Download PDF
Thromb Haemost 2020; 120(04): 607-619
DOI: 10.1055/s-0040-1708480
Coagulation and Fibrinolysis
Georg Thieme Verlag KG Stuttgart · New York

Effects of Native Fucosylated Glycosaminoglycan, Its Depolymerized Derivatives on Intrinsic Factor Xase, Coagulation, Thrombosis, and Hemorrhagic Risk

Lutan Zhou
1   Key Laboratory of Medicinal Chemistry for Natural Resource, Ministry of Education and Yunnan Province, School of Chemical Science and Technology, Yunnan University, Kunming, China
2   State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
3   University of Chinese Academy of Sciences, Beijing, China
,
Na Gao   *
4   School of Pharmaceutical Sciences, South Central University for Nationalities, Wuhan, China
,
Huifang Sun
2   State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
3   University of Chinese Academy of Sciences, Beijing, China
,
Chuang Xiao
5   School of Pharmaceutical Science and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming, China
,
Lian Yang
2   State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
,
Lisha Lin
2   State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
3   University of Chinese Academy of Sciences, Beijing, China
,
Ronghua Yin
2   State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
,
Zi Li
2   State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
,
Hongbin Zhang
1   Key Laboratory of Medicinal Chemistry for Natural Resource, Ministry of Education and Yunnan Province, School of Chemical Science and Technology, Yunnan University, Kunming, China
,
Xu Ji
1   Key Laboratory of Medicinal Chemistry for Natural Resource, Ministry of Education and Yunnan Province, School of Chemical Science and Technology, Yunnan University, Kunming, China
6   Suzhou Banbutang Biological Technology Co., LTD, Suzhou, China
,
Jinhua Zhao
2   State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
4   School of Pharmaceutical Sciences, South Central University for Nationalities, Wuhan, China
› Author Affiliations Funding This work was funded in part by the National Natural Science Foundation of China (81773737, 81872774, 81703374, 81673330 and 31900926), and Yunnan Provincial Science and Technology Department in China (2018HC012 and 2019ZF011–2). Also, it was supported by the grants from the Dongwu Leading Talent Plan of Scientific-and-Technological Innovation and Entrepreneurship.
Further Information

Publication History

02 September 2019

23 January 2020

Publication Date:
14 April 2020 (online)

    Permissions and Reprints

Abstract

A native fucosylated glycosaminoglycan from sea cucumber Holothuria fuscopunctata (nHG), mainly branched with Fuc3S4S, exhibited potent anticoagulant activity by intrinsic tenase iXase (FIXa-FVIIIa complex) and antithrombin-dependent factor IIa (FIIa) inhibition, but also had the effects of FXII activation and platelet aggregation. For screening a selective iXase inhibitor, depolymerized nHG (dHG-1 ∼ –6) and a pure octasaccharide (oHG-8) were prepared. Like nHG, dHG-1 ∼ –6 and oHG-8 could potently inhibit iXase, and competitive binding assay indicated that dHG-5 and oHG-8 could bind to FIXa. Nevertheless, dHG-5 and oHG-8 had no effects on FXII and platelet activation. nHG, dHG-5, and oHG-8 could significantly prolong the activated partial thromboplastin time of human, rat, and rabbit plasma. In the rat deep venous thrombosis model, dHG-5 and oHG-8 showed potent antithrombotic effects in a dose-dependent manner, while the thrombus inhibition rate of nHG at high dose was markedly reduced. Additionally, dHG-5 and oHG-8 did not increase bleeding at the doses up to 10-fold of the effectively antithrombotic doses compared with nHG and low molecular weight heparin in the mice tail-cut model. Considering that dHG-5 possesses strong anti-iXase and antithrombotic activities, and its preparation process is simpler and its yield is higher compared with oHG-8, it might be a promising antithrombotic candidate.

Keywords

dHG-5 - iXase - anticoagulant - antithrombotic - hemorrhagic risk

* These authors contributed equally to this work.


