Investigation of CGRP receptors and peptide pharmacology in human coronary arteries. Characterization with a nonpeptide antagonist

Philip Hasbak; Ole Saetrum Opgaard; Karen Eskesen; Søren Schifter; Henrik Arendrup; Jenny Longmore; Lars Edvinsson

doi:10.1124/jpet.102.037754

Investigation of CGRP receptors and peptide pharmacology in human coronary arteries. Characterization with a nonpeptide antagonist

Philip Hasbak, Ole Saetrum Opgaard, Karen Eskesen, Søren Schifter, Henrik Arendrup, Jenny Longmore, Lars Edvinsson

Department of Clinical Experimental Research

48 Citations (Scopus)

Abstract

Calcitonin gene-related peptide (CGRP), adrenomedullin (AM), and amylin are structurally related peptides mediating vasorelaxation in the coronary circulation possibly via CGRP receptors (subtypes 1 or 2). Functional CGRP1 receptors appear to consist of at least three different kinds of proteins: the calcitonin receptor-like receptor (CRLR), receptor-activity-modifying proteins (RAMPs) and the receptor component protein (RCP). No CGRP2 receptor has yet been cloned. Using reverse transcriptase - polymerase chain reaction, the presence of mRNA sequences encoding CRLR, RCP and RAMPs was demonstrated in human coronary arteries. Relaxant responses were studied on isolated segments of coronary arteries after precontraction with U46619 (9,11-dideoxy-11alpha,9alpha-epoxymethano-prostaglandin F(2alpha)). The human peptides alphaCGRP, AM, and amylin induced relaxation with mean pEC50 values of 8.6, 6.8, and 6.3 M, respectively. Preincubation with alphaCGRP(8-37) (10(-7) -10(-5) M) and a novel nonpeptide CGRP antagonist "Compound 1" (WO98/11128) (10(-7)-10(-5) M) caused a dose-dependent rightward shift of the concentration-response curves for alphaCGRP with pA(2) values of 7.0 and 7.1, respectively. Preincubation with alphaCGRP(8-37) (10(-6) M) and Compound 1 (10(-6) M) caused significant rightward shift of the concentration-response curves for AM and amylin as well with pK B values between 6.6 and 7.5. Preincubation with AM(22-52) had no antagonistic effect on the AM and amylin response, neither did diacetoamidomethyl cysteine CGRP cause any concentration dependent (10(-11)-10(-6) M) dilatation. In conclusion, mRNA for the components forming CGRP1 and AM receptors was detected in the human left anterior descending coronary arteries. alphaCGRP, AM, and amylin mediated vasorelaxation via the CGRP1 receptor. Compound 1 acted as a nonpeptide antagonist at the CGRP1 receptor and could thus become a tool for the study of CGRP-mediated functional responses in human tissue.

Original language	English
Journal	The Journal of pharmacology and experimental therapeutics
Volume	304
Issue number	1
Pages (from-to)	326-33
Number of pages	8
ISSN	0022-3565
DOIs	https://doi.org/10.1124/jpet.102.037754
Publication status	Published - Jan 2003

Keywords

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid
Adrenomedullin
Adult
Amyloid
Calcitonin Gene-Related Peptide
Calcitonin Receptor-Like Protein
Coronary Vessels
Female
Humans
In Vitro Techniques
Intracellular Signaling Peptides and Proteins
Islet Amyloid Polypeptide
Male
Membrane Proteins
Middle Aged
Muscle Contraction
Muscle Relaxation
Muscle, Smooth, Vascular
Peptide Fragments
Peptides
Piperazines
Piperidines
RNA, Messenger
Receptor Activity-Modifying Proteins
Receptors, Calcitonin
Receptors, Calcitonin Gene-Related Peptide

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10.1124/jpet.102.037754

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Investigation of CGRP receptors and peptide pharmacology in human coronary arteries. Characterization with a nonpeptide antagonist. / Hasbak, Philip; Saetrum Opgaard, Ole; Eskesen, Karen et al.
In: The Journal of pharmacology and experimental therapeutics, Vol. 304, No. 1, 01.2003, p. 326-33.

