Ikuta Hiroyuki, Shimada Hiroaki, Sakamoto Kenjiro, Nakamura Rena, Kawase Atsushi, Iwaki Masahiro
Faculty of Pharmacy, Kindai University, Osaka, Japan.
Pharmaceutical Research and Technology Institute, Kindai University, Osaka, Japan.
Xenobiotica. 2022 Jul;52(7):653-660. doi: 10.1080/00498254.2022.2131484. Epub 2022 Oct 10.
Acyl glucuronides (AGs) are known as one of the causes of idiosyncratic drug toxicity (IDT). Although AGs can be enzymatically hydrolysed by β-glucuronidase and esterase, much information on their characteristics and species differences is lacking. This study was aimed to clarify species differences in AG hydrolysis between human and rat liver microsomes (HLM and RLM).To evaluate the AG hydrolysis profile, and the contribution of β-glucuronidase and esterase towards AG hydrolysis in HLM and RLM, nonsteroidal anti-inflammatory drugs (NSAIDs) were used. AGs were incubated with 0.1 M Tris-HCl buffer (pH 7.4) and 0.3 mg/mL HLM or RLM in the absence or presence of β-glucuronidase inhibitor, D-saccharic acid 1,4-lactone (D-SL) and esterase inhibitor, phenylmethylsulfonyl fluoride (PMSF).AGs of mefenamic acid (MEF-AG) and etodolac (ETO-AG) showed significantly higher AG hydrolysis rates in RLM than in HLM. Esterases were found to serve as AG hydrolases dominantly in HLM, whereas both esterases and β-glucuronidase equally contribute to AG hydrolysis in RLM. However, MEF-AG and ETO-AG were hydrolysed only by β-glucuronidase.We demonstrated for the first time that the activity of AG hydrolases towards NSAID-AGs differs between humans and rats.
酰基葡萄糖醛酸（AGs）被认为是特异质性药物毒性（IDT）的原因之一。尽管AGs可被β-葡萄糖醛酸酶和酯酶进行酶促水解，但关于它们的特性和种属差异仍缺乏很多信息。本研究旨在阐明人和大鼠肝微粒体（HLM和RLM）之间AG水解的种属差异。为评估AG水解情况以及β-葡萄糖醛酸酶和酯酶对HLM和RLM中AG水解的作用，使用了非甾体抗炎药（NSAIDs）。在不存在或存在β-葡萄糖醛酸酶抑制剂D-糖二酸1,4-内酯（D-SL）和酯酶抑制剂苯甲基磺酰氟（PMSF）的情况下，将AGs与0.1 M Tris-HCl缓冲液（pH 7.4）以及0.3 mg/mL的HLM或RLM一起孵育。甲芬那酸（MEF-AG）和依托度酸（ETO-AG）的AGs在RLM中的AG水解速率显著高于HLM。发现酯酶在HLM中主要作为AG水解酶起作用，而在RLM中酯酶和β-葡萄糖醛酸酶对AG水解的作用相当。然而，MEF-AG和ETO-AG仅被β-葡萄糖醛酸酶水解。我们首次证明了人和大鼠中AG水解酶对NSAID-AGs的活性存在差异。
