LC-MS/MS analysis of the plasma concentrations of a cocktail of 5 cytochrome P450 and P-glycoprotein probe substrates and their metabolites using subtherapeutic doses
| Autor: | Vera Lucia Lanchote, Adriana Rocha, Gabriela Campos de Oliveira Filgueira, Monica A. C. T. Cintra, Diego Alberto Ciscato Cusinato, Eduardo Barbosa Coelho |
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| Rok vydání: | 2019 |
| Předmět: |
Adult
Male CYP2D6 Spectrometry Mass Electrospray Ionization ATP Binding Cassette Transporter Subfamily B Midazolam Clinical Biochemistry Pharmaceutical Science CYP2C19 Pharmacology 01 natural sciences Losartan Analytical Chemistry Pharmacokinetics Oral administration In vivo Tandem Mass Spectrometry Caffeine Drug Discovery medicine Humans Volunteer CYP2C9 Spectroscopy Chromatography High Pressure Liquid Cytochrome P-450 CYP2C9 Fexofenadine 010405 organic chemistry Chemistry 010401 analytical chemistry Reproducibility of Results ESPECTROMETRIA DE MASSAS Healthy Volunteers 0104 chemical sciences Cytochrome P-450 CYP2C19 Isoenzymes Biological Variation Population Cytochrome P-450 CYP2D6 Terfenadine Omeprazole medicine.drug Metoprolol |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| Popis: | Drug transporters and CYP enzymes are important sources of pharmacokinetics (PK) variability in drug responses and can cause various pharmacological and toxicological consequences, leading to either toxicity or an insufficient pharmacological effect. In recent years, the cocktail approach was developed to determine in vivo CYP and transporters activities, but these approaches are somewhat limited. We described the development and validation of three sensitive and specific LC–MS/MS assays for the determination of P-gp and major human CYP isoenzyme activities following oral administration of a drug cocktail of subtherapeutic doses (lower than 10 times) of caffeine (CAF), omeprazole (OME), losartan (LOS), midazolam (MDZ), metoprolol (METO) and fexofenadine (FEX) in healthy volunteers. The three validated methods were selective for all tested analytes. No interference or matrix effect was observed for the mass transition and retention times for all compounds monitored. Additionally, assays were linear over a wide range, and limits of quantification varied between 0.01–5 ng/mL plasma. The coefficients of variation obtained in the precision studies and the inter- and intra-assay accuracies were less than 15%, guaranteeing the reproducibility and repeatability of the results. All substrates and metabolites were stable in plasma during freeze-thaw cycles. Three healthy volunteers were selected based on genotyping for CYP2C9, CYP2C19 and CYP2D6. One volunteer was genotyped as an extensive metabolizer (EM) for all tested CYP isoforms, one volunteer was genotyped as a poor metabolizer (PM) for the CYP2C9 isoform (CYP2C9*3/*3), and one volunteer was genotyped as a PM for the CYP2D6 isoform (CYP2D6*4/*4). The methods allowed the quantification of all analytes over the entire sampling period (12 h) in all studied genotypes. Thus, the analytical methods described here were sufficiently sensitive for use in low-dose pharmacokinetic studies. |
| Databáze: | OpenAIRE |
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