Download PDF
Thromb Haemost 2002; 87(02): 238-244
DOI: 10.1055/s-0037-1612979
Letters to the Editor
Schattauer GmbH

Effect of New Synthetic Heparin Mimetics on Whole Blood Thrombin Generation In Vivo and In Vitro in Rats

J.P. Hérault
1   Sanofi∼Synthélabo Recherche, Toulouse, France
,
A. Bernat
1   Sanofi∼Synthélabo Recherche, Toulouse, France
,
C. Gaich
1   Sanofi∼Synthélabo Recherche, Toulouse, France
,
J.M. Herbert
1   Sanofi∼Synthélabo Recherche, Toulouse, France
› Author Affiliations
Further Information

Publication History

Received 01 January 2001

Accepted after resubmission 15 November 2001

Publication Date:
13 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

The effect of new heparin mimetics (synthetic oligosaccharides) was studied in vitro with regard to thrombin generation (TG) in rat platelet rich plasma (PRP) and whole blood (WB) and in vivo on stasis-induced venous thrombosis in the rat.

TG in PRP and in WB was highly dependent on platelet count and strongly influenced by the haematocrit. The peak of TG appeared to be significantly higher in WB than in PRP whereas the endogenous thrombin potential (ETP) was not significantly different under either condition.

The effect of hirudin, the synthetic pentasaccharide SR90107/ Org31540 (SP) and heparin were measured on TG in PRP and WB. We then compared the effect of two new synthetic heparin mimetics (SR121903A and SanOrg123781) with potent and comparable antithrombin (AT) mediated activity against factor Xa and thrombin. These two compounds were made of a pentasaccharide with a high affinity to AT, prolonged at the non-reducing end by an oligosaccharide chain recognised by thrombin. In SR121903A, the charge density and charge distribution was analogous to that of heparin whereas in SanOrg123781 the charges were only located on the last 5 saccharides of the non-reducing end of the molecule. In PRP and in WB, SR121903A acted on the lag time and on the AUC whereas SanOrg123781 inhibited thrombin formation with no effect on the lag time. SanOrg123781 was more potent in inhibiting TG than SR121903A. This difference was due to the structures of the compounds that differed in their ability to be neutralised by platelet factor 4. The antithrombotic effect of the two compounds was examined in a venous thrombosis model in rats. We observed that SanOrg123781 was more active than SR121903A and heparin.

Taken together, these results indicate that the activity of oligosaccharides is greatly influenced by the global charge density of the molecule and show that SanOrg123781 is a potent and promising antithrombotic drug candidate.

