WIN 55，212-2）的浓度依赖性的方式，这种拮抗剂的作用是不模仿S1 P。FTY 720和鞘氨醇还抑制了在表达CB 1受体的完整中国仓鼠卵巢（CHO）细胞中由WIN 55，212-2激活的细胞外信号调节激酶1和2以及Akt，并减弱了在CHO细胞中由WIN 55，212-2刺激的荧光标记的CB 1受体的内化。此外，FTY 720和鞘氨醇产生的浓度-效应曲线的大麻素激动剂G-蛋白激活的100倍的位移，表明它们作为竞争性的CB 1拮抗剂。这些结果表明，CB 1受体可能是FTY 720的新靶点，鞘氨醇可能是内源性CB 1拮抗剂。
Sphingosine-1-phosphate (S1P) and cannabinoid receptors are G-protein-coupled receptors that mediate the effects of S1P and endocannabinoids, respectively. Cannabinoid receptors also mediate the effects of Delta(9)-tetrahydrocannabinol, the primary psychoactive ingredient in marijuana, whereas S1P receptors contribute to the immunosuppressant effects of 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol (FTY720). FTY720 is a sphingosine analog that can prevent renal graft rejections and suppress a variety of autoimmune disorders in animal models and clinical trials. We now report that both FTY720 and sphingosine interact with CB 1 but not CB 2 cannabinoid receptors. FTY720 and sphingosine inhibited the binding of the CB1-selective antagonist [H-3] N-(piperidinyl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide ([H-3] SR141716A) and the cannabinoid agonist [H-3](-)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans4-(3-hydroxypropyl) cyclohexanol ([H-3]CP55,940) in a concentration-dependent manner in both CB1-expressing cell lines and mouse cerebellum. However, these compounds did not significantly alter [ 3H] CP55,940 binding to CB 2 receptors. In G-protein activation assays, FTY720 and sphingosine inhibited the maximal stimulation of guanosine 5'-O-(3-[S-35] thio) triphosphate binding by the cannabinoid agonist R-(+)-[2,3-dihydro-5-methyl-3-[( morpholinyl) methyl] pyrrolo[ 1,2,3-de]1,4-benzoxazinyl]-(1-naphthalenyl) methanone mesylate (WIN55,212-2) in a concentration-dependent manner, and this antagonist effect was not mimicked by S1P. FTY720 and sphingosine also inhibited activation of extracellular signal-regulated kinases 1 and 2 and Akt by WIN55,212-2 in intact Chinese hamster ovary (CHO) cells expressing CB1 receptors and attenuated WIN55,212-2-stimulated internalization of a fluorescence-tagged CB1 receptor in CHO cells. Moreover, both FTY720 and sphingosine produced rightward shifts in the concentration-effect curves of cannabinoid agonists for G-protein activation, indicating that they act as competitive CB1 antagonists. These results suggest that the CB1 receptor could be a novel target of FTY720 and that sphingosine could be an endogenous CB1 antagonist.