Active site for heparin cofactor II in low molecular weight dermatan sulfate. Contribution to the antithrombotic activity of the high HCII-affinity fractions

In order to investigate the influence of charge distribution and chain length on the biological properties of sulfated polysaccharides, additional sulfate groups were introduced into the galactosaminoglycans, chondriotin sulfate and dermatan sulfate. Using a flexible method (with sulfuric acid and chlorosulfonic acid) for concurrent sulfation and controlled depolymerization, numerous products were obtained and characterized by chemical, enzymatic and nuclear magnetic resonance spectroscopic methods. The biologic actions of these products were profiled in both in vitro and in vivo assays for antithrombotic activity. Despite a weaker in vitro anticoagulant activity, low molecular weight over sulfated galactosaminoglycans produced significant dose-dependent antithrombotic actions in animal models which were similar to the actions observed with oversulfated low molecular weight heparins. These results suggest that a significant antithrombotic activity can be elicited through non-specific interactions of polysulfates with cellular and plasma components, and that clusters of sulfate groups such as the 4-6 disulfate group on D-galactosaminoglycan residues may be important for these interactions. Furthermore, these results, also suggest that supersulfation of glycosaminogly-cans results in products with biologic activity distinct from the native material.

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Effects of Dermatan Sulfate, a Heparin Cofactor II Mediated Thrombin Inhibitor, on the Endotoxin-Induced Disseminated Intravascular Coagulation Model in the Rat: Comparison With Low-Molecular Weight Heparin, Nafamostat Mesilate and Argathroban

The Japanese Journal of Pharmacology ◽

10.1254/jjp.76.397 ◽

1998 ◽

Vol 76 (4) ◽

pp. 397-404 ◽

Cited By ~ 10

Author(s):

Junichi Onaya ◽

Mamoru Kyogashima ◽

Atsuko Sunose ◽

Satoshi Miyauchi ◽

Syoji Mizuno ◽

...

Keyword(s):

Molecular Weight ◽

Disseminated Intravascular Coagulation ◽

Low Molecular Weight Heparin ◽

Dermatan Sulfate ◽

Thrombin Inhibitor ◽

Low Molecular Weight ◽

Nafamostat Mesilate ◽

Heparin Cofactor Ii ◽

Intravascular Coagulation

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The Additive Effect of Low Molecular Weight Heparins on Thrombin Inhibition by Dermatan Sulfate

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649602 ◽

1993 ◽

Vol 70 (03) ◽

pp. 443-447 ◽

Cited By ~ 6

Author(s):

Benilde Cosmi ◽

Giancarlo Agnelli ◽

Edward Young ◽

Jack Hirsh ◽

Jeffrey Weitz

Keyword(s):

Molecular Weight ◽

Antithrombin Iii ◽

Dermatan Sulfate ◽

Anticoagulant Activity ◽

Additive Effect ◽

Low Molecular Weight ◽

Low Molecular Weight Heparins ◽

Heparin Cofactor Ii ◽

Thrombin Inhibition ◽

Sds Page

SummaryThe aim of this study was to investigate the mechanism by which the anticoagulant activity of dermatan sulfate (DS) is increased by low molecular weight heparin (LMWH). In platelet poor plasma, LMWH enhances the effect of DS on thrombin (IIa) inhibition as determined by thrombin clotting times and with a chromogenic substrate assay. Analysis of the results of the chromogenic assays using either the algebraic fractional or the graphic isobole method suggests that LMWH has an additive effect on the anti-IIa activity of DS. This additive effect was lost when the experiments were repeated in plasma immunodepleted of antithrombin III (ATIII), indicating that the anti-IIa activity of LMWH is ATIII-dependent. To further explore the mechanism of the interaction between LMWH and DS, 125I-labeled IIa was added to plasma in the presence or absence of DS and/or LMWH and the formation of IIa-inhibitor complexes was assessed using SDS-PAGE followed by autoradiography. DS addition selectively increases the formation of heparin cofactor II (HCII)-IIa complexes, whereas LMWH enhances ATIII-IIa complex generation. Compared to plasma containing DS alone, the formation of ATIII-IIa complexes also is increased when the combination of DS and LMWH is added. These findings suggest that the additive effect of LMWH on the anti-IIa activity of DS reflects their different modes of IIa inhibition; DS potentiates IIa inhibition by HCII, while LMWH catalyses ATIII-dependent IIa inactivation. The potential clinical significance of these findings requires further investigation.