Keywords

Oligosaccharides - thrombin generation - PF4 - heparin - thrombin - factor Xa
 

  • References

  • 1 Hemker HC. Thrombin generation, an essential Stepp in haemostasis and thrombosis. In: Haemostasis (3rd edition). Bloom AL, Forbes CD, Thomas DP, Tuddenham EGD. (eds) London, UK: Churchill Livingstone; 1994: 477-89.
    Search in Google Scholar
  • 2 Biggs R, Mac RGFarlane. Human blood coagulation and its disorders. Oxford: Blackwell Scientific Publication; 1953: 54-68.
    Search in Google Scholar
  • 3 Walsh PM, Schmaier AH. Platelet-coagulant protein interactions. In: Hemostasis & Thrombosis: Basic principles and clinical practice. 3rd ed. Colman RW, Hirsh J, Marder VJ, Salzman EW. (eds) JB Lippicott Co: Philadelphia; 1994: 629-51.
    Search in Google Scholar
  • 4 Bevers EM, Comfurius P, Van Rijn JLML, Hemker C, Zwaal RFA. Generation of Prothrombin-converting activity and the exposure of phosphatidyl serine at the outer surface of platelets. Eur J Biochem 1982; 122: 429-36.
    CrossrefPubMedSearch in Google Scholar
  • 5 Bevers EM, Comfurius P, Zwaal RFA. Mechanismc involved in platelets procoagulant response. In: Mechanism of Platelet Activation and Control. Authi KS. et al. (eds) Plenum Publishing corp; New York: 1994: 195-207.
    Search in Google Scholar
  • 6 Tracy PB, Allen DH, Worfolk LA, Lawler RR. Monocytes/Macrophages regulation of coagulant events. Haemostas 1996; 26 (Suppl. 06) 45.
    PubMedSearch in Google Scholar
  • 7 Tijburg PNM, DE Groot P. Endothelial cell-mediated thrombin formation. Semin Thromb Haemostas 1992; 18: 256-64.
    Thieme ConnectPubMedSearch in Google Scholar
  • 8 Helley D, Eldor A, Girot R, Ducrocq R, Guillin MC, Bezeaud A. Increased procoagulant activity of red blood cells from patients with homozygous sickle cell-disease and beta-thalassemia. Thromb Haemost 1996; 76: 322-7.
    Thieme ConnectPubMedSearch in Google Scholar
  • 9 Kuypers FA, Lewis RA, Hua M, Schott MA, Discher D, Ernst JD, Lubin B. Detection of altered membrane phospholipid asymmetry in subpopulations of human red blood cells using fluorescently labelled annexin V. Blood 1996; 87: 1179-87.
    PubMedSearch in Google Scholar
  • 10 Wood BL, Gibson Df, Tait JF. Increased erthrocyte phosphatidylserine exposure in sickle cell disease: Flow-cytometric measurement and clinical associations. Blood 1996; 88: 1873-80.
    PubMedSearch in Google Scholar
  • 11 Ruf A, Pick M, Deutsch V, Patscheke H, Goldfarb A, Rachmilewitz EA, Guillin MC, Eldor A. In-vivo platelet activation correlates with red cell anionic phospholipid exposure in patients with beta-thalassemia major. Br J Haematol 1997; 98: 51-6.
    CrossrefPubMedSearch in Google Scholar
  • 12 Kessels H, Béguin S, Andree H, Hemker HC. Measurement of thrombin generation in whole blood - the effect of heparin ad aspirin. Thromb Haemost 1994; 72: 78-83.
    Thieme ConnectPubMedSearch in Google Scholar
  • 13 Peyrou V, Lormeau JC, Hérault JP, Boneu B, Herbert JM. Long-term persistence of low-molecular-weight material with anti-factor Xa activity contributes to anticoagulant and antithrombotic effects after subcutaneous injection of CY 216 in the rabbit. Blood Coagul Fibrinol 1997; 08 (03) 175-84.
    CrossrefPubMedSearch in Google Scholar
  • 14 Peyrou V, Lormeau JC, Hérault JP, Gaich C, Pfliegger AM, Herbert JM. Contribution of erythrocytes to thrombin generation in whole blood. Thromb Haemost 1999; 81 (03) 400-6.
    Thieme ConnectPubMedSearch in Google Scholar
  • 15 Herbert JM, Petitou M, Lormeau JC, Cariou R, Necciari J, Magnani HM, Zandberg P, van Amsterdam RGM, van Boeckel DG, Meulman CAA. SR90107A/Org31540, a novel anti-factor Xa antithrombotic agent. Cardiovascular Drug Reviews 1997; 01: 1-26.
    PubMedSearch in Google Scholar
