Stereoselective interaction of ketamine with recombinant μ, κ, and δ opioid receptors expressed in Chinese hamster ovary cells

Kazuyoshi Hirota, Hirobumi Okawa, Balraj L. Appadu, David Grandy, Lakshmi A. Devi, David G. Lambert

    Research output: Contribution to journalArticle

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    Abstract

    Background: The authors examined the interaction of ketamine with recombinant μ, κ, and δ opioid receptors and recombinant orphan opioid receptors expressed in Chinese hamster ovary cells (CHO-μ, CHO-κ, CHO-δ, and CHOORL1, respectively). Methods: CHO-μ, CHO-κ, and CHO-δ membranes were incubated with the opioid receptor radioligand [3H]diprenorphine at room temperature. Ketamine (racemic, R(-) and S(+)) was included at concentrations covering the clinical range. CHOORL1 membranes were incubated with [125I]Tyr14nociceptin and racemic ketamine at room temperature. The effects of racemic ketamine and selective opioid receptor agonists (μ: [D-Ala2, MePhe4, Gly(ol)5] enkephalin (DAMGO); κ: spiradoline or δ: [D-pen2, D-pen5] enkephalin (DPDPE)) on forskolin-stimulated cyclic adenosine monophosphate formation also were examined. Data are mean ± SEM. Results: Racemic ketamine increased the radioligand equilibrium dissociation constant for [3H]diprenorphine from 85 ± 5 to 273 ± 11, 91 ± 6 to 154 ± 16, and 372 ± 15 to 855 ± 42 pM in CHO-μ, CHO-κ, and CHO-δ, respectively. The concentration of radioligand bound at saturation was unaffected. In CHO-μ and CHO-κ cells, racemic ketamine did not slow the rate of naloxone-induced [3H]diprenorphine dissociation. Ketamine and its isomers also displaced [3H]diprenorphine binding to μ, κ, and δ receptors in a dose-dependent manner, with pKi values for racemic ketamine of 4.38 ± 0.02, 4.55 ± 0.04, and 3.57 ± 0.02, respectively. S(+)-ketamine was two to three times more potent than R(-)-ketamine at μ and κ receptors. Racemic ketamine displaced [125I]Tyr14nociceptin with an estimated affinity constant of 0.5 mM. Racemic ketamine inhibited the formation of cyclic adenosine monophosphatc (naloxone insensitive) in a dose-dependent manner (concentration producing 50% inhibition ∼ 2 mM) in all cell lines, including untransfected CHO cells. Ketamine (100 μM) reversed DAMGO (μ) and spiradoline (κ) inhibition of formation of cyclic adenosine monophosphate. Conclusions: Ketamine interacts stereoselectively with recombinant μ and κ opioid receptors.

    Original languageEnglish (US)
    Pages (from-to)174-182
    Number of pages9
    JournalAnesthesiology
    Volume90
    Issue number1
    StatePublished - Jan 1999

    Fingerprint

    Ketamine
    Opioid Receptors
    Cricetulus
    Ovary
    Diprenorphine
    CHO Cells
    Ala(2)-MePhe(4)-Gly(5)-enkephalin
    Naloxone
    Cyclic AMP
    D-Penicillamine (2,5)-Enkephalin
    Temperature
    Membranes
    Enkephalins
    Colforsin
    Adenosine

    Keywords

    • Adenylyl cyclase
    • Opioid receptor antagonist
    • Radioligand binding
    • Radioreceptor assay

    ASJC Scopus subject areas

    • Anesthesiology and Pain Medicine

    Cite this

    Hirota, K., Okawa, H., Appadu, B. L., Grandy, D., Devi, L. A., & Lambert, D. G. (1999). Stereoselective interaction of ketamine with recombinant μ, κ, and δ opioid receptors expressed in Chinese hamster ovary cells. Anesthesiology, 90(1), 174-182.

    Stereoselective interaction of ketamine with recombinant μ, κ, and δ opioid receptors expressed in Chinese hamster ovary cells. / Hirota, Kazuyoshi; Okawa, Hirobumi; Appadu, Balraj L.; Grandy, David; Devi, Lakshmi A.; Lambert, David G.

    In: Anesthesiology, Vol. 90, No. 1, 01.1999, p. 174-182.

