Development of a Specific Substrate-Inhibitor Panel (Liver-on-a-Chip) for Evaluation of Cytochrome P450 Activity

A A Zakhariants; O A Burmistrova; M Y Shkurnikov; A A Poloznikov; D A Sakharov

doi:10.1007/s10517-016-3567-z

Development of a Specific Substrate-Inhibitor Panel (Liver-on-a-Chip) for Evaluation of Cytochrome P450 Activity

Bull Exp Biol Med. 2016 Nov;162(1):170-174. doi: 10.1007/s10517-016-3567-z. Epub 2016 Nov 23.

Authors

A A Zakhariants¹, O A Burmistrova², M Y Shkurnikov³, A A Poloznikov², D A Sakharov²

Affiliations

¹ Biocilicum Research and Production Center, Moscow, Russia. zakhariants@bioclinicum.com.
² Biocilicum Research and Production Center, Moscow, Russia.
³ P. A. Hertsen Moscow Oncology Research Institute, Moscow, Russia.

PMID: 27882460
DOI: 10.1007/s10517-016-3567-z

Abstract

We developed a cytochrome P450 substrate-inhibitor panel for preclinical in vitro evaluation of drugs in a 3D histotypical microfluidic cell model of human liver (liver-on-a-chip technology). The concentrations of substrates and inhibitors were optimized to ensure reliable detection of the principal metabolites by HPLC-mass-spectroscopy. The selected specific substrate-inhibitor pairs, namely bupropion/2-phenyl-2-(1-piperidinyl)propane) for evaluation of CYP2B6B activity, tolbutamide/sulfaphenazole for CYP2C9, omeprazole/(+)-N-benzylnirvanol for CYP2C19, and testosterone/ketoconazole for CYP3A4, enable reliable evaluation of the drug metabolism pathway. In contrast to animal models characterized by species-specific expression profile and activity of cytochrome P450 isoforms, our in vitro model reflects the metabolism of human hepatocytes in vivo.

Keywords: HepaRG; biotransformation; liver cell model; liver-on-a-chip; preclinical in vitro studies.

MeSH terms

Bupropion / metabolism
Chromatography, High Pressure Liquid
Cytochrome P-450 CYP2B6 / analysis
Cytochrome P-450 CYP2B6 / metabolism*
Cytochrome P-450 CYP2C19 / analysis
Cytochrome P-450 CYP2C19 / metabolism*
Cytochrome P-450 CYP2C9 / analysis
Cytochrome P-450 CYP2C9 / metabolism*
Cytochrome P-450 CYP3A / analysis
Cytochrome P-450 CYP3A / metabolism*
Cytochrome P-450 Enzyme Inhibitors / pharmacology
Humans
Ketoconazole / pharmacology
Lab-On-A-Chip Devices*
Liver / drug effects
Liver / enzymology
Mass Spectrometry
Mephenytoin / analogs & derivatives
Mephenytoin / pharmacology
Microsomes, Liver / drug effects
Microsomes, Liver / enzymology
Omeprazole / metabolism
Phencyclidine / analogs & derivatives
Phencyclidine / pharmacology
Substrate Specificity
Sulfaphenazole / pharmacology
Testosterone / metabolism
Tolbutamide / metabolism

Substances

2-phenyl-2-(1-piperidinyl)propane
Cytochrome P-450 Enzyme Inhibitors
N-3-benzylnirvanol
Bupropion
Sulfaphenazole
Testosterone
Tolbutamide
CYP2C9 protein, human
Cytochrome P-450 CYP2C9
CYP2B6 protein, human
CYP2C19 protein, human
Cytochrome P-450 CYP2B6
Cytochrome P-450 CYP2C19
Cytochrome P-450 CYP3A
CYP3A4 protein, human
Phencyclidine
Omeprazole
Mephenytoin
Ketoconazole