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Thromb Haemost 1997; 77(05): 1025-1033
DOI: 10.1055/s-0038-1656097
Animal models
Schattauer GmbH Stuttgart

Comparison of Dose Regimens for the Administration of Recombinant Pro-urokinase in a Canine Thrombosis Model

Sandra E Burke
1   The Department of General Pharmacology, Abbott Laboratories, Abbott Park, Illinois, USA and
,
Nathan L Lubbers
1   The Department of General Pharmacology, Abbott Laboratories, Abbott Park, Illinois, USA and
,
Richard A Nelson
1   The Department of General Pharmacology, Abbott Laboratories, Abbott Park, Illinois, USA and
,
Jack Henkin
2   The Department of Thrombolytic Venture, Abbott Laboratories, Abbott Park, Illinois, USA
› Author Affiliations
Further Information

Publication History

Received 30 January 1996

Accepted after revision 11 December 1996

Publication Date:
11 July 2018 (online)

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Summary

Pro-urokinase represents an important addition to the array of thrombolytic drugs currently available for clinical use because of its high clot specificity but distinctly different mechanism compared with that of t-PA. Recombinant pro-urokinase (r-proUK) is a single-chain precursor of high molecular weight urokinase which has been expressed in a mouse myeloma cell line. The present study was conducted to determine the dosing regimen which would produce optimal clot lysis and restoration of blood flow 2 h after treatment with r-proUK, using a dog model of arterial thrombosis. Efficacy was indicated by lysis of a radiolabelled clot which was formed in the heat-damaged femoral arteries of 39 male beagle dogs. The animals were divided into six heparinized treatment groups, each receiving one of five dosing regimens or the vehicle for r-proUK. The total dose (80,000 U/kg) was divided into an initial loading bolus, followed by either a second bolus or by infusions for various time periods, as shown below:

It was concluded that optimal clot lysis and restoration of femoral flow was accomplished using a regimen in which 50% of the dose was given as a bolus, followed immediately by the remaining 50% given as a 30 min intravenous infusion (Group 5). At the dose used in this study, r-proUK did not produce degradation of fibrinolytic or hemostatic plasma proteins.

 

