药学学报, 2019, 54(8): 1484-1492
引用本文:
徐叶, 杜江波, 冯慧瑾, 左建平, 许红涛, 李援朝, 钟大放. 雷公藤甲素衍生物雷腾舒的体外代谢研究[J]. 药学学报, 2019, 54(8): 1484-1492.
XU Ye, DU Jiang-bo, FENG Hui-jin, ZUO Jian-ping, XU Hong-tao, LI Yuan-chao, ZHONG Da-fang. Studies on the metabolism of a triptolide derivative (5R)-5-hydroxytriptolide in vitro[J]. Acta Pharmaceutica Sinica, 2019, 54(8): 1484-1492.

雷公藤甲素衍生物雷腾舒的体外代谢研究
徐叶1,2, 杜江波1, 冯慧瑾1, 左建平1, 许红涛1, 李援朝1, 钟大放1,2
1. 中国科学院上海药物研究所新药研究国家重点实验室, 上海 201203;
2. 中国科学院大学, 北京 100049
摘要:
本课题的目的是研究雷公藤甲素衍生物雷腾舒的体外代谢特征。将雷腾舒分别与人、猴、犬、大鼠或小鼠的肝细胞一起孵育,共鉴定出4个代谢物,分别为环氧水解开环代谢物(M1)、谷胱甘肽结合代谢物(M2)和单氧化并谷胱甘肽结合代谢物(M3-1和M3-2)。在人或大鼠肝微粒体孵育体系中,共鉴定出7个代谢物,分别为脱氢代谢物(M4)和单氧化代谢物(M5-1~M5-6)。通过化学半合成和大鼠原代肝细胞孵育的方法获得代谢物的对照品,并与以上代谢物相比对,确认了5个代谢物的结构,分别为12,13-环氧水解开环代谢物M1、12-谷胱甘肽结合代谢物M2、(16S)-单羟基化代谢物M5-1、(2R)-单羟基化代谢物M5-4和(19R)-单羟基化代谢物M5-5。体外活性评价显示,仅(2R)-羟基化代谢物表现出弱的免疫抑制活性,其活性不足母体药物的十分之一,同时毒性也显著降低。提示雷腾舒可能在体内经历代谢失活和减毒。
关键词:    雷腾舒      药物代谢      代谢物鉴定      肝细胞      肝微粒体     
Studies on the metabolism of a triptolide derivative (5R)-5-hydroxytriptolide in vitro
XU Ye1,2, DU Jiang-bo1, FENG Hui-jin1, ZUO Jian-ping1, XU Hong-tao1, LI Yuan-chao1, ZHONG Da-fang1,2
1. State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China
Abstract:
The purpose of current study is to investigate the metabolic profile of a triptolide derivative (5R)-5-hydroxytriptolide in vitro. (5R)-5-Hydroxytriptolide was incubated with the hepatocytes of human, monkey, dog, rat or mouse, respectively. Compared with inactivated hepatocytes, four metabolites were identified in hepatocytes from all five species:oxidative ring-opening metabolite (M1), glutathione-conjugating metabolite (M2), and monooxidative combined with glutathione-conjugating metabolites (M3-1 and M3-2), respectively. In human or rat liver microsomes, seven metabolites of (5R)-5-hydroxytriptolide were found, dehydrogenated metabolite (M4) and monooxidative metabolites (M5-1-M5-6), respectively. Reference standards for the metabolites were obtained either through chemical semisynthesis or biotransformation through rat primary hepatocytes. The structures of five metabolites were confirmed, which were 12,13-epoxy ring-opening metabolite M1, 12-glutathione-conjugating metabolite M2, (16S)-, (2R)- and (19R)-monohydroxylated metabolites M5-1, M5-4, and M5-5, respectively. In vitro activity assay revealed that only (2R)-hydroxylated metabolite exhibited weak immunosuppressive activity with less than one-tenth the activity of its parent drug, and a significant decrease in toxicity was observed. It is suggested that (5R)-5-hydroxytriptolide might undergo metabolic inactivation and detoxification in vivo.
Key words:    (5R)-5-hydroxytriptolide    metabolism    metabolite confirmation    hepatocytes    liver microsomes   
收稿日期: 2019-04-24
DOI: 10.16438/j.0513-4870.2019-0327
基金项目: 国家自然科学基金资助项目(81373479).
通讯作者: 钟大放,Tel:86-21-50800738,E-mail:dfzhong@simm.ac.cn;李援朝,E-mail:ycli@mail.shcnc.ac.cn;许红涛,E-mail:xuht@shanghaitech.edu.cn
Email: dfzhong@simm.ac.cn;ycli@mail.shcnc.ac.cn;xuht@shanghaitech.edu.cn
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参考文献:
[1] Chen BJ. Triptolide, a novel immunosuppressive and anti-inflammatory agent purified from a Chinese herb Tripterygium wilfordii Hook F[J]. Leuk Lymphoma, 2001, 42:253-265.
