Oral 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. In 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. The 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.

中文翻译：

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通过哺乳动物人工染色体表达四个CYP的Caco-2细胞的发育。

口服给药是将药物输送到全身循环或靶器官的最常用方法。口服药物在肠道吸收，在肠道和肝脏代谢。在药物开发的早期阶段，准确预测首过代谢以选择具有高生物利用度的候选药物非常重要。源自结肠直肠癌的Caco-2细胞系被广泛用作评估药物膜通透性的肠道模型。但是，由于主要的药物代谢酶（例如细胞色素P450（CYP））在Caco-2细胞中的表达极低，因此很难预测肠道代谢，这是预测口服药物生物利用度的重要因素。以前，我们构建了一个小鼠人工染色体载体，该载体带有CYP（CYP2C9，CYP2C19，CYP2D6，和CYP3A4）和P450氧化还原酶（POR）（4CYPs-MAC）基因，并通过该载体的转移增加了HepG2细胞的CYP表达和代谢活性。在当前的研究中，为了通过考虑代谢改善Caco-2细胞测定模型，我们试图通过将4CYPs-MAC转移到Caco-2细胞中来增加CYP表达。携带4CYPs-MAC的Caco-2细胞显示出较高的CYP mRNA表达和活性。此外，还证实了其高代谢活性，渗透性测试的可行性以及评估药物相互作用的潜力。已建立的具有4CYPs-MAC的Caco-2细胞预计比亲本的Caco-2细胞能够更准确地预测人肠中的吸收和代谢。哺乳动物人工染色体载体系统将为药物开发提供有用的模型。