Stereoselective Metabolism of Prasugrel in Humans Using a Novel Chiral Liquid Chromatography-Tandem Mass Spectrometry Method

Wickremsinhe, Enaksha; Tian, Ye; Ruterbories, Kenneth J.; Verburg, Elizabeth M.; Weerakkody, Govinda J.; Kurihara, Atsushi; Farid, Nagy A.

doi:10.1124/dmd.106.014530

Cited by 46 publications

(27 citation statements)

References 10 publications

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“…In humans, greater concentrations of the R-configuration at the 4-position of R-138727 (Wickremsinhe et al, 2007) were observed in plasma. This in vivo stereoselectivity may be explained in part by the observed in vitro stereoselectivity in the P450-mediated oxidation of the thiolactone to R-138727 by cDNA-expressed P450 isoforms (Rehmel et al, 2006;Baker et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

Biotransformation of Prasugrel, a Novel Thienopyridine Antiplatelet Agent, to the Pharmacologically Active Metabolite

Hagihara

Kazui²,

Kurihara³

et al. 2010

Drug Metab Dispos

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ABSTRACT:Prasugrel, a novel thienopyridine antiplatelet agent, undergoes rapid hydrolysis in vivo to a thiolactone, R-95913, which is further converted to its thiol-containing, pharmacologically active metabolite, R-138727, by oxidation via cytochromes P450 (P450). We trapped a sulfenic acid metabolite as a mixed disulfide with 2-nitro-5-thiobenzoic acid in an incubation mixture containing the thiolactone R-95913, expressed CYP3A4, and NADPH. Further experiments investigated one possible mechanism for the conversion of the sulfenic acid to the active thiol metabolite in vitro. A mixed disulfide form of R-138727 with glutathione was found to be a possible precursor of R-138727 in vitro when glutathione was present. The rate constant for the reduction of the glutathione conjugate of R-138727 to R-138727 was increased by addition of human liver cytosol to the human liver microsomes. Thus, one possible mechanism for the ultimate formation of R-138727 in vitro can be through formation of a sulfenic acid mediated by P450s followed possibly by a glutathione conjugation to a mixed disulfide and reduction of the disulfide to the active metabolite R-138727. Prasugrel [Effient (Eli Lilly and Company, Indianapolis, IN) in the United States and Efient (Eli Lilly and Company) in the EuropeanUnion], clopidogrel [Plavix (Sanofi-Aventis, Paris, France)/Iscover (Bristol-Myers Squibb, New York, NY)], and ticlopidine (Ticlid; Sanofi-Aventis) are thienopyridine antiplatelet agents. Prasugrel has been shown to reduce the rate of thrombotic cardiovascular events and stent thrombosis in patients with acute coronary syndrome that are undergoing percutaneous coronary intervention (Wiviott et al., 2007) (Effient package insert). The thienopyridines are prodrugs that are converted in vivo to their pharmacologically active metabolites that possess a thiol group via a corresponding thiolactone metabolite (Farid et al., 2010). However, with the exception of an oxidation step catalyzed by cytochrome P450 (Savi et al., 1994;Rehmel et al., 2006), the mechanism for the active metabolite formation from thienopyridines remained unknown until recently reported for ticlopidine and clopidogrel (Dansette et al., 2009). In the case of prasugrel, the active metabolite R-138727 was not detected when the thiolactone metabolite R-95913 was incubated with liver homogenates or microsomes in the absence of cofactors, but it was detected when NADPH and a reducing agent such as glutathione were added to the system (Kazui et al., 2000). A sulfenic acid, which is a likely intermediate for the activation of prasugrel, reacts readily with a thiol compound, typically glutathione in vivo, to yield a disulfide metabolite (Decker et al., 1991;Kassahun et al., 2001;Reddy et al., 2005;Dansette et al., 2009). The formed disulfide can be further reduced to provide a thiolcontaining compound. The objective of this study is to investigate the involvement of a sulfenic acid and a glutathione conjugate of R-138727 in the in vitro production of prasugrel's active metabolite (R-1...

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Section: Discussionmentioning

confidence: 99%

Biotransformation of Prasugrel, a Novel Thienopyridine Antiplatelet Agent, to the Pharmacologically Active Metabolite

Hagihara

Kazui²,

Kurihara³

et al. 2010

Drug Metab Dispos

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“…RR isomer .. SS isomer = SR isomer (Hasegawa et al, 2005). In humans, the pharmacologically active RS and RR isomers were detected in plasma at about 5-fold higher levels than the pharmacologically less active SS and SR isomers after dosing of prasugrel (Wickremsinhe et al, 2007). In rats, the RS and RR isomers were the major forms detected in plasma, and the SS and SR isomers were only detected at much lower concentrations in plasma (Kazui et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Hepatic Microsomal Thiol Methyltransferase Is Involved in Stereoselective Methylation of Pharmacologically Active Metabolite of Prasugrel

