Arteriosclerosis, Thrombosis, and Vascular Biology. 1999;19:2007-2011
© 1999 American Heart Association, Inc.
Thrombosis |
Specific Impairment of Human Platelet P2YAC ADP Receptor–Mediated Signaling by the Antiplatelet Drug Clopidogrel
From the Institute of Clinical Biochemistry and Pathobiochemistry (J.G., J.B., P.H.-L., M.E., U.W.) and the Department of Cardiology (P.S.), Medical University Clinic, Würzburg, Germany, and Sanofi Recherche (J.M.H.), Toulouse, France.
Correspondence to Dr U. Walter, Institute of Clinical Biochemistry and Pathobiochemistry, Medical University Clinic/Josef-Schneider Str 2, 97080 Würzburg, Germany. E-mail uwalter{at}klin-biochem.uni-wuerzburg.de
Abstract—Clopidogrel is an effective new antiplatelet agent useful for the treatment of ischemic cerebrovascular, cardiac, and peripheral arterial disease. However, the mechanism of clopidogrel action is not well understood, although it is known to inhibit ADP-evoked platelet aggregation. In the current study, the effect of clopidogrel on recently identified human platelet ADP receptors and their signaling pathways was investigated by using platelets from clopidogrel-treated subjects, 6 healthy volunteers (2 females and 4 males) who received 75 mg of clopidogrel daily for 7 days. Blood was taken and various platelet receptor signaling pathways were analyzed before treatment, after 7 days of medication, and 4 weeks after treatment had ceased. Platelet tests included the analysis of aggregation, rapid calcium influx, calcium mobilization from intracellular stores, adenylyl cyclase, and phosphorylation of vasodilator-stimulated phosphoprotein (VASP). The data indicate that clopidogrel does not affect those platelet ADP receptors coupled to cation influx (P2X1 ADP receptors) or calcium mobilization (P2Y1 ADP receptors). In contrast, clopidogrel treatment specifically impairs the ADP receptor coupled to Gi/adenylyl cyclase (P2YAC ADP receptors). Clopidogrel abolishes the inhibitory P2YAC receptor–mediated ADP effects on prostaglandin E1–stimulated, cAMP-dependent phosphorylation of VASP without affecting epinephrine, thrombin, and thromboxane signaling. VASP phosphorylation is known to be closely correlated with the inhibition of platelet and fibrinogen receptor (glycoprotein IIb/IIIa) activation. Therefore, inhibition of the platelet P2YAC ADP receptor and its intracellular signaling, including decreased VASP phosphorylation, is suggested as a molecular mechanism of clopidogrel action.
Key Words: platelet inhibition • purinergic receptors • vasodilator-stimulated phosphoprotein
This article has been cited by other articles:
 |
|
 |
|
  P. Gladding, H. White, J. Voss, J. Ormiston, J. Stewart, P. Ruygrok, B. Bvaldivia, R. Baak, C. White, and M. Webster Pharmacogenetic Testing for Clopidogrel Using the Rapid INFINITI Analyzer: A Dose-Escalation Study J. Am. Coll. Cardiol. Intv., November 1, 2009; 2(11): 1095 - 1101. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Marin, R. Gonzalez-Conejero, P. Capranzano, T. A. Bass, V. Roldan, and D. J. Angiolillo Pharmacogenetics in cardiovascular antithrombotic therapy. J. Am. Coll. Cardiol., September 15, 2009; 54(12): 1041 - 1057. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. E. Wrishko, C. S. Ernest II, D. S. Small, Y. G. Li, G. J. Weerakkody, J. R. Riesmeyer, W. L. Macias, S. Rohatagi, D. E. Salazar, E. M. Antman, et al. Population Pharmacokinetic Analyses to Evaluate the Influence of Intrinsic and Extrinsic Factors on Exposure of Prasugrel Active Metabolite in TRITON-TIMI 38 J. Clin. Pharmacol., August 1, 2009; 49(8): 984 - 998. