Download PDF
Thromb Haemost 1996; 75(04): 585-589
DOI: 10.1055/s-0038-1650326
Original Article
Schattauer GmbH Stuttgart

Experimental Haemorrhagic Effect of Two-Domain Non-Glycosylated Tissue Factor Pathway Inhibitor Compared to Low Molecular Weight Heparin

Jan Holst
1   The Dept. of Surgery, Helsirrgborg Hospital, Helsingborg, Sweden
2   Dept. of Surgery and Experimental Research, Malmö University Hospital, Malmö, Sweden
,
Bengt Lindblad
2   Dept. of Surgery and Experimental Research, Malmö University Hospital, Malmö, Sweden
,
Stefan E Matthíasson
2   Dept. of Surgery and Experimental Research, Malmö University Hospital, Malmö, Sweden
,
Ulf Stjernquist
2   Dept. of Surgery and Experimental Research, Malmö University Hospital, Malmö, Sweden
,
Mirella Ezban
3   Novo Nordisk A/S, Gentofte, Denmark
,
Per B Østergaard
3   Novo Nordisk A/S, Gentofte, Denmark
,
Ulla Hedner
3   Novo Nordisk A/S, Gentofte, Denmark
› Author Affiliations
Further Information

Publication History

Received 07 July 1995

Accepted after resubmission 18 December 1995

Publication Date:
10 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

The glycosylated multivalent three-domain Kunitz inhibitor TFPI is a natural inhibitor of tissue factor-FVIIa complex in the presence of FXa. TFPI has an experimental antithrombotic capacity indistinguishable from LMWH in a prophylactic dose, regardless of glycosylation and of the third domain. An inherited equilibrium between antithrombosis and haemorrhage exists. The aim of the study was to evaluate whether a two-domain non-glycosylated TFPI (117QTFPI1−161) has a bleeding potential in a rat gastric mucosa model. Groups; placebo, LMWH (tinzaparin) 60 and 250 anti-Xa IU/kg and 117QTFPIM61−161 1.0 and 10.0 mg/kg, given i.v. (bolus injection), randomised double dummy design.

All actively treated groups significantly prolonged both the bleeding volume (493-984 Μl) and the bleeding time (10-20 min) compared to placebo (41 Μl, 2 min). It was not possible to distinguish a difference between the lower dose of LMWH and 117QTFPI1−161 in either parameter (p = 0.23-0.71). The two doses of 117QTFPI1−161 caused elevation of plasma-TFPI, 18 and 150 times baseline value. Both LMWH doses (0.6-3.2 anti-Xa IU/ml) and both 117QTFPI1−161 doses (0.2-2.7 anti-Xa IU/ml), caused significant effect in the anti-Xa assay, however 117QTFPI1−161 significantly less. Only the largest dose of 117QTFPI1−161 caused significant prolongation in the APTT assay (34 s). Both doses of LMWH caused significant prolongation (60-300 s). LMWH was the only substance to prolong the dilute-PT assay.

Non-glycosylated two-domain 1.0 mg/kg TFPI, yielding supra-physiological plasma concentration, has an experimental haemorrhagic potential indistinguishable from LMWH in a prophylactic dose. The effect mediated by this type of TFPI could primarily be due to an inhibition of FXa.

 