et al. 2014

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Prasugrel, a thienopyridine antiplatelet drug, is converted in animals and humans to the pharmacologically active metabolite R-138727 [(2Z)-{1-[(1RS)-2-cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl]-4-sulfanylpiperidin-3-ylidene}ethanoic acid], which has two chiral centers, occurring as a mixture of four isomers. The RS and RR isomers are more active than the SS and SR isomers (RS > RR > > SR = SS). The pharmacologically active metabolite is further metabolized to an S-methylated metabolite that is the major identified inactive metabolite in humans. In rat, dog, and human liver microsomes supplemented with S-adenosyl methione, the SS and SR isomers of the active metabolite were extensively S-methylated while the RS and RR isomers were not. Addition of 2,3-dichloromethyl benzylamine (50 mM) completely inhibited the S-methylation reaction, indicating that the microsomal and cytosolic thiol methyltransferase but not the cytosolic thiopurine S-methyltransferase is involved in the methylation. The hepatic intrinsic clearance values for methylation of the RS, RR, SS, and SR isomers (ml/min/kg) were 0, 0, 40.4, and 37.6, respectively, in rat liver microsomes, 0, 0, 11.6, and 2.5, respectively, in dog liver microsomes, and 0, 0, 17.3, and 17.7, respectively, in human liver microsomes, indicating that the RS and RR isomers are not methylated in vitro and that the methylation of SS and SR isomers is high with rat > human > dog. This finding in vitro agreed well with the in vivo observation in rats and dogs, where the S-methylated SS and SR isomers were the major metabolites in the plasma whereas negligible amounts of S-methylated RS and RR isomers were detected after intravenous administration of the pharmacologically active metabolites.

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“…Several authors demonstrated that the acidic and basic mobile phase additives can have a significant impact in addition to the classical factors such as temperature, pH and type of organic modifier, which can influence the enantio separation in LC (Matthijs et al, 2006, Perrin et al, 2002, Ye Y et al, 2001, Levin et al, 1993, Khatuna et al, 2015, Gübitz et al, 2006. These studies mostly carried by normal-phase chromatography, as these additives were supposed to minimize the non-specific interactions between the analytes and the free silanol groups of the CSP.…”

Section: Optimization Of Chromatographic Conditionsmentioning

confidence: 99%

“…One method discusses about Stereo selective Metabolism of Prasugrel in Humans Using a Novel Chiral Liquid Chromatography-Tandem Mass Spectrometry Method (Enaksha et al, 2007). Another work reveals about Electrospray ionization LC-MS/MS validated method for the determination of the active metabolite (R-138727) of prasugrel in human plasma and its application to a bioequivalence study (Ojikumar et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

“…Another work reveals about Electrospray ionization LC-MS/MS validated method for the determination of the active metabolite (R-138727) of prasugrel in human plasma and its application to a bioequivalence study (Ojikumar et al, 2011). Few more research articles discusses about impurity profiling and comparison of forced degradation behavior of prasugrel by HPLC and UPLC (Singh et al, 2015, Malati et al, 2012, Kapendra et al, 2011. But, till date no work represents enantiomeric separation of prasugrel in active pharmaceutical ingredient's perspective, which basically add core value to manufacturing sector.…”

Section: Introductionmentioning

confidence: 99%

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Chiral recognition of polysaccharide based CSP for separation of enantiomers and regio isomers of Prasugrel and its related impurities

Nagireddy¹,

Krishna²,

Malati³

et al. 2017

J App Pharm Sci

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The current study focuses on the efficiency of chiral chromatographic conditions for separation of critical enantiomers and regio-isomeric pairs of Prasugrel, an antiplatelet drug and its related impurities. Totally, 10 chiral compounds were screened and well resolved using Phenominex-LUX-2-Amylose based chiral HPLC column (polysaccharide CSP). Chiral separations were achieved using Isopropanol, ethanol as organic modifiers and trimethylamine (TEA), trifluoro acetic acid (TFA) as polar modifier. Impact assessment for selection of superior chromatographic conditions to achieve best resolution was evaluated by varying column temperatures and concentrations of organic and polar modifiers. Significant enantiomeric separations were achieved with low alcohol and high TFA concentrations as modifiers in mobile phase. With optimized conditions, not only Prasugrel's enantiomers, two process related impurities (3-floro and 4-floro prasugrel) and major degradant i.e., Des acetyl impurity which exhibits keto-enol tautomerism in solution state were also separated into respective individual enantiomeric forms. Hence, the current work can be implemented for evaluation of chiral impurities of Prasugrel during synthesis as well as in Active Pharmaceutical Ingredient.

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Stereoselective Metabolism of Prasugrel in Humans Using a Novel Chiral Liquid Chromatography-Tandem Mass Spectrometry Method

Cited by 46 publications

References 10 publications

Biotransformation of Prasugrel, a Novel Thienopyridine Antiplatelet Agent, to the Pharmacologically Active Metabolite

Biotransformation of Prasugrel, a Novel Thienopyridine Antiplatelet Agent, to the Pharmacologically Active Metabolite

Hepatic Microsomal Thiol Methyltransferase Is Involved in Stereoselective Methylation of Pharmacologically Active Metabolite of Prasugrel

Chiral recognition of polysaccharide based CSP for separation of enantiomers and regio isomers of Prasugrel and its related impurities

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