2. Academic Validation
  3. Discovery of 7, 4'-dimethoxy-3-hydroxyflavone as a protease-activated receptor 4 antagonist with antithrombotic activity and less bleeding tendency in mice

Discovery of 7, 4'-dimethoxy-3-hydroxyflavone as a protease-activated receptor 4 antagonist with antithrombotic activity and less bleeding tendency in mice

  • Biochem Pharmacol. 2022 Aug:202:115152. doi: 10.1016/j.bcp.2022.115152.
Ying-Ting Lin 1,  Yu Li 2,  Hui-Ching Hsu 2,  Ju-Ying Tsai 2,  Jia-Hau Lee 2,  Chi-Jung Tai 3,  Ming-Jung Wu 4,  Chin-Chung Wu 5
Affiliations

Affiliations

  • 1 Department of Biotechnology, College of Life Science, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
  • 2 Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
  • 3 Department of Family Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
  • 4 Department of Chemistry, National Sun Yat-Sen University, Kaohsiung 804, Taiwan.
  • 5 Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan; Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan; Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung 807, Taiwan. Electronic address: ccwu@kmu.edu.tw.
PMID: 35752281 DOI: 10.1016/j.bcp.2022.115152
Abstract

There is growing evidence of the importance of Protease-activated Receptor 4 (PAR4), one of Thrombin receptors, as a therapeutic target in thrombotic cardiovascular diseases. In the present study, we utilized ligand-based virtual screening, bioassay, and structure-activity relationship study to discover PAR4 antagonists with new chemical scaffolds from natural origin, and examined their application as antiplatelet agents. By using these approaches, we have identified a flavonoid, 7, 4'-dimethoxy-3-hydroxyflavone, that exhibits anti-PAR4 activity. 7, 4'-Dimethoxy-3-hydroxyflavone inhibited PAR4-mediated human platelet aggregation, GPIIb/IIIa activation, and P-selectin secretion. Also, it inhibited PAR4 downstream signaling pathways, including CA2+/protein kinase C, Akt, and MAP kinases ERK and p38, in human platelets, and suppressed PAR4-mediated β-arrestin recruitment in CHO-K1 cells exogenously expressed human PAR4. In a microfluidic system, 7, 4'-dimethoxy-3-hydroxyflavone reduced thrombus formation on collagen-coated chambers at an arterial shear rate in recalcified whole blood. Furthermore, mice treated with 7, 4'-dimethoxy-3-hydroxyflavone were significantly protected from FeCl3-induced carotid arterial occlusions, without significantly affecting tail bleeding time. In conclusion, 7, 4'-dimethoxy-3-hydroxyflavone represents a new class of nature-based PAR4 Antagonist, it shows effective in vivo antithrombotic properties with less bleeding tendency, and could be a potential candidate for developing new antiplatelet agents.

Keywords

Antiplatelet drugs; Flavonoids; Protease-activated receptor 4; Thrombosis; Virtual screening.

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