PDF herunterladen
Thromb Haemost 2006; 96(01): 7-13
DOI: 10.1160/TH06-02-0105
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Antithrombotic properties of Ixolaris, a potent inhibitor of the extrinsic pathway of the coagulation cascade

Rômulo A. Nazareth
1   Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
,
Luana S. Tomaz
1   Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
,
Susana Ortiz-Costa
2   Instituto de Nutrição, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
,
Geórgia C. Atella
1   Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
,
José M. C. Ribeiro
3   Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA
,
Ivo M. B. Francischetti
3   Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA
,
Robson Q. Monteiro
1   Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
› InstitutsangabenFinancial support: This research was supported in part by a grant from “Programa Interinstituciona de Ensino, Pesquisa e Extensão em Biologia do Câncer” by Fundação Ary Frauzino par Pesquisa e Controle do Câncer (FAF) and Fundação Educacional Charles Darwin (FECD). Additional support was provided by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro Carlos Chagas Filho (FAPERJ), Fundação Universitária José Bonifácio and fro the Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, USA.
Weitere Informationen

Publikationsverlauf

Received 21. Februar 2006

Accepted after revision 11. Juni 2006

Publikationsdatum:
29. November 2017 (online)

Auch verfügbar auf
    Lizenzen und Reprints

Summary

Ixolaris is a two-Kunitz tick salivary gland protein identified in Ixodes scapularis that presents extensive sequence homology t TFPI.It binds to FXa or FX as scaffolds and inhibits tissue factor/ FVIIa complex (extrinsic Xnase). Differently from TFPI, ixolaris does not bind to the active site cleft of FXa. Instead, comple formation is mediated by the FXa heparin-binding exosite,which may also results in decreased FXa activity into the prothrombi nase complex.In this report,we show that recombinant 125I-ixo laris interacts with rat and human FX in plasma and prolongs the prothrombin time (PT) and activated partial thromboplastin time (aPTT) in vitro.We have also investigated the effects of ixo laris in vivo, using a venous thrombosis model. Subcutaneous (s.c.) or intravenous (i.v.) administration of ixolaris in rats caused a dose-dependent reduction in thrombus formation, with complete inhibition attained at 20 µg/kg and 10 µg/kg, re spectively. Antithrombotic effects were observed 3 h after s.c. administration of ixolaris and lasted for 24 h thereafter. Ex vivo experiments showed that ixolaris (up to 100 µg/kg) did not affect the aPTT,while the PT was increased by ∼0.4-fold at the hig hest ixolaris concentration. Remarkably, effective antithrom botic doses of ixolaris (20 µg/kg) was not associated with bleed ing which was significant only at higher doses of the anticoagulant (40 µg/kg).Our experiments demonstrate that ixolaris is an effective and possibly safe antithrombotic agen in viv .

Keywords

Anticoagulant - ixolaris - venous thrombosis - factor Xa - tissue factor - tick saliva - Ixodes scapularis
 