  • References

  • 1 Gurewich V, Pannell R, Louie S, Kelley P, Suddith RL, Greenlee R. Effective and fibrin-specific clot lysis by a zymogen precursor form of urokinase (pro-urokinase). A study in vitro and in two animal species. J Clin Invest 1984; 73: 1731-1739
    CrossrefPubMedGoogle Scholar
  • 2 Collen D, Stassen JM, Blaber M, Winkler M, Verstraete M. Biological and thrombolytic properties of proenzyme and active forms of human urokinase - III. Thrombolytic properties of natural and recombinant urokinase in rabbits with experimental jugular vein thrombosis. Thromb Haemost 1984; 52: 27-30
    PubMedGoogle Scholar
  • 3 Pannell R, Gurewich V. Pro-urokinase, a study of its stability in plasma and of a mechanism for its selective fibrinolytic effect. Blood 1986; 67: 1215-1223
    PubMedGoogle Scholar
  • 4 Flameng W, Vanhaecke J, Stump DC, van de WerfF, Holmes W, Guenzler WA, Flohe L, Collen D. Coronary thrombolysis by intravenous infusion of recombinant single chain urokinase-type plasminogen activator of recombinant urokinase in baboons: effect on regional blood flow, infarct size and hemostasis. J Am Coll Cardiol 1986; 8: 118-124
    CrossrefPubMedGoogle Scholar
  • 5 Burke SE, Lubbers NL, Nelson RA, Henkin J. Recombinant pro-urokinase requires heparin for optimal clot lysis and restoration of blood flow in a canine femoral artery thrombosis model. Thromb Haemost 1993; 4: 375-380
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Gurewich V. Pro-urokinase: physiochemical properties and promotion of its fibrinolytic activity by urokinase and by tissue plasminogen activator with which it has a complementary action. Semin Thromb Hemost 1988; 14: 110-115
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Kirschenbaum JM, Bahr RD, Flaherty JT, Gurewich V, Levine HJ, Loscalzo J, Schumacher RR, Topol EJ, Wahr DW, Braunwald E. Clot-selective coronary thrombolysis with low-dose synergistic combinations of single-chain urokinase-type plasminogen activator and recombinant tissue-type plasminogen activator. The Pro-Urokinase for Myocardial Infarction Study Group. Am J Cardiol 1991; 68: 1564-1569
    CrossrefPubMedGoogle Scholar
  • 8 Kasper W, Hohnloser SH, Engler H, Meinertz T, Wilkens J, Roth E, Lang K, Limbourg P, Just H. Coronary reperfusion studies with pro-urokinase in acute myocardial infarction: evidence for synergism of low dose urokinase. J Am Coll Cardiol 1990; 16: 733-738
    CrossrefPubMedGoogle Scholar
  • 9 Weaver WD, Hartmann JR, Anderson JL, Reddy PS, Sobolski JC, Sasa-hara AA. New recombinant glycosylated Prourokinase for treatment of patients with acute myocardial infarction. J Am Coll Cardiol 1994; 24: 1242-1248
    CrossrefPubMedGoogle Scholar
  • 10 Bode C, Nordt TK, Runge MS. Thrombolytic therapy in acute myocardial infarction – Selected recent developments. Ann Hematol 1994; 69: 35-40
    CrossrefPubMedGoogle Scholar
  • 11 Sasahara St AA, Martin CC, Henkin J, Barker WM. Approach to the patient with venous thromboembolism: Treatment with thrombolytic agents. Hematol Oncol Clin North Am 1992; 6: 1141-1159
    CrossrefPubMedGoogle Scholar
  • 12 Van de Werf F, Nobuhara M, Collen D. Coronary thrombolysis with human single-chain, urokinase-type plasminogen activator (pro-urokinase) in patients with acute myocardial infarction. Ann Intern Med 1986; 104: 345-348
    CrossrefPubMedGoogle Scholar
  • 13 Trubestein G, Popov S, Wolf H, Welzel D. Effects of pro-urokinase and urokinase on the fibrinolytic system in man. Haemost 1987; 17: 238-244
    CrossrefPubMedGoogle Scholar
  • 14 Kambara H, Kawai C, Kajiwara N, Niitani H, Sasayama S, Kanmatsuse K, Kodama K, Sato H, Nobuyoshi M, Nakashima M. et al. Randomized double-blinded multicenter study. Comparison of intracoronary single-chain urokinase-type plasminogen activator pro-urokinase (GE-0943), and intracoronary urokinase in patients with acute myocardial infarction. Circulation 1988; 78: 899-905
    CrossrefPubMedGoogle Scholar
  • 15 Loscalzo J, Wharton TP, Kirshenbaum JM, Levine HJ, Flaherty JT, Topol EJ, Ramaswamy K, Kosowsky BD, Salem DN, Ganz P. et al. Clot-selective coronary thrombolysis with pro-urokinase. Circulation 1989; 79: 776-782
    CrossrefPubMedGoogle Scholar
  • 16 PRIMI Trial Study Group. Randomised double-blind trial of recombinant pro-urokinase against streptokinase in acute myocardial infarction. Lancet 1989; 1: 863-868
    PubMedGoogle Scholar
  • 17 Bode C, Schuler G, Nordt T, Schonermark S, Baumann H, Richardt G, Dietz R, Gurewich V, Kubler W. Intravenous thrombolytic therapy with a combination of single-chain urokinase-type plasminogen activator and recombinant tissue-type plasminogen activator in acute myocardial infarction. Circulation 1990; 81: 907-913
    CrossrefPubMedGoogle Scholar
  • 18 Kasper W, Meinertz T, Hohnloser S, Engler H, Hasler C, Rossler W, Wolf H, Welzel D, Just H, Gurewich V. Coronary thrombolysis in man with prourokinase: improved efficacy with low dose urokinase. Klin Wochenschr 1988; 66: 109-114
    PubMedGoogle Scholar
  • 19 Gulba DC, Bode C, Sen S, Topp J, Fischer K, Wolf H, Hecker H. Multicenter dose-finding trial for thrombolysis with urokinase preactivated prourokinase (TCL 598) in acute myocardial infarction. German Preactivated Pro-Urokinase Study Group. Cathet Cardiovasc Diagn 1992; 26: 177-184
    CrossrefPubMedGoogle Scholar
  • 20 Badylak SF, Poehlman E, Williams C, Klabunde RE, Henkin J, Simple canin. model of arterial thrombosis with endothelial injury suitable for investigation of thrombolytic agents. J Pharmacol Methods 1988; 19: 293-304
    CrossrefPubMedGoogle Scholar
  • 21 Sutherland JW, Banick DW, Schmakel CO. Stabilized procedure for the determination of urokinase fibrinolytic activity potency. Anal Biochem 1991; 194: 121-129
    CrossrefPubMedGoogle Scholar
  • 22 Friberger P, Knӧs M. Plasminogen determination in human plasma. In: Peptide Substrates: Chemical and Clinical Usage. Scully MF, Kakkar VV. eds. Edinburgh: Churchill Livingstone; 1979. pp 128
    Google Scholar
  • 23 Edy J, De Cock F, Collen D. Inhibition of plasmin by normal and antiplas- min-depleted human plasma. Thromb Res 1976; 8: 513-518
    CrossrefPubMedGoogle Scholar
  • 24 Vermylen C, De Vreker RA, Verstraete M. A rapid enzymatic method for assay of fibrinogen fibrin polymerization time (FPT test). Clin Chemica Acta 1963; 8: 418-424
    PubMedGoogle Scholar
  • 25 Lo K-M, Gillies SD. High level expression of human proteins in murine hybridoma cells: induction by methotrexate in the absence of gene amplification. Biochimica et Biophysica Acta 1991; 1088: 217-224
    CrossrefPubMedGoogle Scholar
  • 26 Credo RB, Burke SE. Fibrinolytic mechanism, biochemistry, and pre-clinical pharmacology of recombinant prourokinase. J Vase Intervent Radiol 1995; 6: 08-18
    PubMedGoogle Scholar
  • 27 Lenich C, Pannell R, Henkin J, Gurewich V. The influence of glycosylation on the catalytic and fibrinolytic properties of pro-urokinase. Thromb Haemost 1992; 68: 539-544
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Tukey JW, Ciminera JL, Heyse JF. Testing the statistical certainty of a response to increasing doses of a drug. Biometrics 1985; 4: 295-301
    PubMedGoogle Scholar