title: A quantitative approach to hepatic clearance prediction of metabolism by aldehyde oxidase using custom pooled hepatocytes.
abstract: We describe the usability of human pooled hepatocytes for non-CYP metabolism evaluation and an in vivo-in vitro correlation analysis for aldehyde oxidase (AO) substrate compounds using pooled hepatocytes. By comparing intrinsic clearance values of 18 compounds primarily metabolized by AO, UDP-glucuronosyltransferase, carbonyl/aldo-keto reductase, flavin-containing monooxygenase, and monoamineoxidase in individual hepatocytes and pooled hepatocytes from the same individual donors, intrinsic clearance in the pooled hepatocytes was ± 30% of the average clearance value in individuals for 15 of 18 compounds, suggesting that pooled hepatocytes maintained the average activity of the individual hepatocytes. Although the results of an in vivo-in vitro correlation analysis for AO substrate compounds showed a trend toward under-prediction, the underestimation ratios for all AO substrates were nevertheless comparable (7.2- to 14.9-fold), suggesting that hepatic clearance prediction for these compounds can be quantified using empirical scaling. These observations enabled us to obtain specific pooled hepatocytes that showed the expected non-CYP enzyme activities by pre-characterization and to quantify hepatic clearance prediction for AO compounds using an empirical scaling factor.
authors.affiliation: Astellas Research Institute of America LLC, Skokie, IL, USA.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Akabane
authors.forename: Nicolas
authors.initials: N
authors.lastname: Gerst
authors.forename: Jeffrey N
authors.initials: JN
authors.lastname: Masters
authors.forename: Kouichi
authors.initials: K
authors.lastname: Tamura
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism; D042931:Aldehyde Oxidase / Q000378:metabolism*; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D015925:Cryopreservation; D014453:Glucuronosyltransferase / Q000378:metabolism; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D008657:Metabolic Clearance Rate / Q000502:physiology*; D015394:Molecular Structure; D008995:Monoamine Oxidase / Q000378:metabolism; D010105:Oxygenases / Q000378:metabolism; D014461:Ultracentrifugation; D015262:Xenobiotics / Q000097:blood / Q000378:metabolism*; D000099148:UGT1A1 Enzyme
chemical_list: D000429:Alcohol Oxidoreductases; D042931:Aldehyde Oxidase; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D014453:Glucuronosyltransferase; D008995:Monoamine Oxidase; D010105:Oxygenases; D015262:Xenobiotics; D000099148:UGT1A1 Enzyme; C020066:dimethylaniline monooxygenase (N-oxide forming)