Synergistic Antithrombotic Properties of G4120, a RGD-Containing Synthetic Peptide, and Argatroban, a Synthetic Thrombin Inhibitor, in a Hamster Femoral Vein Platelet-Rich Thrombosis Model

Yoshimi Imura; Jean-Marie Stassen; Ingrid Vreys; Emmanuel Lesaffre; Herman K Gold; Désiré Collen

doi:10.1055/s-0038-1656376

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1992; 68(03): 336-340
DOI: 10.1055/s-0038-1656376

Original Article

Schattauer GmbH Stuttgart

Synergistic Antithrombotic Properties of G4120, a RGD-Containing Synthetic Peptide, and Argatroban, a Synthetic Thrombin Inhibitor, in a Hamster Femoral Vein Platelet-Rich Thrombosis Model

Yoshimi Imura

The Center for Thrombosis and Vascular Research, Belgium

,

Jean-Marie Stassen

The Center for Thrombosis and Vascular Research, Belgium

,

Ingrid Vreys

The Center for Thrombosis and Vascular Research, Belgium

,

Emmanuel Lesaffre

¹The Division of Epidemiology University of Leuven, Belgium

,

Herman K Gold

²The Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

,

Désiré Collen

The Center for Thrombosis and Vascular Research, Belgium

› Institutsangaben

Abstract

als PDF herunterladen

Summary

The synergistic antithrombotic properties of G4120, a synthetic Arg-Gly-Asp (RGD) containing peptide which strongly inhibits platelet aggregation, and of Argatroban, a synthetic thrombin inhibitor, were examined in a reproducible quantitative hamster femoral vein platelet-rich mural thrombosis model. Bolus injections of G4120 and Argatroban inhibit thrombus formation in a dose-dependent way; 50% inhibition (ID₅₀) is obtained with 11 µg/kg G4120 and with 2 mg/kg Argatroban. Combined bolus injections of 3 µg/kg G4120 with 0.5, 0.75 or 1 mg/kg Argatroban and of 1 mg/kg Argatroban with 1.5 or 3 µg/kg G4120 caused linear dose-dependent inhibition of thrombus formation, whereas 3 µg/kg G4120 or 1 mg/kg Argatroban alone had very little effect (<20% inhibition). ID₅₀ was obtained with the combination of 3 µg/kg G4120 and 0.5 mg/kg Argatroban, corresponding to an equi-effective fractional combination of 0.62 with a 95% confidence interval of 0.50 to 0.74. Alternatively the ID₅₀ was obtained with the combination of 1 mg/kg Argatroban and 1.3 µg/kg G4120, corresponding to an equi-effective fractional combination of 0.52 with a 95% confidence interval of 0.18 to 0.86. In both instances these results are indicative of a significant synergistic interaction. Bolus injection of 10 mg/kg aspirin, 100 U/kg heparin or the combination did not inhibit thrombus formation.

The synergistic effect of the combination of platelet inhibiting RGD-peptides and synthetic thrombin inhibitors could be useful in the prevention of arterial occlusion with platelet-rich thrombus in patients with ischemic heart disease following thrombolytic therapy or angioplasty, although this combination is not expected to reverse platelet thrombus formation.

PDF (1336 kb)

