Laboratory of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan.
New Business Solutions Department, Innovative Solutions Center for Information & Communication Technology, Mitsui Chemicals, Inc., Chiba, Japan.
Drug Metab Pharmacokinet. 2023 Oct;52:100511. doi: 10.1016/j.dmpk.2023.100511. Epub 2023 Apr 26.
Drug-induced liver injury (DILI) is a major factor underlying drug withdrawal from the market. Therefore, it is important to predict DILI during the early phase of drug discovery. Metabolic activation and mitochondrial toxicity are good indicators of the potential for DILI. However, hepatocyte function, including drug-metabolizing enzyme activity and mitochondrial function, reportedly decreases under conventional culture conditions; therefore, these conditions fail to precisely detect metabolic activation and mitochondrial toxicity-induced cell death. To resolve this issue, we employed a newly developed cell culture plate with high oxygen permeability and low drug sorption (4-polymethyl-1-pentene [PMP] plate). Under PMP plate conditions, cytochrome P450 (CYP) activity and mitochondrial function were increased in primary rat hepatocytes. Following l-buthionine-sulfoximine-induced glutathione depletion, acetaminophen-induced cell death significantly increased under PMP plate conditions. Additionally, 1-aminobenzotriazole reduced cell death. Moreover, mitochondrial toxicity due to mitochondrial complex inhibitors (ketoconazole, metformin, and phenformin) increased under PMP plate conditions. In summary, PMP plate conditions could improve CYP activity and mitochondrial function in primary rat hepatocytes and potentially detect metabolic activation and mitochondrial toxicity.
药物性肝损伤（DILI）是导致药物从市场上撤出的主要因素。因此，在药物发现的早期阶段预测 DILI 非常重要。代谢激活和线粒体毒性是 DILI 潜在性的良好指标。然而，据报道，在常规培养条件下，肝细胞功能（包括药物代谢酶活性和线粒体功能）下降；因此，这些条件无法准确检测代谢激活和线粒体毒性诱导的细胞死亡。为了解决这个问题，我们使用了一种新开发的具有高氧气渗透性和低药物吸附性的细胞培养板（4-聚甲基-1-戊烯[PMP]板）。在 PMP 板条件下，原代大鼠肝细胞中的细胞色素 P450（CYP）活性和线粒体功能增加。在用 L-丁硫氨酸亚砜诱导谷胱甘肽耗竭后，在 PMP 板条件下，对乙酰氨基酚诱导的细胞死亡显著增加。此外，1-氨基苯并三唑降低了细胞死亡。此外，线粒体毒性抑制剂（酮康唑、二甲双胍和苯乙双胍）在 PMP 板条件下也会增加。总之，PMP 板条件可以改善原代大鼠肝细胞中的 CYP 活性和线粒体功能，并可能检测到代谢激活和线粒体毒性。
