豆彩霞;邱翔;万丽;
DOU Cai-xia;QIU Xiang;WAN Li;School of Pharmacy,Chengdu University of Traditional Chinese Medicine;State Key Laboratory of Biotherapy,West China Hospital,Sichuan University;

• 1:成都中医药大学药学院
• 2:四川大学华西医院肿瘤生物治疗研究室

摘要(Abstract)：

应用体外肝微粒体孵育体系,采用超快速液相色谱质谱联用(UFLC-MS/MS)检测方法,研究海南崖豆藤抗炎活性成分6-methoxy-8,8-dimethyl-3-(2,4,5-trimethoxyphenyl)-4H,8H-pyrano[2,3-f]chromen-4-one(HN-1)在大鼠、小鼠、恒河猴、Beagle犬和人5个种属肝微粒体中的代谢稳定性,比较代谢种属间的差异;采用化学抑制剂法确定其在人肝微粒体中的代谢表型。采用UPLC-Q-TOF-MS/MS检测方法,通过比较体外孵育0,60 min的样品,初步推断各种属体外代谢产物;比较粪便、尿液、血浆空白和给药后的生物样品,推测HN-1在SD大鼠体内的代谢产物。结果表明HN-1在各种属肝微粒体中的代谢均较为稳定,其中犬与人的肝微粒体代谢性质最相近;在人肝微粒体中主要经CYP1A1,CYP2D6,CYP3A4 3种同工酶催化代谢;初步推测出HN-1在体内体外的代谢产物,其中包括3个体外代谢产物和5个体内代谢产物,研究结果为HN-1进一步的药理药效研究奠定基础。
Ultra-fast performance liquid chromatography-mass spectrometry( UFLC-MS/MS) was used to study the anti-inflammatory active ingredient of Millettia pachyloba,6-methoxy-8,8-dimethyl-3-( 2,4,5-trimethoxyphenyl)-4 H,8 H-pyrano[2,3-f]chromen-4-one( HN-1),in liver microsomes of rats,mice,rhesus monkeys,Beagle dogs and humans metabolic stability,and compare the metabolic differences between different species. The metabolic phenotype in human liver microsomes was determined by chemical inhibitor method. Using UPLC-Q-TOF-MS/MS detection method,the in vitro metabolites of various liver microsomes were preliminarily inferred by comparing the samples incubated for 0 min and 60 min in vitro. The metabolites of HN-1 in SD rats were presumed by comparing feces,urine,plasma blanks and samples after administration. The results showed that the metabolism of HN-1 in various liver microsomes was stable,and the metabolic properties of dog and human liver microsomes were the closest. It is mainly catabolized by CYP1 A1,CYP2 D6 and CYP3 A4 isoenzymes in human liver microsomes. The metabolites of HN-1 in vitro and in vivo,including 3 in vitro metabolites and5 in vivo metabolites,were preliminarily estimated. The results laid the foundation for further pharmacological studies of HN-1.

关键词(KeyWords)： 海南崖豆藤;肝微粒体;代谢稳定性;代谢产物
Millettia pachyloba;liver microsomes;metabolic stability;metabolites

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81673653);国家自然科学基金青年基金项目(81703344)

作者(Authors): 豆彩霞;邱翔;万丽;
DOU Cai-xia;QIU Xiang;WAN Li;School of Pharmacy,Chengdu University of Traditional Chinese Medicine;State Key Laboratory of Biotherapy,West China Hospital,Sichuan University;