  • References

  • 1 Lindahl AK, Sandset PM, Abildgaard U. The present status of tissue factor pathway inhibitor. Blood Coag Fibrinol 1992; 3: 439-449
    CrossrefPubMedGoogle Scholar
  • 2 Broze Jr GJ. Tissue factor pathway inhibitor. Thromb Haemost 1995; 74: 90-93
    PubMedGoogle Scholar
  • 3 Mann KG, Krishnaswamy S, Lawson JH. Surface-dependent hemostasis. Semin Hematol 1992; 29: 213-226
    PubMedGoogle Scholar
  • 4 Rapaport SI, Rao LVM. The tissue factor pathway; how it has become a “prima ballerina”. Thromb Haemost 1995; 74: 7-17
    PubMedGoogle Scholar
  • 5 Holst J, Lindblad B, Bergqvist D, Nordfang O, Østergaard PB. Is EPI as antithrombotic agent comparable to LMWH? - A pilot double-dummy experimental study. Thromb Haemost 1991; 6: 951
    PubMedGoogle Scholar
  • 6 Holst J, Lindblad B, Bergqvist D, Nordfang O, Østergaard PB, Petersen J, Nielsen G, Hedner U. Antithrombotic effect of recombinant truncated Tissue Factor Pathway Inhibitor (TFPI1-161) in experimental venous thrombosis - a comparison with low molecular weight heparin. Thromb Haemost 1994; 71: 214-219
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Holst J, Lindblad B, Nordfang O, Østergaard PB, Hedner U. Does the glycosylation influence the experimental antithrombotic effect of two-domain Tissue Factor Pathway Inhibitor?. Haemostasis 1996; 26: 23-30
    PubMedGoogle Scholar
  • 8 Holst J, Lindblad B, Westerlund G, Bregengaard C, Ezban M, Østergaard PB, Nordfang O, Hedner U. Pharmacokinetics and delayed antithrombotic effect of two-domain non-glycosylated Tissue Factor Pathway Inhibitor (TFPI). Thromb Res (accepted).
    PubMedGoogle Scholar
  • 9 Warn-Cramer BJ, Rao LVM, Maki SL, Rapaport SI. Modifications of extrinsic pathway inhibitor (EPI) and factor Xa that effect their ability to interact and to inhibit factor VH-tissue factor: evidence for a two-step model of inhibition. Thromb Haemost 1988; 60: 453-456
    Article in Thieme ConnectPubMedGoogle Scholar
  • 10 Mersereau WA, Hinchey EJ. Effect of gastric acidity on gastric ulceration induced by hemorrhage in rat, utilizing a gastric chamber technique. Gastroenterology 1973; 64: 1130-1135
    PubMedGoogle Scholar
  • 11 Whittle BJR, Kaufman Jr GL, Moncada S. Hemostatic mechanism, independent of platelet aggregation, arrest gastric mucosal bleeding. Proc Natl Acad Sci USA 1986; 83: 5683-5687
    CrossrefPubMedGoogle Scholar
  • 12 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI). Thromb Res 1988; 50: 803-813
    CrossrefPubMedGoogle Scholar
  • 13 Petersen JGL, Meyn G, Rasmussen JS, Christiansen L, Petersen J, Bjørn SE, Jonassen I, Nordfang O. Chararacterization of human tissue factor pathway inhibitor variants expressed in Saccharomyces cerevisae. J Biol Chem 1993; 268: 13344-13351
    PubMedGoogle Scholar
  • 14 Sandset PM, Abildgaard U, Pettersen M. A sensitive assay of extrinsic coagulation pathway inhibitor (EPI) in plasma and plasma fractions. Thromb Res 1987; 47: 389-400
    CrossrefPubMedGoogle Scholar
  • 15 Pedersen AH, Nordfang O, Norris F, Wiberg FC, Christensen M, Moeller KB, Meidahl-Pedersen J, Beck TC, Norris K, Hedner U, Kiesel W. Recombinant Human Extrinsic Pathway Inhibitor. Production, isolation, and characterization of its inhibitory activity on tissue factor-initiated coagulation reactions. J Biol Chem 1990; 265: 16786-16793
    PubMedGoogle Scholar
  • 16 Mätzsch T, Bergqvist D, Hedner U. Østergaard PB. Effects of an enzymatically depolymerized heparin as compared with conventional heparin in healthy volunteers. Thromb Haemost 1987; 57: 97-101
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Berstad A, Bang CJ, Talstad I. A new model to assess haemorrhagic potential of various heparin preparations. Acta Chir Scand 1990; 556: 62-67
    PubMedGoogle Scholar
  • 18 Broze GJ, Likert K, Higuchi D. Inhibition of Factor Vila/tissue factor by antithrombin III and tissue factor pathway inhibitor. Blood 1993; 82: 1679-1680
    PubMedGoogle Scholar