Research output: Contribution to journal › Journal article › Research › peer-review

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title = "Investigation of CGRP receptors and peptide pharmacology in human coronary arteries. Characterization with a nonpeptide antagonist",

abstract = "Calcitonin gene-related peptide (CGRP), adrenomedullin (AM), and amylin are structurally related peptides mediating vasorelaxation in the coronary circulation possibly via CGRP receptors (subtypes 1 or 2). Functional CGRP1 receptors appear to consist of at least three different kinds of proteins: the calcitonin receptor-like receptor (CRLR), receptor-activity-modifying proteins (RAMPs) and the receptor component protein (RCP). No CGRP2 receptor has yet been cloned. Using reverse transcriptase - polymerase chain reaction, the presence of mRNA sequences encoding CRLR, RCP and RAMPs was demonstrated in human coronary arteries. Relaxant responses were studied on isolated segments of coronary arteries after precontraction with U46619 (9,11-dideoxy-11alpha,9alpha-epoxymethano-prostaglandin F(2alpha)). The human peptides alphaCGRP, AM, and amylin induced relaxation with mean pEC50 values of 8.6, 6.8, and 6.3 M, respectively. Preincubation with alphaCGRP(8-37) (10(-7) -10(-5) M) and a novel nonpeptide CGRP antagonist {"}Compound 1{"} (WO98/11128) (10(-7)-10(-5) M) caused a dose-dependent rightward shift of the concentration-response curves for alphaCGRP with pA(2) values of 7.0 and 7.1, respectively. Preincubation with alphaCGRP(8-37) (10(-6) M) and Compound 1 (10(-6) M) caused significant rightward shift of the concentration-response curves for AM and amylin as well with pK B values between 6.6 and 7.5. Preincubation with AM(22-52) had no antagonistic effect on the AM and amylin response, neither did diacetoamidomethyl cysteine CGRP cause any concentration dependent (10(-11)-10(-6) M) dilatation. In conclusion, mRNA for the components forming CGRP1 and AM receptors was detected in the human left anterior descending coronary arteries. alphaCGRP, AM, and amylin mediated vasorelaxation via the CGRP1 receptor. Compound 1 acted as a nonpeptide antagonist at the CGRP1 receptor and could thus become a tool for the study of CGRP-mediated functional responses in human tissue.",

keywords = "15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid, Adrenomedullin, Adult, Amyloid, Calcitonin Gene-Related Peptide, Calcitonin Receptor-Like Protein, Coronary Vessels, Female, Humans, In Vitro Techniques, Intracellular Signaling Peptides and Proteins, Islet Amyloid Polypeptide, Male, Membrane Proteins, Middle Aged, Muscle Contraction, Muscle Relaxation, Muscle, Smooth, Vascular, Peptide Fragments, Peptides, Piperazines, Piperidines, RNA, Messenger, Receptor Activity-Modifying Proteins, Receptors, Calcitonin, Receptors, Calcitonin Gene-Related Peptide",

author = "Philip Hasbak and \{Saetrum Opgaard\}, Ole and Karen Eskesen and S{\o}ren Schifter and Henrik Arendrup and Jenny Longmore and Lars Edvinsson",

year = "2003",

month = jan,

doi = "10.1124/jpet.102.037754",

language = "English",

volume = "304",

pages = "326--33",

journal = "The Journal of pharmacology and experimental therapeutics",

issn = "0022-3565",

publisher = "Elsevier Inc.",

number = "1",

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TY - JOUR

T1 - Investigation of CGRP receptors and peptide pharmacology in human coronary arteries. Characterization with a nonpeptide antagonist

