Download PDF
Thromb Haemost 1992; 67(06): 686-691
DOI: 10.1055/s-0038-1648523
Original Articles
Schattauer GmbH Stuttgart

G4120, an Arg-Gly-Asp Containing Pentapeptide, Enhances Arterial Eversion Graft Recanalization with Recombinant Tissue-Type Plasminogen Activator in Dogs

Hua Rong Lu
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Herman K Gold
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Zaomin Wu
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Tsunehiro Yasuda
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Patrick Pauwels
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Hansjörg J Rapold
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Mary Napier
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Stuart Bunting
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
,
Désiré Collen
The Center for Thrombosis and Vascular Research, the Laboratory of Experimental Cardiology and the Department of Histopathology, University of Leuven, Belgium, the Cardiac Unit, Massachusetts General Hospital, Harvard Medical School, Boston MA, USA and the Department of Cardiovascular Research, Genentech Inc., South San Francisco, CA, USA
› Author Affiliations
Further Information

Publication History

Received 11 October 1991

Accepted after revision 18 December 1991

Publication Date:
03 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

The effects of G4120, a cyclic Arg-Gly-Asp (RGD) containing peptide which inhibits fibrinogen binding to the platelet receptor GPIIb/IIIa, on thrombolysis with recombinant tissue-type plasminogen activator (rt-PA) were investigated in a combined arterial and venous thrombosis model in heparinized dogs. The arterial thrombus model consisted of a 3 cm everted (inside-out) carotid arterial segment inserted into a transsected femoral artery which occludes within 30 min with platelet-rich material and which is resistant to recanalization with 0.5 mg/kg rt-PA. The venous thrombus was a 125I-fibrin labeled whole blood clot produced in the contralateral femoral vein.

In 5 dogs given an intravenous bolus of 0.05 mg/kg G4120 followed by a continuous infusion of 0.05 mg/kg per hour for 3 h (group I), arterial occlusion persisted throughout a 4 h observation period and was still present at 24 h in all dogs; the extent of venous clot lysis after 120 min was 27 ± 7%. In 5 dogs given the same infusion of G4120 in combination with 0.5 mg/kg rt-PA over 60 min, recanalization of the arterial graft occurred in all dogs, within 13 ± 2 min and persisted throughout the observation period of 4 h (p = 0.01 versus G4120 or rt-PA alone); at 24 h, however, all grafts were occluded. Venous clot lysis in this group was 75 ± 8% (p = 0.002 versus G4120 alone andp NS versus rt-PA alone). Pathologic analysis revealed platelet-rich or mixed thrombus with platelet-rich and erythrocyte-rich zones. The last 6 dogs were given a reduced dose of G4120 consisting either of a 0.05 mg/kg bolus followed by an infusion of 0.05 mg/kg over 1 h in 3 dogs (group III) or of a single 0.05 mg/kg bolus in 3 dogs (group IV), both given in combination with 0.5 mg/kg rt-PA infused over 60 min. These protocols produced recanalization within 15 ± 2 and 34 ± 8 min, respectively, which was maintained throughout the 4 h observation period. Venous lysis in these groups was 63 ± 4 and 97 ± 1% respectively. Bleeding times prolonged from 1 to 2 min to >30 min with G4120, but returned towards baseline within 2 h after the end of the infusion. Platelet aggregation with ADP was completely inhibited with G4120 but partially recovered within 1 h after the end of the infusion. No fibrinogen breakdown was observed in association with the rt-PA infusion.

Thus, G4120, a synthetic GPIIb/IIIa receptor antagonist, enhances and accelerates lysis of platelet-rich arterial thrombosis with rt-PA and prevents reocclusion during and within 3 h after the infusion. It may be useful for the conjunctive use with thrombolytic agents in patients with arterial thromboembolic disease.

 