[2] Lin N, Liu CF, Xiao C, et al. Triptolide, a diterpenoid triepoxide, suppresses inflammation and cartilage destruction in collagen-induced arthritis mice[J]. Biochem Pharmacol, 2007, 73:136-146.
[3] Li XJ, Jiang ZZ, Zhang LY. Triptolide:progress on research in pharmacodynamics and toxicology[J]. J Ethnopharmacol, 2014, 155:67-79.
[4] Xi C, Peng SJ, Wu ZP, et al. Toxicity of triptolide and the molecular mechanisms involved[J]. Biomed Pharmacother, 2017, 90:531-541.
[5] Cui J, Chen X, Su JC. Advanced progress of main pharmacology activities of triptolide[J]. China J Chin Mater Med (中国中药杂志), 2017, 42:2655-2658.
[6] Zhou ZL, Yang YX, Ding J, et al. Triptolide:structural modifications, structure-activity relationships, bioactivities, clinical development and mechanisms[J]. Nat Prod Rep, 2012, 29:457-475.
[7] Tang W, Zuo JP. Immunosuppressant discovery from Tripterygium wilfordii Hook F:the novel triptolide analog (5R)-5-hydroxytriptolide (LLDT-8)[J]. Acta Pharmacol Sin, 2012, 33:1112-1118.
[8] Wang L, Xu YP, Fu L, et al. (5R)-5-Hydroxytriptolide (LLDT-8), a novel immunosuppressant in clinical trials, exhibits potent antitumor activity via transcription inhibition[J]. Cancer Lett, 2012, 324:75-82.
[9] Xu Y, Zhang YF, Chen XY, et al. CYP3A4 inducer and inhibitor strongly affect the pharmacokinetics of triptolide and its derivative in rats[J]. Acta Pharmacol Sin, 2018, 39:1386-1392.
[10] Xu HT, Tang HY, Yang Z, et al. Synthesis and biological evaluation of 20-hydroxytriptonide and its analogues[J]. Tetrahedron, 2014, 70:3107-3115.
[11] Xu HT, Tang HY, Feng HJ, et al. Divergent total synthesis of triptolide, triptonide, tripdiolide, 16-hydroxytriptolide, and their analogues[J]. J Org Chem, 2014, 79:10110-10122.
[12] Naritomi Y, Terashita S, Kimura S, et al. Prediction of human hepatic clearance from in vivo animal experiments and in vitro metabolic studies with liver microsomes from animals and humans[J]. Drug Metab Dispos, 2001, 29:1316-1324.
[13] Davies B, Morris T. Physiological-parameters in laboratoryanimals and humans[J]. Pharm Res, 1993, 10:1093-1095.
[14] Zhou L, Pang XY, Xie C, et al. Chemical and enzymatic transformations of nimesulide to GSH conjugates through reductive and oxidative mechanisms[J]. Chem Res Toxicol, 2015, 28:2267-2277.
[15] Crincoli CM, Patel NN, Harvison PJ. Formation of glutathione (GSH) conjugates from 3-(3,5-dichlorophenyl)-2,4-thiazolidinedione (DCPT) by rat liver microsomes[J]. FASEB J, 2007, 21:A813.
[16] Deng P, Zhan Y, Chen XY, et al. Derivatization methods for quantitative bioanalysis by LC-MS/MS[J]. Bioanalysis, 2012, 4:49-69.
[17] Liu J, Chen XY, Zhang YF, et al. Derivatization of (5R)-hydroxytriptolide from benzylamine to enhance mass spectrometric detection:application to a phase I pharmacokinetic study in humans[J]. Anal Chim Acta, 2011, 689:69-76.
[18] Kerns EH, Di L. Part 3:Disposition, Metabolism, and Safety, Drug-like Properties:Concepts, Structure Design and Methods[M]. Oxford, UK:Elsevier, 2008.
[19] Li W, Liu Y, He YQ, et al. Characterization of triptolide hydroxylation by cytochrome P450 in human and rat liver microsomes[J]. Xenobiotica, 2008, 38:1551-1565.
[20] Liu J, Li L, Zhou X, et al. Metabolite profiling and identification of triptolide in rats[J]. J Chromatogr B, 2013, 939:51-58.
[21] Ye XC, Li WY, Yan Y, et al. Effects of cytochrome P4503A inducer dexamethasone on the metabolism and toxicity of triptolide in rat[J]. Toxicol Lett, 2010, 192:212-220.
[22] Tai T, Huang X, Su YW, et al. Glycyrrhizin accelerates the metabolism of triptolide through induction of CYP3A in rats[J]. J Ethnopharmacol, 2014, 152:358-363.
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