title: Functional estimation of endothelin-1 receptor antagonism by bosentan, macitentan and ambrisentan in human pulmonary and radial arteries in vitro.
abstract: BACKGROUND
  Endothelin receptor antagonists are approved for pulmonary arterial hypertension. Development of selective ETA-receptor antagonists over mixed or dual receptor antagonists has depended on a range of receptor binding assays, second messenger assays and functional blood vessel assays. This study compared the 3 clinically-approved endothelin receptor antagonists in assays of human isolated pulmonary and radial arteries in vitro.
  
  
  METHODS
  Human isolated pulmonary (i.d. 5.5mm) and human radial (i.d. 3.23mm) artery ring segments were mounted in organ baths for isometric force measurement. Single concentration-contraction curves to endothelin-1 were constructed in the absence or presence of bosentan (1-10µM), macitentan (0.03-0.3µM) or ambrisentan (0.1-1µM).
  
  
  RESULTS
  All 3 endothelin antagonists caused competitive rightward shifts in the endothelin-1 concentration-response curves in both arteries. The Clark plot and analysis gave the following pKBvalues: bosentan, pulmonary artery 6.28±0.13 and radial artery 6.04±0.10; macitentan, pulmonary artery 8.02±0.13 and radial artery 7.49±0.08; and ambrisentan, pulmonary artery 7.38±0.13 and radial artery 6.96±0.10.
  
  
  CONCLUSIONS
  Noting the maximum plasma levels attained from recommended oral doses of each antagonist in volunteers, the pKBfindings here show that there would be significant antagonism of endothelin-1 contraction in the pulmonary and radial arteries at therapeutic plasma levels. This functional assay confirms in human tissue that much higher plasma concentrations of endothelin-1 receptor antagonists are required to be effective than those predicted from binding or other biochemical assays.
authors.affiliation: Cardiovascular Therapeutics Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia. Electronic address: jamesaa@unimelb.edu.au.
authors.forename: James A
authors.initials: JA
authors.lastname: Angus
authors.affiliation: Cardiovascular Therapeutics Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia. Electronic address: paulsoeding@bigpond.com.
authors.forename: Paul F
authors.initials: PF
authors.lastname: Soeding
authors.affiliation: Cardiovascular Therapeutics Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia. Electronic address: richard.hughes@uqconnect.edu.au.
authors.forename: Richard J A
authors.initials: RJA
authors.lastname: Hughes
authors.affiliation: Cardiovascular Therapeutics Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia. Electronic address: cewright@unimelb.edu.au.
authors.forename: Christine E
authors.initials: CE
authors.lastname: Wright
mesh_terms: D000077300:Bosentan; D004305:Dose-Response Relationship, Drug; D065128:Endothelin Receptor Antagonists / Q000494:pharmacology* / Q000627:therapeutic use; D019332:Endothelin-1 / Q000378:metabolism; D006801:Humans; D006976:Hypertension, Pulmonary / Q000188:drug therapy; D010666:Phenylpropionates / Q000494:pharmacology / Q000627:therapeutic use; D011651:Pulmonary Artery / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology*; D011724:Pyridazines / Q000494:pharmacology / Q000627:therapeutic use; D011743:Pyrimidines / Q000494:pharmacology / Q000627:therapeutic use; D017534:Radial Artery / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology*; D044022:Receptor, Endothelin A / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology / Q000627:therapeutic use; D014022:Tissue Survival / Q000187:drug effects; D014661:Vasoconstriction / Q000187:drug effects
chemical_list: D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1; D010666:Phenylpropionates; D011724:Pyridazines; D011743:Pyrimidines; D044022:Receptor, Endothelin A; D013449:Sulfonamides; C467894:ambrisentan; D000077300:Bosentan; C533860:macitentan
keywords: Ambrisentan; Ambrisentan (PubChem CID: 6918493); Bosentan; ET(1)-receptors; Endothelin-1; Human pulmonary artery; Human radial artery; Macitentan; and macitentan (PubChem CID: 16004692); bosentan sodium (PubChem CID: 44387533); endothelin-1 (PubChem CID: 16132423)