Effects of antiplatelet agents on platelet-induced thrombin generation.
OBJECTIVE
The influence of antiplatelet agents on platelet-induced thrombin generation may increase the risk of bleeding. Assessment of the endogenous thrombin potential (ETP), is therefore a parameter deserving attention in early pharmacodynamic studies with antiplatelet drugs. The aim ofthis study was to assess whether an automated ETP-assay can be used to determine possible inhibitory effects of antiplatelet drugs on platelet-associated thrombin generation.


METHODS
We first characterized the in vitro dose-response relationship of several platelet agonists (ADP, collagen, U46619, TRAP (amino acid sequence: SFLLRNP) and tissue factor (TF) using the generation of ETP. One submaximal concentration of each agonist was then used to assess the influence of in vivo treatment with aspirin (single oral dose of 500 mg as inhibitor of thromboxane synthesis) and clopidogrel (given orally for 6 days, as an inhibitor of the purinergic P2Y12-receptor on platelets) and in vitro treatment with abciximab (which inhibits the platelet glycoprotein IIb/IIIa-receptor for fibrinogen), on the ETP.


RESULTS
The effect of TF and the other platelet inducers on thrombin generation was dose-dependent. Repeat measurements on samples from the same subject, with the same inducer concentration on 2 different occasions showed a variability of approx. 22% (absolute difference between 2 measurements as % of mean). The coefficient on variation of repeat measurements of one sample varied between 7% and 17%, depending on the inducer. After a single dose of aspirin, ETP was reduced by 25-40%, depending on the platelet activating agent used. The reduction in ETP with abciximab in vitro was more pronounced. In contrast, TF-induced ETP was not influenced by aspirin or abciximab. Clopidogrel, administered for 6 days, reduced the ETP by 60% when platelets were stimulated using 20 microM ADP, whereas collagen-induced ETP and TF-induced ETP remained unchanged.


CONCLUSIONS
The ETP-method is a sensitive and reproducible method for the detection of drug effects on platelet-induced thrombin generation of high throughput, and can be recommended for studies on the pharmacodynamic profile of drugs interfering with platelet function.
Wegert
W
W
Institute for Clinical Pharmacology, University Hospital, Frankfurt am Main, Germany.
Graff
J
J
Kaiser
D
D
Breddin
H K
HK
Klinkhardt
U
U
Harder
S
S
D000328:Adult; D001331:Automation; D001681:Biological Assay; D001792:Blood Platelets / Q000502:physiology*; D006470:Hemorrhage / Q000503:physiopathology; D006801:Humans; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013917:Thrombin / Q000032:analysis / Q000096:biosynthesis* / Q000494:pharmacology
D010975:Platelet Aggregation Inhibitors; D013917:Thrombin