Development and Validation of a Higher-Throughput Cytochrome P450 Inhibition Assay with the Novel Cofactor-Supplemented Permeabilized Cryopreserved Human Hepatocytes (MetMax Human Hepatocytes).
Cell Culture Techniques
/ methods
Cryopreservation
Culture Media
/ chemistry
Cytochrome P-450 Enzyme Inhibitors
/ pharmacology
Cytochrome P-450 Enzyme System
/ metabolism
Drug Interactions
Enzyme Assays
/ methods
Hepatocytes
High-Throughput Screening Assays
/ methods
Humans
Inhibitory Concentration 50
Isoenzymes
/ antagonists & inhibitors
Microsomes, Liver
Journal
Drug metabolism and disposition: the biological fate of chemicals
ISSN: 1521-009X
Titre abrégé: Drug Metab Dispos
Pays: United States
ID NLM: 9421550
Informations de publication
Date de publication:
10 2019
10 2019
Historique:
received:
06
06
2019
accepted:
22
07
2019
pubmed:
4
8
2019
medline:
2
6
2020
entrez:
4
8
2019
Statut:
ppublish
Résumé
Here, we report the application of a novel hepatocyte system, the cofactor-supplemented permeabilized cryopreserved human hepatocytes [MetMax human hepatocytes (MMHHs)] in a higher-throughput 384-well plate assay for the evaluation of cytochrome P450 (P450) inhibition. The assay was created to develop physiologically relevant P450 inhibition information, taking advantage of the complete organelle composition and their associated drug-metabolizing enzymes of the MMHH but with the ease of use of human liver microsomes, including storage at -80°C instead of in liquid nitrogen, and thaw and use without centrifugation and microscopic evaluation as required for intact hepatocytes. Nine key P450 isoforms for drug metabolism (CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4) were evaluated using multiple isoform-selective inhibitors. Results with MMHH were found to be comparable to those obtained with intact cryopreserved human hepatocytes (CHHs). Isoform-selective drug-metabolizing enzyme pathways evaluated were phenacetin
Identifiants
pubmed: 31375472
pii: dmd.119.088237
doi: 10.1124/dmd.119.088237
doi:
Substances chimiques
Culture Media
0
Cytochrome P-450 Enzyme Inhibitors
0
Isoenzymes
0
Cytochrome P-450 Enzyme System
9035-51-2
Types de publication
Journal Article
Validation Study
Langues
eng
Sous-ensembles de citation
IM
Pagination
1032-1039Informations de copyright
Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.