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Gladding, M. Webster, I. Zeng, H. Farrell, J. Stewart, P. Ruygrok, J. Ormiston, S. El-Jack, G. Armstrong, P. Kay, et al. The Pharmacogenetics and Pharmacodynamics of Clopidogrel Response: An Analysis From the PRINC (Plavix Response in Coronary Intervention) Trial J. Am. Coll. Cardiol. Intv., December 1, 2008; 1(6): 620 - 627. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Berger, C. B. Frye, Q. Harshaw, F. H. Edwards, S. R. Steinhubl, and R. C. Becker Impact of Clopidogrel in Patients With Acute Coronary Syndromes Requiring Coronary Artery Bypass Surgery: A Multicenter Analysis J. Am. Coll. Cardiol., November 18, 2008; 52(21): 1693 - 1701. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A M Veitch, T P Baglin, A H Gershlick, S M Harnden, R Tighe, and S Cairns Guidelines for the management of anticoagulant and antiplatelet therapy in patients undergoing endoscopic procedures Gut, September 1, 2008; 57(9): 1322 - 1329. [Full Text] [PDF]
|
|
|
|
 |
|
 M. Dittrich, I. Birschmann, S. Mietner, A. Sickmann, U. Walter, and T. Dandekar Platelet Protein Interactions: Map, Signaling Components, and Phosphorylation Groundstate Arterioscler Thromb Vasc Biol, July 1, 2008; 28(7): 1326 - 1331. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. S. Kleiman Will Measuring Vasodilator-Stimulated Phosphoprotein Phosphorylation Help Us Optimize the Loading Dose of Clopidogrel? J. Am. Coll. Cardiol., April 8, 2008; 51(14): 1412 - 1414. [Full Text] [PDF]
|
|
|
|
|
|
P. A. Gurbel, W. C. Lau, and U. S. Tantry Omeprazole: A Possible New Candidate Influencing the Antiplatelet Effect of Clopidogrel J. Am. Coll. Cardiol., January 22, 2008; 51(3): 261 - 263. [Full Text] [PDF]
|
|
|
|
 |
|
 C. Gachet and B. Aleil Testing antiplatelet therapy Eur. Heart J. Suppl., January 1, 2008; 10(suppl_A): A28 - A34. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. L. Graciano, A. Nishiyama, K. Jackson, D. M. Seth, R. M. Ortiz, M. C. Prieto-Carrasquero, H. Kobori, and L. G. Navar Purinergic receptors contribute to early mesangial cell transformation and renal vessel hypertrophy during angiotensin II-induced hypertension Am J Physiol Renal Physiol, January 1, 2008; 294(1): F161 - F169. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Gurbel, R. C. Becker, K. G. Mann, S. R. Steinhubl, and A. D. Michelson Platelet Function Monitoring in Patients With Coronary Artery Disease J. Am. Coll. Cardiol., November 6, 2007; 50(19): 1822 - 1834. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. E. Eckert and S. L. Jones Regulation of VASP serine 157 phosphorylation in human neutrophils after stimulation by a chemoattractant J. Leukoc. Biol., November 1, 2007; 82(5): 1311 - 1321. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. A. Schumacher, J. S. Bostwick, M. L. Ogletree, A. B. Stewart, T. E. Steinbacher, J. Hua, L. A. Price, P. C. Wong, and R. P. Rehfuss Biomarker Optimization to Track the Antithrombotic and Hemostatic Effects of Clopidogrel in Rats J. Pharmacol. Exp. Ther., July 1, 2007; 322(1): 369 - 377. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. O'Donoghue and S. D. Wiviott Clopidogrel Response Variability and Future Therapies: Clopidogrel: Does One Size Fit All? Circulation, November 28, 2006; 114(22): e600 - e606. [Full Text] [PDF]