Supplementary Material

 

  • References

  • 1 Konstantinides S, Torbicki A. Management of venous thrombo-embolism: an update. Eur Heart J 2014; 35 (41) 2855-2863
    CrossrefPubMedGoogle Scholar
  • 2 Mackman N. Triggers, targets and treatments for thrombosis. Nature 2008; 451 (7181): 914-918
    PubMedGoogle Scholar
  • 3 Eikelboom JW, Weitz JI. New anticoagulants. Circulation 2010; 121 (13) 1523-1532
    CrossrefPubMedGoogle Scholar
  • 4 Schulman S, Beyth RJ, Kearon C, Levine MN. Hemorrhagic complications of anticoagulant and thrombolytic treatment: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition). Chest 2008; 133 (6, Suppl): 257S-298S
    CrossrefPubMedGoogle Scholar
  • 5 Fareed J, Hoppensteadt DA, Fareed D. , et al. Survival of heparins, oral anticoagulants, and aspirin after the year 2010. Semin Thromb Hemost 2008; 34 (01) 58-73
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Salmela B, Joutsi-Korhonen L, Armstrong E, Lassila R. Active online assessment of patients using new oral anticoagulants: bleeding risk, compliance, and coagulation analysis. Semin Thromb Hemost 2012; 38 (01) 23-30
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Renné T. Safe(r) anticoagulation. Blood 2010; 116 (22) 4390-4391
    PubMedGoogle Scholar
  • 8 Eriksson BI, Quinlan DJ, Eikelboom JW. Novel oral factor Xa and thrombin inhibitors in the management of thromboembolism. Annu Rev Med 2011; 62: 41-57
    CrossrefPubMedGoogle Scholar
  • 9 Fareed J, Thethi I, Hoppensteadt D. Old versus new oral anticoagulants: focus on pharmacology. Annu Rev Pharmacol Toxicol 2012; 52: 79-99
    CrossrefPubMedGoogle Scholar
  • 10 Colman RW. Are hemostasis and thrombosis two sides of the same coin?. J Exp Med 2006; 203 (03) 493-495
    CrossrefPubMedGoogle Scholar
  • 11 Howard EL, Becker KCD, Rusconi CP, Becker RC. Factor IXa inhibitors as novel anticoagulants. Arterioscler Thromb Vasc Biol 2007; 27 (04) 722-727
    CrossrefPubMedGoogle Scholar
  • 12 Eikelboom JW, Zelenkofske SL, Rusconi CP. Coagulation factor IXa as a target for treatment and prophylaxis of venous thromboembolism. Arterioscler Thromb Vasc Biol 2010; 30 (03) 382-387
    CrossrefPubMedGoogle Scholar
  • 13 Mackman N. New insights into the mechanisms of venous thrombosis. J Clin Invest 2012; 122 (07) 2331-2336
    CrossrefPubMedGoogle Scholar
  • 14 Plow EF, Wang Y, Simon DI. The search for new antithrombotic mechanisms and therapies that may spare hemostasis. Blood 2018; 131 (17) 1899-1902
    CrossrefPubMedGoogle Scholar
  • 15 Müller F, Gailani D, Renné T. Factor XI and XII as antithrombotic targets. Curr Opin Hematol 2011; 18 (05) 349-355
    CrossrefPubMedGoogle Scholar
  • 16 He R, Chen D, He S. Factor XI: hemostasis, thrombosis, and antithrombosis. Thromb Res 2012; 129 (05) 541-550
    CrossrefPubMedGoogle Scholar
  • 17 Büller HR, Bethune C, Bhanot S. , et al; FXI-ASO TKA Investigators. Factor XI antisense oligonucleotide for prevention of venous thrombosis. N Engl J Med 2015; 372 (03) 232-240
    CrossrefPubMedGoogle Scholar
  • 18 Yau JW, Liao P, Fredenburgh JC. , et al. Selective depletion of factor XI or factor XII with antisense oligonucleotides attenuates catheter thrombosis in rabbits. Blood 2014; 123 (13) 2102-2107
    CrossrefPubMedGoogle Scholar
  • 19 Eriksson BI, Dahl OE, Lassen MR. , et al; Fixit Study Group. Partial factor IXa inhibition with TTP889 for prevention of venous thromboembolism: an exploratory study. J Thromb Haemost 2008; 06 (03) 457-463
    CrossrefPubMedGoogle Scholar
