A long-acting PAI-1 inhibitor reduces thrombus formation

Shuangzhou Peng; Guangpu Xue; Lihu Gong; Chao Fang; Jingfei Chen; Cai Yuan; Zhuo Chen; Lishan Yao; Bruce Furie; Mingdong Huang

doi:10.1160/TH16-11-0891

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2017; 117(07): 1338-1347
DOI: 10.1160/TH16-11-0891

Coagulation and Fibrinolysis

Schattauer GmbH

A long-acting PAI-1 inhibitor reduces thrombus formation

Shuangzhou Peng

¹State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, China

²College of life sciences, Fujian Agriculture and Forestry University, Fuzhou, Fujian, China

,

Guangpu Xue

¹State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, China

⁵College of Life science, Fujian Normal University, Fuzhou, Fujian, China

,

Lihu Gong

¹State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, China

,

Chao Fang

⁶Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

⁷School of Basic Medicine, Tongji Medical College, Huazhong University of Science & Technology, Huazhong, China

,

Jingfei Chen

⁴Qingdao Institute of Biomass Energy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China

,

Cai Yuan

³Fuzhou University,Fuzhou, Fujian, China

,

Zhuo Chen

¹State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, China

,

Lishan Yao

⁴Qingdao Institute of Biomass Energy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China

,

Bruce Furie

⁶Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

,

Mingdong Huang

¹State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, China

³Fuzhou University,Fuzhou, Fujian, China

› Institutsangaben

Abstract

Summary

Plasminogen activator inhibitor 1 (PAI-1) is the main inhibitor of tissue-type and urokinase-type plasminogen activators (t/uPA) and plays an important role in fibrinolysis. Inhibition of PAI-1 activity prevents thrombosis and accelerates fibrinolysis, indicating that PAI-1 inhibitors may be used as effective antithrombotic agents. We previously designed a PAI-1 inhibitor (PAItrap) which is a variant of inactivated urokinase protease domain. In the present study, we fused PAItrap with human serum albumin (HSA) to develop a long-acting PAI-1 inhibitor. Unfortunately, the fusion protein PAItrap-HSA lost some potency compared to PAItrap (33 nM vs 10 nM). Guided by computational method, we carried out further optimisation to enhance inhibitory potency for PAI-1. The new PAItrap, denominated PAItrap(H37R)-HSA, which was the H37R variant of PAItrap fused to HSA, gave a six-fold improvement of IC₅₀ (5 nM) for human active PAI-1 compared to PAItrap-HSA, and showed much longer plasma half-life (200-fold) compared to PAItrap. We further demonstrated that the PAItrap(H37R)-HSA inhibited exogenous or endogenous PAI-1 to promote fibrinolysis in fibrin-clot lysis assay. PAItrap(H37R)-HSA inhibits murine PAI-1 with IC₅₀ value of 12 nM, allowing the inhibitor to be evaluated in murine models. Using an intravital microscopy, we demonstrated that PAItrap(H37R)-HSA blocks thrombus formation and platelet accumulation in vivo in a laser-induced vascular injury mouse model. Additionally, mouse tail bleeding assay showed that PAItrap(H37R)-HSA did not affect the global haemostasis. These results suggest that PAItrap(H37R)-HSA have the potential benefit to prevent thrombosis and accelerates fibrinolysis.

Keywords

PAI-1 inhibitor - human serum albumin (HSA) - thrombosis - clot lysis

Volltext

Referenzen

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