An improved method for specific and quantitative determination of the clopidogrel active metabolite isomers in human plasma

Tuffal, Gilles; Roy, Sébastien; Lavisse, M.; Brasseur, Denis; Schofield, Jill; Touchard, Nathalie Delesque; Savi, Pierre; Brémond, Nicolas; Rouchon, Marie-Claude; Hurbin, Fabrice; Sultan, Eric

doi:10.1160/th10-09-0582

Cited by 82 publications

(16 citation statements)

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“…The second step in generating the physiologically active metabolite involves another CYP450 catalyzed step in the presence of glutathione to generate the active metabolite. The ability to clarify the metabolism was dependent on sophisticated liquid chromatography/mass spectrometric analyses including derivatization of the microsomal generated metabolites [83,84]. The metabolite referred to as H4 was shown to be the pharmacologically relevant metabolite with the H2 metabolite having approximately half the biological activity measured in vitro [84].…”

Section: Targets Of Paraoxonase Hydrolysismentioning

confidence: 99%

“…The ability to clarify the metabolism was dependent on sophisticated liquid chromatography/mass spectrometric analyses including derivatization of the microsomal generated metabolites [83,84]. The metabolite referred to as H4 was shown to be the pharmacologically relevant metabolite with the H2 metabolite having approximately half the biological activity measured in vitro [84]. Hydrolysis of a metabolic intermediate with an ‘endo’ double bond by PON1 leads to an inactive metabolite present at lower levels than the active metabolite H4 [83].…”

Section: Targets Of Paraoxonase Hydrolysismentioning

confidence: 99%

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Pharmacogenetics of Paraoxonase Activity: Elucidating the Role of High-Density Lipoprotein in Disease

et al. 2013

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PON1 is a key component of high-density lipoproteins (HDLs) and is at least partially responsible for HDL's antioxidant/atheroprotective properties. PON1 is also associated with numerous human diseases, including cardiovascular disease, Parkinson's disease and cancer. In addition, PON1 metabolizes a broad variety of substrates, including toxic organophosphorous compounds, statin adducts, glucocorticoids, the likely atherogenic l-homocysteine thiolactone and the quorum-sensing factor of Pseudomonas aeruginosa. Numerous cardiovascular and antidiabetic pharmacologic agents, dietary macronutrients, lifestyle factors and antioxidant supplements affect PON1 expression and enzyme activity levels. Owing to the importance of PON1 to HDL function and its individual association with diverse human diseases, pharmacogenomic interactions between PON1 and the various factors that alter its expression and activity may represent an important therapeutic target for future investigation.

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Section: Targets Of Paraoxonase Hydrolysismentioning

confidence: 99%

Section: Targets Of Paraoxonase Hydrolysismentioning

confidence: 99%

Pharmacogenetics of Paraoxonase Activity: Elucidating the Role of High-Density Lipoprotein in Disease

et al. 2013

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“…Only H4 and H2 have antiplatelet effects, with H4 exhibiting about twice the activity of H2 (10). In the human body, only H3 and H4 isomers can be detected (11, 12). As a result, the H4 isomer is the only active circulating form of CAM in humans (10).…”

Section: Introductionmentioning

confidence: 99%

“…The H4 isomer selectively and irreversibly blocks adenosine diphosphate (ADP) binding to the P2Y12 receptor thereby inhibiting ADP-induced platelet aggregation. In humans, about 15% of the parent clopidogrel undergoes bioactivation to both the H3 and H4 CAM isomers (11). The remaining 85% is metabolized in the liver to an inactive clopidogrel carboxylic acid (Fig.…”

Section: Introductionmentioning

confidence: 99%

Validation of a method for quantitation of the clopidogrel active metabolite, clopidogrel, clopidogrel carboxylic acid, and 2-oxo-clopidogrel in feline plasma

