Methods of measuring inhibition of platelet aggregation by thrombin receptor antagonists

A technology for thrombin receptor, platelet aggregation, used in the field of diagnostic assays

Inactive Publication Date: 2010-03-17
ACCUMETRICS INC
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  • Abstract
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  • Application Information

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Problems solved by technology

Phase II clinical study showed no increase in bleeding time and no prolongation of clotting time in patients treated with SCH 530348

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  • Methods of measuring inhibition of platelet aggregation by thrombin receptor antagonists

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Embodiment

[0068] Using VERIFYNOW™ IIb / IIIa Assay (Accumetrics, Inc., San Diego, Calif.) and using PAR-1 Thrombin Receptor Activating Peptide (TRAP-1) as a platelet activator, experiments were performed to determine the PAR-1 antagonist E5555 ( Eisai Ltd.) dose-dependent inhibition of platelets.

[0069] Dose response tests were performed using 3.7 [mu]M TRAP-1. E5555 was used at final concentrations of 30 ng / ml, 10 ng / ml, 5.0 ng / ml and 1.0 ng / ml. 20 [mu]L of each stock solution was added to 2.0 ml of whole blood (1:100 dilution) and mixed carefully to avoid hemolysis. After mixing, blood samples were incubated at 37°C for 30 minutes prior to the platelet inhibition assay.

[0070] Blood samples from four human donors were run in duplicate at baseline and each of the four E5555 concentrations. As expected, the level of platelet inhibition measured at a given E5555 concentration varied from donor to donor. Actual results for the four donors were on average slightly lower than expected...

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Abstract

A method is provided for measuring inhibition of platelet aggregation by a thrombin receptor antagonist. First, a blood sample is obtained from a patient treated with a thrombin receptor antagonist. The blood sample is mixed in combination with particles including an immobilized GPIIb / IIIa receptor ligand and a thrombin receptor activator. The combination is then incubated under conditions suitable for agglutinating the particles, and platelet-mediated agglutination is assessed in the mixture. The absence of agglutination indicates that the patient has reduced ability to form platelet thrombiin response to the thrombin receptor antagonist treatment. Also provided is a kit for measuring inhibition of platelet aggregation by a thrombin receptor antagonist that includes a GPIIb / IIIa receptorligand immobilized on a particle, a thrombin receptor activator, an anticoagulant, and a buffer to maintain the anticoagulated blood in a condition suitable for platelet aggregation.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Serial No. 60 / 915,820, filed May 3, 2007, which is hereby incorporated by reference in its entirety. technical field [0003] The present invention relates to the field of diagnostic assays, in particular to measuring the functional activity of platelets in blood samples to study the effects of antiplatelet compositions, and more particularly to platelet activators such as thrombin receptor activating peptide ( TRAP)) for measuring platelet function with enhanced sensitivity thrombin receptor inhibitors including E5555 (Eisai) and SCH 530348 (Schering-Plough). Background technique [0004] Platelets play a very diverse role in mammalian physiology, but their main role is to promote hemostasis. In many instances, it is desirable to evaluate the ability of blood to coagulate, a parameter often governed by the ability of platelets to adhere and / or aggregate. Therefore, it is of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86
CPCG01N33/86C12Q1/56
Inventor 丹尼斯·德宾
Owner ACCUMETRICS INC
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