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ACTIVE SITE FOR HEPARIN COFACTOR II IN LOW MOLECULAR MASS DERMATAN SULFATE. CONTRIBUTION TO THE ANTITHROMBOTIC ACTIVITY OF FRACTIONS WITH HIGH AFFINITY FOR HEPARIN COFACTOR II

Thrombosis Research ◽

10.1016/0049-3848(96)00158-2 ◽

1996 ◽

Vol 84 (1) ◽

pp. 21-32 ◽

Cited By ~ 18

Author(s):

Giuseppe Mascellani ◽

Lino Liverani ◽

Bruna Parma ◽

GianLuca Bergonzini ◽

Pietro Bianchini

Keyword(s):

Molecular Mass ◽

Active Site ◽

Dermatan Sulfate ◽

Heparin Cofactor Ii ◽

Antithrombotic Activity ◽

High Affinity

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Circulating Activities During Constant Infusion of Heparin or a Low Molecular Weight Derivative (Enoxaparine): Failure to Demonstrate any Circadian Variations

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646058 ◽

1987 ◽

Vol 58 (04) ◽

pp. 1068-1072 ◽

Cited By ~ 10

Author(s):

P Toulon ◽

J F Vitoux ◽

C Leroy ◽

T Lecomte ◽

M Roncato ◽

...

Keyword(s):

Molecular Weight ◽

Circadian Rhythm ◽

Biological Activities ◽

Constant Infusion ◽

Low Molecular Weight ◽

Circadian Variations ◽

Heparin Cofactor Ii ◽

Heparin Infusion ◽

Chromogenic Assay ◽

Heparin Affinity

SummaryWe compared in six patients successively treated with an unfractionated heparin (UFH) and a low molecular weight heparin (LMWH) the variations in plasma anti-Xa activity, measured in a chromogenic assay, during a 36 h constant infusion. The values varied in a wider range during UHF infusion, but remained in the therapeutic range except once in one patient. No circadian rhythm could be demonstrated in our six patients. LMWH infusion yielded very constant anti-Xa circulating activities. In both cases, there were no significant modifications of three proteins with high heparin affinity (antithrombin III, heparin cofactor II, histidine-rich glycoprotein).Our results suggest that the circadian rhythm of the biological activities previously observed in patients treated with constant heparin infusion using clotting method is due to other factors than heparin itself.

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The Venous Antithrombotic Profile of Naroparcil in the Rabbit

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648977 ◽

1994 ◽

Vol 72 (06) ◽

pp. 874-879 ◽

Cited By ~ 11

Author(s):

Jean Millet ◽

Jocelyne Theveniaux ◽

Neil L Brown

Keyword(s):

Oral Administration ◽

Bleeding Time ◽

Dermatan Sulfate ◽

Factor Xa ◽

Bleeding Risk ◽

Thrombin Time ◽

Heparin Cofactor Ii ◽

Antithrombotic Activity ◽

Thrombin Inhibition ◽

Maximal Reduction

SummaryThe venous antithrombotic profile of naroparcil or (4-[4-cyanoben-zoyl]-phenyl)-1.5-dithio-β-D-xylopyranoside was investigated in the rabbit following single i. v. and oral administration. Naroparcil attenuated thrombus development in a Wessler stasis model of venous thrombosis (jugular vein) employing bovine factor Xa as a thrombogenic stimulus giving ED50 values of 21.9 mg/kg and 36.0 mg/kg after respectively i. v. and oral administration. Venous antithrombotic activity was maximal 2-3 h after i. v. administration and 4-8 h after oral administration. Four hours after the oral administration of maximal antithrombotic (Wessler model, factor Xa) doses (100 and 400 mg/kg), naroparcil had no significant effect on bleeding time. In platelet poor plasma obtained from animals treated 4 h previously with various doses (25 to 400 mg/kg) of naroparcil, there was no detectable anti-factor Xa nor antithrombin activity. Similarly, naroparcil had no effect on APTT nor on thrombin time. A sensitized thrombin time (to about 35 s) was modestly but significantly increased following oral administration of the compound at 400 mg/kg. However, thrombin generation by the intrinsic pathway was reduced in a dose-related manner, maximal reduction being 65% at 400 mg/kg. The same doses of naroparcil enhanced the formation of thrombin/heparin cofactor II complexes at the expense of thrombin/antithrombin III complexes in plasma incubated with (125I)-human a-thrombin and induced the appearance of dermatan sulfate-like material in the plasma of treated rabbits, as measured by a heparin cofactor II-mediated thrombin inhibition assay. The results suggest that naroparcil could have a safe venous antithrombotic profile following oral administration (antithrombotic effect compared to bleeding risk). It is probable that part of the mechanism of action of the β-D-xyloside, naroparcil, is due to the induction of chondroitin sulfate-like glycosaminoglycan biosynthesis, this material being detectable in the plasma.

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Heparin-Associated Thrombocytopenia Treatment with Low Molecular Weight Heparin

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661442 ◽

1986 ◽

Vol 55 (01) ◽

pp. 037-039 ◽

Cited By ~ 56

Author(s):

Jean-F Vitoux ◽

Jean-F Mathieu ◽

Martine Roncato ◽

Jean-N Fiessinger ◽

Martine Aiach

Keyword(s):

Molecular Weight ◽

Clinical Improvement ◽

Low Molecular Weight Heparin ◽

Low Molecular Weight ◽

Antithrombotic Activity ◽

Heparin Derivative

SummaryEight patients with heparin associated thrombocytopenia (HAT) were treated by a low molecular weight heparin derivative (LMW). Biological and clinical improvement occurred in all patients. This efficiency confirms the antithrombotic activity of LMW and allows its use in patients with HAT.

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