Quantitative ADME Proteomics – CYP and UGT Enzymes in the Beagle Dog Liver and Intestine

• Published in: Pharmaceutical research Vol. 32; no. 1; pp. 74 - 90
• Main Authors: Heikkinen, Aki T., Friedlein, Arno, Matondo, Mariette, Hatley, Oliver J. D., Petsalo, Aleksanteri, Juvonen, Risto, Galetin, Aleksandra, Rostami-Hodjegan, Amin, Aebersold, Ruedi, Lamerz, Jens, Dunkley, Tom, Cutler, Paul, Parrott, Neil
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.01.2015; Springer Nature B.V; American Association of Pharmaceutical Scientists
• Subjects: Animals; Biochemistry; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Biomedicine; Colon - enzymology; Cytochrome P-450 Enzyme System - analysis; Cytochrome P-450 Enzyme System - metabolism; Dogs; Enzymes; Female; Glucuronosyltransferase - analysis; Glucuronosyltransferase - metabolism; Humans; Intestine, Small - enzymology; Life Sciences; Liver; Liver - enzymology; Male; Mass Spectrometry; Medical Law; Microsomes - enzymology; Models, Biological; Pharmaceutical sciences; Pharmacology/Toxicology; Pharmacy; Proteomics; Proteomics - methods; Research Paper; Small intestine; Species Specificity; Drug metabolism; Protein quantitation; extrapolation; -; Interspecies differences; Canine; In vitro-in vivo extrapolation
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-014-1446-8

Purpose Beagle dogs are used to study oral pharmacokinetics and guide development of drug formulations for human use. Since mechanistic insight into species differences is needed to translate findings in this species to human, abundances of cytochrome P450 (CYP) and uridine diphosphate glucuronosyltransferase (UGT) drug metabolizing enzymes have been quantified in dog liver and intestine. Methods Abundances of enzymes were measured in Beagle dog intestine and liver using selected reaction monitoring mass spectrometry. Results Seven and two CYPs were present in the liver and intestine, respectively. CYP3A12 was the most abundant CYP in both tissues. Seven UGT enzymes were quantified in the liver and seven in the intestine although UGT1A11 and UGT1A9 were present only in the intestine and UGT1A7 and UGT2B31 were found only in the liver. UGT1A11 and UGT1A2 were the most abundant UGTs in the intestine and UGT2B31 was the most abundant UGT in the liver. Summed abundance of UGT enzymes was similar to the sum of CYP enzymes in the liver whereas intestinal UGTs were up to four times more abundant than CYPs. The estimated coefficients of variation of abundance estimates in the livers of 14 donors were separated into biological and technical components which ranged from 14 to 49% and 20 to 39%, respectively. Conclusions Abundances of canine CYP enzymes in liver and intestine have been confirmed in a larger number of dogs and UGT abundances have been quantified for the first time. The biological variability in hepatic CYPs and UGTs has also been estimated.