AU - Hasbak, Philip

AU - Saetrum Opgaard, Ole

AU - Eskesen, Karen

AU - Schifter, Søren

AU - Arendrup, Henrik

AU - Longmore, Jenny

AU - Edvinsson, Lars

PY - 2003/1

Y1 - 2003/1

N2 - Calcitonin gene-related peptide (CGRP), adrenomedullin (AM), and amylin are structurally related peptides mediating vasorelaxation in the coronary circulation possibly via CGRP receptors (subtypes 1 or 2). Functional CGRP1 receptors appear to consist of at least three different kinds of proteins: the calcitonin receptor-like receptor (CRLR), receptor-activity-modifying proteins (RAMPs) and the receptor component protein (RCP). No CGRP2 receptor has yet been cloned. Using reverse transcriptase - polymerase chain reaction, the presence of mRNA sequences encoding CRLR, RCP and RAMPs was demonstrated in human coronary arteries. Relaxant responses were studied on isolated segments of coronary arteries after precontraction with U46619 (9,11-dideoxy-11alpha,9alpha-epoxymethano-prostaglandin F(2alpha)). The human peptides alphaCGRP, AM, and amylin induced relaxation with mean pEC50 values of 8.6, 6.8, and 6.3 M, respectively. Preincubation with alphaCGRP(8-37) (10(-7) -10(-5) M) and a novel nonpeptide CGRP antagonist "Compound 1" (WO98/11128) (10(-7)-10(-5) M) caused a dose-dependent rightward shift of the concentration-response curves for alphaCGRP with pA(2) values of 7.0 and 7.1, respectively. Preincubation with alphaCGRP(8-37) (10(-6) M) and Compound 1 (10(-6) M) caused significant rightward shift of the concentration-response curves for AM and amylin as well with pK B values between 6.6 and 7.5. Preincubation with AM(22-52) had no antagonistic effect on the AM and amylin response, neither did diacetoamidomethyl cysteine CGRP cause any concentration dependent (10(-11)-10(-6) M) dilatation. In conclusion, mRNA for the components forming CGRP1 and AM receptors was detected in the human left anterior descending coronary arteries. alphaCGRP, AM, and amylin mediated vasorelaxation via the CGRP1 receptor. Compound 1 acted as a nonpeptide antagonist at the CGRP1 receptor and could thus become a tool for the study of CGRP-mediated functional responses in human tissue.

AB - Calcitonin gene-related peptide (CGRP), adrenomedullin (AM), and amylin are structurally related peptides mediating vasorelaxation in the coronary circulation possibly via CGRP receptors (subtypes 1 or 2). Functional CGRP1 receptors appear to consist of at least three different kinds of proteins: the calcitonin receptor-like receptor (CRLR), receptor-activity-modifying proteins (RAMPs) and the receptor component protein (RCP). No CGRP2 receptor has yet been cloned. Using reverse transcriptase - polymerase chain reaction, the presence of mRNA sequences encoding CRLR, RCP and RAMPs was demonstrated in human coronary arteries. Relaxant responses were studied on isolated segments of coronary arteries after precontraction with U46619 (9,11-dideoxy-11alpha,9alpha-epoxymethano-prostaglandin F(2alpha)). The human peptides alphaCGRP, AM, and amylin induced relaxation with mean pEC50 values of 8.6, 6.8, and 6.3 M, respectively. Preincubation with alphaCGRP(8-37) (10(-7) -10(-5) M) and a novel nonpeptide CGRP antagonist "Compound 1" (WO98/11128) (10(-7)-10(-5) M) caused a dose-dependent rightward shift of the concentration-response curves for alphaCGRP with pA(2) values of 7.0 and 7.1, respectively. Preincubation with alphaCGRP(8-37) (10(-6) M) and Compound 1 (10(-6) M) caused significant rightward shift of the concentration-response curves for AM and amylin as well with pK B values between 6.6 and 7.5. Preincubation with AM(22-52) had no antagonistic effect on the AM and amylin response, neither did diacetoamidomethyl cysteine CGRP cause any concentration dependent (10(-11)-10(-6) M) dilatation. In conclusion, mRNA for the components forming CGRP1 and AM receptors was detected in the human left anterior descending coronary arteries. alphaCGRP, AM, and amylin mediated vasorelaxation via the CGRP1 receptor. Compound 1 acted as a nonpeptide antagonist at the CGRP1 receptor and could thus become a tool for the study of CGRP-mediated functional responses in human tissue.

KW - 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid

KW - Adrenomedullin

KW - Adult

KW - Amyloid

KW - Calcitonin Gene-Related Peptide

KW - Calcitonin Receptor-Like Protein

KW - Coronary Vessels

KW - Female

KW - Humans

KW - In Vitro Techniques

KW - Intracellular Signaling Peptides and Proteins

KW - Islet Amyloid Polypeptide

KW - Male

KW - Membrane Proteins

KW - Middle Aged

KW - Muscle Contraction

KW - Muscle Relaxation

KW - Muscle, Smooth, Vascular

KW - Peptide Fragments

KW - Peptides

KW - Piperazines

KW - Piperidines

KW - RNA, Messenger

KW - Receptor Activity-Modifying Proteins

KW - Receptors, Calcitonin

KW - Receptors, Calcitonin Gene-Related Peptide

UR - https://www.scopus.com/pages/publications/0037216736

U2 - 10.1124/jpet.102.037754

DO - 10.1124/jpet.102.037754

M3 - Journal article

C2 - 12490608

SN - 0022-3565

VL - 304

SP - 326

EP - 333

JO - The Journal of pharmacology and experimental therapeutics

JF - The Journal of pharmacology and experimental therapeutics

IS - 1

ER -

Investigation of CGRP receptors and peptide pharmacology in human coronary arteries. Characterization with a nonpeptide antagonist

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Keywords

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