  • 20 Wheeler AP, Gailani D. The intrinsic pathway of coagulation as a target for antithrombotic therapy. Hematol Oncol Clin North Am 2016; 30 (05) 1099-1114
    CrossrefPubMedGoogle Scholar
  • 21 Fay PJ. Activation of factor VIII and mechanisms of cofactor action. Blood Rev 2004; 18 (01) 1-15
    CrossrefPubMedGoogle Scholar
  • 22 Brandstetter H, Bauer M, Huber R, Lollar P, Bode W. X-ray structure of clotting factor IXa: active site and module structure related to Xase activity and hemophilia B. Proc Natl Acad Sci U S A 1995; 92 (21) 9796-9800
    CrossrefPubMedGoogle Scholar
  • 23 Sheehan JP, Walke EN. Depolymerized holothurian glycosaminoglycan and heparin inhibit the intrinsic tenase complex by a common antithrombin-independent mechanism. Blood 2006; 107 (10) 3876-3882
    CrossrefPubMedGoogle Scholar
  • 24 Buyue Y, Sheehan JP. Fucosylated chondroitin sulfate inhibits plasma thrombin generation via targeting of the factor IXa heparin-binding exosite. Blood 2009; 114 (14) 3092-3100
    PubMedGoogle Scholar
  • 25 Pomin VH. Holothurian fucosylated chondroitin sulfate. Mar Drugs 2014; 12 (01) 232-254
    CrossrefPubMedGoogle Scholar
  • 26 Li JZ, Lian ECY. Aggregation of human platelets by acidic mucopolysaccharide extracted from Stichopus japonicus Selenka. Thromb Haemost 1988; 59 (03) 435-439
    Article in Thieme ConnectPubMedGoogle Scholar
  • 27 Li JZ, Lian ECY. Mechanism of rabbit platelet agglutination induced by acidic mucopolysaccharide extracted from Stichopus japonicus Selenka. Thromb Haemost 1988; 59 (03) 432-434
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Fonseca RJC, Santos GRC, Mourão PAS. Effects of polysaccharides enriched in 2,4-disulfated fucose units on coagulation, thrombosis and bleeding. Practical and conceptual implications. Thromb Haemost 2009; 102 (05) 829-836
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Suzuki N, Kitazato K, Takamatsu J, Saito H. Antithrombotic and anticoagulant activity of depolymerized fragment of the glycosaminoglycan extracted from Stichopus japonicus Selenka. Thromb Haemost 1991; 65 (04) 369-373
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Kitazato K, Kitazato KT, Nagase H, Minamiguchi K. DHG, a new depolymerized holothurian glycosaminoglycan, exerts an antithrombotic effect with less bleeding than unfractionated or low molecular weight heparin, in rats. Thromb Res 1996; 84 (02) 111-120
    CrossrefPubMedGoogle Scholar
  • 31 Wu M, Huang R, Wen D. , et al. Structure and effect of sulfated fucose branches on anticoagulant activity of the fucosylated chondroitin sulfate from sea cucumber Thelenata ananas . Carbohydr Polym 2012; 87 (01) 862-868
    CrossrefPubMedGoogle Scholar
  • 32 Gao N, Lu F, Xiao C. , et al. β-Eliminative depolymerization of the fucosylated chondroitin sulfate and anticoagulant activities of resulting fragments. Carbohydr Polym 2015; 127: 427-437
    CrossrefPubMedGoogle Scholar
  • 33 Zhao L, Wu M, Xiao C. , et al. Discovery of an intrinsic tenase complex inhibitor: pure nonasaccharide from fucosylated glycosaminoglycan. Proc Natl Acad Sci U S A 2015; 112 (27) 8284-8289
    CrossrefPubMedGoogle Scholar
  • 34 Wu M, Wen D, Gao N. , et al. Anticoagulant and antithrombotic evaluation of native fucosylated chondroitin sulfates and their derivatives as selective inhibitors of intrinsic factor Xase. Eur J Med Chem 2015; 92: 257-269
    CrossrefPubMedGoogle Scholar
  • 35 Shang F, Mou R, Zhang Z. , et al. Structural analysis and anticoagulant activities of three highly regular fucan sulfates as novel intrinsic factor Xase inhibitors. Carbohydr Polym 2018; 195: 257-266
