2023 Fiscal Year Final Research Report
Establishment of a culture system of alternative human hepatocytes with high expression of drug-metabolizing enzyme functions for exploratory drug metabolism and pharmacokinetic studies
| Project/Area Number |
20K07161
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 47060:Clinical pharmacy-related
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| Research Institution | Nihon Pharmaceutical University |
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Principal Investigator |
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| Project Period (FY) |
2020-04-01 – 2024-03-31
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| Keywords | 代替ヒト肝細胞 / シトクロムP450 / グルクロン酸転移酵素 / 硫酸転移酵素 / HepG2細胞 / ヒト肝癌由来細胞 / DNAメチル基転移酵素阻害剤 / Huh-7細胞 |
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| Outline of Final Research Achievements |
We focused on the fact that epigenomic treated of human hepatoblastoma-derived HepG2 cells with 5-azacytidine (5-Aza), a DNA methyltransferase inhibitor, resulted in high expression of drug-metabolizing enzyme activity, and investigated a culture method to further increase the expression of drug-metabolizing enzyme function. By adding low-molecular-weight compound-X, which suppresses Snail gene expression, to the epigenome-treated medium, higher enzyme activity and CYP inducibility were observed in a shorter period of culture than with 5-Aza alone treatment. Furthermore, higher drug-metabolizing enzyme activity was observed by decreasing the glucose concentration in the epigenome-treated medium, suggesting that HepG2 cells epigenomic treated with the newly constructed medium in this study may be an alternative human hepatocyte that can be used for exploratory studies of drug metabolism and pharmacokinetics.
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| Free Research Field |
薬物動態
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| Academic Significance and Societal Importance of the Research Achievements |
ヒト肝細胞は、創薬での薬物動態研究においては必須のツールである。しかし薬物代謝機能を高発現しているヒト肝細胞の入手経路は限られており、国内において入手することはほぼ不可能なために海外からの入手に依存している。そのために安定的な入手が困難かつ高額である。そこで、創薬研究でルーチン的に活用できる安価で安定供給可能な代替ヒト肝細胞の創製が望まれていることから、増殖可能なヒト肝癌細胞株HepG2細胞を5-アザシチジンでエピゲノム処理することにより薬物代謝酵素活性を高発現することに注目し、更なる高活性を発現するためのエピゲノム処理法を構築し、薬物動態研究に資する代替ヒト肝細胞を創製した。
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