Abstract
BackgroundOral administration is the most common way to deliver drugs to the systemic circulation or target organs. Orally administered drugs are absorbed in the intestine and metabolized in the intestine and liver. In the early stages of drug development, it is important to predict first-pass metabolism accurately to select candidate drugs with high bioavailability. The Caco-2 cell line derived from colorectal cancer is widely used as an intestinal model to assess drug membrane permeability. However, because the expression of major drug-metabolizing enzymes, such as cytochrome P450 (CYP), is extremely low in Caco-2 cells, it is difficult to predict intestinal metabolism, which is a significant factor in predicting oral drug bioavailability. Previously, we constructed a mouse artificial chromosome vector carrying the CYP (CYP2C9, CYP2C19, CYP2D6, and CYP3A4) and P450 oxidoreductase (POR) (4CYPs-MAC) genes and increased CYP expression and metabolic activity in HepG2 cells via transfer of this vector.ResultsIn the current study, to improve the Caco-2 cell assay model by taking metabolism into account, we attempted to increase CYP expression by transferring the 4CYPs-MAC into Caco-2 cells. The Caco-2 cells carrying the 4CYPs-MAC showed higher CYP mRNA expression and activity. In addition, high metabolic activity, availability for permeation test, and the potential to assess drug–drug interactions were confirmed.ConclusionsThe established Caco-2 cells with the 4CYPs-MAC are expected to enable more accurate prediction of the absorption and metabolism in the human intestine than parental Caco-2 cells. The mammalian artificial chromosome vector system would provide useful models for drug development.
Highlights
Oral administration is the most common way to deliver drugs to the systemic circulation or target organs
Caco-2 cells express a variety of efflux and uptake transporters, they have an absence or low levels of cytochrome P450 (CYP) isoforms, such as CYP3A4 and CYP2C, that are typically expressed in the human intestinal epithelium [2]
To examine whether the CYP and P450 oxidoreductase (POR) genes were introduced into the obtained clones, genomic PCR analyses were performed
Summary
Oral administration is the most common way to deliver drugs to the systemic circulation or target organs. Because the expression of major drug-metabolizing enzymes, such as cytochrome P450 (CYP), is extremely low in Caco-2 cells, it is difficult to predict intestinal metabolism, which is a significant factor in predicting oral drug bioavailability. Oral drug bioavailability is often limited by metabolizing enzymes and efflux transporters in the gut [1]. The Caco-2 cell line derived from human colon carcinoma is a commonly used model for estimating the intestinal absorption of new drug candidates. Caco-2 cells express a variety of efflux and uptake transporters, they have an absence or low levels of cytochrome P450 (CYP) isoforms, such as CYP3A4 and CYP2C, that are typically expressed in the human intestinal epithelium [2]. To predict the intestinal absorption of drugs more accurately, it is necessary to modify Caco-2 cells to increase their expression of CYP isoforms
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