Development and validation of a fast and sensitive UPLC–MS/MS method for the quantification of six probe metabolites for the in vitro determination of cytochrome P450 activity

Bock, Lies De; Boussery, Koen; Colin, Pieter; Smet, Julie; T’jollyn, Huybrecht; Bocxlaer, Jan Van

doi:10.1016/j.talanta.2011.11.083

Cited by 39 publications

(26 citation statements)

References 22 publications

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“…To pellet the proteins, samples were centrifuged at 20,000g for 10 minutes at 4°C. The formed metabolites were quantified using a previously described ultra high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method (De Bock et al, 2012a), and activities were expressed as picomoles of metabolite formed/(minute Â mg of protein).…”

Section: Methodsmentioning

confidence: 99%

In Vitro Cytochrome P450 Activity Decreases in Children with High Pediatric End-Stage Liver Disease Scores

et al. 2012

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To improve the modeling and simulation of pharmacokinetics in pediatric patients, research into developmental and disease-specific determinants is needed. This article describes the evaluation of the activity of in vitro cytochrome P450 (P450), an important enzyme family in drug metabolism, in children with hepatic dysfunction. The activity of six P450 isoforms (CYP1A2, 2C9, 2C19, 2D6, 2E1, and 3A4) was evaluated in 31 patients with different pathologies, predominantly biliary atresia (n = 23). Hypervariable activity was observed for all the isoforms. Compared with average adult activity, low activity levels were seen for CYP1A2, 2C19, 2E1, and 3A4. For CYP2E1 and 3A4, a positive correlation between activity and abundance was observed. Age, comedication, and genotype could not be used as predictors for P450 activity in this patient population. In contrast, the pediatric end-stage liver disease score was negatively correlated with the ln(activity). This finding suggests a decrease in P450 activity with deteriorating hepatic function. Moreover, the activity of all isoforms was correlated, demonstrating a concomitant decrease of all isoforms in young patients with liver disease. To our knowledge, this is the first study to evaluate P450 activity in children with hepatic impairment. The presented data may provide support in the further optimization of a diseasespecific model in this patient population.

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Section: Methodsmentioning

confidence: 99%

In Vitro Cytochrome P450 Activity Decreases in Children with High Pediatric End-Stage Liver Disease Scores

et al. 2012

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“…Chromatographic and MS conditions were optimized as previously described. [23][24][25] Chromatographic separations were used a Waters Acquity BEHC18 column (2.1×50 mm, particle size 1.7 µm, Waters, Wexford, Ireland) with an isocratic elution. Mobile phase was water with 0.1% formic acid/acetonitrile (70/30) at a flow rate of 0.2 mL/min.…”

Section: Methodsmentioning

confidence: 99%

Effects of Oridonin on Hepatic Cytochrome P450 Expression and Activities in PXR-Humanized Mice

Zhang

Bao

Lou

et al. 2018

Biological & Pharmaceutical Bulletin

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Oridonin, the major terpene found in Rabdosia rubescens, is widely used as dietary supplement or therapeutic drug, while the effects of oridonin on CYP450 were still unclear. The pregnane X receptor (PXR) is an important regulatory factor for major drug metabolism enzyme CYPs, and it has been reported to have species-specific differences. Therefore, this study has employed more reliable models PXR-humanized mouse to investigate the influence of oridonin on PXR and downstream metabolism enzyme. Eight-week-old male PXR-humanized mice were treated with oridonin by orally (0, 25, 50, 100, 200 mg/kg) for 15 d. The effects of oridonin on major downstream CYPs of PXR were examined at both the mRNA and enzyme activity levels by RT-PCR and HPLC-MS/MS. In general, there was no significant toxic reaction in liver of PXR-humanized mice. The mRNA expression of CYPs and cytochrome P450 oxidoreductase (POR) were increased with oridonin treatment in a dose-dependent manner. CYP2c and CYP3a family catalytic activity were increased significantly in two higher doses groups. These results indicate that oridonin induced the expression and activation of CYP2c and CYP3a family, which might contribute to potential drug-drug interactions and appear to be a risk when co-administered with other clinical drugs.

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“…The reaction was initiated by the addition of NADPH to the reaction mixture containing the microsomes, KCl, phosphate buffer and CZ. After 15 min, reactions were stopped using a mixture of formic acid, acetonitrile, and water (3:55:42 (v:v:v)), and the amount of hydroxychlorzoxazone (HCZ) was quantified using UPLC-MS/MS [12].…”

Section: Proof Of Concept: Application Of the Methods And Cyp Activitymentioning

confidence: 99%

Quantification of cytochrome 2E1 in human liver microsomes using a validated indirect ELISA

Bock¹,

Colin²,

Boussery³

et al. 2014

Journal of Pharmaceutical and Biomedical Analysis

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Development and validation of a fast and sensitive UPLC–MS/MS method for the quantification of six probe metabolites for the in vitro determination of cytochrome P450 activity

Cited by 39 publications

References 22 publications

In Vitro Cytochrome P450 Activity Decreases in Children with High Pediatric End-Stage Liver Disease Scores

In Vitro Cytochrome P450 Activity Decreases in Children with High Pediatric End-Stage Liver Disease Scores

Effects of Oridonin on Hepatic Cytochrome P450 Expression and Activities in PXR-Humanized Mice

Quantification of cytochrome 2E1 in human liver microsomes using a validated indirect ELISA

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