ABSTRACT
Purpose
Physiologically based models, when verified in pre-clinical species, optimally predict human pharmacokinetics. However, modeling of intestinal metabolism has been a gap. To establish in vitro/in vivo scaling factors for metabolism, the expression and activity of CYP enzymes were characterized in the intestine and liver of beagle dog.
Methods
Microsomal protein abundance in dog tissues was determined using testosterone-6β-hydroxylation and 7-hydroxycoumarin-glucuronidation as markers for microsomal protein recovery. Expressions of 7 CYP enzymes were estimated based on quantification of proteotypic tryptic peptides using multiple reaction monitoring mass spectrometry. CYP3A12 and CYP2B11 activity was evaluated using selective marker reactions.
Results
The geometric mean of total microsomal protein was 51 mg/g in liver and 13 mg/cm in intestine, without significant differences between intestinal segments. CYP3A12, followed by CYP2B11, were the most abundant CYP enzymes in intestine. Abundance and activity were higher in liver than intestine and declined from small intestine to colon.
Conclusions
CYP expression in dog liver and intestine was characterized, providing a basis for in vitro/in vivo scaling of intestinal and hepatic metabolism.
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Abbreviations
- CYP:
-
cytochrome P450
- DIM:
-
dog intestinal microsomes
- DLM:
-
dog liver microsomes
- LCMS:
-
liquid chromatography – mass spectrometry
- MRM:
-
multiple reaction monitoring
- PBPK:
-
physiologically based pharmacokinetics
- PK:
-
pharmacokinetics
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ACKNOWLEDGMENTS & DISCLOSURES
The authors thank Pascal Schenk and Paul Schmid for their contribution in setting up the analytical methods for diazepam metabolites.
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Heikkinen, A.T., Friedlein, A., Lamerz, J. et al. Mass Spectrometry-Based Quantification of CYP Enzymes to Establish In Vitro/In Vivo Scaling Factors for Intestinal and Hepatic Metabolism in Beagle Dog. Pharm Res 29, 1832–1842 (2012). https://doi.org/10.1007/s11095-012-0707-7
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DOI: https://doi.org/10.1007/s11095-012-0707-7