In vitro pharmacological characterization of vorapaxar, a novel platelet thrombin receptor antagonist

Brian E. Hawes, Ying Zhai, David Hesk, Mark Wirth, Huijun Wei, Madhu Chintala, Dietmar Seiffert

Research output: Contribution to journalArticle

Abstract

Abstract Vorapaxar is a novel protease-activated receptor-1 (PAR1) antagonist recently approved for the reduction of thrombotic cardiovascular events in patients with a history of myocardial infarction or with peripheral arterial disease. The present study provides a comprehensive in vitro pharmacological characterization of vorapaxar interaction with the PAR1 receptor on human platelets. Similar studies were performed with a metabolite of vorapaxar (M20). Vorapaxar and M20 were competitive PAR1 antagonists that demonstrated concentration-dependent, saturable, specific, and slowly reversible binding to the receptor present on intact human platelets. The affinities of vorapaxar and M20 for the PAR1 receptor were in the low nanomolar range, as determined by saturation-, kinetic- and competitive binding studies. The calculated Kd and Ki values for vorapaxar increased in the presence of plasma, indicating a decrease in the free fraction available for binding to the PAR1 receptor on human platelets. Vorapaxar was also evaluated in functional assays using thrombin or a PAR1 agonist peptide (SFLLRN). Vorapaxar and M20 completely blocked thrombin-stimulated PAR1/β-arrestin association in recombinant cells and abolished thrombin-stimulated calcium influx in washed human platelets and vascular smooth muscle cells. Moreover, vorapaxar and M20 inhibited PAR1 agonist peptide-mediated platelet aggregation in human platelet rich plasma with a steep concentration response relationship. Vorapaxar exhibited high selectivity for inhibition of PAR1 over other platelet GPCRs. In conclusion, vorapaxar is a potent PAR1 antagonist exhibiting saturable, reversible, selective binding with slow off-rate kinetics and effectively inhibits thrombin's PAR1-mediated actions on human platelets.

Original languageEnglish (US)
Article number70010
Pages (from-to)221-228
Number of pages8
JournalEuropean Journal of Pharmacology
Volume762
Issue number1
DOIs
StatePublished - 2015

Keywords

  • In vitro pharmacology
  • Myocardial infarction
  • PAR1 antagonist
  • Peripheral arterial disease
  • Platelets
  • Vorapaxar

ASJC Scopus subject areas

  • Pharmacology

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