Tailoring naringenin conjugates with amplified and triple antiplatelet activity profile: Rational design, synthesis, human plasma stability and in vitro evaluation

Biochim Biophys Acta Gen Subj. 2017 Nov;1861(11 Pt A):2609-2618. doi: 10.1016/j.bbagen.2017.08.018. Epub 2017 Aug 24.

Abstract

Background: The current standard-of-care antiplatelet therapy in cardiovascular disease patients is consisted of cyclooxygenase-1 (COX-1) inhibitor aspirin, along with a platelet receptor P2Y12 antagonist. Recently, the triple antiplatelet therapy with aspirin, a P2Y12 receptor antagonist and a protease activated receptor-1 (PAR-1) antagonist, has been suggested for the secondary prevention of atherothrombotic events, however presented an increased risk of bleeding. Therefore, the quest for novel antiplatelet agents simultaneously targeting the three pathways with improved efficacy/safety profile is of immense importance. Flavonoids as pre-validated ligands for numerous targets could serve as scaffolds targeting the three platelet activation pathways.

Methods: Computational methods, Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS) plasma stability and in vitro platelet aggregation experiments were used to establish the antiplatelet activity of the flavonoid naringenin and its conjugates.

Results: In silico studies indicated that naringenin could bear a potent triple antiplatelet activity by inhibiting different platelet aggregation mechanisms. However, we found that in human platelets naringenin has diminished activity. We rationally designed and synthesized different naringenin conjugates aiming to amplify the antiplatelet activity of the parent compound. UHPLC-MS/MS revealed a slow degradation rate for a docosahexaenoic acid (DHA) - naringenin conjugate in human plasma. The antiplatelet profile of the new analogues was evaluated against in vitro platelet aggregation induced by several platelet agonists.

Conclusions: The DHA - naringenin hybrid presented triple antiplatelet activity simultaneously targeting PAR-1, P2Y12 and COX-1 platelet activation pathways.

General significance: Natural products could offer a rich source for novel bioactives as a powerful alternative to the current combinatorial use of three different antiplatelet drugs.

Keywords: Antiplatelet; Cardiovascular diseases; DHA; Naringenin; Plasma stability; Triple quadrupole.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspirin / therapeutic use
  • Cardiovascular Diseases / blood
  • Cardiovascular Diseases / drug therapy*
  • Computer Simulation
  • Cyclooxygenase 1 / drug effects
  • Cyclooxygenase 1 / metabolism*
  • Docosahexaenoic Acids / administration & dosage
  • Docosahexaenoic Acids / chemistry
  • Flavanones / administration & dosage*
  • Flavanones / chemical synthesis
  • Flavonoids / administration & dosage
  • Flavonoids / chemical synthesis
  • Hemorrhage / blood
  • Hemorrhage / chemically induced
  • Hemorrhage / prevention & control
  • Humans
  • Platelet Activation / drug effects
  • Platelet Aggregation / drug effects
  • Platelet Aggregation Inhibitors / administration & dosage*
  • Platelet Aggregation Inhibitors / chemical synthesis
  • Receptor, PAR-1 / antagonists & inhibitors
  • Receptor, PAR-1 / metabolism*
  • Receptors, Purinergic P2Y12 / drug effects
  • Receptors, Purinergic P2Y12 / metabolism*
  • Signal Transduction / drug effects
  • Standard of Care
  • Tandem Mass Spectrometry

Substances

  • Flavanones
  • Flavonoids
  • P2RY12 protein, human
  • Platelet Aggregation Inhibitors
  • Receptor, PAR-1
  • Receptors, Purinergic P2Y12
  • Docosahexaenoic Acids
  • Cyclooxygenase 1
  • PTGS1 protein, human
  • naringenin
  • Aspirin