DOI: 10.19540/j.cnki.cjcmm.20190620.201

参考文献(References)：

• [1] Shi X,Liu D,Zhang J,et al. Extraction and purification of total flavonoids from pine needles of Cedrus deodara,contribute to anti-tumor in vitro[J]. BMC Complement Altern Med,2016,16(1):245.
• [2] Li Qinglin,Xin Wenxiu,Zhong Like,et al. A study on the antitumor mechanism of total flavonoids from Radix Tetrastigmae against additional cell line based on COX-2-mediated Wnt/β-catenin signaling pathway[J]. Oncotarget, 2017, 8(33):54304.
• [3] Cushnie T P T,Lamb A J. Recent advances in understanding the antibacterial properties of flavonoids[J]. Int J Antimicrob Agents,2011,38(2):99.
• [4] Wan L,Jiang J G. Protective effects of plant-derived flavonoids on hepatic injury[J]. J Funct Foods,2018,44:283.
• [5] Garcíalafuente A,Guillamón E,Villares A,et al. Flavonoids as anti-inflammatory agents:implications in cancer and cardiovascular disease[J]. Inflamm Res,2009,58(9):537.
• [6] Zeinali M,Rezaee S A,Hosseinzadeh H. An overview on immunoregulatory and anti-inflammatory properties of chrysin and flavonoids substances[J]. Biomed Pharmacother,2017,92:998.
• [7] Teresa V,Rodríguez-Nogales Alba,Francesca A,et al. Flavonoids in inflammatory bowel disease:a review[J]. Nutrients,2016,8(4):211.
• [8] Fang S C,Hsu C L,Lin H T,et al. Anticancer effects of flavonoid derivatives isolated from Milletti areticulata Benth in SKHep-1 human hepatocellular carcinoma cells[J]. J Agr Food Chem,2010,58(2):814.
• [9]许梦莹，郭日新，张晓，等．沙苑子化学成分研究[J]．中国中药杂志，2018,43(7):1459.
• [10]周北斗，张项林，牛海渊，等．蔓九节枝叶中化学成分研究[J]．中国中药杂志，2018,43(24):4878.
• [11]王小俊，邓玉环，张丽萍，等．UPLC-DAD-MS定性和定量分析蕲艾中的酚酸和黄酮类成分[J]．中国中药杂志，2019,44(5):983.
• [12]郑雪花，姜赞，王小雨，等．黄酮类化合物对AKR1C3抑制作用的研究[J]．今日药学，2018,28(8):505.
• [13]邵葵阳，张璇，焦文君，等．珍珠梅黄酮纳米粒抑制脂多糖诱发肝癌细胞的炎性作用[J]．药学学报，2017,52(10):1549.
• [14] Ye H,Wu W,Liu Z,et al. Bioactivity-guided isolation of antiinflammation flavonoids from the stems of Millettia dielsiana Harms.[J]. Fitoterapia,2014,95(2):154.
• [15] Aggarwal B B,Gupta S C,Sung B. Curcumin:an orally bioavailable blocker of TNF and other pro-inflammatory biomarkers[J]. Bri J Pharm,2013,169(8):1672.
• [16] Kollias G. TNF pathophysiology in murine models of chronic inflammation andautoimmunity[J]. Semin Arthritis Rheum,2005,34(5):3.
• [17] Ting G,Dong-Xiao W,Ruo-Yun C,et al. Novel benzil and isoflavone derivatives from Milletti adielsiana[J]. Planta Med,2009,75(3):236.
• [18] Cohen L H,Remley M J,Raunig D,et al. In vitro drug interaction of cytochrome P450:an evaluation of fluorogenic to conventional substrants[J]. Drug Metab Dispos,2003,31(8):1005.
• [19] Liu Z J,Fu D X,Tang G. FDA drug interaction research guide(draft)interpretation of 2006 edition[J]. J Int Pharm Res,2008,35(1):50.
• [20] Mohutsky M A,Chienv J Y,Ring B J,et al. Predictions of the in vivo clearance of drugs from rate of loss using human liver microsomes for phase I and phaseⅡbiotransformations[J]. Pharm Res,2006,23(4):654.
• [21] Li C Z,Lin Q H,Zhang X M,et al. In vitro O-demethylation of rotundine by recombinant human CYP isoenzyme[J]. Acta Pharm Sin,2010,45(3):307.
• [22]中国药典．四部[S].2015:363.
• [23]常立娟，李佐静，李清，等．雷公藤致大鼠肾毒性血清代谢组学分析[J]．中国实验方剂学杂志，2016,22(24):89.