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Effect of chronic treatment of ohmefentanyl stereoisomers on cyclic AMP formation in Sf9 insect cells expressing human mu-opioid receptors

机译:慢性芬欧芬太尼立体异构体处理对表达人μ阿片受体的Sf9昆虫细胞中环AMP形成的影响

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The binding affinity of ohmefentanyl stereoisomers for mu-opioid receptors and the effect of chronic ohmefentanyl stereoisomers pretreatments on intracellular cAMP formation were investigated in Sf9 insect cells expressing human mu-opioid receptors (Sf9-mu cells). Competitive assay of [H-3]ohmefentanyl binding revealed that these isomers had high affinity for mu-opioid receptors in Sf9-mu cells. Isomer F9204 had the highest affinity for mu-opioid receptors with the K-i value of 1.66 +/- 0.28 nM. After pretreated Sf9-mu cells with increasing concentrations of these isomers for 6 h, addition of naloxone (1 muM) precipitated an overshoot of foskolin-stimulated cAMP accumulation. The ability of these isomers to induce cAMP overshoot differed greatly with the order of F9202 > F9205 > F9208 > F9206 > F9204 > F9207. Of these isomers, F9202 was 2.7-fold less potent than F9204 in receptor binding affinity, but 71.5-fold more potent in ability to induce cAMP overshoot. These results suggested that there was a significant stereo-structural difference among ohmefentanyl stereoisomers in ability to induce naloxone-precipitated cAMP overshoot in Sf9-mu cells. (C) 2004 Published by Elsevier Inc.
机译:在表达人μ阿片受体的Sf9昆虫细胞(Sf9-mu细胞)中研究了欧姆芬太尼立体异构体对μ阿片受体的结合亲和力和慢性欧姆芬太尼立体异构体预处理对胞内cAMP形成的影响。竞争性检测[H-3]芬太尼的结合表明,这些异构体对Sf9-mu细胞中的μ阿片受体具有很高的亲和力。异构体F9204对μ阿片受体的亲和力最高,K-i值为1.66 +/- 0.28 nM。在用这些异构体的浓度增加的Sf9-mu细胞进行预处理6小时后,添加纳洛酮(1μM)会导致福考林刺激的cAMP积累过高。这些异构体引起cAMP过冲的能力差异很大,依次为F9202> F9205> F9208> F9206> F9204> F9207。在这些异构体中,在受体结合亲和力方面,F9202的效力比F9204低2.7倍,但在诱导cAMP超调的能力上却高71.5倍。这些结果表明,奥芬太尼立体异构体之间在诱导Sf9-mu细胞中纳洛酮沉淀的cAMP超调能力方面存在显着的立体结构差异。 (C)2004由Elsevier Inc.出版

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