SR123781A, a Synthetic Heparin Mimetic

Summary

SR123781A, a synthetic hexadecasaccharide comprising an anti-thrombin (AT) binding domain, a thrombin binding domain, and a neutral methylated hexasaccharide sequence, was obtained from glucose through a convergent synthesis. SR123781A showed high affinity for human AT (Kd = 58 ± 22 nM) and was a potent catalyst of its inhibitory effect with regard to factor Xa (IC₅₀ = 77 ± 5 ng/ml – 297 ± 13 U/mg) and thrombin (IC₅₀ = 4.0 ± 0.5 ng/ml – 150 ± 30 U/mg). SR123781A which acted exclusively via AT (no effect via heparin cofactor II at a concentration of 6 g/ml) inhibited thrombin generation occurring via both the extrinsic and intrinsic pathways in vitro in human plasma. SR123781A did not compete for ³H-heparin binding to PF4 and did not activate platelets in the presence of plasma from patients with heparin-induced thrombocytopenia. After intravenous or subcutaneous administration to rats, rabbits or baboons, SR123781A displayed prolonged anti-factor Xa and anti-factor IIa activity ex vivo. After intravenous injection to baboons, decreases of the anti-factor Xa and anti-thrombin activities were parallel and disappeared with the same pharmacodynamics. Intravenous administrations of SR123781A strongly inhibited thrombus formation in an experimental model of thromboplastin-induced venous thrombosis in rats with an ED₅₀ value of 18 ± 0.1 g/kg (vs 77 ± 3 g/kg for heparin). SR123781A inhibited arterial thrombus formation induced on a silk thread in an arterio-venous shunt and in the vena cava (ED₅₀ values of 225 ± 10 and 27 ± 8 g/kg, respectively). Compared to standard and low molecular weight heparin and to presently used drugs, SR123781A exhibited a highly favourable anti-thrombotic/bleeding ratio therefore showing that it might be considered as a promising compound in the treatment and prevention of various thrombotic diseases.

