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Thromb Haemost 1986; 56(02): 160-164
DOI: 10.1055/s-0038-1661632
Original Article
Schattauer GmbH Stuttgart

D-Phe-Pro-Arg-Chloromethylketone: Its Potential Use in Inhibiting the Formation of In Vitro Artifacts in Blood Collected During Tissue-Type Plasminogen Activator Thrombolytic Therapy

M A Mohler
The Department of Genentech, Inc., South San Francisco, CA, USA
,
C J Refino
The Department of Genentech, Inc., South San Francisco, CA, USA
,
S A Chen
The Department of Genentech, Inc., South San Francisco, CA, USA
,
A B Chen
*   The Departments of Pharmacological Sciences and Medicinal and Analytical Chemistry, South San Francisco, CA, USA
,
A J Hotchkiss
The Department of Genentech, Inc., South San Francisco, CA, USA
› Author Affiliations
Further Information

Publication History

Received 25 February 1986

Accepted 02 July 1986

Publication Date:
20 July 2018 (online)

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Summary

In vitro artifacts due to proteolysis may occur in blood samples containing recombinant tissue-type plasminogen activator (rt-PA) due to continued activation of plasminogen to plasmin by rt-PA. The aim of this study was to identify a rapid inhibitor of rt-PA that would not interfere in assays designed to monitor thrombolytic events.

When rt-PA was added at 5 μg/ml to whole blood and incubated at 25° C, fibrinogen decreased 50 percent, plasminogen levels decreased 90 percent and α2-antiplasmin decreased below detectable levels. If D-Phe-Pro-Arg-chloromethylketone (PPACK) or aprotinin were added before the addition of rt-PA there was no significant loss of fibrinogen. Only PPACK completely inhibited changes in fibrin degradation products, plasminogen and α2-antiplasmin. PPACK was also found to inhibit the binding of rt-PA to plasma protease inhibitors in vitro.

Rhesus monkeys were infused with rt-PA and blood samples were taken with either PPACK or aprotinin in the collection syringe. There was a significant increase in the recovery of immunoreactive rt-PA and consistent measures of fibrinogen, FDPs, plasminogen, and α2-antiplasmin in the PPACK group as compared to the aprotinin group which indicates that PPACK will prevent the in vitro formation of artifacts due to the presence of active rt-PA

Key words

Tissue-type plasminogen activator - Tripeptide chloromethyl-ketone inhibitor - Thrombolytic and coagulation assays
 

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