Thrombolytic and Haemorrhagic Effects of Bolus Doses of Tissue-Type Plasminogen Activator and a Hybrid Plasminogen Activator with Prolonged Plasma Half-Life (K2tu-PA: CGP 42935)

Giancarlo Agnelli; Claudia Pascucci; Giuseppe G Nenci; Antonio Mele; Rolf Bürgi; Jutta Heim

doi:10.1055/s-0038-1649569

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1993; 70(02): 294-300
DOI: 10.1055/s-0038-1649569

Original Articles

Fibrinolysis

Schattauer GmbH Stuttgart

Thrombolytic and Haemorrhagic Effects of Bolus Doses of Tissue-Type Plasminogen Activator and a Hybrid Plasminogen Activator with Prolonged Plasma Half-Life (K₂tu-PA: CGP 42935)

Giancarlo Agnelli

¹The Institute of Internal and Vascular Medicine, University of Perugia, Italy

,

Claudia Pascucci

¹The Institute of Internal and Vascular Medicine, University of Perugia, Italy

,

Giuseppe G Nenci

¹The Institute of Internal and Vascular Medicine, University of Perugia, Italy

,

Antonio Mele

²The Menarini Ricerche Sud, Rome, Italy

,

Rolf Bürgi

³The Biotechnology Department, Ciba Geigy, Basel, Switzerland

,

Jutta Heim

³The Biotechnology Department, Ciba Geigy, Basel, Switzerland

› Institutsangaben

Abstract

als PDF herunterladen

Summary

K₂tu-PA is a hybrid plasminogen activator linking the kringle 2 domain of tissue-type plasminogen activator (t-PA) to the catalytic protease domain of single-chain urokinase-type plasminogen activator (scu-PA). K₂tu-PA, as t-PA has high affinity for fibrin and is activated by fibrin but has a longer plasma half-life (over 30 min). The aim of this study was to compare the effects of bolus doses of recombinant t-PA (rt-PA) and K₂tu-PA, on: 1) lysis of preformed thrombi (fibrinolysis), 2) accretion of new fibrin on pre-existing thrombi during fibrinolysis (thrombus growth), 3) thrombolysis as assessed by reduction of thrombus weight and 4) systemic plasma proteolysis and blood loss from a standard wound. A jugular vein thrombosis model and an ear bleeding model were adopted in rabbits. Saline produced 11 ± 2% fibrinolysis. rt-PA, 0.2 mg/kg, 0.4 mg and 0.8 mg/kg produced 35 ± 4%, 54 ± 4% and 78 ± 6% fibrinolysis, respectively. K₂tu-PA, at the same doses, produced 39 ± 5%, 57 ± 6% and 83 ± 6% fibrinolysis, respectively. Thus, no differences in the fibrinolytic activity of rt-PA and K₂tu-PA were observed. Injection of saline was followed by an accretion of 56.4 ± 5.9 μg of radioactive new fibrin on the thrombi. The injection of the three increasing doses of rt-PA was followed by an accretion of 54.9 ± 5.3 μg, 49.1 ± 6.1 μg and 47.2 ± 4.8 μg. The injection of three increasing doses of K₂tu-PA was followed by an accretion of 38.1 ± 3.4 μg, 29.6 ± 2.5 μg and 17.1 ± 3.4 μg. At each of the three doses, K₂tu-PA was more effective than rt-PA in reducing the accretion of new fibrin on the thrombi (p <0.01) and, as a consequence, in reducing thrombus weight (p <0.01). The two lower doses of rt-PA and K₂tu-PA did not produce systemic proteolysis and bleeding. The highest dose of K₂tu-PA produced a statistically significant more intense systemic proteolysis and bleeding than the highest dose of rt-PA.

This study demonstrates that bolus doses of K₂tu-PA and rt-PA produce a similar degree of fibrinolysis. Due to its longer half-life K₂tu-PA is more efficient than rt-PA in inhibiting accretion of new fibrin on the thrombi during thrombolysis so that the thrombus size is more efficiently reduced. As a consequence the concomitant use of heparin might not be necessary. The potential increased risk of bleeding with bolus of high doses of K₂tu-PA has to be seen in view of the advantage of avoiding the concomitant use of heparin.

