Structure-activity relationship in heparin: A synthetic pentasaccharide with high affinity for antithrombin III and eliciting high anti-factor Xa activity

Choay, J; Petitou, Maurice; Lormeau, J C; Sînaÿ, Pierre; Casu, Benito; Gatti, Giuseppe

doi:10.1016/0006-291x(83)90550-8

Cited by 680 publications

(339 citation statements)

References 18 publications

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“…The recent chemical synthesis of heparin oligosaccharides by Choay and co-workers (Choay et al, 1983;Sinay et al, 1984;Petitou, 1984) has allowed a more critical examination of the precise molecular determinants of the antiproliferative capacity. A pentasaccharide (Fig.…”

Section: Discussionmentioning

confidence: 99%

Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. II. Evidence for a pentasaccharide sequence that contains a 3-O-sulfate group.

Castellot¹,

Choay²,

Lormeau³

et al. 1986

The Journal of Cell Biology

155

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Abstract. Earlier work from our laboratory demonstrated that heparin inhibited the proliferation of vascular smooth muscle cells in vivo and in vitro. Both anticoagulant and non-anticoagulant heparin species were equally effective as antiproliferative agents. Previous structure-function studies indicated that hexasaccharide and larger fragments retained antiproliferative activity, whereas tetra-and disaccharides were inactive. These experiments also suggested that both Nand O-sulfates of heparin were necessary for growth inhibitory capacity. In this paper, we have further analyzed the structural determinants of the antiproliferative activity of heparin. These experiments were done using synthetically prepared and therefore chemically defined heparin oligosaccharides. We present evidence that a pentasaccharide fragment retains antiproliferative activity, and that the 3-O-sulfate on the internal glucosamine residue is critical for growth inhibitory capacity of the pentasaccharide. We also show that heparins obtained from different manufacturers differ significantly in their ability to suppress smooth muscle cell proliferation.T HE proliferation of vascular smooth muscle cells (SMC) ~ after endothelial injury is thought to be a key step in the pathogenesis of arteriosclerosis (Ross and Glomset, 1973). Although much attention has been focused on factors that stimulate SMC proliferation (Schwartz et al., 1982), very little is known about the mechanisms involved in maintaining these cells in a quiescent growth state in healthy blood vessels, and in re-establishing the quiescent growth state of SMC after their proliferative response to intimal injury.Earlier work from our laboratory demonstrated that heparin inhibited the proliferation of SMC in vivo and in vitro (Clowes and Karnovsky, 1977;Guyton et al., 1980;Hoover et al., 1980; Castellot et al., 1981). Both anticoagulant and non-anticoagulant heparin species were equally effective as SMC antiproliferative agents. These studies suggested both a pharmacological basis for the use ofhepafin after vessel injury, and a possible physiological role for heparin-like species as regulators of SMC growth in the vasculature.Previous structure-function studies (Castellot et al., 1984) indicated that hexasaccharide and larger fragments retained antiproliferative activity, whereas tetra-and disaccharides were inactive against SMC growth. These experiments also indicated that both the N-and O-sulfates of heparin were necessary for antiproliferative activity, although the N-sulfates could be replaced with acetyl groups without significant loss of activity. Both the hexasaccharide fragment and the Ndesulfated, re-N-acetylated heparin are devoid of anticoagulant activity (Kazatchkine et al., 1981;Oosta et al., 1981), thus providing a chemical basis for the difference between the antiproliferative and anticoagulant activities of this glycosaminoglycan.In this paper, we have further analyzed the structural determinants of the antiproliferative effect of heparin. These experiments were ...

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Section: Discussionmentioning

confidence: 99%

Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. II. Evidence for a pentasaccharide sequence that contains a 3-O-sulfate group.

Castellot¹,

Choay²,

Lormeau³

et al. 1986

The Journal of Cell Biology

155

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“…I' In addition, with fibronectin, heparin acts as an opsonin, and may facilitate clearance of endotoxin by the mononuclear phagocytic system. 1 13, ' 14 …”