Keywords

Heparin mimetic - SR123781A - Factor Xa - thrombin - thrombosis

• References

• 1 Lane DA, Lindahl U. Heparin. Eds. Edward Arnold London: 1989
• 2 Casu B, Oreste P, Torri G, Zoppetti G, Choay J, Lormeau JC, Petitou M, Sinaÿ P. The structure of heparin oligosaccharide fragments with anti-factor Xa activity containing the minimal antithrombin III-binding sequence. Biochem J 1981; 197: 599-609.
• 3 Lormeau JC, Herault JP. The effect of the synthetic pentasaccharide SR 90107/Org31540 on thrombin generation ex vivo is uniquely due to AT-III-mediated neutralization of factor Xa. Thromb Haemost 1995; 75: 1474-7.
• 4 Choay J, Petitou M, Lormeau JC, Sinaÿ P, Casu B, Gatti G. Structure-activity relationship in heparin: a synthetic pentasaccharide with high affinity for antithrombin III and eliciting high anti-factor Xa Activity. Biochem Biophys Res Commun 1983; 116: 492-9.
• 5 Casu B. Structure and biological activity of heparin. Adv Carbohydr Chem Biochem 1985; 43: 51-134.
• 6 Stubbs MT, Bode W. The clot thickens: clues provided by thrombin structure. Trends Biochem Sci 1995; 20: 23-8.
• 7 Olson ST, Björk I. Regulation of thrombin activity by antithrombin and heparin. Semin Thromb Hemost 1994; 20: 373-409.
• 8 Stuckey JA, St Charles R, Edwards BFP. Proteins: Structure, Function and Genetics. 1992; 14: 277-87.
• 9 Warkentin TE, Chong BH, Greinacher A. Heparin-induced thrombocytopenia: Towards consensus. Thromb Haemost 1998; 79: 1-7.
• 10 Van Boeckel CAA, Petitou M. The unique antithrombin binding domain of heparin: a lead to new synthetic antithrombotics. Angew Chem Int Ed Engl 1993; 32: 1671-90.
• 11 Béguin S, Choay J, Hemker HC. The action of a synthetic pentasaccharide on thrombin generation in whole plasma. Thromb Haemost 1989; 61: 397-401.
• 12 Vogel GM, Meuleman DG, Bourgondiën FG, Hobbelen M. Comparison of two experimental thrombosis models in rats: Effect of four glycosaminoglycans. Thromb Res 1989; 54: 399-410.
• 13 Hobbelen PMJ, Van Dinther TG, Vogel GMT, Van Boeckel CAA, Moelker HCT, Meuleman DG. Pharmacological profile of the chemically synthetized antithrombin III binding fragment of heparin (pentasaccharide) in rats. Thromb Haemost 1990; 63: 265-72.
• 14 Walenga JM, Fareed J, Petitou M, Samama M, Lormeau JC, Choay J. Intravenous antithrombotic activity of a synthetic heparin pentasaccharide in a human serum induced stasis thrombosis model. Thromb Res 1986; 43: 243-8.
• 15 Walenga JM, Petitou M, Lormeau JC, Samama M, Fareed J, Choay J. Anti-thrombotic activity of a synthetic heparin pentasaccharide in a rabbit stasis thrombosis model using different thrombogenic challenges. Thromb Res 1987; 46: 187-98.
• 16 Walenga JM, Petitou M, Samama M, Fareed J, Choay J. Importance of a 3-O-sulphate group in a heparin pentasaccharide for antithrombotic activity. Thromb Res 1988; 52: 553-63.
• 17 Cadroy Y, Hansson SR, Harker LA. Antithrombotic effects of synthetic pentasaccharide with high affinity for plasma antithrombin III in non-human primates. Thromb Haemost 1993; 70: 631-5.
• 18 Boneu B, Necciari J, Cariou R, Sié P, Gabaig AM, Kieffer G, Dickinson J, Lamond G, Moelker H, Mant T, Magnani H. Pharmacokinetics and tolerance of the natural pentasaccharide (SR90107A/Org 31540) with high affinity for antithrombin III in man. Thromb Haemost 1995; 74: 1468-74.
• 19 Herbert JM, Petitou M, Lormeau JC, Carriou R, Necciari J, Magnani HM, Zandberg P, Van Amsterdam RGM, Van Boeckel CAA, Meuleman DG. SR 90107/Org 31540, a novel anti-factor Xa antithrombotic agent. Cardiovasc Drug Rev 1997; 15: 1-26.
• 20 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.
• 21 Olson ST, Halvorson HR, Björk I. Quantitative characterization of the thrombin-heparin interaction. Discrimination between specific and non-specific binding models. J Biol Chem 1991; 266: 6342-52.
• 22 Grootenhuis PDJ, Westerduin P, Meuleman D, Petitou M, Van Boeckel CAA. Rational design of synthetic heparin analogues with tailor-made coagulation factor inhibitory activity. Nature Struct Biol 1995; 2: 736-9.