    Research output: Contribution to journalArticle

    Hirota, K, Okawa, H, Appadu, BL, Grandy, D, Devi, LA & Lambert, DG 1999, 'Stereoselective interaction of ketamine with recombinant μ, κ, and δ opioid receptors expressed in Chinese hamster ovary cells', Anesthesiology, vol. 90, no. 1, pp. 174-182.
    Hirota, Kazuyoshi ; Okawa, Hirobumi ; Appadu, Balraj L. ; Grandy, David ; Devi, Lakshmi A. ; Lambert, David G. / Stereoselective interaction of ketamine with recombinant μ, κ, and δ opioid receptors expressed in Chinese hamster ovary cells. In: Anesthesiology. 1999 ; Vol. 90, No. 1. pp. 174-182.
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    title = "Stereoselective interaction of ketamine with recombinant μ, κ, and δ opioid receptors expressed in Chinese hamster ovary cells",
    abstract = "Background: The authors examined the interaction of ketamine with recombinant μ, κ, and δ opioid receptors and recombinant orphan opioid receptors expressed in Chinese hamster ovary cells (CHO-μ, CHO-κ, CHO-δ, and CHOORL1, respectively). Methods: CHO-μ, CHO-κ, and CHO-δ membranes were incubated with the opioid receptor radioligand [3H]diprenorphine at room temperature. Ketamine (racemic, R(-) and S(+)) was included at concentrations covering the clinical range. CHOORL1 membranes were incubated with [125I]Tyr14nociceptin and racemic ketamine at room temperature. The effects of racemic ketamine and selective opioid receptor agonists (μ: [D-Ala2, MePhe4, Gly(ol)5] enkephalin (DAMGO); κ: spiradoline or δ: [D-pen2, D-pen5] enkephalin (DPDPE)) on forskolin-stimulated cyclic adenosine monophosphate formation also were examined. Data are mean ± SEM. Results: Racemic ketamine increased the radioligand equilibrium dissociation constant for [3H]diprenorphine from 85 ± 5 to 273 ± 11, 91 ± 6 to 154 ± 16, and 372 ± 15 to 855 ± 42 pM in CHO-μ, CHO-κ, and CHO-δ, respectively. The concentration of radioligand bound at saturation was unaffected. In CHO-μ and CHO-κ cells, racemic ketamine did not slow the rate of naloxone-induced [3H]diprenorphine dissociation. Ketamine and its isomers also displaced [3H]diprenorphine binding to μ, κ, and δ receptors in a dose-dependent manner, with pKi values for racemic ketamine of 4.38 ± 0.02, 4.55 ± 0.04, and 3.57 ± 0.02, respectively. S(+)-ketamine was two to three times more potent than R(-)-ketamine at μ and κ receptors. Racemic ketamine displaced [125I]Tyr14nociceptin with an estimated affinity constant of 0.5 mM. Racemic ketamine inhibited the formation of cyclic adenosine monophosphatc (naloxone insensitive) in a dose-dependent manner (concentration producing 50{\%} inhibition ∼ 2 mM) in all cell lines, including untransfected CHO cells. Ketamine (100 μM) reversed DAMGO (μ) and spiradoline (κ) inhibition of formation of cyclic adenosine monophosphate. Conclusions: Ketamine interacts stereoselectively with recombinant μ and κ opioid receptors.",
    keywords = "Adenylyl cyclase, Opioid receptor antagonist, Radioligand binding, Radioreceptor assay",
    author = "Kazuyoshi Hirota and Hirobumi Okawa and Appadu, {Balraj L.} and David Grandy and Devi, {Lakshmi A.} and Lambert, {David G.}",
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    T1 - Stereoselective interaction of ketamine with recombinant μ, κ, and δ opioid receptors expressed in Chinese hamster ovary cells

    AU - Hirota, Kazuyoshi

    AU - Okawa, Hirobumi

    AU - Appadu, Balraj L.

    AU - Grandy, David

    AU - Devi, Lakshmi A.

    AU - Lambert, David G.