  • 19 Rao LVM, Rapaport SI, Hoang AD. Binding of factor Vila to tissue factor permits rapid antithrombin Ill/heparin inhibition of factor Vila. Blood 1993; 81: 2600-2607
    PubMedGoogle Scholar
  • 20 Rao LV, Nordfang O, Hoang AD, Pendurthi UR. Mechanism of antithrombin III inhibition of factor Vila/tissue factor activity on cell surfaces. Comparison with tissue factor pathway inhibitor/factor Xa-induced inhibition of factor Vila/tissue factor activity. Blood 1995 85. 121-129
    PubMedGoogle Scholar
  • 21 Enjyoji K, Emi M, Mukai T, Kato H. cDNA cloning and expression of rat tissue factor pathway inhibitor (TFPI). J Biochem 1992; 111: 681-687
    CrossrefPubMedGoogle Scholar
  • 22 Janson TL, Stormorken H, Prydz H. Species specificity of tissue thromboplastin. Haemostasis 1984; 14: 440-444
    PubMedGoogle Scholar
  • 23 Warn-Cramer BJ, Maki SL, Rapaport SI. A sulfated rabbit endothelial cell glycoprotein that inhibits factor Vila/tissue factor is functionally and immunogenetically identical to rabbit extrinsic pathway inhibitor (EPI). ThrombRes 1991; 61: 515-527
    PubMedGoogle Scholar
  • 24 Matthiasson SE, Lindblad B, Stjernquist U, Bergqvist D. The haemorrhagic effect of low molecular weight heparins, dermatan sulphate and hirudin. Haemostasis 1995; 25: 203-211
    PubMedGoogle Scholar
  • 25 Nordfang O, Bjørn SE, Valentin S, Nielsen LS, Wildgoose P, Beck TC, Hedner U. The C-terminus of tissue factor pathway inhibitor is essential to its anticoagulant activity. Biochem 1991; 30: 10371-10376
    CrossrefPubMedGoogle Scholar
  • 26 Haskel EJ, Torr KC, Day KC, Palmier MO, Wun TC, Sobel BE, Abend-schein DR. Prevention of reocclusion after thrombolysis with recombinant lipoprotein associated coagulation inhibitor. Circulation 1991; 84: 821-827
    CrossrefPubMedGoogle Scholar
  • 27 Spokas EG, Wun TC. Venous thrombosis produced in the vena cava of rabbits by vascular damage and stasis. J Pharmacol Toxicol Methods 1992; 27: 225-232
    CrossrefPubMedGoogle Scholar
  • 28 Khouri RK, Koudsi B, Kaiding F, Ornberg RL, Wun T-C. Prevention of thrombosis by topical application of tissue factor pathway inhibitor in a rabbit model of vascular trauma. Ann Plast Surg 1993; 30: 398-404
    CrossrefPubMedGoogle Scholar
  • 29 Arnljots B, Bergqvist D, Dahlbäck B. Inhibition of microarterial thrombosis by activated protein C in a rabbit model. Thromb Haemost 1994; 72: 415-420
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Novotny WF, Girard TJ, Miletich JP, Broze Jr. GJ. Purification and charactarization of the lipoprotein-associated coagulation inhibitor from human plasma. J Biol Chem 1989; 264: 18832-18837
    CrossrefPubMedGoogle Scholar
  • 31 Novotny WF, Brown SG, Girard TJ, Miletich JP, Broze Jr. GJ. Plasma LACI antigen in patient samples. Blood 1991; 78: 387-393
    PubMedGoogle Scholar
  • 32 Llobet D, Mateo J, Vallvé C, Martinez E, Fontcuberta Borrell M. TFPI deficiency in two siblings with thrombophilia. Thromb Haemost 1995; 73: 945
    PubMedGoogle Scholar
  • 33 Holst J, Lindblad B, Wedeberg E, Bergqvist D, Nordfang O, Østergaard PB, Hedner U. Tissue Factor Pathway Inhibitor levels in patients with spontaneous deep vein thrombosis. Thromb Res 1993; 72: 467-470
    CrossrefPubMedGoogle Scholar
  • 34 Novotny WF, Girard TJ, Miletich JP, Broze Jr GJ. Platelets secrete a coagulation inhibitor functionally and antigenetically similar to the lipoprotein associated coagulation inhibitor. Blood 1988; 72: 2020-2025
    PubMedGoogle Scholar
  • 35 Callander NS, Rao MVL, Nordfang O, Warn-Cramer B, Rapaport SI. Mechanism of binding of recombinant extrinsic pathway inhibitor (rEPI) to cultured cell surfaces. J Biol Chem 1992; 267: 876-882
    PubMedGoogle Scholar
  • 36 Bauer KA, Kass BL, Cate ten H, Bednarek MA, Hawinger JJ. Rosenberg RD Detection of factor X activation in humans. Blood 1989; 74: 2007-2015
    PubMedGoogle Scholar
  • 37 Esmon CT. The role of protein C and thrombomodulin in the regulation of blood coagulation. J Bio Chem 1989; 264: 4743-4746
    PubMedGoogle Scholar