• 23 Hemker HC, Willems GM, Béguin S. A computer assisted method to obtain the prothrombin activation velocity in whole plasma independent of thrombin decay processes. Thromb Haemost 1986; 56: 9-17.
• 24 Lormeau JC, Herault JP. Comparative inhibition of extrinsic and intrinsic thrombin generation by standard heparin, a low molecular weight heparin and the synthetic ATIII-binding pentasaccharide. Thromb Haemost 1993; 69: 152-6.
• 25 Atha DH, Lormeau JC, Petitou M, Rosenberg RD, Choay J. Contribution of 3-O- and 6-O-Sulfated Glucosamine Residues in the Heparin-Induced Conformational Change in Antithrombin III. Biochemistry 1987; 26: 6454-61.
• 26 Born GVR, Cross MJ. The aggregation of blood platelets. J Physiol 1963; 168: 178-95.
• 27 Stringer SE, Gallagher JT. Specific binding of the chemokine platelet factor 4 to heparan sulfate. J Biol Chem 1994; 272 (Suppl. 33) 20508-14.
• 28 Sheridan D, Carter C, Kelton JG. A diagnostic test for heparin-thrombocytopenia. Blood 1986; 67: 27-30.
• 29 Umetsu T, Sanai K. Effect of KC-6141, an anti-aggregating compound, on experimental thrombosis in rats. Thromb Haemost 1978; 39: 74-83.
• 30 Kumada T, Ishihara M, Ogawa H, Abiko Y. Experimental model of venous thrombosis in rats and effects of some agents. Thromb Res 1980; 18: 189-203.
• 31 Van Amsterdan RGM, Vogel GMT, Visser A, Kop WJ, Buiting MT, Meuleman DG. Synthetic analogues of the antithrombin III-binding pentasaccharide sequence of heparin – Prediction of the in vivo residence times, Arterioscler. Thromb Vasc Biol 1995; 15: 495-503.
• 32 Verstraete M. Pharmacotherapeutic aspects of unfractionnated and low molecular weight heparins. Drugs 1990; 40: 498-530.
• 33 Thomas DP. Does low molecular weight heparin cause less bleeding?. Thromb Haemost 1997; 78: 1422-5.
• 34 Paulsen H. Advances in selective chemical syntheses of complex oligosaccharides. Angew Chem Int Ed Engl 1982; 21: 155-73.
• 35 Herbert JM, Hérault JP, Bernat A, van Amsterdam RGM, Vogel GMT, Lormeau JC, Petitou M, Meuleman DG. Biochemical and pharmacological properties of Sanorg32701. Comparison with the “synthetic pentasaccharide” (SR90107/Org31540) and standard heparin. Circ Res 1996; 76: 590-600.
• 36 Herbert JM, Hérault JP, Bernat A, Van Amsterdam RGM, Lormeau JC, Petitou M, Van Boeckel V, Hoffmann Meuleman DG. Biochemical and pharmacological properties of Sanorg34006, a potent and long-acting synthetic pentasaccharide. Blood 1998; 91: 4197-205.
• 37 Tollefsen DM. Insight into the mechanism of action of heparin cofactor II. Thromb Haemost 1995; 74: 1209-14.
• 38 Visentin GP, Ford SE, Scott JP, Aster RH. Antibodies from patients with heparin-induced thrombocytopenia/thrombosis are specific for platelet factor 4 complexed with heparin or bound to endothelial cells. J Clin Invest 1994; 93: 81-8.
• 39 Carrie D, Caranobe C, Saivin S, Houin G, Petitou M, Lormeau JC, Van Boeckel C, Meuleman D, Boneu B. Pharmacokinetic and antithrombotic properties of two pentasaccharides with high affinity to antithrombin III in the rabbit: Comparison with CY 216. Blood 1994; 84: 2571-7.
• 40 Freund M, Cazenave JP, Courtney M, Degryse E, Roitsch C, Bernat A, Delebassee D, Defreyn G, Maffrand JP. Inhibition by recombinant hirudins of experimental venous thrombosis and disseminated intravascular coagulation induced by tissue factor in rats. Thromb Haemost 1990; 63: 187-92.
• 41 Just M, Tripier D, Seiffge D. Antithrombotic effects of recombinant hirudin in different animal models. Haemost 1991; 21: 80-7.
• 42 Talbot MD, Ambler J, Butler KD, Findlay VS, Mitchell KA, Peters RF, Tweed MF, Wallis RB. Recombinant desulphatohirudin (CGP39393). Anticoagulant and antithrombotic properties in vivo. Thromb Haemost 1989; 61: 77-80.
• 43 Jackson CV, Crowe G, Frank JD, Wilson HC, Coffman WJ, Utterback BG, Jakubowshi JA, Smith GF. Pharmacological assessment of antithrombotic activity of the peptide inhibitor, D-methyl-phenylalanyl-prolyl-arginyl (GYKI-14766), in a canine model of coronary artery thrombosis. J Pharmacol Exp Ther 1992; 261: 546-52.