Abstract
The small intestine plays a critical role in the absorption and metabolism of orally administered drugs. Therefore, a model capable of evaluating drug absorption and metabolism in the small intestine would be useful for drug discovery. Patients with genotype UGT1A1*6 (exon 1, 211G > A) treated with the antineoplastic drug SN-38 have been reported to exhibit decreased glucuronide conjugation and increased incidence of intestinal toxicity and its severe side effects, including severe diarrhea. To ensure the safety of drugs, we must develop a drug metabolism and toxicity evaluation model which considers UGT1A1*6. In this study, we generated CYP3A4·POR·UGT1A1 KI- and CYP3A4·POR·UGT1A1*6 KI-Caco-2 cells for pharmaceutical research using a PITCh system. The CYP3A4·POR·UGT1A1 KI-Caco-2 cells were shown to express functional CYP3A4 and UGT1A1. The CYP3A4·POR·UGT1A1*6 KI-Caco-2 cells were sensitive to SN-38-induced intestinal toxicity. We thus succeeded in generating CYP3A4·POR·UGT1A1 KI- and CYP3A4·POR·UGT1A1*6 KI-Caco-2 cells, which can be used in pharmaceutical research. We also developed an intestinal epithelial cell model of patients with UGT1A1*6 and showed that it was useful as a tool for drug discovery.
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The authors declare that all the data related to this study are available within the paper or can be obtained from the authors upon reasonable request.
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Acknowledgements
This research was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI (Grant Number 21K15264), Japan Agency for Medical Research and Development (AMED) (Grant Number 21mk0101214), The Nakatomi Foundation, The Promotion and Mutual Aid Corporation for Private Schools of Japan (PMAC) and the Research Promotion Program for Acquiring Grants-in-Aid-for Scientific Research of the Ritsumeikan University.
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RN conceived and supervised the study. RN and NY designed the experiments. RN, NY and KW performed the experiments. RN, NY, KW, YK and TF analyzed the data. RN wrote the manuscript.
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Negoro, R., Yamada, N., Watanabe, K. et al. Generation of Caco-2 cells stably expressing CYP3A4·POR·UGT1A1 and CYP3A4·POR·UGT1A1*6 using a PITCh system. Arch Toxicol 96, 499–510 (2022). https://doi.org/10.1007/s00204-021-03175-0
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DOI: https://doi.org/10.1007/s00204-021-03175-0