|
|
|
|
|
|
B. Ibanez, G. Vilahur, and J. J. Badimon Pharmacology of thienopyridines: rationale for dual pathway inhibition Eur. Heart J. Suppl., October 1, 2006; 8(suppl_G): G3 - G9. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. J. Angiolillo, A. Fernandez-Ortiz, E. Bernardo, C. Ramirez, U. Cavallari, E. Trabetti, M. Sabate, R. Hernandez, R. Moreno, J. Escaned, et al. Contribution of Gene Sequence Variations of the Hepatic Cytochrome P450 3A4 Enzyme to Variability in Individual Responsiveness to Clopidogrel Arterioscler Thromb Vasc Biol, August 1, 2006; 26(8): 1895 - 1900. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Gurbel, K. P. Bliden, W. Samara, J. A. Yoho, K. Hayes, M. Z. Fissha, and U. S. Tantry Clopidogrel Effect on Platelet REactivity in Patients With Stent Thrombosis: Results of the CREST Study J. Am. Coll. Cardiol., November 15, 2005; 46(10): 1827 - 1832. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Geiger, L. Teichmann, R. Grossmann, B. Aktas, U. Steigerwald, U. Walter, and R. Schinzel Monitoring of Clopidogrel Action: Comparison of Methods Clin. Chem., June 1, 2005; 51(6): 957 - 965. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. A. Nguyen, J. G. Diodati, and C. Pharand Resistance to clopidogrel: A review of the evidence J. Am. Coll. Cardiol., April 19, 2005; 45(8): 1157 - 1164. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Lepantalo, K. S Virtanen, J. Heikkila, U. Wartiovaara, and R. Lassila Limited early antiplatelet effect of 300 mg clopidogrel in patients with aspirin therapy undergoing percutaneous coronary interventions Eur. Heart J., March 2, 2004; 25(6): 476 - 483. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Kim, J. Jin, and S. P. Kunapuli Akt Activation in Platelets Depends on Gi Signaling Pathways J. Biol. Chem., February 6, 2004; 279(6): 4186 - 4195. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. C. Lau, P. A. Gurbel, P. B. Watkins, C. J. Neer, A. S. Hopp, D. G.M. Carville, K. E. Guyer, A. R. Tait, and E. R. Bates Contribution of Hepatic Cytochrome P450 3A4 Metabolic Activity to the Phenomenon of Clopidogrel Resistance Circulation, January 20, 2004; 109(2): 166 - 171. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Massberg, S. Gruner, I. Konrad, M. I. Garcia Arguinzonis, M. Eigenthaler, K. Hemler, J. Kersting, C. Schulz, I. Muller, F. Besta, et al. Enhanced in vivo platelet adhesion in vasodilator-stimulated phosphoprotein (VASP)-deficient mice Blood, January 1, 2004; 103(1): 136 - 142. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. W. Chan, D. J. Moliterno, P. B. Berger, G. W. Stone, P. M. DiBattiste, S. L. Yakubov, S. K. Sapp, K. Wolski, D. L. Bhatt, E. J. Topol, et al. Triple antiplatelet therapy during percutaneous coronary intervention is associated withimproved outcomes including one-year survival: Results from the do tirofiban and reoprogive similar efficacy outcome trial (TARGET) J. Am. Coll. Cardiol., October 1, 2003; 42(7): 1188 - 1195. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. Tcheng and M. E. Campbell Platelet inhibition strategies in percutaneous coronary intervention: Competition or coopetition? J. Am. Coll. Cardiol., October 1, 2003; 42(7): 1196 - 1198. [Full Text] [PDF]
|
|
|
|
|
|