  • 16 Herbert JM, Hérault JP, Bernat A, van Amsterdam RGM, Lormeau JC, Petitou M, van Boeckel C, Hofmann P, Meulman DG. Biochemical and pharmacological properties of SANORG 34006, a potent and long-acting synthetic pentasaccharide. Blood 1998; 91 (11) 4197-205.
    PubMedSearch in Google Scholar
  • 17 Petitou M, Duchaussoy P, Driguez PA, Jaurand G, Hérault JP, Lormeau JC, van Boeckel C, Herbert JM. First synthetic carbohydrates with the full anticoagulant properties of heparin. Angew Chem Int Ed Engl 1998; 37: 3009-14.
    CrossrefPubMedSearch in Google Scholar
  • 18 Petitou M, Hérault JP, Bernat A, Driguez PA, Duchaussoy P, Lormeau JC, Herbert JM. Synthesis of thrombin-inhibiting heparin mimetics without side effects. Nature 1999; 398: 417-22.
    CrossrefPubMedSearch in Google Scholar
  • 19 Petitou M, Duchaussoy P, Driguez PA, Hérault JP, Lormeau JC, Herbert JM. New synthetic heparin mimetics able to inhibit thrombin and factor Xa. Biorg Chem Med Lett 1999; 09: 1155-60.
    PubMedSearch in Google Scholar
  • 20 Petitou M, Driguez PA, Duchaussoy P, Hérault JP, Lormeau JC, Herbert JM. Synthetic oligosaccharides having different functional domains: potent and safe heparin mimetics. Biorg Chem Med Lett 1999; 09: 1161-6.
    PubMedSearch in Google Scholar
  • 21 Herbert JM, Herault JP, Bernat A, Schaeffer P, Driguez PA, Duchaussoy P, Petitou M. SR123781A, a synthetic heparin mimetic. Thromb Haemost 2001; 85: 852-60.
    Thieme ConnectPubMedSearch in Google Scholar
  • 22 Hemker HC, Willems GM, Béguin S. A computer assisted method to obtain the prothrombin activation velocity in whole plasma independant of thrombin decay process. Thromb Haemost 1986; 56: 9-17.
    Thieme ConnectPubMedSearch in Google Scholar
  • 23 Beguin S, Lindhout T, Hemker HC. The effect of trace amounts of tissue factor on thrombin generation in platelet rich plasma, its inhibition by heparin. Thromb Haemost 1989; 61: 25-9.
    Thieme ConnectPubMedSearch in Google Scholar
  • 24 Hemker HC, Beguin S. Thrombin generation in plasma: its assessment via the endogenous thrombin potential. Thromb Haemost 1995; 74: 134-8.
    Thieme ConnectPubMedSearch in Google Scholar
  • 25 Nieuwenhuys CMA, Feijge MAH, Beguin S, Heemskerk JWM. Monitoring hypocoagulant conditions in rat plasma: Factors determining the endogenous thrombin potential of tissue factor-activated plasma. Thromb Haemost 2000; 84: 1045-50.
    Thieme ConnectPubMedSearch in Google Scholar
  • 26 Teien AN, Lie M. Evaluation of an amidolytic heparin assay method: Increased sensitivity by adding purified antithrombin III. Thromb Res 1978; 13: 285-8.
    CrossrefPubMedSearch in Google Scholar
  • 27 Vogel GM, Meuleman DG, Bourgoudieu FG, Hobbelen PM. Comparaison of two experimental thromosis modles in rats: effects of four glycosaminoglycan. Thromb Res 1989; 54: 399-410.
    CrossrefPubMedSearch in Google Scholar
  • 28 Reverter JC, Beguin S, Kessels R, Hemker HC, Coller BS. Inhibition of platelet-mediated, tissue factor-induced thrombin generation by the mouse/human chimeric 7E3 antibody Potential implications for the effect of c7E3 Fab treatment on acute thrombosis and “clinical restenosis”. J Clin Invest 1996; 98 (03) 863-74.
    CrossrefPubMedSearch in Google Scholar
  • 29 Hérault JP, Peyrou V, Savi P, Bernat A, Herbert JM. Effect of SR121566A, a potent GP IIb-IIIa antagonist on platelet-mediated thrombin generation in vitro and in vivo. Thromb Haemost 1998; 79 (02) 383-8.
    Thieme ConnectPubMedSearch in Google Scholar
  • 30 Hérault JP, Bernat A, Pflieger AM, Lormeau JC, Herbert JM. Comparative effects of two direct and indirect factor Xa inhibitors on free and clot-bound prothrombinase. J Pharmacol Exp Ther 1997; 283 (01) 16-22.
    PubMedSearch in Google Scholar