  • References

  • 1 Collen D, Stump DC, Gold HK. Thrombolytic therapy. Annu Rev Med 1988; 39: 405-423
    CrossrefPubMedGoogle Scholar
  • 2 Braunwald E. Myocardial reperfusion, limitation of infarct size, reduction of left ventricular dysfunction, and improved survival. Should the paradigm be expanded?. Circulation 1989; 79: 441-444
    CrossrefPubMedGoogle Scholar
  • 3 Ohman EM, Califf RM. Thrombolytic therapy: overview of clinical trials. Coron Art Dis 1990; 1: 23-33
    CrossrefPubMedGoogle Scholar
  • 4 Neuhaus KL, Feuerer W, Jeep-Tebbe S, Niederer W, Vogt A, Tebbe U. Improved thrombolysis with a modified dose regimen of recombinant tissue-type plasminogen activator. J Am Coll Cardiol 1989; 14: 1566-1569
    CrossrefPubMedGoogle Scholar
  • 5 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
    PubMedGoogle Scholar
  • 6 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
    PubMedGoogle Scholar
  • 7 Yasuda T, Gold HK, Fallon JT, Leinbach RC, Guerrero JL, Scudder LE, Kanke M, Shealy D, Ross MJ, Collen D, Coller BS. Monoclonal antibody against the platelet glycoprotein (GP)IIb/IIIa receptor prevents coronary artery reocclusion after reperfusion with recombinant tissue-type plasminogen activator in dogs. J Clin Invest 1988; 81: 1284-1291
    CrossrefPubMedGoogle Scholar
  • 8 Mickelson JK, Simpson PJ, Cronin M, Homeister JW, Laywell E, Kitzen J, Lucchesi BR. Antiplatelet antibody [7E3F(ab’)2] prevents rethrombosis after recombinant tissue-type plasminogen activator-induced coronary artery thrombolysis in a canine model. Circulation 1990; 81: 617-627
    CrossrefPubMedGoogle Scholar
  • 9 Yasuda T, Gold HK, Leinbach RC, Saito T, Guerrero JL, Jang IK, Holt R, Fallon JT, Collen D. Lysis of plasminogen activator-resistant platelet-rich coronary artery thrombus with combined bolus injection of recombinant tissue-type plasminogen activator and anti-platelet GPIIb/IIIa antibody. J Am Coll Cardiol 1990; 16: 1728-1735
    CrossrefPubMedGoogle Scholar
  • 10 Lu HR, Gold HK, Wu Z, De Cock F, Jang IK, Pauwels P, Collen D. Acceleration and persistence of rt-PA induced arterial eversion graft recanalization with a single bolus injection of F(ab’)2 fragments of the antiplatelet GPIIb/IIIa antibody 7E3. Coron Art Dis 1991; 2: 1039-1046
    PubMedGoogle Scholar
  • 11 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 recombinant tissue-type plasminogen activator and monoclonal antiplatelet GPIIb/IIIa antibody in a canine preparation. Circulation 1988; 77: 670-677
    CrossrefPubMedGoogle Scholar
  • 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
    CrossrefPubMedGoogle Scholar
  • 13 Ruoslahti E, Pierschbacher MD. Arg-Gly-Asp: a versatile cell recognition signal. Cell 1986; 44: 517-518
    CrossrefPubMedGoogle Scholar
  • 14 Ouyang C, Huang TF. Potent platelet aggregation inhibitor from trimeresurus gramineus snake venom. Biochim Biophys Acta 1983; 757: 332-341
    CrossrefPubMedGoogle Scholar
  • 15 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
    PubMedGoogle Scholar
  • 16 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
    PubMedGoogle Scholar
  • 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
    CrossrefPubMedGoogle Scholar
  • 18 Nichols A, Vasko J, Koster P, Smith J, Barone F, Nelson A, Stadel J, Powers J, Rhodes G, Miller-Stein C, Boppana V, Bennet D, Berry D, Romoff T, Calvo R, Ali F, Sorenson E, Samanen J. SK&F 106760, a novel GPIIb/IIIa antagonist: antithrombotic activity and potentiation of streptokinase-mediated thrombolysis. Eur J Pharmacol 1990; 183: 2019 (abstr)
    CrossrefPubMedGoogle Scholar
  • 19 Barker PL, Burnier JP, Gadek T, Thorsett ED. Small cyclic peptide aggregation inhibitor. PCT Inti Appl W091 01331, February 7 1991
    PubMedGoogle Scholar
  • 20 Rapold HJ, Gold HK, Wu Z, Napier M, Bunting S, Collen D. Effects of G4120, a Arg-Gly-Asp containing synthetic platelet glycoprotein IIb/IIIa receptor antagonist on arterial and venous thrombolysis with recombinant tissue-type plasminogen activator in dogs. (Submitted)
    PubMedGoogle Scholar
  • 21 Jang IK, Gold HK, Ziskind AA, Fallon JT, Holt RE, Leinbach RC, May JW, Collen D. Differential sensitivity of erythrocyte-rich and platelet-rich arterial thrombi to lysis with recombinant tissue-type plasminogen activator. A possible explanation for resistance to coronary thrombolysis. Circulation 1989; 79: 920-928
    CrossrefPubMedGoogle Scholar
  • 22 Giles AR. Guidelines for the use of animals in biomedical research. Thromb Haemostas 1987; 58: 1078-1084
    PubMedGoogle Scholar
  • 23 Kominger C, Matsuo O, Suy R, Stassen JM, Collen D. Thrombolysis with human extrinsic (tissue-type) plasminogen activator in dogs with femoral vein thrombosis. J Clin Invest 1982; 69: 573-580
    CrossrefPubMedGoogle Scholar
  • 24 Clauss A. Gerinnungsphysiologische Schnellmethode zur Bestim-mung des Fibrinogens. Acta Haematol 1957; 17: 237-246
    CrossrefPubMedGoogle Scholar
  • 25 Vermylen J, De Vreker R, Verstraete M. A rapid enzymatic method for assay of fibrinogen. Fibrin polymerization time (FPT-test). Clin Chim Acta 1963; 8: 418-424
    CrossrefPubMedGoogle Scholar
  • 26 Edy J, Collen D, Verstraete M. Quantitation of the plasma protease inhibitor, antiplasmin with the chromogenic substrate S-2251. In: Progress in Chemical Fibrinolysis and Thrombolysis. Vol 3. Davidson JF. et al (eds). Raven Press; New York: 1978. pp 315-322
    Google Scholar
  • 27 Holvoet P, Cleemput H, Collen D. Assay of human tissue-type plasminogen activator (t-PA) with an enzyme-linked immunosorbent assay (ELISA) based on three murine monoclonal antibodies to t-PA. Thromb Haemostas 1985; 54: 684-687
    PubMedGoogle Scholar
  • 28 Badimon L, Lassila R, Badimon J, Vallabhajosula S, Chesebro JH, Fuster V. Residual thrombus is more thrombogenic than severely damaged vessel wall. Circulation 1988; 78 Suppl II 11-119
    PubMedGoogle Scholar
  • 29 Plow EF, Ginsberg MH. Cellular adhesion: GPIIb/IIIa as a prototypic adhesion receptor. Prog Hemostas Thromb 1989; 9: 117-156
    PubMedGoogle Scholar
  • 30 Spriggs D, Gold HK, Hashimoto Y, Vanhoutte E, Vermylen J, Collen D. Absence of potentiation with murine antiplatelet GPIIb/IIIa antibody of thrombolysis with recombinant tissue-type plasminogen activator (rt-PA) in a canine venous thrombosis model. Thromb Haemostas 1989; 61: 93-96
    PubMedGoogle Scholar