Biomol Ther.  2015 Sep;23(5):486-492. 10.4062/biomolther.2015.041.

Measurement of Human Cytochrome P450 Enzyme Induction Based on Mesalazine and Mosapride Citrate Treatments Using a Luminescent Assay

• Kim YH
• Bae YJ
• Kim HS
• Cha HJ
• Yun JS
• Shin JS
• Seong WK
• Lee YM
• Han KM

• KMID: 2380299
• DOI: http://doi.org/10.4062/biomolther.2015.041

Abstract

Drug metabolism mostly occurs in the liver. Cytochrome P450 (CYP) is a drug-metabolizing enzyme that is responsible for many important drug metabolism reactions. Recently, the US FDA and EU EMA have suggested that CYP enzyme induction can be measured by both enzymatic activity and mRNA expression. However, these experiments are time-consuming and their inter-assay variability can lead to misinterpretations of the results. To resolve these problems and establish a more powerful method to measure CYP induction, we determined CYP induction by using luminescent assay. Luminescent CYP assays link CYP enzyme activity to firefly luciferase luminescence technology. In this study, we measured the induction of CYP isozymes (1A2, 2B6, 2C9, and 3A4) in cryopreserved human hepatocytes (HMC424, 478, and 493) using a luminometer. We then examined the potential induction abilities (unknown so far) of mesalazine, a drug for colitis, and mosapride citrate, which is used as an antispasmodic drug. The results showed that mesalazine promotes CYP2B6 and 3A4 activities, while mosapride citrate promotes CYP1A2, 2B6, and 3A4 activities. Luminescent CYP assays offer rapid and safe advantages over LC-MS/MS and qRT-PCR methods. Furthermore, luminescent CYP assays decrease the interference between the optical properties of the test compound and the CYP substrates. Therefore, luminescent CYP assays are less labor intensive, rapid, and can be used as robust tools for high-throughput CYP screening during early drug discovery.

Keyword

CYP; Human hepatocytes; Luminescent assay; Mesalazine; Mosapride citrate

MeSH Terms

Citric Acid*
Colitis
Cytochrome P-450 CYP1A2
Cytochrome P-450 Enzyme System*
Cytochromes*
Drug Discovery
Enzyme Induction*
Fireflies
Hepatocytes
Humans*
Isoenzymes
Liver
Luciferases
Luminescence
Luminescent Measurements*
Mass Screening
Mesalamine*
Metabolism
RNA, Messenger
Citric Acid
Cytochrome P-450 CYP1A2
Cytochrome P-450 Enzyme System
Cytochromes
Isoenzymes
Luciferases
Mesalamine
RNA, Messenger