  • References

  • 1 Furie B, Furie BC. The molecular basis of blood coagulation. Cell 1988; 53: 505-18.
    CrossrefPubMedSuche in Google Scholar
  • 2 Mann KG. Biochemistry and physiology of blood coagulation. Thromb Haemost 1999; 82: 165-74.
    Thieme ConnectPubMedSuche in Google Scholar
  • 3 Monroe DM, Hoffman M, Roberts HR. Platelets and thrombin generation. Arterioscler Thromb Vasc Biol 2002; 22: 1381-9.
    CrossrefPubMedSuche in Google Scholar
  • 4 Gresele P, Agnelli G. Novel approaches to the treat ment of thrombosis. Trends Pharmacol Sci 2002; 23: 25-32.
    CrossrefPubMedSuche in Google Scholar
  • 5 Weitz JI, Bates SM. New anticoagulants. J Thromb Hæmost 2005; 03: 1843-53.
    PubMedSuche in Google Scholar
  • 6 Broze Jr GJ. Tissue factor pathway inhibitor and the revised theory of coagulation. AnnuRev Med 1995; 46: 103-12.
    PubMedSuche in Google Scholar
  • 7 Bajaj MS, Birktoft JJ, Steer SA. et al. Structure and biology of tissue factor pathway inhibitor. Thromb Haemost 2001; 86: 959-72.
    Thieme ConnectPubMedSuche in Google Scholar
  • 8 Law JH, Ribeiro JM, Wells MA. Biochemical in sights derived from insect diversity. Annu Rev Bio chem 1992; 61: 87-111.
    PubMedSuche in Google Scholar
  • 9 Ribeiro JMC, Francischetti IMB. Roleofarthropod saliva in blood feeding: sialome and post-sialome per spectives. Ann Rev Entomol 2003; 48: 73-88.
    CrossrefPubMedSuche in Google Scholar
  • 10 Francischetti IM, Valenzuela JG, Andersen JF. et al. Ixolaris, a novel recombinant tissue factor pathway in hibitor (TFPI) from the salivary gland of the tick, Ixodes scapularis: identification of factor X and factor Xa as scaffolds for the inhibition of factor VIIa/tissue factor complex. Blood 2002; 99: 3602-12.
    CrossrefPubMedSuche in Google Scholar
  • 11 Monteiro RQ, Rezaie AR, Ribeiro JM. et al. Ixola ris: a factor Xa heparin-binding exosite inhibitor. Bio chem J 2005; 387: 871-7.
    PubMedSuche in Google Scholar
  • 12 Vogel GM, Meuleman DG, Bourgondien FG. et al. Comparison of two experimental thrombosis models in rats: effects of four glycosaminoglycans. Thromb Res 1989; 54: 399-410.
    CrossrefPubMedSuche in Google Scholar
  • 13 Farias WR, Nazareth RA, Mourao PA. Dual effects of sulfated D-galactans from the red algæ Botryocladia occidentalis preventing thrombosis and inducing pla telet aggregation. ThrombHaemost 2001; 86: 1540-6.
    PubMedSuche in Google Scholar
  • 14 Herbert JM, Héralt JP, Bernat A. et al. Biochemical and pharmacological properties of SANORG 32701 Comparison with the “synthetic pentasacharide (SR90107/org 31549)” and standard heparin. Circ Res 1996; 79: 590-600.
    CrossrefPubMedSuche in Google Scholar
  • 15 Rezaie AR. Identification of basic residues in the heparin binding exosite of factor Xa critical for heparin and factorVa binding. J Biol Chem 2000; 275: 3320-7.
    CrossrefPubMedSuche in Google Scholar
  • 16 Manithody C, Rezaie AR. Functional mapping of charged residues of the 82-116 sequence in factor Xa evidence that lysine 96 is a factor Va independent re cognition site for prothrombin in the prothrombinase complex. Biochemistry 2005; 44: 10063-70.
    CrossrefPubMedSuche in Google Scholar
  • 17 Pacheco RG, Vicente CP, Zancan P. et al. Different antithrombotic mechanisms among glycosaminoglyc-ans revealed with a new fucosylated chondroitin sulfate from an echinoderm. Blood Coagul Fibrinolysis 2000; 11: 563-73.
    CrossrefPubMedSuche in Google Scholar
  • 18 Vicente CP, Zancan P, Peixoto LL. et al. Unbalanced effects of dermatan sulfates with different sul-fation patterns on coagulation, thrombosis and bleeding. Thromb Haemost 2001; 86: 1215-20.
    Thieme ConnectPubMedSuche in Google Scholar
  • 19 Finkle CD, St Pierre A, Leblond L. et al. BCH2763, a novel potent parenteral thrombin inhibi tor, is an effective antithrombotic agent in rodent models of arterial and venous thrombosis comparisons with heparin, r-hirudin, hirulog, inogatran and argatroban. Thromb Haemost 1998; 79: 431-8.
    Thieme ConnectPubMedSuche in Google Scholar
  • 20 Roberts HR, Monroe III DM, Hoffman M. Molecular biology and biochemistry of the coagulation factors and pathways of hemostasis. In: Williams Hema tology; Mc Graw Hill: 2001: 1409-34.
    Suche in Google Scholar
  • 21 Moons AH, Peters RJ, Bijsterveld NR. et al. Rec ombinant nematode anticoagulant protein c2, an in hibitor of the tissue factor/factor VIIa complex, in patients undergoing elective coronary angioplasty. J Am Coll Cardiol 2003; 41: 2147-53.
    CrossrefPubMedSuche in Google Scholar
  • 22 Vlasuk GP, Bradbury A, Lopez-Kinninger L. et al. Pharmacokinetics and anticoagulant properties of the factor VIIa-tissue factor inhibitor recombinant nema-tode anticoagulant protein c2 following subcutaneous administration in man Dependence on the stoichio-metric binding to circulating factor X. Thromb Hae-most 2003; 90: 803-12.
    Thieme ConnectPubMedSuche in Google Scholar
  • 23 Belting M, Ahamed J, Ruf W. Signaling of the tissue factor coagulation pathway in angiogenesis and cancer Arterioscler Thromb Va sc Biol. 2005; 25: 1545-50.
    PubMedSuche in Google Scholar
  • 24 White RJ, Galaria II, Harvey J. et al. Tissue factor is induced in a rodent model of severe pulmonary hyper tension characterized by neointimal lesions typical of human disease. Chest 2005; 128: 612S-613S.
    PubMedSuche in Google Scholar
  • 25 Levi M, van der Poll T, Te n Cate H. Tissue factor in infection and severe inflammation. Semin Thromb He-most 2006; 32: 33-9.
    Thieme ConnectPubMedSuche in Google Scholar