Comparative Thrombolytic Properties of Tissue-Type Plasminogen Activator and of a Plasminogen Activator Inhibitor-1 -Resistant Glycosylation Variant, in a Combined Arterial and Venous Thrombosis Model in the Dog

Désiré Collen; Jean-Marie Stassen; Tsunehiro Yasuda; Canio Refino; Nicholas Paoni; Bruce Keyt; Tania Roskams; J Luis Guerrero; Henri R Lijnen; Herman K Gold; William F Bennett

doi:10.1055/s-0038-1648819

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1994; 72(01): 098-104
DOI: 10.1055/s-0038-1648819

Original Article

Schattauer GmbH Stuttgart

Comparative Thrombolytic Properties of Tissue-Type Plasminogen Activator and of a Plasminogen Activator Inhibitor-1 -Resistant Glycosylation Variant, in a Combined Arterial and Venous Thrombosis Model in the Dog

Désiré Collen

¹The Center for Molecular and Vascular Biology, University of Leuven, Belgium

,

Jean-Marie Stassen

¹The Center for Molecular and Vascular Biology, University of Leuven, Belgium

,

Tsunehiro Yasuda

²Division of Cardiology, Massachusetts General Hospital Harvard Medical School, Boston, MA, USA

,

Canio Refino

³Department of Cardiovascular Pharmacology, Genentech Inc, South San Francisco, CA, USA

,

Nicholas Paoni

³Department of Cardiovascular Pharmacology, Genentech Inc, South San Francisco, CA, USA

,

Bruce Keyt

³Department of Cardiovascular Pharmacology, Genentech Inc, South San Francisco, CA, USA

,

Tania Roskams

⁴Department of Pathology, University of Leuven, Belgium

,

J Luis Guerrero

²Division of Cardiology, Massachusetts General Hospital Harvard Medical School, Boston, MA, USA

,

Henri R Lijnen

¹The Center for Molecular and Vascular Biology, University of Leuven, Belgium

,

Herman K Gold

²Division of Cardiology, Massachusetts General Hospital Harvard Medical School, Boston, MA, USA

,

William F Bennett

³Department of Cardiovascular Pharmacology, Genentech Inc, South San Francisco, CA, USA

› Author Affiliations

Abstract

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Summary

rt-PA-K, a variant of recombinant tissue-type plasminogen activator (rt-PA) with substitution of amino acids 296 to 299 with alanine (KHRR296-299AAAA) has increased fibrin-specificity and reduced sensitivity to plasminogen activator inhibitor-1; rt-PA-T, with threonine 103 replaced by asparagine has an additional glycosylation site and a reduced clearance; and rt-PA-N, with asparagine 117 mutagen-ized to glutamine lacks the high mannose carbohydrate side chain. We have investigated whether combination of these properties in a single molecule might yield an improved thrombolytic agent.

The thrombolytic potency and fibrin-specificity of the combination mutant rt-PA-TNK was compared with that of rt-PA in a combined venous whole blood clot model and platelet-rich arterial eversion graft thrombosis model in dogs given intravenous heparin and aspirin. Infusion of 0.125 to 1.0 mg/kg over 60 min in groups of 4 to 5 dogs produced dose-dependent fibrin-specific venous clot lysis. The thrombolytic potency (percent lysis per mg compound administered per kg body weight) of rt-PA-TNK was significantly higher than that of rt-PA as evidenced by a higher maximal rate of lysis of 480 ± 100% versus 140 ± 40% within the 2 h observation period per mg of compound administered per kg body weight (mean ± SEM, p = 0.004) and a significantly lower dose of 0.08 ± 0.01 versus 0.21 ± 0.04 mg/kg body weight at which the maximal rate of lysis was obtained (p = 0.004). This higher thrombolytic potency was the result of a significantly reduced clearance (240 ± 32 versus 540 ± 49 ml/min, p = 0.002) and a similar specific thrombolytic activity (percent lysis per |ig/ml plasma antigen level). Arterial reflow was obtained with 1 mg/kg rt-PA and with 0.5 mg/kg rt-PA-TNK, but with each agent recanalization was consistently associated with cyclic reocclusion and reflow. The frequency of arterial recanalization was somewhat higher with rt-PA-TNK (10/12) than with rt-PA (4/12) (p = 0.07) but the total patency times during a 2 h observation period were similar.

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References

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