Lyngby

Court

Lee

2017

Journal of Veterinary Cardiology

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Objectives To validate a new method for stabilization and quantitation of the clopidogrel active metabolite (CAM), clopidogrel, and inactive clopidogrel carboxylic acid and 2-oxo-clopiodgrel. Animals Feline plasma was used for assay validation. A pilot pharmacokinetic study was conducted with 2 healthy cats. Methods CAM stabilization was achieved by adding 2-bromo-3′methoxyacetophenone to blood tubes to form a derivatized CAM (CAM-D). CAM-D was quantified using high performance liquid chromatography and tandem mass spectrometry. Validation of the methodology included evaluation of calibration curve linearity, accuracy and precision, and stability using quality control samples spiked with CAM-D, clopidogrel, clopidogrel carboxylic acid, and 2-oxo-clopidogrel. In vivo utility of this assay was evaluated by conducting a pharmacokinetic study in cats receiving a single oral dose of 18.75mg clopidogrel. Results The 2-oxo-clopidogrel metabolite was unstable. CAM-D, clopidogrel, and clopidogrel carboxylic acid appear stable for 1 week at room temperature and 9 months at −80°C. Standard curves showed linearity for CAM- D, clopidogrel, and clopidogrel carboxylic acid (r>0.99). Between assay accuracy and precision was ≤2.6% and ≤2.8% for CAM-D and ≤17.9% and ≤11.3% for clopidogrel and clopidogrel carboxylic acid. Within assay precision for all three compounds was ≤7%. All three compounds were detected in plasma from healthy cats receiving clopidogrel. Conclusions This methodology is accurate and precise for simultaneous quantitation of CAM-D, clopidogrel, and clopidogrel carboxylic acid in feline plasma but unsuitable for 2-oxo-clopidogrel. Validation of this assay is the first step to more fully understanding the use of clopidogrel in cats in both a population and individual setting.

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“…Among the various CYP enzymes that catalyse the oxidative activation of clopidogrel, the CYP2C19 polymorphic variants *2 and *3 are mostly responsible for reduced exposure to CTM, leading to the decreased antiplatelet effect of clopidogrel in some patients [9]. CTM is a chiral compound and may exist as four isomers (H1–H4), of which only H3 and H4 were present in the obtained clinical samples [10]. In vitro studies have confirmed that the H4 isomer can be considered as the only active circulating isomer of CTM [9, 11].…”

Section: Introductionmentioning

confidence: 99%

Clinical Pharmacokinetics of Clopidogrel and Its Metabolites in Patients with Cardiovascular Diseases

et al. 2013

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Background and ObjectiveApproximately 5–40 % of patients treated with clopidogrel do not display an adequate antiplatelet response. Clopidogrel resistance may be caused by insufficient drug absorption or impaired metabolic activation of the drug. The aim of this study was to evaluate the pharmacokinetics of clopidogrel and its metabolites in plasma samples from patients treated with high and low doses of clopidogrel, to obtain a possible explanation for antiplatelet resistance.MethodsThe study included patients receiving either a single 300 mg loading dose of clopidogrel (n = 17) or a 75 mg dose (n = 45) for at least 7 days before sample collection. The concentrations of clopidogrel and its metabolites—the inactive H3 and the pharmacologically active H4 isomers of the thiol metabolite and the inactive carboxylic acid metabolite—in plasma samples (stabilized with 2-bromo-3′-methoxyacetophenone) from three patients after 300 mg and from 41 patients after 75 mg of the drug were determined using a validated high-performance liquid chromatography method with tandem mass spectrometry. The non-stabilized samples from the remaining patients were analysed using a validated capillary electrophoresis method. The calculated concentrations were used to determine the pharmacokinetic parameters of the analytes. The pharmacodynamic response to clopidogrel treatment, expressed as adenosine diphosphate-induced platelet aggregation, was measured using a Multiplate analyser.ResultsThe pharmacokinetic parameter values for the H3 and H4 isomers determined in the studied group of patients treated with clopidogrel 75 mg (maximum plasma concentration [Cmax] 5.29 ± 5.54 and 7.13 ± 6.32 ng/mL for H3 and H4, respectively; area under the plasma concentration-time curve from time zero to time t [AUCt] 7.37 ± 6.71 and 11.30 ± 9.58 ng·h/mL for H3 and H4, respectively) were lower than those reported in healthy volunteers, according to the literature data. Platelet aggregation measured with a Multiplate analyser ranged between 37 and 747 AU·min. A significant correlation was found between the Cmax of the active H4 isomer and platelet aggregation (p = 0.025).ConclusionThe Cmax of the active H4 isomer and platelet aggregation measured by the Multiplate analyser may serve as markers of the patient response to clopidogrel therapy.

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An improved method for specific and quantitative determination of the clopidogrel active metabolite isomers in human plasma

Cited by 82 publications

References 20 publications

Pharmacogenetics of Paraoxonase Activity: Elucidating the Role of High-Density Lipoprotein in Disease

Pharmacogenetics of Paraoxonase Activity: Elucidating the Role of High-Density Lipoprotein in Disease

Validation of a method for quantitation of the clopidogrel active metabolite, clopidogrel, clopidogrel carboxylic acid, and 2-oxo-clopidogrel in feline plasma

Clinical Pharmacokinetics of Clopidogrel and Its Metabolites in Patients with Cardiovascular Diseases

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