Section: Hemodynamic Alterations Associated With Endotoxemiamentioning

confidence: 99%

Heparin: A Review of its Pharmacology and Therapeutic Use in Horses

Moore

Hinchcliff

1994

Veterinary Internal Medicne

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Heparin is used clinically in horses to treat hemostatic abnormalities associated with severe gastrointestinal disease, septicemia, and endotoxemia. The primary anticoagulant effect of heparin is through the suppression of thrombin-dependent amplification of the coagulation cascade, and inhibition of thrombin-mediated conversion of fibrinogen to fibrin. Heparin may be of benefit in preventing the complications associated with hypercoagulable states such as jugular vein thrombosis, laminitis, and organ failure. Heparin may also be beneficial in the prevention of intraabdominal adhesions after gastrointestinal surgery, and in amelioration of hernodynamic abnormalities associated with endotoxic shock. Because a sequential rise in serum unique compound with anticoagulant activity was ac-A cidentally discovered in 19 16 by McClean, a medical student, while he was attempting to identify a procoagulant in the liver.' The substance was named heparin because of its abundance in hepatic tissue.' In 1939, Brinkhous discovered that a plasma cofactor was necessary for the anticoagulant activity of h e~a r i n .~ Abildgaard named the cofactor antithrombin 111 (AT 111), and the specific mechanism of action of this cofactor was elucidated shortly thereafter. [4][5][6] Heparin is composed of a family of straight-chain, highly sulfated, anionic polysaccharides, specifically glycosaminoglycuronan sulfate esters, with molecular weights varying from 4,000 to 40,000 d.7-9 The structural unit of this complex molecule is a two hexose sugar that, in 8 to 15 repeating disaccharide subunits, forms a polymer of differing length^.^.' The structure of heparin is markedly heterogenous and varies with respect to the sequence and ratio of the different hexose units, the extent and type of sulfation, and the extent of N-a~etylation.~ The heterogeneity of heparin is not an artifact of preparation, but occurs because of the randomness of the biosynthetic process. lo Endogenous heparin is produced within mast cells of the lung, liver, and intestine.' ' Commercially-available heparin is derived from the mast cells of porcine intestinal mucosa and bovine lung."A small and distinct fraction of the heparin molecule is responsible for its anticoagulant effect. l 3 The active site of the heparin molecule contains a unique glucosamine unit with a specific pentasaccharide ~equence.'~.'~ This distinct fraction of the heparin molecule binds to AT 111, an endog-

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“…Fondaparinux, a methoxy form of the natural pentasaccharide sequence, is also a selective antithrombin (AT)‐dependent FXa inhibitor. However, in contrast to rivaroxaban, fondaparinux does not inhibit prothrombin‐bound or clot‐associated FXa 10, 11, 12. Dabigatran acts as a reversible direct thrombin inhibitor that specifically binds to both free and fibrin‐bound thrombin 13, 14.…”

Section: Introductionmentioning

confidence: 99%

Comparison of antithrombin‐dependent and direct inhibitors of factor Xa or thrombin on the kinetics and qualitative characteristics of blood clots

Salta

Papageorgiou

Larsen

et al. 2018

Research and Practice in Thrombosis and Haemostasis

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BackgroundVenous thromboembolism (VTE) is associated with significant morbidity and mortality.ObjectivesWe investigated the impact of direct and AT‐dependent FXa or thrombin inhibitors on thrombus formation.MethodsWhole blood thromboelastometry and thrombin generation were assessed after triggering the TF pathway. Clinically relevant concentrations of rivaroxaban, fondaparinux, dabigatran or tinzaparin and an association of rivaroxaban and dabigatran were examined.ResultsAll agents delayed thrombus formation in a concentration‐dependent manner, as documented by the prolongation of the clotting time (CT) and clot formation time (CFT). Rivaroxaban did not significantly alter the α‐angle or maximum clot firmness (MCF). In contrast, dabigatran and fondaparinux altered the process of clot structure by decreasing the α‐angle, but did not modify clot firmness. The later property was significantly affected only by tinzaparin that also reduced the MCF. The association of rivaroxaban and dabigatran did not affect the MCF, although it amplified the effect on CFT and α‐angle.ConclusionsAll agents delayed thrombus formation. However, the compounds differed substantially with respect to fibrin polymerization rate and clot firmness. Comparison of the data obtained by thrombin generation assessment with those obtained by the thromboelastometric study shows that the delay in clot formation is principally associated with prolongation of the initiation phase of thrombin formation as well as a reduction of the propagation phase. Tinzaparin was much more potent than the other agents both with regard to suppression of thrombin generation and by delay in clot formation.

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Structure-activity relationship in heparin: A synthetic pentasaccharide with high affinity for antithrombin III and eliciting high anti-factor Xa activity

Cited by 680 publications

References 18 publications

Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. II. Evidence for a pentasaccharide sequence that contains a 3-O-sulfate group.

Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. II. Evidence for a pentasaccharide sequence that contains a 3-O-sulfate group.

Heparin: A Review of its Pharmacology and Therapeutic Use in Horses

Comparison of antithrombin‐dependent and direct inhibitors of factor Xa or thrombin on the kinetics and qualitative characteristics of blood clots

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