Human delta opioid receptor

Functional studies on stably transfected Chinese hamster ovary cells after acute and chronic treatment with the selective nonpeptidic agonist SNC-80

Ewa Malatynska, Yu Wang, Richard J. Knapp, Sue Waite, Silvia Calderon, Kenner Rice, Victor J Hruby, Henry I. Yamamura, William R Roeske

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Abstract

The SNC-80 series of nonpeptidic agonists for the δ-opioid receptor are being developed as potential analgesic drugs. It is important to understand their acute and chronic effects at human δ-opioid receptors. Thus, we measured the ability of SNC-80 and [D-Pen2,4'-Cl-Phe4,D-Pen5]enkephalin to inhibit forskolin-stimulated adenylyl cyclase activity in recombinant Chinese hamster ovary cells stably expressing the cloned human δ-opioid receptor. The calculated EC50 values for [D-Pen2,4'-Cl-Phe4,D-Pen5]enkephalin and SNC-80 were 0.6 ± 0.1 nM and 6.3 ± 0.1 nM, respectively. Pretreatment of these cells with SNC-80 (100 nM) for 24 hr produced 1) a time-dependent reduction of δ receptor density, as measured by radioligand binding studies with [3H]naltrindole; 2) a shift in the EC50 value of SNC-80 from 7.7 ± 4.2 nM to 44.1 ± 12 nM, as measured by the cyclic AMP assay; 3) a reduction in the maximum inhibition of adenylyl cyclase activity from 86% to 48%; 4) a marked increase in the forskolin stimulation of basal cyclic AMP accumulation by nearly 100% (from 442 pmol/mg of protein to 824 pmol/mg of protein); and 5) a 5-fold increase in forskolin-stimulated cyclic AMP accumulation after addition of naltrindole. These studies showed that SNC-80 produced desensitization and down-regulation of human δ-opioid receptors in recombinant Chinese hamster ovary cells after chronic treatment and that this effect was associated with an increase in adenylyl cyclase activity.

Original languageEnglish (US)
Pages (from-to)1083-1089
Number of pages7
JournalJournal of Pharmacology and Experimental Therapeutics
Volume278
Issue number3
StatePublished - Sep 1996

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delta Opioid Receptor
Cricetulus
Ovary
naltrindole
Opioid Receptors
Colforsin
Adenylyl Cyclases
Cyclic AMP
Analgesics
4-(alpha-(4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl)-N,N-diethylbenzamide
Proteins
Down-Regulation

ASJC Scopus subject areas

  • Pharmacology

Cite this

Human delta opioid receptor : Functional studies on stably transfected Chinese hamster ovary cells after acute and chronic treatment with the selective nonpeptidic agonist SNC-80. / Malatynska, Ewa; Wang, Yu; Knapp, Richard J.; Waite, Sue; Calderon, Silvia; Rice, Kenner; Hruby, Victor J; Yamamura, Henry I.; Roeske, William R.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 278, No. 3, 09.1996, p. 1083-1089.

Research output: Contribution to journalArticle

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abstract = "The SNC-80 series of nonpeptidic agonists for the δ-opioid receptor are being developed as potential analgesic drugs. It is important to understand their acute and chronic effects at human δ-opioid receptors. Thus, we measured the ability of SNC-80 and [D-Pen2,4'-Cl-Phe4,D-Pen5]enkephalin to inhibit forskolin-stimulated adenylyl cyclase activity in recombinant Chinese hamster ovary cells stably expressing the cloned human δ-opioid receptor. The calculated EC50 values for [D-Pen2,4'-Cl-Phe4,D-Pen5]enkephalin and SNC-80 were 0.6 ± 0.1 nM and 6.3 ± 0.1 nM, respectively. Pretreatment of these cells with SNC-80 (100 nM) for 24 hr produced 1) a time-dependent reduction of δ receptor density, as measured by radioligand binding studies with [3H]naltrindole; 2) a shift in the EC50 value of SNC-80 from 7.7 ± 4.2 nM to 44.1 ± 12 nM, as measured by the cyclic AMP assay; 3) a reduction in the maximum inhibition of adenylyl cyclase activity from 86{\%} to 48{\%}; 4) a marked increase in the forskolin stimulation of basal cyclic AMP accumulation by nearly 100{\%} (from 442 pmol/mg of protein to 824 pmol/mg of protein); and 5) a 5-fold increase in forskolin-stimulated cyclic AMP accumulation after addition of naltrindole. These studies showed that SNC-80 produced desensitization and down-regulation of human δ-opioid receptors in recombinant Chinese hamster ovary cells after chronic treatment and that this effect was associated with an increase in adenylyl cyclase activity.",
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