Investigation of the stereoselective in vitro metabolism of the chiral antiasthmatic/antiallergic drug flezelastine by high-performance liquid chromatography and capillary zone electrophoresis

Paris, Stéphane; Blaschke, Gottfried; Locher, Mathias; Borbe, Harald O.; Engel, Jeffrey A.

doi:10.1016/s0378-4347(96)00480-x

Cited by 15 publications

(3 citation statements)

References 4 publications

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“…Using preparations with rat liver microsomes, CZE with sulfobutyl-b-CD as chiral selector was employed to study the metabolism of the anthelminthic drug praziquantel [10] and the metabolism of the sedative drug thalidomide was investigated with carboxymethyl-b-CD [11] and the combined use of sulfobutylb-CD and b-CD [12]. Incubations of flezelastine with human, rat, bovine, and porcine liver microsomes were analyzed by CZE in the presence of b-CD [13] and the stereoselectivity of the mephenytoin (MEPH) metabolism in an incubation with human liver microsomes (HLM) was monitored by MEKC using taurodeoxycholic acid and b-CD as chiral buffer additives [14]. The number of achiral applications reported in the literature is not much larger [15] and all these incubations were performed with liver microsomes from animals or humans, i.e., preparations which contain multiple CYPs.…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis to assess drug metabolism induced in vitro using single CYP450 enzymes (Supersomes™): Application to the chiral metabolism of mephenytoin and methadone

Prost

Thormann

2003

Electrophoresis

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Capillary electrophoresis (CE) with multiwavelength absorbance detection is demonstrated to be an effective tool for the assessment of in vitro drug metabolism studies using microsomes containing single human cytochrome P450 enzymes (CYPs) expressed in baculovirus-infected insect cells (Supersomes). Mephenytoin (MEPH), dextromethorphan, diclofenac, caffeine, and methadone (MET) were successfully applied as test substrates for CYP2C19, CYP2D6*1, CYP2C9*1, CYP1A2, and CYP3A4, respectively. For each system, the CE-based assay could be shown to permit the simultaneous analysis of the parent drug and its targeted metabolite. Using a chiral micellar electrokinetic capillary chromatography assay, the aromatic hydroxylation of MEPH catalyzed by CYP2C19 could thereby be confirmed to be highly stereoselective, an aspect that is in agreement with data obtained via urinary analysis after intake of racemic MEPH by extensive metabolizer phenotypes. The MET to 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) conversion was investigated with a chiral zone electrophoresis assay. Incubation of racemic and nonracemic MET with CYP3A4 revealed no stereoselectivity for the transformation to EDDP, whereas no EDDP formation was observed with CYP1A2. CYP2C9 and CYP2C19 provided enhanced formation of R-EDDP and CYP2D6 incubation resulted in the preferential conversion to S-EDDP. Investigations using racemic MET and human liver microsomes revealed a modest stereoselectivity with an R/S EDDP ratio < 1 which is similar to the in vivo findings in urine.

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Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis to assess drug metabolism induced in vitro using single CYP450 enzymes (Supersomes™): Application to the chiral metabolism of mephenytoin and methadone

Prost

Thormann

2003

Electrophoresis

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“…Other first reports of this technology encompass work dealing with stereoselective drug metabolism in vitro (hydroxylation of mephenytoin in an incubation with human liver microsomes [33]), determination of drug stereoisomers of toxicological and forensic interest (racemethorphan and racemorphan in urine [34]), analysis of phase-II metabolites (ciprofibrate glucuronides in urine [35]), and monitoring of drug enantiomers in tissues rather than fluids (3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine in hair of ecstasy users [36]). Early work with CE-based chiral assays provided the first data on the stereoselective metabolism of flezelastine [37], clenbuterol [38], MDMA [39] and haloperidol [40]. Our interest in that field commenced with a collaborative effort in which the stereoselectivities of the aromatic hydroxylation of mephenytoin and phenytoin were assessed via MECC analysis of the enantiomers of the hydroxylated metabolites in human urine [41].…”

Section: Introductionmentioning

confidence: 99%

Stereoselective determination of drugs and metabolites in body fluids, tissues and microsomal preparations by capillary electrophoresis (2000–2010)

Caslavska

Thormann

2011

Journal of Chromatography A

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“…Chiral CE methods can readily be used to identifiy the stereochemistry of the enzymatic transformation, since simultaneous separation of several enantiomeric pairs with similar chemical structure can be achieved because of the high efficiency provided by these methods. Recently, several papers on in vitro drug metabolism studies using chiral CE have been published [22][23][24][25]. The first CE method for the analysis of N-oxide enantio- [23].…”

Section: Introductionmentioning

confidence: 99%

Assessment of the N‐oxidation of deprenyl, methamphetamine, and amphetamine enantiomers by chiral capillary electrophoresis: An in vitro metabolism study

et al. 2004

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A chiral capillary electrophoresis method using hydroxypropyl-beta-cyclodextrin as chiral selector was developed and validated for the quantification of the N-oxygenated metabolites of deprenyl, methamphetamine, and amphetamine enantiomers, formed in vitro. The influence of various parameters (selector concentration, buffer pH, temperature, polymer additive, etc.) on the simultaneous separation of the optical isomers of the parent drugs and their metabolites has been evaluated. The buffer pH had the greatest impact on the separation selectivity of the N-oxygenated compounds. Linear calibration curves were obtained over the concentration range of 2.5-50 microM for the enantiomers of amphetamine-hydroxylamine, methamphetamine-hydroxylamine, and deprenyl-N-oxide. The inter- and intra-assay precision and accuracy varied by less than 15% for all analytes at concentrations of 5, 10, and 30 microM, and less than 20% at the lower limit of quantitation (2.5 microM). The sample extraction recovery ranged between 109 and 129% at the three concentration levels. The drug enantiomers were incubated with recombinant human flavin-containing monooxygenase enzymes (FMO3 and FMO1), and human liver microsomes, respectively. The enantioselectivity of the substrate preference, as well as the stereoselective formation of the new chiral center upon the oxidation of the prochiral tertiary nitrogen of deprenyl were assessed. FMO1, the extrahepatic form of the enzyme in man, was shown to be more active in the N-oxygenation of both deprenyl and methamphetamine isomers than FMO3. Deprenyl enantiomers and S-methamphetamine were substrates of human recombinant FMO3. Conversion of amphetamine to its hydroxylamine derivative could not be observed on incubation with either FMO1 or FMO3. Formation of the new chiral center on the nitrogen, during N-oxidation of the tertiary amine deprenyl, was found stereoselective. The two FMO isoforms have shown opposite preference in the formation of this chiral center. Methamphetamine-hydroxylamine formed from methamphetamine was further transformed by FMO, amphetamine-hydroxylamine was identified as the product of a demethylation reaction.

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Investigation of the stereoselective in vitro metabolism of the chiral antiasthmatic/antiallergic drug flezelastine by high-performance liquid chromatography and capillary zone electrophoresis

Cited by 15 publications

References 4 publications

Capillary electrophoresis to assess drug metabolism induced in vitro using single CYP450 enzymes (Supersomes™): Application to the chiral metabolism of mephenytoin and methadone

Capillary electrophoresis to assess drug metabolism induced in vitro using single CYP450 enzymes (Supersomes™): Application to the chiral metabolism of mephenytoin and methadone

Stereoselective determination of drugs and metabolites in body fluids, tissues and microsomal preparations by capillary electrophoresis (2000–2010)

Assessment of the N‐oxidation of deprenyl, methamphetamine, and amphetamine enantiomers by chiral capillary electrophoresis: An in vitro metabolism study

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