Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2017; 117(01): 44-56
DOI: 10.1160/TH16-05-0416

Coagulation and Fibrinolysis

Schattauer GmbH

A novel hirudin derivative inhibiting thrombin without bleeding for subcutaneous injection

Bing Zhao

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Yanling Zhang

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Yinong Huang

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Jinchao Yu

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Yaran Li

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Qi Wang

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Yixin Ma

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Hou-Yan Song

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

³Collaborative Innovation Center for Biotherapy, Sichuan University, Huaxi Campus, Chengdu, China

,

Min Yu

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Wei Mo

¹Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China

²Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

› Author Affiliations

Summary

Currently, anticoagulants would be used to prevent thrombosis. Thrombin is an effector enzyme for haemostasis and thrombosis. We designed a direct thrombin inhibitor peptide (DTIP) using molecular simulation and homology modelling and demonstrated that the C-terminus of DTIP interacts with exosite I, and N-terminus with the activity site of thrombin, respectively. DTIP interfered with thrombin-mediated coagulation in human, rat and mouse plasma (n=10 per group) and blocked clotting in human whole blood in vitro. When administered subcutaneously, DTIP showed potent and dose-dependent extension of aPTT, PT, TT and CT in rats (n=10 per group). The antithrombotic dose of DTIP induced significantly less bleeding than bivalirudin determined by transecting distal tail assay in rats. Furthermore, DTIP reached peak blood concentration in 0.5–1 hour and did not cause increased bleeding after five days of dosing compared to dabigatran etexilate. The antithrombotic effect of DTIP was evaluated in mice using lethal pulmonary thromboembolism model and FeCl₃-induced mesenteric arteriole thrombus model. DTIP (1.0 mg/kg, sc) prevented deep venous thrombosis and increased the survival rate associated with pulmonary thromboembolism from 30 % to 80 %. Intravital microscopy showed that DTIP (1.0 mg/kg, sc) decelerated mesenteric arteriole thrombosis caused by FeCl₃ injury. These data establish that DTIP is a novel antithrombotic agent that could be used to prevent thrombosis without conferring an increased bleeding risk.

Supplementary Material to this article is available at www.thrombosis-online.com.

Keywords

Keywords

Thrombin - r-RGD-hirudin - thrombosis - subcutaneous injection - direct thrombin inhibitor peptide (DTIP)

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Supplementary Material

Supplementary Material

Online Supplementary Material (PDF)