Engineering Factor Xa Inhibitor with Multiple Platelet-Binding Sites Facilitates its Platelet Targeting
Targeted delivery of antithrombotic drugs centralizes the effects in the thrombosis site and reduces the hemorrhage side effects in uninjured vessels.We have recently reported that the platelet-targeting factor Xa(FXa)inhibitors,constructed by engineering one Arg-Giy-Asp(RGD)motif into Ancylostoma caninum anticoagulant peptide 5(AcAPS),can reduce the risk of systemic bleeding than non-targeted AcAP5 in mouse arterial injury model.Increasing the number of platelet-binding sites of FXa inhibitors may facilitate their adhesion to activated platelets,and further lower the bleeding risks.For this purpose,we introduced three RGD motifs into AcAP5 to generate a variant NR4 containing three platelet-binding sites.NR4 reserved its inherent anti-FXa activity.Protein-protein docking showed that all three RGD motifs were capable of binding to platelet receptor αⅡbβ3.Molecular dynamics simulation demonstrated that NR4 has more opportunities to interact with αⅡbβ3than single-RGD-containing NR3.Flow cytometry analysis and rat arterial thrombosis model further confirmed that NR4 possesses enhanced platelet targeting activity.Moreover,NR4-treated mice showed a trend toward Iess tail bleeding time than NR3-treated mice in carotid artery endothelium injury model.Therefore,our data suggest that engineering multiple binding sites in one recombinant protein is a useful tool to improve its platelet-targeting efficiency.
Yuanjun Zhu Ruyi Li Yuan Lin Mengyang Shui Xiaoyan Liu Huan Chen Yinye Wang
Department of Molecular and Cellular Pharmacology,Peking University School of Pharmaceutical Science State Key Laboratory of Bioactive Substances and Function of Natural Medicine,Institute of Materia M
国内会议
大连
英文
369-379
2016-09-24（万方平台首次上网日期，不代表论文的发表时间）