Biotransformation of diclofenac and effects on the metabolome of primary human hepatocytes upon repeated dose exposure.
Biotransformation of diclofenac and effects on the metabolome of primary human hepatocytes upon repeated dose exposure.
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双氯芬酸的生物转化以及重复剂量暴露对原代人肝细胞代谢组的影响
DOI：
10.1016/j.ejps.2012.01.014
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Noor F.
中科院分区：
文献类型：
--
作者：
Mueller D;Müller-Vieira U;Biemel K;Tascher G;Nussler AK;Noor F.
用于评估慢性药物诱导效应的体外重复剂量测试是临床前药物开发中的巨大挑战。慢性毒性会导致治疗中止或上市后停药，尽管进行了体内临床前筛选。在肝毒性的情况下，由于原代肝细胞的长期活力和功能有限，难以检测慢性肝脏效应。在本研究中，我们将原代人肝细胞在无血清培养基中培养超过3周，并分析了肝细胞的生理、活力和药物代谢能力。此外，我们评估了浓度范围（10-1000μM）的双氯芬酸急性（24 h）毒性。还检测了一种临床相关浓度和另一种较高浓度（6.4和100μM）下的慢性（9次重复给药）毒性。我们研究了重复给药暴露后双氯芬酸的I期和II期代谢，并分析了对细胞外代谢组的影响。急性24小时评估显示仅最高试验浓度（1 mM）具有毒性。重复给药暴露后，即使在低临床相关浓度（6.4μM）下也观察到毒性作用。生物转化途径活跃3周，检测到双甘酯-酰基葡萄糖醛酸苷为主要代谢产物。在9次重复给药后，观察到对外泌代谢组的剂量依赖性双氢叶酸诱导效应，如对乳酸和3-羟基丁酸的产生以及葡萄糖和半乳糖代谢的影响。总之，我们表明，对原代人肝细胞的长期功能培养物进行重复剂量试验可用于评估临床前筛选中的长期毒性作用，并可能有助于替代/减少体内动物试验。
In vitro repeated dose testing for the assessment of chronic drug-induced effects is a huge challenge in preclinical pharmaceutical drug development. Chronic toxicity results in discontinuation of therapy or post-marketing withdrawal of drugs despite in vivo preclinical screening. In case of hepatotoxicity, due to limited long term viability and functionality of primary hepatocytes, chronic hepatic effects are difficult to detect. In this study, we maintained primary human hepatocytes in a serum-free cultivation medium for more than 3weeks and analyzed physiology, viability and drug metabolizing capacities of the hepatocytes. Moreover, we assessed acute (24h) diclofenac toxicity in a range of (10–1000μM) concentrations. The chronic (9 repeated doses) toxicity at one clinically relevant and another higher concentration (6.4 and 100μM) was also tested. We investigated phase I and II metabolism of diclofenac upon repeated dose exposure and analyzed effects on the cellular exometabolome. Acute 24h assessment revealed toxicity only for the highest tested concentration (1mM). Upon repeated dose exposure, toxic effects were observed even at a low, clinically relevant concentration (6.4μM). Biotransformation pathways were active for 3weeks and diclofenac-acylglucuronide was detected as the predominant metabolite. Dose dependent diclofenac-induced effects on exometabolome, such as on the production of lactate and 3-hydroxybutyric acid as well as glucose and galactose metabolism, were observed upon nine repeated doses. Summarizing, we show that repeated dose testing on long-term functional cultures of primary human hepatocytes may be included for the assessment of long term toxic effects in preclinical screening and can potentially help replace/reduce in vivo animal testing.
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影响因子：
9.3
作者：
Amacher, DE;Adler, R;Townsend, RR
通讯作者：
Townsend, RR
如 13C 代谢流分析所示，Doxorubicin 会增加 HL-1 心肌细胞的氧化代谢。
DOI：
10.1093/toxsci/kfr298
发表时间：
2012
期刊：
Toxicological sciences : an official journal of the Society of Toxicology
影响因子：
--
作者：
Alexander Strigun;J. Wahrheit;J. Niklas;E. Heinzle;F. Noor
通讯作者：
F. Noor
影响因子：
3.2
作者：
Beckers, Simone;Noor, Fozia;Heinzle, Elmar
通讯作者：
Heinzle, Elmar
双氯芬酸钠（扶他林）在动物和人体中的生物转化。
DOI：
--
发表时间：
1979
期刊：
Xenobiotica; the fate of foreign compounds in biological systems
影响因子：
--
作者：
H. Stierlin;J. Faigle
通讯作者：
J. Faigle
3D 中空纤维生物反应器中原代人肝细胞的深入生理特征
影响因子：
3.3
作者：
Daniel Mueller;G. Tascher;U. Müller‐Vieira;D. Knobeloch;A. Nuessler;K. Zeilinger;E. Heinzle;F. Noor
通讯作者：
F. Noor