Referenzen

References
1 Friedman M, Van den Bovenkamp GJ. The pathogenesis of a coronary thrombus. J Am Pathol 1966; 48: 19-44
2 Davies MJ, Thomas AC. Plaque fissuring – the cause of acute myocardial infarction, sudden ischaemic death, and crescendo angina. Br Heart J 1985; 53: 363-373
3 Fuster V, Chesebro JH. Mechanisms of unstable angina. N Engl J Med 1986; 315: 1023-1025
4 Willerson JT, Hillis LD, Winniford M, Buja LM. Speculation regarding mechanisms responsible for acute ischemic heart disease syndromes. J Am Coll Cardiol 1986; 8: 245-250
5 Collen D, Gold HK. Fibrin-specific thrombolytic agents and new approaches to coronary arterial thrombolysis. Thrombolysis in Cardiovascular Disease Julian D, Kübler W, Norris RM, Swan MJC, Collen D, Verstraete M. Marcel Dekker Inc., New York: 1989: 45-67
6 Marguerie GA, Plow EF, Edgington TS. Human platelets possess an inducible and saturable receptor specific for fibrinogen. J Biol Chem 1979; 254: 5357-5363
7 Plow EF, Ginsberg MH. Cellular adhesion: GPIIb/IIIa as a prototypic adhesion receptor. Prog Hemostas Thromb 1989; 9: 117-156
8 Coller BS, Scudder LE. Inhibition of dog platelet function by in vivo infusion of F(ab′)₂ fragments of a monoclonal antibody to the platelet glycoprotein IIb/IIIa receptor. Blood 1985; 66: 1456-1459
9 Huang TF, Holt JC, Kirby EP, Niewiarowski S. Trigramin: primary structure and its inhibition of von Willebrand factor binding to glycoprotein IIb/IIIa complex on human platelets. Biochemistry 1989; 28: 661-666
10 Gan ZR, Gould RJ, Jacobs JW, Friedman PA, Polokoff MA. Echistatin: a potent platelet aggregation inhibitor from the venom of the viper-echis carinatus. J Biol Chem 1988; 263: 19827-19832
11 Dennis MS, Henzel WJ, Pitti RM, Lipari MT, Napier MA, Deisher TA, Bunting S, Lazarus RA. Platelet glycoprotein GPIIb/IIIa protein antagonists from snake venoms: evidence for a family of platelet aggregation inhibitors. Proc Natl Acad Sci USA 1989; 87: 2471-2475
12 Plow EF, Pierschbacher MD, Ruoslahti E, Marguerie GA, Ginsberg MH. The effect of Arg-Gly-Asp-containing peptides on fibrinogen and von Willebrand factor binding to platelets. Proc Natl Acad Sci USA 1985; 82: 8057-8061
13 Ruoslahti E, Pierschbacher MD. Arg-Gly-Asp: a versatile cell recognition signal. Cell 1986; 44: 517-518
14 Coller BS, Folts JD, Scudder LE, Smith SR. Antithrombotic effect of a monoclonal antibody to the platelet glycoprotein IIb/IIIa receptor in an experimental animal model. Blood 1986; 68: 783-786
15 Gold HK, Coller BS, Yasuda T, Saito T, Fallon JT, Guerrero JL, Leinbach RC, Ziskind AA, Collen D. Rapid and sustained coronary artery recanalization with combined bolus injection of tissue-type plasminogen activator and monoclonal antiplatelet GPIIb/IIIa antibody in a canine preparation. Circulation 1988; 77: 670-677
16 Shebuski RJ, Berry DE, Bennett DB, Romoff T, Storer BL, Ali F, Samanen J. Demonstration of Ac-Arg-Gly-Asp-Ser-NH₂ as an antiaggregatory agent in the dog by intracoronary administration. Thromb Haemostas 1989; 61: 183-188
17 Yasuda T, Gold HK, Leinbach RC, Yaoita H, Fallon JT, Guerrero L, Napier MA, Bunting S, Collen D. Kistrin, a polypeptide platelet GPIIb/IIIa receptor antagonist, enhances and sustains coronary arterial thrombolysis with recombinant tissue-type plasminogen activator in a canine preparation. Circulation 1991; 83: 1038-1047
18 Fenton JW. Thrombin. Ann NY Acad Sci 1986; 485: 05-15
19 Harmon JT, Jamieson GA. Platelet activation by alpha-thrombin is a receptor-mediated event. Ann Ny Acad Sci 1986; 485: 387-395
20 Eidt JF, Allison P, Noble S, Ashton J, Golino P, Monatt J, Buja LM, Willerson JT. Thrombin is an important mediator of platelet aggregation in stenosed canine coronary arteries with endothelial injury. J Clin Invest 1989; 84: 18-27
21 Heras M, Chesebro JH, Webster MWI, Mruk JS, Grill DE, Penney WJ, Bowie W, Badimon L, Fuster V. Hirudin, heparin, and placebo during deep arterial injury in the pig. The in vivo role of thrombin in platelet-mediated thrombosis. Circulation 1990; 82: 1476-1484
22 Jang IK, Gold HK, Ziskind AA, Leinbach RC, Fallon JT, Collen D. Prevention of platelet-rich arterial thrombosis by selective thrombin inhibition. Circulation 1990; 81: 219-225
23 Stockmans F, Deckmyn H, Gruwez J, Vermylen J, Acland R. Continuous quantitative monitoring of moral, platelet-dependent, thrombus kinetics in the crushed rat femoral vein. Thromb Haemostas 1991; 65: 425-431
24 Imura Y, Stassen JM, Bunting S, Stockmans F, Collen D. Antithrombotic properties of G4120, a cyclic Arg-Gly-Asp-containing pentapeptide, in a hamster platelet-rich femoral vein thrombosis model. Blood (submitted)
25 Imura Y, Stassen JM, Collen D. Comparative antithrombotic effects of heparin, recombinant hirudin, and argatroban in a hamster femoral vein platelet-rich mural thrombosis model. J Pharm Exp Med (in press)
26 Kikumoto R, Tamao Y, Tezuka T, Tonomura S, Hara H, Ninomiya K, Hijikata A, Okamoto S. Selective inhibition of thrombin by (2R,4R)-4-methyl-1-[N²-[(3-methyl-1,2,3,4-tetrahydro-8-quinolinyl) sulfonyl]-L-arginyl)]-2-piperidine carboxylic acid. Biochemistry 1984; 23: 85-90
27 SAS/STAT User’s Guide, Version 6, 4th Edition. SAS Institute: Cary, NC: 1990
28 Berenbaum MC. The expected effect of a combination of agents: the general solution. J Theor Biol 1985; 114: 413-431
29 Collen D. Synergism of thrombolytic agents: investigational procedures and clinical potential. Circulation 1988; 77: 731-735
30 Rao CR. Linear Statistical Inference and its Applications. 2nd edition. John Wiley & Sons: New York: 1972
31 Seber GAF Linear Regression Analysis. John Wiley & Sons: New York: 1977
32 Fitzgerald DG, Fragetta J, FitzGerald GA. Prostaglandin endoperoxides modulate the response to thromboxane synthase inhibition during coronary thrombosis. J Clin Invest 1988; 85: 1708-1713
33 Gresele P, Deckmyn H, Nenci GG, Vermylen J. Thromboxane synthase inhibitors, thromboxane receptor antagonists and dual blockers in thrombotic disorders. TiPS 1991; 12: 158-163
34 Radomski MW, Palmer RM, Moncada S. The anti-aggregating properties of vascular endothelium: interactions between prostacyclin and nitric oxide. Br J Pharmacol 1989; 92: 639-646
35 Gold HK. Conjunctive antithrombotic and thrombolytic therapy for coronary-artery occlusion. N Engl J Med 1990; 323: 1483-1485
36 Hanson SA, Harker LA. Interruption of acute platelet-dependent thrombosis by the synthetic antithrombin D-phenylalanyl-L-prolyl-L-arginyl chloromethylketone. Proc Natl Acad Sci USA 1988; 85: 3184-3188
37 Church FC, Phillips JE, Woods JL. Chimeric antithrombin peptide: characterization of an Arg-Gly-Asp (RGD)- and hirudin carboxyl terminus-containing synthetic peptide. J Biol Chem 1991; 266: 11975-11979