    PY - 1999/1

    Y1 - 1999/1

    N2 - Background: The authors examined the interaction of ketamine with recombinant μ, κ, and δ opioid receptors and recombinant orphan opioid receptors expressed in Chinese hamster ovary cells (CHO-μ, CHO-κ, CHO-δ, and CHOORL1, respectively). Methods: CHO-μ, CHO-κ, and CHO-δ membranes were incubated with the opioid receptor radioligand [3H]diprenorphine at room temperature. Ketamine (racemic, R(-) and S(+)) was included at concentrations covering the clinical range. CHOORL1 membranes were incubated with [125I]Tyr14nociceptin and racemic ketamine at room temperature. The effects of racemic ketamine and selective opioid receptor agonists (μ: [D-Ala2, MePhe4, Gly(ol)5] enkephalin (DAMGO); κ: spiradoline or δ: [D-pen2, D-pen5] enkephalin (DPDPE)) on forskolin-stimulated cyclic adenosine monophosphate formation also were examined. Data are mean ± SEM. Results: Racemic ketamine increased the radioligand equilibrium dissociation constant for [3H]diprenorphine from 85 ± 5 to 273 ± 11, 91 ± 6 to 154 ± 16, and 372 ± 15 to 855 ± 42 pM in CHO-μ, CHO-κ, and CHO-δ, respectively. The concentration of radioligand bound at saturation was unaffected. In CHO-μ and CHO-κ cells, racemic ketamine did not slow the rate of naloxone-induced [3H]diprenorphine dissociation. Ketamine and its isomers also displaced [3H]diprenorphine binding to μ, κ, and δ receptors in a dose-dependent manner, with pKi values for racemic ketamine of 4.38 ± 0.02, 4.55 ± 0.04, and 3.57 ± 0.02, respectively. S(+)-ketamine was two to three times more potent than R(-)-ketamine at μ and κ receptors. Racemic ketamine displaced [125I]Tyr14nociceptin with an estimated affinity constant of 0.5 mM. Racemic ketamine inhibited the formation of cyclic adenosine monophosphatc (naloxone insensitive) in a dose-dependent manner (concentration producing 50% inhibition ∼ 2 mM) in all cell lines, including untransfected CHO cells. Ketamine (100 μM) reversed DAMGO (μ) and spiradoline (κ) inhibition of formation of cyclic adenosine monophosphate. Conclusions: Ketamine interacts stereoselectively with recombinant μ and κ opioid receptors.

    AB - Background: The authors examined the interaction of ketamine with recombinant μ, κ, and δ opioid receptors and recombinant orphan opioid receptors expressed in Chinese hamster ovary cells (CHO-μ, CHO-κ, CHO-δ, and CHOORL1, respectively). Methods: CHO-μ, CHO-κ, and CHO-δ membranes were incubated with the opioid receptor radioligand [3H]diprenorphine at room temperature. Ketamine (racemic, R(-) and S(+)) was included at concentrations covering the clinical range. CHOORL1 membranes were incubated with [125I]Tyr14nociceptin and racemic ketamine at room temperature. The effects of racemic ketamine and selective opioid receptor agonists (μ: [D-Ala2, MePhe4, Gly(ol)5] enkephalin (DAMGO); κ: spiradoline or δ: [D-pen2, D-pen5] enkephalin (DPDPE)) on forskolin-stimulated cyclic adenosine monophosphate formation also were examined. Data are mean ± SEM. Results: Racemic ketamine increased the radioligand equilibrium dissociation constant for [3H]diprenorphine from 85 ± 5 to 273 ± 11, 91 ± 6 to 154 ± 16, and 372 ± 15 to 855 ± 42 pM in CHO-μ, CHO-κ, and CHO-δ, respectively. The concentration of radioligand bound at saturation was unaffected. In CHO-μ and CHO-κ cells, racemic ketamine did not slow the rate of naloxone-induced [3H]diprenorphine dissociation. Ketamine and its isomers also displaced [3H]diprenorphine binding to μ, κ, and δ receptors in a dose-dependent manner, with pKi values for racemic ketamine of 4.38 ± 0.02, 4.55 ± 0.04, and 3.57 ± 0.02, respectively. S(+)-ketamine was two to three times more potent than R(-)-ketamine at μ and κ receptors. Racemic ketamine displaced [125I]Tyr14nociceptin with an estimated affinity constant of 0.5 mM. Racemic ketamine inhibited the formation of cyclic adenosine monophosphatc (naloxone insensitive) in a dose-dependent manner (concentration producing 50% inhibition ∼ 2 mM) in all cell lines, including untransfected CHO cells. Ketamine (100 μM) reversed DAMGO (μ) and spiradoline (κ) inhibition of formation of cyclic adenosine monophosphate. Conclusions: Ketamine interacts stereoselectively with recombinant μ and κ opioid receptors.

    KW - Adenylyl cyclase

    KW - Opioid receptor antagonist

    KW - Radioligand binding

    KW - Radioreceptor assay

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