Inhibition of endothelin A receptor by a novel, selective receptor antagonist enhances morphine-induced analgesia: Possible functional interaction of dimerized endothelin A and μ-opioid receptors.
Analgesics, Opioid
/ administration & dosage
Animals
Dose-Response Relationship, Drug
Endothelin A Receptor Antagonists
/ administration & dosage
HEK293 Cells
Humans
Male
Mice
Mice, Inbred C57BL
Morphine
/ administration & dosage
Pain Measurement
/ drug effects
Peptides, Cyclic
/ administration & dosage
Protein Multimerization
/ drug effects
Receptor, Endothelin A
/ metabolism
Receptors, Opioid, mu
/ metabolism
Analgesic
BQ-123 sodium salt (PubChem CID: 52943236)
BQ-788 sodium salt (PubChem CID: 23693553)
Endothelin A receptor antagonist
G protein-coupled receptor
Morphine
Pain
Receptor heterodimerization
and Bosentan Monohydrate (PubChem CID: 185462)
included endothelin-1 (PubChem CID: 16212950)
morphine hydrochloride (PubChem CID: 5464110)
Journal
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie
ISSN: 1950-6007
Titre abrégé: Biomed Pharmacother
Pays: France
ID NLM: 8213295
Informations de publication
Date de publication:
Sep 2021
Sep 2021
Historique:
received:
01
03
2021
revised:
13
05
2021
accepted:
24
05
2021
pubmed:
28
6
2021
medline:
16
12
2021
entrez:
27
6
2021
Statut:
ppublish
Résumé
The misuse of opioids has led to an epidemic in recent times. The endothelin A receptor (ETAR) has recently attracted attention as a novel therapeutic target to enhance opioid analgesia. We hypothesized that endothelin A receptors may affect pain mechanisms by heterodimerization with μ opioid receptors. We examined the mechanisms of ETAR-mediated pain and the potential therapeutic effects of an ETAR antagonist, Compound-E, as an agent for analgesia. Real-time in vitro effect of Compound-E on morphine response was assessed in HEK293 cells expressing both endothelin A and μ opioid receptors through CellKey™ and cADDis cAMP assays. Endothelin A/μ opioid receptor dimerization was assessed by immunoprecipitation and live cell imaging. The in vivo effect of Compound-E was evaluated using a morphine analgesia mouse model that observed escape response behavior, body temperature, and locomotor activity. In CellKey™ and cAMP assays, pretreatment of cells with endothelin-1 attenuated morphine-induced responses. These responses were improved by Compound-E, but not by BQ-123 nor by bosentan, an ETAR and endothelin B receptor antagonist. Dimerization of ETARs and μ opioid receptors was confirmed by Western blot and total internal reflection fluorescence microscopy in live cells. In vivo, Compound-E potentiated and prolonged the analgesic effects of morphine, enhanced hypothermia, and increased locomotor activity compared to morphine alone. The results suggest that attenuation by endothelin-1 of morphine analgesia may be caused by dimerization of Endothelin A/μ opioid receptors. The novel ETAR antagonist Compound-E could be an effective adjunct to reduce opioid use.
Sections du résumé
BACKGROUND
BACKGROUND
The misuse of opioids has led to an epidemic in recent times. The endothelin A receptor (ETAR) has recently attracted attention as a novel therapeutic target to enhance opioid analgesia. We hypothesized that endothelin A receptors may affect pain mechanisms by heterodimerization with μ opioid receptors. We examined the mechanisms of ETAR-mediated pain and the potential therapeutic effects of an ETAR antagonist, Compound-E, as an agent for analgesia.
METHODS
METHODS
Real-time in vitro effect of Compound-E on morphine response was assessed in HEK293 cells expressing both endothelin A and μ opioid receptors through CellKey™ and cADDis cAMP assays. Endothelin A/μ opioid receptor dimerization was assessed by immunoprecipitation and live cell imaging. The in vivo effect of Compound-E was evaluated using a morphine analgesia mouse model that observed escape response behavior, body temperature, and locomotor activity.
RESULTS
RESULTS
In CellKey™ and cAMP assays, pretreatment of cells with endothelin-1 attenuated morphine-induced responses. These responses were improved by Compound-E, but not by BQ-123 nor by bosentan, an ETAR and endothelin B receptor antagonist. Dimerization of ETARs and μ opioid receptors was confirmed by Western blot and total internal reflection fluorescence microscopy in live cells. In vivo, Compound-E potentiated and prolonged the analgesic effects of morphine, enhanced hypothermia, and increased locomotor activity compared to morphine alone.
CONCLUSION
CONCLUSIONS
The results suggest that attenuation by endothelin-1 of morphine analgesia may be caused by dimerization of Endothelin A/μ opioid receptors. The novel ETAR antagonist Compound-E could be an effective adjunct to reduce opioid use.
Identifiants
pubmed: 34175819
pii: S0753-3322(21)00582-5
doi: 10.1016/j.biopha.2021.111800
pii:
doi:
Substances chimiques
Analgesics, Opioid
0
Endothelin A Receptor Antagonists
0
Peptides, Cyclic
0
Receptor, Endothelin A
0
Receptors, Opioid, mu
0
Morphine
76I7G6D29C
cyclo(Trp-Asp-Pro-Val-Leu)
S2A8YZM151
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
111800Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Masson SAS.. All rights reserved.