4′‑O‑MethylbavachalconeB Targeted 14–3–3ζ Blocking the Integrin β3 Early Outside-In Signal to Inhibit Platelet Aggregation and Thrombosis

• Published in: Journal of agricultural and food chemistry 2024-04, Vol.72 (13), p.7043-7054
• Main Authors: Tang, Ziqi, Lin, Fanqi, Chen, Zhiwen, Yu, Boyang, Liu, Ji-Hua, Liu, Xiufeng
• Format: Article
• Language: English
• Subjects: adhesion; amino acids; Bioactive Constituents, Metabolites, and Functions; food chemistry; integrins; ligands; mass spectrometry; mutation; phosphorylation; platelet aggregation; therapeutics; thrombosis
• ISSN: 0021-8561; 1520-5118; 1520-5118
• DOI: 10.1021/acs.jafc.3c05211

14–3–3ζ protein, the key target in the regulation and control of integrin β3 outside-in signaling, is an attractive new strategy to inhibit thrombosis without affecting hemostasis. In this study, 4′-O-methylbavachalconeB (4-O-MB) in Psoraleae Fructus was identified as a 14–3–3ζ ligand with antithrombosis activity by target fishing combined with ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) analysis. The competitive inhibition analysis showed that 4-O-MB targeted 14–3–3ζ and blocked the 14–3–3ζ/integrin β3 interaction with inhibition constant (Ki ) values of 9.98 ± 0.22 μM. Molecular docking and amino acid mutation experiments confirmed that 4-O-MB specifically bound to 14–3–3ζ through LSY9 and SER28 to regulate the 14–3–3ζ/integrin β3 interaction. Besides, 4-O-MB affected the integrin β3 early outside-in signal by inhibiting AKT and c-Src phosphorylation. Meanwhile, 4-O-MB could inhibit ADP-, collagen-, or thrombin-induced platelet aggregation function but had no effect on platelet adhesion to collagen-coated surfaces in vivo. Administration of 4-O-MB could significantly inhibit thrombosis formation without disturbing hemostasis in mice. These findings provide new prospects for the antithrombotic effects of Psoraleae Fructus and the potential application of 4-O-MB as lead compounds in the therapy of thrombosis by targeting 14–3–3ζ.