PDF (1870 kb)

Referenzen

References
1 Agnelli G, Buchanan MR, Fernandez F, Boneu B, Van Ryn J, Hirsh J, Collen D. A comparison of the thrombolytic and haemorrhagic effects of tissue-type plasminogen activator and streptokinase in rabbits. Circulation 1985; 72: 178-182
2 Rao AK, Pratt CP, Berke A, Jaffe A, Ockene I, Schreiber TL, Bell WR, Knatterud G, Robertson TL, Terin ML. Thrombolysis in myocardial infarction (TIMI) trial - phase I: The fibrinolytic system in patients treated with recombinant tissue plasminogen activator and streptokinase. J Am Coll Cardiol 1988; 11: 1-11
3 Bovill EG, Terrin ML, Stump DC, Berke AD, Frederick M, Collen D, Feit F, Gore JM, Hillis LD, Lambrew CT, Leiboff R, Mann KG, Markis JE, Pratt CM, Sharkey SW, Sopko G, Tracy RP, Chesebro JH. Haemorrhagic events during therapy with recombinant tissue-type plasminogen activator, heparin, and aspirin for acute myocardial infarction. Ann Intern Med 1991; 115: 256-265
4 Agnelli G, Buchanan MR, Fernandez F, Van Ryn J, Hirsh J. Sustained thrombolysis with DNA-recombinant tissue type plasminogen activator in rabbits. Blood 1985; 66: 399-401
5 Agnelli G, Buchanan MR, Fernandez F, Hirsh J. The thrombolytic and haemorrhagic effects of tissue type plasminogen activator: influence of dosage regimens in rabbits. Thromb Res 1985; 40: 769-777
6 Clozel JP, Tscopp T, Luedin E, Holvoet P. Time course of thrombolysis induced by intravenous bolus or infusion of tissue plasminogen activator in a rabbit jugular vein thrombosis model. Circulation 1989; 79: 125-133
7 Shiffman F, Ducas J, Hollett P, Israels E, Greenberg D, Cook R, Prewitt RM. Treatment of canine embolic pulmonary hypertension with recombinant tissue plasminogen activator: efficacy of dosing regimens. Circulation 1988; 78: 214-220
8 Prewitt RM, Shiffman F, Greenburg D, Cook R, Ducas J. Recombinant tissue-type plasminogen activator in canine embolic pulmonary hypertension: effects of bolus versus short-term administration on dynamics of thrombolysis and on pulmonary vascular pressure-flow characteristics. Circulation 1989; 79: 929-938
9 Asselbergs FAM, Bürgi R, Heim J, van OostrumJ, Agnelli G. Biochemical and biological characterization of K₂tu-PA, a hybrid plasminogen activator. Thromb Haemostas 1991; 65: 779
10 Agnelli G, Pascucci C, Colucci M, Nenci GG, Mele A, Bürgi R, Heim J. Thrombolytic activity of two chimeric recombinant plasminogen activators (FK₂tu-PA and K₂tu-PA) in rabbits. Thromb Haemostas 1992; 68: 331-335
11 Giles AR. Guidelines for the use of animals in biomedical research. Thromb Haemostas 1987; 58: 1078-1084
12 Carter CJ, Kelton JG, Hirsh J. Comparison of the haemorrhagic effects of porcine and bovine heparin in rabbits. Thromb Res 1979; 15: 581-587
13 Clauss A. Gerinnungsphysiologische Schnellmethode zur Bestimmung des Fibrinogens. Acta Haematol 1957; 17: 237-246
14 Friberger P, Knos M. Plasminogen determination in human plasma. In Chromogenic Peptide Substances: Chemistry and Clinical Use. Scully M, Kakkar W. (eds) Edinburgh: Churchill Livingstone; 1979: 128-140
15 Teger-Nilsson AC, Friberger P, Gyzander E. Determination of a new rapid plasmin inhibitor in human blood by means of a plasmin specific tripeptide substrate. Scand J Lab Invest 1977; 37: 403-409
16 Colucci M, Cavallo LG, Agnelli G, Mele A, Bürgi R, Heim J, Semeraro N. Properties of chimeric (tissue-type/urokinase-type) plasminogen activators obtained by fusion at the plasmin cleavage site. Thromb Haemostas 1993; 69: 466-472
17 Gruppo Italiano per lo Studio della Streptochinasi nell’Infarto Miocardiaco (GISSI). Effectiveness of intravenous thrombolytic treatment in acute myocardiac infarction. Lancet 1986; 1: 397-402
18 ISIS-2 (Second International Study on Infarct Survival) Collaborative Group. Randomised trial of intravenous streptokinase, oral aspirin, both, or neither among 17,187 cases of suspected acute myocardial infarction: ISIS-2. Lancet 1988; 2: 349-360
