Monitoring of clopidogrel action: comparison of methods.
BACKGROUND
Clopidogrel is a potent drug for prevention of adverse effects during and after coronary intervention. Increasing experience indicates that a significant proportion of patients do not respond adequately to clopidogrel. Because failure of antiplatelet therapy can have severe consequences, there is need for a reliable assay to quantify the effectiveness of clopidogrel treatment.


METHODS
Of 24 healthy volunteers admitted to the study, 18 were treated for 1 week with clopidogrel (300-mg loading dose and 75-mg maintenance dose), and 6 with placebo. Platelet function was monitored by 2 assays, based on flow cytometry and enzyme immunoassay, that measure the phosphorylation status of vasodilator-stimulated phosphoprotein (VASP) and by aggregometry, flow cytometry of P-selectin, and the platelet function analyzer at baseline, on days 1-5, and on day 9 of treatment.


RESULTS
Aggregometry and VASP phosphorylation revealed a loss of platelet response to ADP within 12 h after clopidogrel intake. The phosphorylation status of VASP correlated with the inhibition of platelet aggregation. In contrast, neither P-selectin expression nor PFA-100 closure time was a clear indicator of clopidogrel effects on platelets.


CONCLUSIONS
VASP phosphorylation assays are reliable for quantifying clopidogrel effects. Because the VASP assay directly measures the function of the clopidogrel target, the P2Y12 receptor, the assay is selective for clopidogrel effects rather than effects of other platelet inhibitors commonly in use.
Geiger
Jörg
J
Institut für Klinische Biochemie und Pathobiochemie und Zentrallabor, Universität Würzburg, Würzburg, Germany. Geiger@klin-biochem.uni-wuerzburg.de
Teichmann
Lino
L
Grossmann
Ralf
R
Aktas
Barsom
B
Steigerwald
Udo
U
Walter
Ulrich
U
Schinzel
Reinhard
R
D000328:Adult; D001241:Aspirin / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D001760:Bleeding Time; D015815:Cell Adhesion Molecules / Q000378:metabolism; D000077144:Clopidogrel; D016903:Drug Monitoring / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D008297:Male; D008840:Microfilament Proteins; D019007:P-Selectin / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000009:adverse effects*; D013988:Ticlopidine / Q000009:adverse effects / Q000031:analogs & derivatives*; D013997:Time Factors; D000098662:Vasodilator-Stimulated Phosphoprotein
D015415:Biomarkers; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D019007:P-Selectin; D010750:Phosphoproteins; D010975:Platelet Aggregation Inhibitors; D000098662:Vasodilator-Stimulated Phosphoprotein; D000077144:Clopidogrel; D013988:Ticlopidine; D001241:Aspirin