P. Fontana, A. Dupont, S. Gandrille, C. Bachelot-Loza, J.-L. Reny, M. Aiach, and P. Gaussem Adenosine Diphosphate-Induced Platelet Aggregation Is Associated With P2Y12 Gene Sequence Variations in Healthy Subjects Circulation, August 26, 2003; 108(8): 989 - 995. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Ding, S. Kim, R. T. Dorsam, J. Jin, and S. P. Kunapuli Inactivation of the human P2Y12 receptor by thiol reagents requires interaction with both extracellular cysteine residues, Cys17 and Cys270 Blood, May 15, 2003; 101(10): 3908 - 3914. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Matsagas, I. A. Jagroop, G. Geroulakos, and D. P. Mikhailidis The Effect of a Loading Dose (300 mg) of Clopidogrel on Platelet Function in Patients with Peripheral Arterial Disease Clinical and Applied Thrombosis/Hemostasis, April 1, 2003; 9(2): 115 - 120. [Abstract] [PDF]
|
|
|
|
|
|
M. A. van Gestel, J. W.M. Heemskerk, D. W. Slaaf, V. V.T. Heijnen, R. S. Reneman, and M. G.A. oude Egbrink In Vivo Blockade of Platelet ADP Receptor P2Y12 Reduces Embolus and Thrombus Formation but Not Thrombus Stability Arterioscler Thromb Vasc Biol, March 1, 2003; 23(3): 518 - 523. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Aktas, A. Utz, P. Hoenig-Liedl, U. Walter, and J. Geiger Dipyridamole Enhances NO/cGMP-Mediated Vasodilator-Stimulated Phosphoprotein Phosphorylation and Signaling in Human Platelets: In Vitro and In Vivo/Ex Vivo Studies Stroke, March 1, 2003; 34(3): 764 - 769. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. C. Lau, L. A. Waskell, P. B. Watkins, C. J. Neer, K. Horowitz, A. S. Hopp, A. R. Tait, D. G.M. Carville, K. E. Guyer, and E. R. Bates Atorvastatin Reduces the Ability of Clopidogrel to Inhibit Platelet Aggregation: A New Drug-Drug Interaction Circulation, January 7, 2003; 107(1): 32 - 37. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J.J. Wood When Increased Therapeutic Benefit Comes at Increased Cost N. Engl. J. Med., June 6, 2002; 346(23): 1819 - 1821. [Full Text] [PDF]
|
|
|
|
|
|
S. Goto, N. Tamura, K. Eto, Y. Ikeda, and S. Handa Functional Significance of Adenosine 5'-Diphosphate Receptor (P2Y12) in Platelet Activation Initiated by Binding of von Willebrand Factor to Platelet GP Ib{alpha} Induced by Conditions of High Shear Rate Circulation, May 28, 2002; 105(21): 2531 - 2536. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Leon, M. Freund, C. Ravanat, A. Baurand, J.-P. Cazenave, and C. Gachet Key Role of the P2Y1 Receptor in Tissue Factor-Induced Thrombin-Dependent Acute Thromboembolism : Studies in P2Y1-Knockout Mice and Mice Treated With a P2Y1 Antagonist Circulation, February 6, 2001; 103(5): 718 - 723. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Helft, J. I. Osende, S. G. Worthley, A. G. Zaman, O. J. Rodriguez, E. I. Lev, M. E. Farkouh, V. Fuster, J. J. Badimon, and J. H. Chesebro Acute Antithrombotic Effect of a Front-Loaded Regimen of Clopidogrel in Patients With Atherosclerosis on Aspirin Arterioscler Thromb Vasc Biol, October 1, 2000; 20(10): 2316 - 2321. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J.J. Wood Thrombotic Thrombocytopenic Purpura and Clopidogrel -- A Need for New Approaches to Drug Safety N. Engl. J. Med., June 15, 2000; 342(24): 1824 - 1826. [Full Text]
|
|
|
|
|
|
I. M. B. Francischetti, J. M. C. Ribeiro, D. Champagne, and J. Andersen Purification, Cloning, Expression, and Mechanism of Action of a Novel Platelet Aggregation Inhibitor from the Salivary Gland of the Blood-sucking Bug, Rhodnius prolixus J. Biol. Chem., April 21, 2000; 275(17): 12639 - 12650. [Abstract] [Full Text] [PDF]
|
|