19 Gruppo Italiano per lo Studio della Sopravvivenza nell’Infarto Miocardiaco (Gissi) 2. A factorial randomised trial of alteplase versus streptokinase and heparin versus no heparin among 12,490 patients with acute myocardial infarction. Lancet 1990; 336: 65-71
20 ISIS-3 (Third International Study of Infarct Survival) Collaborative Group. A randomized comparison of streptokinase vs tissue plasminogen activator vs anistreplase and of aspirin plus heparin vs aspirin alone among 41,299 cases of suspected acute myocardial infarction. Lancet 1992; 339: 753-770
21 Agnelli G. Rationale for bolus t-PA therapy to improve efficacy and safety. Chest 1990; 97: 161S-167S
22 Agnelli G, Parise P. Bolus thrombolysis in venous thromboembolism. Chest 1992; 101: 172S-82S299
23 Lijnen HR, Collen D. Strategies for the improvement of thrombolytic agents. Thromb Haemostas 1991; 66: 88-110
24 Gheysen D, Lijnen HR, Pierard L, de Foresta F, Demarsin E, Jacobs P, De Wilde M, Bollen A, Collen D. Characterization of a recombinant fusion protein of the finger domain of tissue-type plasminogen activator with a truncated single chain urokinase-type plasminogen activator. J Biol Chem 1987; 262: 11779-11784
25 Pierard L, Jacobs P, Gheysen D, Hoylaerts M, André B, Topisirovic L, Cravador A, de Foresta F, Herzog A, Collen D, De Wilde M, Bollen A. Mutant and chimeric recombinant plasminogen activators. J Biol Chem 1987; 262: 11771-11778
26 Nelles L, Lijnen HR, Collen D, Holmes WE. Characterization of a fusion protein consisting of amino acids 1 to 263 of tissue-type plasminogen activator and amino acids 144 to 411 of urokinase-type plasminogen activator. J Biol Chem 1987; 262: 10855-10862
27 Lijnen HR, Nelles L, van Hoef B, De CockR, Collen D. Inhibition in purified systems and in human plasma of chimaeric plasminogen activators consisting of the NH₂-terminal region of tissue-type plasminogen activator and the COOH-terminal region of urokinase-type plasminogen activator. Thromb Haemostas 1988; 60: 247-250
28 Wikström K, Mattson C, Sterky C, Pohl G. Tissue plasminogen activator mutants lacking the growth factor domain and first kringle domain: II Enzymatic properties in plasma and in vivo thrombolytic activity and clearance rates in rabbits. Fibrinolysis 1991; 5: 31-41
29 Pohl G, Sterky C, Attersand A, Nyberg E, Löwenadler B, Hansson L. Tissue plasminogen activator mutants lacking the growth factor domain and first kringle domains: I DNA constructions, expressions in mammalian cells, protein structure, fibrin affinity and enzymatic properties. Fibrinolysis 1991; 5: 17-29
30 Collen D, Lijnen HR, Vanlinthout I, Kieckens L, Nelles L, Stassen JM. Thrombolytic and pharmacokinetic properties of human tissue-type plasminogen activator variants, obtained by deletion and/or duplication of structural/functional domains, in a hamster pulmonary embolism model. Thromb Haemostas 1991; 65: 174-180
31 Collen D, Lu HR, Lijnen HR, Nelles L, Stassen JM. Thrombolytic and pharmacokinetic properties of chimeric tissue-type and urokinase-type plasminogen activators. Circulation 1991; 84: 1216-1234
32 Agnelli G, Pascucci C, Cosmi B, Nenci GG. Effects of therapeutic doses of heparin on thrombolysis with tissue-type plasminogen activator in rabbits. Blood 1990; 76: 2030-2036
33 Rapold HJ, Lu HR, Wu Z, Nijs H, Collen D. Requirement of heparin for arterial and venous thrombolysis with recombinant tissue-type plasminogen activator. Blood 1991; 77: 1020-1024
34 Weitz JI, Leslie B, Ginsberg J. Soluble fibrin degradation products potentiate tissue plasminogen activator-induced fibrinogen proteolysis. J Clin Invest 1991; 87: 1-10
35 Oczkowski WJ, Falcone D, Buchanan MR, Hirsh J. Tissue plasminogen activator associated bleeding is increased by the presence of a lysing thrombus. Circulation 1988; 78 (Suppl. 02) 352