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

• Published in: Biochimica et biophysica acta. General subjects Vol. 1861; no. 11; pp. 2609 - 2618
• Main Authors: Stylos, Evgenios, Chatziathanasiadou, Maria V., Tsiailanis, Antonis, Kellici, Tahsin F., Tsoumani, Maria, Kostagianni, Androniki D., Deligianni, Maria, Tselepis, Alexandros D., Tzakos, Andreas G.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.11.2017
• Subjects: antagonists; Antiplatelet; aspirin; Aspirin - therapeutic use; bioactive compounds; blood platelets; Cardiovascular diseases; Cardiovascular Diseases - blood; Cardiovascular Diseases - drug therapy; Computer Simulation; Cyclooxygenase 1 - drug effects; Cyclooxygenase 1 - metabolism; DHA; docosahexaenoic acid; Docosahexaenoic Acids - administration & dosage; Docosahexaenoic Acids - chemistry; Flavanones - administration & dosage; Flavanones - chemical synthesis; Flavonoids - administration & dosage; Flavonoids - chemical synthesis; hemorrhage; Hemorrhage - blood; Hemorrhage - chemically induced; Hemorrhage - prevention & control; Humans; in vitro studies; ligands; liquid chromatography; Naringenin; patients; Plasma stability; Platelet Activation - drug effects; platelet aggregation; Platelet Aggregation - drug effects; Platelet Aggregation Inhibitors - administration & dosage; Platelet Aggregation Inhibitors - chemical synthesis; prostaglandin synthase; proteinases; Receptor, PAR-1 - antagonists & inhibitors; Receptor, PAR-1 - metabolism; Receptors, Purinergic P2Y12 - drug effects; Receptors, Purinergic P2Y12 - metabolism; risk; Signal Transduction - drug effects; Standard of Care; Tandem Mass Spectrometry; therapeutics; Triple quadrupole; AA; RT; PAR-1; Naringenin; COX-1; Triple quadrupole; IS; ΜRΜ; ADP; TRAP-6; LC-MS/MS; Antiplatelet; Plasma stability; DHA; Cardiovascular diseases
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2017.08.018

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. 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. 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. The DHA – naringenin hybrid presented triple antiplatelet activity simultaneously targeting PAR-1, P2Y12 and COX-1 platelet activation pathways. Natural products could offer a rich source for novel bioactives as a powerful alternative to the current combinatorial use of three different antiplatelet drugs. [Display omitted] •The flavonoid naringenin presented diminished antiplatelet activity•A regioselective docosahexaenoic acid–naringenin hybrid was designed and synthesized•The naringenin hybrid showed enhanced stability in human plasma•The naringenin hybrid presented potent antiplatelet activity•This hybrid functions through PAR-1, P2Y12 and COX-1 receptors.