In vitro and in vivo pharmacological characterization of J-113397, a potent and selective non-peptidyl ORL1 receptor antagonist
In vitro and in vivo pharmacological characterization of J-113397, a potent and selective non-peptidyl ORL1 receptor antagonist
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DOI：
10.1016/s0014-2999(00)00520-3
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发表时间：
2000-08-18
影响因子：
5
通讯作者：
Ohta, H
中科院分区：
文献类型：
--
作者：
Ozaki, S;Kawamoto, H;Ohta, H
1-[(3R，4R)-1-cyclooctylmethyl-3-hydroxymethyl-4-piperidyl]3-ethyl-1，3-dihydro-2H-benzimidazol-2-one(J-113397)被发现是第一个有效的非肽基ORL1受体拮抗剂(K-I：克隆人ORL1=1.8nM)，比其他阿片受体具有高选择性(K-I：人Mu-阿片受体1000 nM，人β-阿片受体10,000 nM，人kappa-阿片受体640 nM)。在体外，J-113397可抑制伤害素/孤啡肽FQ刺激的鸟苷5‘-O-(γ-硫代)三磷酸鸟苷与表达ORL1的中国仓鼠卵巢(CHO)细胞(CHO-ORL1)的结合，IC50值为5.3 nM，而J-113397本身对[S-35]GTP-γ-S的结合无影响。用CHO-ORL1对[S-35]GTP-γ进行S结合试验和cAMP分析，结果表明J-113397对ORL1受体具有竞争性拮抗作用。在表达u阿片受体、β阿片受体和kappa阿片受体的CHO细胞中，J-113397对浓度为100nM的[S-35]GTPγ-S无影响，表明该化合物对ORL_1受体有选择性拮抗作用。在体内，皮下注射J-113397可剂量依赖性地抑制侧脑室注射(i.c.v)引起的痛敏反应。在小鼠甩尾试验中给予伤害素/孤儿FQ。用小鼠脑进行的体外结合研究表明，J-113397与小鼠ORL1受体(K-I：1.1nM)以及人的受体都有很高的亲和力。综上所述，J-113397是第一个有效的、选择性的ORL1受体拮抗剂，可能有助于阐明伤害素/孤儿FQ的生理作用。(C)2000 Elsevier Science B.V.保留所有权利。
1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4-piperidyl]3-ethyl-1,3-dihydro-2H-benzimidazol-2-one (J-113397) was found to be the first potent nonpeptidyl ORL1 receptor antagonist (K-i: cloned human ORL1 = 1.8 nM) with high selectivity over other opioid receptors (K-i: 1000 nM for human mu-opioid receptor, > 10,000 nM for human delta-opioid receptor, and 640 nM for human kappa-opioid receptor). In vitro, J-113397 inhibited nociceptin/orphanin FQ-stimulated [S-35]guanosine 5'-O-(gamma-thio)triphosphate (GTP gamma S) binding to Chinese Hamster Ovary (CHO) cells expressing ORL1 (CHO-ORL1) with an IC50 value of 5.3 nM but had no effect on [S-35]GTP gamma S binding by itself. Schild plot analysis of the [S-35]GTP gamma S binding assay and cAMP assay using CHO-ORL1 indicated competitive antagonism of J-113397 on the ORL1 receptor. In CHO cells expressing mu-, delta- or kappa-opioid receptors, J-113397 had no effects on [S-35]GTP gamma S binding up to a concentration of 100 nM, indicating selective antagonism of the compound on the ORL1 receptor. In vivo, J-113397, when administered subcutaneously (s.c.), dose-dependently inhibited hyperalgesia elicited by intracerebroventricular (i.c.v.) administration of nociceptin/orphanin FQ in a tail-flick test with mice. An in vitro binding study using mouse brains indicated that J-113397 possesses high affinity for the mouse ORL1 receptor(K-i: 1.1 nM) as well as the human receptor. In summary, J-113397 is the first potent, selective ORL1 receptor antagonist that may be useful in elucidating the physiological roles of nociceptin/orphanin FQ. (C) 2000 Elsevier Science B.V. All rights reserved.