TY - JOUR
T1 - Unexpected antinociceptive potency of cyclic [D-Tca1]CTAP
T2 - potential for a novel mechanism of action
AU - Horan, Peter J.
AU - Wild, Kenneth D.
AU - Kazmierski, Wieslaw W.
AU - Ferguson, Ronald
AU - Hruby, Victor J
AU - Weber, Steven J.
AU - Davis, Thomas P.
AU - Fang, Lei
AU - Knapp, Richard J.
AU - Yamamura, Henry I.
AU - Kramer, Thomas H.
AU - Burks, Thomas F.
AU - Bowen, Wayne D.
AU - Takemori, A. E.
AU - Porreca, Frank
N1 - Funding Information:
This work as supported by USPHS Grant DA 06284 from the National Institute on Drug Abuse
PY - 1993/3/16
Y1 - 1993/3/16
N2 - This study tested the hypothesis that compounds which may bind simultaneously to δ and μ receptors may be more potent antinociceptive agents than would be predicted from their bindings affinities at individual μ and δ opioid receptors. D-Tca-Cys- Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 ([D-Tca1]CTAP) where D-Ta is a cyclic D-trytophan analogue) was synthesized and evaluated in radioligand competition assays, opioid bioassays, and in an antinociceptive assay (the tail-flick test in mice). Additionally, the metabolic stability of [D-Tca1]CTAP was evaluated in striatal and cerebellar tissue slices. In the rat brain in vitro, [D-Tca1]CTAP competed weakly for sites labelled by [3H]D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH2 ([3H]CTOP) (μ-ligand), and [3H][D-Pen2, pCl-Phe5]enkephalin (δ-ligand); [D-Pen2, D-Pen5]enkephalin (DPDPE) (δ-agonist) was 6.5-fold less and 230-fold more potent, respectively, against these ligands. Additionally, in mouse isolated vas deferens and guinea pig isolated ileum smooth muscle preparations, [D-Tca1]CTAP proved to be weak as either a δ(IC50 of approximately 2 μM) or μ (IC50 > 8 μM) receptor agonist. Surprisingly, however, i.c.v. [D-Tca1]CTAP produced antinociception with potency no similar to DPDPE. The antinociceptive actions of [D-Tca1]CTAP were apparently not due to a metabolite or the release of endogeneous opiods, as this compound proved stable in both striatal and cerebellar tissue slices and its antinociceptive actions were not enhance by the 'enkephalinase' inhibitor thiorphan. The suggestion that [D-Tca1]CTAP might be acting by binding simultaneously go μ and δ receptors to produced its antinociceptive effect is supported by the demonstrated antagonism resulting from μ receptor blockade with either β-funaltrexamine (β-FNA) or naloxonazine, or by δ receptor blockade by ICI 174,864 ([N,N-dially-Tyr1, Aib2,3,Leu5] enkephalin). Furthermore, the antinociceptive properties of [D-Tca1]CTAP were antagonized by (naltrindole-5′-isothiocyanate) (5′-NTII), an antagonist at the δ2 opioid receptor subtype, but not by the δ1 antagonist [D-Ala2, D-Leu5, Cys6]enkephalin (DALCE). Additionally, no antagonism was produced by nor-binaltorphimine (nor-BNI), a κ antagonist. From these data, [D-Tca1]CTAP appears to bind to μ, and 5′-NTII-sensitive δ2, opioid receptors, and may represent the first of a class of compounds which may act at an opioid recptor complex via 'self-potentiation'.
AB - This study tested the hypothesis that compounds which may bind simultaneously to δ and μ receptors may be more potent antinociceptive agents than would be predicted from their bindings affinities at individual μ and δ opioid receptors. D-Tca-Cys- Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 ([D-Tca1]CTAP) where D-Ta is a cyclic D-trytophan analogue) was synthesized and evaluated in radioligand competition assays, opioid bioassays, and in an antinociceptive assay (the tail-flick test in mice). Additionally, the metabolic stability of [D-Tca1]CTAP was evaluated in striatal and cerebellar tissue slices. In the rat brain in vitro, [D-Tca1]CTAP competed weakly for sites labelled by [3H]D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH2 ([3H]CTOP) (μ-ligand), and [3H][D-Pen2, pCl-Phe5]enkephalin (δ-ligand); [D-Pen2, D-Pen5]enkephalin (DPDPE) (δ-agonist) was 6.5-fold less and 230-fold more potent, respectively, against these ligands. Additionally, in mouse isolated vas deferens and guinea pig isolated ileum smooth muscle preparations, [D-Tca1]CTAP proved to be weak as either a δ(IC50 of approximately 2 μM) or μ (IC50 > 8 μM) receptor agonist. Surprisingly, however, i.c.v. [D-Tca1]CTAP produced antinociception with potency no similar to DPDPE. The antinociceptive actions of [D-Tca1]CTAP were apparently not due to a metabolite or the release of endogeneous opiods, as this compound proved stable in both striatal and cerebellar tissue slices and its antinociceptive actions were not enhance by the 'enkephalinase' inhibitor thiorphan. The suggestion that [D-Tca1]CTAP might be acting by binding simultaneously go μ and δ receptors to produced its antinociceptive effect is supported by the demonstrated antagonism resulting from μ receptor blockade with either β-funaltrexamine (β-FNA) or naloxonazine, or by δ receptor blockade by ICI 174,864 ([N,N-dially-Tyr1, Aib2,3,Leu5] enkephalin). Furthermore, the antinociceptive properties of [D-Tca1]CTAP were antagonized by (naltrindole-5′-isothiocyanate) (5′-NTII), an antagonist at the δ2 opioid receptor subtype, but not by the δ1 antagonist [D-Ala2, D-Leu5, Cys6]enkephalin (DALCE). Additionally, no antagonism was produced by nor-binaltorphimine (nor-BNI), a κ antagonist. From these data, [D-Tca1]CTAP appears to bind to μ, and 5′-NTII-sensitive δ2, opioid receptors, and may represent the first of a class of compounds which may act at an opioid recptor complex via 'self-potentiation'.
KW - Antinociception
KW - Metabolism
KW - Opioid receptor antagonists
KW - δ-Opioid receptors
KW - μ-Opioid receptors
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U2 - 10.1016/0014-2999(93)90348-L
DO - 10.1016/0014-2999(93)90348-L
M3 - Article
C2 - 8386089
AN - SCOPUS:0027466444
SN - 0014-2999
VL - 233
SP - 53
EP - 62
JO - European Journal of Pharmacology
JF - European Journal of Pharmacology
IS - 1
ER -