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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
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Yanling Zhang
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Yinong Huang
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Jinchao Yu
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Yaran Li
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Qi Wang
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Yixin Ma
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Hou-Yan Song
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
3   Collaborative Innovation Center for Biotherapy, Sichuan University, Huaxi Campus, Chengdu, China
,
Min Yu
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
,
Wei Mo
1   Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Fudan University, Shanghai, China
2   Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China
› Author Affiliations Financial support: This study was supported by the National Natural Science Foundation of China (NSFC 81673498) and the Science and Technology Commission of Shanghai Municipality (STCSM 16431904600).
Further Information

Publication History

Received: 28 May 2016

Accepted after major revision: 17 September 2016

Publication Date:
14 November 2017 (online)

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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

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

 

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