P2Y12 receptor blockade synergizes strongly with nitric oxide and prostacyclin to inhibit platelet activation

Br J Clin Pharmacol. 2016 Apr;81(4):621-33. doi: 10.1111/bcp.12826. Epub 2016 Feb 10.


Aims: In vivo platelet function is a product of intrinsic platelet reactivity, modifiable by dual antiplatelet therapy (DAPT), and the extrinsic inhibitory endothelial mediators, nitric oxide (NO) and prostacyclin (PGI2 ), that are powerfully potentiated by P2Y12 receptor blockade. This implies that for individual patients endothelial mediator production is an important determinant of DAPT effectiveness. Here, we have investigated this idea using platelets taken from healthy volunteers treated with anti-platelet drugs.

Methods: Three groups of male volunteers (n = 8) received either prasugrel (10 mg), aspirin (75 mg) or DAPT (prasugrel + aspirin) once daily for 7 days. Platelet reactivity in the presence of diethylammonium (Z)-1-(N,N-diethylamino)diazen-1-ium-1,2-diolate (DEA/NONOate) and PGI2 was studied before and following treatment.

Results: Ex vivo, PGI2 and/or DEA/NONOate had little inhibitory effect on TRAP-6-induced platelet reactivity in control conditions. However, in the presence of DAPT, combination of DEA/NONOate + PGI2 reduced platelet aggregation (74 ± 3% to 19 ± 6%, P < 0.05). In vitro studies showed even partial (25%) P2Y12 receptor blockade produced a significant (67 ± 2% to 39 ± 10%, P < 0.05) inhibition when DEA/NONOate + PGI2 was present.

Conclusions: We have demonstrated that PGI2 and NO synergize with P2Y12 receptor antagonists to produce powerful platelet inhibition. Furthermore, even with submaximal P2Y12 blockade the presence of PGI2 and NO greatly enhances platelet inhibition. Our findings highlight the importance of endothelial mediator in vivo modulation of P2Y12 inhibition and introduces the concept of refining ex vivo platelet function testing by incorporating an assessment of endothelial function to predict thrombotic outcomes better and adjust therapy to prevent adverse outcomes in individual patients.

Keywords: blood platelets; endothelium; epoprostenol; nitric oxide; purinergic P2Y receptor antagonists.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aspirin / administration & dosage
  • Aspirin / pharmacology*
  • Blood Platelets / drug effects
  • Drug Synergism
  • Epoprostenol / administration & dosage
  • Epoprostenol / metabolism
  • Epoprostenol / pharmacology*
  • Healthy Volunteers
  • Humans
  • In Vitro Techniques
  • Male
  • Nitric Oxide / administration & dosage
  • Nitric Oxide / metabolism
  • Nitric Oxide / pharmacology*
  • Nitric Oxide Donors / pharmacology
  • Platelet Activation / drug effects*
  • Platelet Aggregation / drug effects
  • Prasugrel Hydrochloride / administration & dosage
  • Prasugrel Hydrochloride / pharmacology*
  • Purinergic P2Y Receptor Antagonists / administration & dosage
  • Purinergic P2Y Receptor Antagonists / pharmacology*
  • Young Adult


  • Nitric Oxide Donors
  • Purinergic P2Y Receptor Antagonists
  • Nitric Oxide
  • Epoprostenol
  • Prasugrel Hydrochloride
  • Aspirin