    CrossrefPubMedGoogle Scholar
  • 36 Yin R, Zhou L, Gao N. , et al. Oligosaccharides from depolymerized fucosylated glycosaminoglycan: structures and minimum size for intrinsic factor Xase complex inhibition. J Biol Chem 2018; 293 (36) 14089-14099
    CrossrefPubMedGoogle Scholar
  • 37 Wu M, Xu S, Zhao J. , et al. Physicochemical characteristics and anticoagulant activities of low molecular weight fractions by free-radical depolymerization of a fucosylated chondroitin sulphate from sea cucumber Thelenata ananas . Food Chem 2010; 122 (03) 716-723
    CrossrefPubMedGoogle Scholar
  • 38 Shang F, Gao N, Yin R. , et al. Precise structures of fucosylated glycosaminoglycan and its oligosaccharides as novel intrinsic factor Xase inhibitors. Eur J Med Chem 2018; 148: 423-435
    CrossrefPubMedGoogle Scholar
  • 39 Zheng W, Zhou L, Lin L. , et al. Physicochemical characteristics and anticoagulant activities of the polysaccharides from sea cucumber Pattalus mollis . Mar Drugs 2019; 17 (04) E198
    CrossrefPubMedGoogle Scholar
  • 40 Xiao C, Zhao L, Gao N, Wu M, Zhao J. Nonasaccharide inhibits intrinsic factor Xase complex by binding to factor IXa and disrupting factor IXa-factor VIIIa interactions. Thromb Haemost 2019; 119 (05) 705-715
    Article in Thieme ConnectPubMedGoogle Scholar
  • 41 Hojima Y, Cochrane CG, Wiggins RC, Austen KF, Stevens RL. In vitro activation of the contact (Hageman factor) system of plasma by heparin and chondroitin sulfate E. Blood 1984; 63 (06) 1453-1459
    CrossrefPubMedGoogle Scholar
  • 42 Fonseca RJ, Oliveira SN, Pomin VH, Mecawi AS, Araujo IG, Mourão PA. Effects of oversulfated and fucosylated chondroitin sulfates on coagulation. Challenges for the study of anticoagulant polysaccharides. Thromb Haemost 2010; 103 (05) 994-1004
    Article in Thieme ConnectPubMedGoogle Scholar
  • 43 Walsh PN, Mills DC, White JG. Metabolism and function of human platelets washed by albumin density gradient separation. Br J Haematol 1977; 36 (02) 287-296
    PubMedGoogle Scholar
  • 44 Born GV. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 1962; 194: 927-929
    PubMedGoogle Scholar
  • 45 Xiao C, Lian W, Zhou L. , et al. Interactions between depolymerized fucosylated glycosaminoglycan and coagulation proteases or inhibitors. Thromb Res 2016; 146: 59-68
    CrossrefPubMedGoogle Scholar
  • 46 Olson ST, Halvorson HR, Björk I. Quantitative characterization of the thrombin-heparin interaction. Discrimination between specific and nonspecific binding models. J Biol Chem 1991; 266 (10) 6342-6352
    CrossrefPubMedGoogle Scholar
  • 47 Yoshida K, Minami Y, Nemoto H. , et al. Structure of DHG, a depolymerized glycosaminoglycan from sea cucumber, Stichopus Japonicus . Tetrahedron Lett 1992; 33 (34) 4959-4962
    CrossrefPubMedGoogle Scholar
  • 48 Mizurini DM, Francischetti IMB, Andersen JF, Monteiro RQ. Nitrophorin 2, a factor IX(a)-directed anticoagulant, inhibits arterial thrombosis without impairing haemostasis. Thromb Haemost 2010; 104 (06) 1116-1123
    Article in Thieme ConnectPubMedGoogle Scholar
  • 49 Vieira RP, Mourão PA. Occurrence of a unique fucose-branched chondroitin sulfate in the body wall of a sea cucumber. J Biol Chem 1988; 263 (34) 18176-18183
    CrossrefPubMedGoogle Scholar
  • 50 Ustyuzhanina NE, Bilan MI, Dmitrenok AS. , et al. Structure and biological activity of a fucosylated chondroitin sulfate from the sea cucumber Cucumaria japonica. Glycobiology 2016; 26 (05) 449-459
    CrossrefPubMedGoogle Scholar