Feidt, Diana M. and Klein, Kathrin and Hofmann, Ute and Riedmaier, Stephan and Knobeloch, Daniel and Thasler, Wolfgang E. and Weiss, Thomas S. and Schwab, Matthias and Zanger, Ulrich M. (2010) Profiling Induction of Cytochrome P450 Enzyme Activity by Statins Using a New Liquid Chromatography-Tandem Mass Spectrometry Cocktail Assay in Human Hepatocytes. DRUG METABOLISM AND DISPOSITION, 38 (9). pp. 1589-1597. ISSN 0090-9556, 1521-009X
Full text not available from this repository. (Request a copy)Abstract
Human hepatocytes in primary culture are a very useful model to directly assess induction of gene expression by xenobiotics. We developed a cytochrome P450 (P450) activity cocktail assay using model substrates for the seven important P450s 1A2 (phenacetin), 2B6 (bupropion), 2C8 (amodiaquine), 2C9 (tolbutamide), 2C19 (S-mephenytoin), 2D6 (propafenone), and 3A4 (atorvastatin). Metabolite formation was determined by liquid chromatography-tandem mass spectrometry in hepatocyte culture supernatants. Atorvastatin has not been previously assessed as a CYP3A probe drug. We demonstrate highly selective atorvastatin ortho-hydroxylation by CYP3A4 by recombinant P450s. In human liver microsomes ortho-hydroxyatorvastatin formation was highly correlated with CYP3A4 protein content (r(s) = 0.78, p < 0.0001, n = 150). We profiled induction of these P450 activities in primary human hepatocytes after treatment with 30 mu M atorvastatin, lovastatin, pravastatin, rosuvastatin, and simvastatin for 24 to 72 h. Except for pravastatin, all statins induced P450 activities to various degrees, approximately in the order atorvastatin > simvastatin > lovastatin > rosuvastatin. Inducibility of P450s followed the order CYP2C8 > CYP3A4 > CYP2C9 > CYP2B6 > CYP2C19 similar to CYP2D6 > CYP1A2. The strongest induction was observed for amodiaquine N-desalkylation, which was induced approximately 20-fold. Quantitative reverse transcription-polymerase chain reaction confirmed corresponding changes on the mRNA level with even more dramatic induction up to almost 100-fold. These data suggest a broader inducing effect of statins on cytochrome P450s and possibly other absorption, distribution, metabolism, and excretion genes than previously known, thus further emphasizing their drug-drug interaction potential. Our cocktail assay should be helpful for economical use of human hepatocytes in the assessment of P450 induction by drugs and drug candidates.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CONSTITUTIVE ANDROSTANE RECEPTOR; PREGNANE-X-RECEPTOR; IN-VITRO COCKTAIL; HUMAN LIVER; REDUCTASE INHIBITORS; DRUG-INTERACTIONS; RAT HEPATOCYTES; EXPRESSION; METABOLISM; CYP2B6; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Chirurgie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 13 Jul 2020 12:36 |
| Last Modified: | 13 Jul 2020 12:36 |
| URI: | https://pred.uni-regensburg.de/id/eprint/24211 |
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