title: Binding and functional pharmacological characteristics of gepant-type antagonists in rat brain and mesenteric arteries.
abstract: The neuropeptide calcitonin gene-related peptide (CGRP) is found in afferent sensory nerve fibers innervating the resistance arteries and plays a pivotal role in a number of neurovascular diseases such as migraine and subarachnoid bleedings. The present study investigates the binding and antagonistic characteristics of small non-peptide CGRP receptor antagonists (i.e. gepants) in isolated rat brain and mesenteric resistance arteries.
  
  
  
  The antagonistic behavior of gepants was investigated in isolated rat mesenteric arteries using a wire myograph setup while binding of gepants to CGRP receptors was investigated in rat brain membranes using a radioligand competitive binding assay. Furthermore, the histological location of the key components of CGRP receptor (RAMP1 and CLR) was assessed by immunohistochemistry.
  
  
  
  Our functional studies clearly show that all gepants are reversible competitive antagonists producing Schild plot slopes not significantly different from unity and thus suggesting presence of a uniform CGRP receptor population in the arteries. A uniform receptor population was also confirmed by radioligand competitive binding studies showing similar affinities for the gepants in rat brain and mesenteric arteries, the exception being rimegepant which had 50-fold lower affinity in brain than mesenteric arteries. CLR and RAMP1 were shown to be located in both vascular smooth muscle and endothelial cells of rat mesenteric arteries by immunohistochemistry.
  
  
  
  The present results indicate that, despite species differences in the CGRP receptor affinity, the antagonistic nature of these gepants, the distribution pattern of CGRP receptor components and the mechanism behind CGRP-induced vasodilation seem to be similar in resistance-sized arteries of human and rats.
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark. Electronic address: mash@sund.ku.dk.
authors.forename: Majid
authors.initials: M
authors.lastname: Sheykhzade
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
authors.forename: Nilofar
authors.initials: N
authors.lastname: Amandi
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
authors.forename: Monica Vidal
authors.initials: MV
authors.lastname: Pla
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
authors.forename: Bahareh
authors.initials: B
authors.lastname: Abdolalizadeh
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Glostrup University Hospital, DK-2600 Glostrup, Denmark.
authors.forename: Anette
authors.initials: A
authors.lastname: Sams
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Glostrup University Hospital, DK-2600 Glostrup, Denmark.
authors.forename: Karin
authors.initials: K
authors.lastname: Warfvinge
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Glostrup University Hospital, DK-2600 Glostrup, Denmark.
authors.forename: Lars
authors.initials: L
authors.lastname: Edvinsson
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
authors.forename: Darryl S
authors.initials: DS
authors.lastname: Pickering
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001381:Azepines / Q000494:pharmacology*; D001667:Binding, Competitive; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology*; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism / Q000494:pharmacology; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D058286:Calcitonin Receptor-Like Protein / Q000378:metabolism; D002531:Cerebellum / Q000098:blood supply*; D002536:Cerebral Arteries / Q000187:drug effects* / Q000378:metabolism; D004151:Dipeptides / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D066298:In Vitro Techniques; D008024:Ligands; D008297:Male; D008638:Mesenteric Arteries / Q000187:drug effects* / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism; D009213:Myography; D010879:Piperazines; D010880:Piperidines / Q000494:pharmacology*; D011485:Protein Binding; D011725:Pyridines / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology*; D011869:Radioligand Assay; D017207:Rats, Sprague-Dawley; D058262:Receptor Activity-Modifying Protein 1 / Q000378:metabolism; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000378:metabolism; D013141:Spiro Compounds / Q000494:pharmacology*; D034421:Sus scrofa; D014664:Vasodilation / Q000187:drug effects; D014665:Vasodilator Agents / Q000378:metabolism / Q000494:pharmacology
chemical_list: C548781:2-(8-(3,5-difluorophenyl)-10-oxo-6,9-diazaspiro(4.5)dec-9-yl)-N-(2'-oxo-1,1',2',3-tetrahydrospiro(indene-2,3'-pyrrolo(2,3-b)pyridin)-5-yl)acetamide; D001381:Azepines; D019086:Bridged Bicyclo Compounds, Heterocyclic; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D058286:Calcitonin Receptor-Like Protein; C545580:Calcrl protein, rat; D004151:Dipeptides; D007093:Imidazoles; D008024:Ligands; D010879:Piperazines; D010880:Piperidines; D011725:Pyridines; D011799:Quinazolines; C545508:Ramp1 protein, rat; D058262:Receptor Activity-Modifying Protein 1; D018015:Receptors, Calcitonin Gene-Related Peptide; D013141:Spiro Compounds; D014665:Vasodilator Agents; C578443:rimegepant sulfate; C525458:telcagepant; D015740:Calcitonin Gene-Related Peptide; C406305:olcegepant
keywords: Binding affinity; Brain; CGRP receptor; Gepants; Mesenteric arteries; Schild plot