液相色谱-串联电喷雾离子阱质谱法研究人尿中埃坡霉素D 的主要代谢产物

中国临床药理学与治疗学 ›› 2009, Vol. 14 ›› Issue (3): 307-312.

液相色谱-串联电喷雾离子阱质谱法研究人尿中埃坡霉素D 的主要代谢产物

张春玲^1,2, 朱晓霞¹, 齐文渊¹, 刘江林^1,2, 孟志云1, 顾若兰¹, 王延琳¹, 吴卓娜¹, 孙国祥², 窦桂芳¹

¹军事医学科学院野战输血研究所, 北京100850;
²沈阳药科大学, 沈阳110016, 辽宁

收稿日期:2009-01-08 修回日期:2009-01-08 发布日期:2020-10-27
通讯作者: 窦桂芳,女, 研究员, 主要从事药物代谢与药物动力学研究。Tel:010-66932951 E-mai l:douguifang @vip163.com
基金资助:
国际科技合作重点项目计划(2005DFA30080)

Study on essential metabolites of Epothilone D in human urine by liquid

ZHANG Chun-ling^1,2, ZHU Xiao-xia¹, QI Wen-yuan¹, LIU Jiang-lin^1,2, MENG Zhi-yun¹, WANG Yan-lin¹, WU Zhuo-na¹, SUN Guo-xiang², DOU Gui-fang¹

¹Institute of Transfusion Medicine, Academy of Military Medical Sciences, Beijing 100850, China;
²Shenyang Pharmaceutical University, Shenyang 110016, Liaoning, China

Received:2009-01-08 Revised:2009-01-08 Published:2020-10-27

摘要/Abstract

摘要： 目的研究埃坡霉素D 在人尿中的主要代谢产物。方法受试者6 名静脉滴注埃坡霉素D,收集0 ~ 72 h 尿样, 经SPE 固相萃取小柱分离纯化后, 采用液相色谱-串联电喷雾离子阱质谱法(LC-MSn) 对样品进行分析, 通过比较空白尿样和给药后的尿样的总离子流色谱图和选择离子扫描色谱图, 分析可能的代谢产物。代谢产物的结构鉴定主要依据代谢产物的各级质谱图与原药的各级质谱图的相关性推断所得。结果在人尿中共检测到11 种主要的代谢产物, 根据所提供的代谢产物标准品鉴定出3 种代谢产物的结构, 代谢方式主要有氧化、还原、水解和结合反应。结论本方法选择性好, 为深入研究埃坡霉素类药物在人体内的代谢规律提供了可靠的方法和可参考的依据。

关键词: 高效液相色谱-串联质谱, 埃坡霉素D, SPE C18固相萃取技术, 代谢产物分析

Abstract: AIM: To investigate the essential metabolites of Epothilone D in human urine. METHODS: The urine samples of six volunteers after intervenous drop infusion of 125mg m2 were collected within 0-72 h, the samples were purified and separated through C18 solid-phase extraction column and analyzed with liquid chromatography-mass spectrometry.The potential metabolites were analyzed through comparing the TIC and SIM chromatograms of the blank urine and sample urine, and the structure of the metabolites were identified through the correlation of the full scan MSn spectra of Epothilone D and the metabolites. RESULTS: Eleven metabolites were found in human urine.There were three kinds of structure of metabolites, the metabolic ways of oxidation, hydrogenise, hydrolization and association reaction. CONCLUSION: The method was proved sensitively and selectively, and it is possible to supply a reliable reference for further study on the metabolites of Epothilones.

Key words: liquid chromatography-mass spectrometry, Epothilone D, solid-phase extraction, metabolite

中图分类号:

R969.1

张春玲, 朱晓霞, 齐文渊, 刘江林, 孟志云, 顾若兰, 王延琳, 吴卓娜, 孙国祥, 窦桂芳. 液相色谱-串联电喷雾离子阱质谱法研究人尿中埃坡霉素D 的主要代谢产物[J]. 中国临床药理学与治疗学, 2009, 14(3): 307-312.

ZHANG Chun-ling, ZHU Xiao-xia, QI Wen-yuan, LIU Jiang-lin, MENG Zhi-yun, WANG Yan-lin, WU Zhuo-na, SUN Guo-xiang, DOU Gui-fang. Study on essential metabolites of Epothilone D in human urine by liquid[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2009, 14(3): 307-312.

参考文献

[1] Julien B, Shah S, Ziermann R, et al.Isolation and characterization of the epothilone biosynthetic gene cluster from Sorangium cellulosum[J].Gene, 2000, 249(1 2):153-160.
[2] Bollag DM, McQueney PA, Zhu J, et al.Epothilones, a new class of microtubule-stabilizing agents with a taxol-like mechanism of action[J].Cancer Res, 1995, 55(11): 2325-2333.
[3] Holfle GH, Bedorf N, Steinemtz H, et al.Epothilone A and B-novel 16-membered macrolides with cytotoxic activi-ty:isolation crystal structure, and confoumation in Solution[J].Angew Chem Int Ed Engl, 1996, 35(13):1567-1569.
[4] 顾勤兰, 王进欣.抗肿瘤药物埃坡霉素生物合成进展[J].中国新药杂志, 2005, 14(10):1148-1150.
[5] 顾勤兰, 尤启冬, 王进欣.抗肿瘤药物埃坡霉素的研究进展[J].江苏药学与临床研究, 2004, 10(12):20-21.
[6] 王进欣, 顾勤兰, 尤启冬, 等.新型抗肿瘤药物埃坡霉素的研究[J].药学进展, 2004, 28(8):354-361.
[7] Lee KW, Briggs JM.Comparative molecular field analysis (coMFA) study of epothilones-tubulin depolymerization inhibitiors: pharmacophore development using 3D QSAR methods[J].J Comput Aided Mol Des, 2001, 15(1):41-55.
[8] Kowalski RJ, Giannakakakou P, Hamel E.Activities of the microtubulestabilizing agents epothilone A and B with purified tubulin and in cells resistant to paclitaxel(Taxol) [J].J Bio Chem, 1997, 272(4):2534-2541.
[9] Nicolaou KC, Roschangar F, Vourloumis D.Chemical biology of epothilones [J].Angew Chem Int Ed, 1998, 37(15):2014-2045.
[10] Stachel SJ, Lee CB, Spassova M, et al.On the interactivity of complex aynthesis and tumor pharmacology in the drug discovery process:total synthesis and comparative in vivo evaluations of the 15-Aza Epothilones[J].J Org Chem, 2001, 66(12):4369-4378.
[11] Humphrey MJ, Smith DA.Role of metabolism and pharmacokinetic studies in the discovery of new drugs-Present and future perspectives[J].Xenobiotica, 1992, 22(7): 743-755.
[12] GumbletonM, Sneader W.Pharmacokinetic considerations in rational drug design[J].Clin Pharmacokinet, 1994, 26(3):161-168.
[13] 潘颖瑜, 俞英, 赖谷仙, 等.高效液相色谱-电喷雾质谱法研究人尿中a-生育酚的主要代谢产物[J].药学学报, 2008, 43(6):634-639.
[14] Li JS.Progress in the vitamin E absorption and metabolism rescarch[J].Chin J Mod Appl Pharm Sin, 2006, 10(41): 1004-1009.
[15] Sindona G.High-throughput screening of tocopherols in natural extracts[J].J Mass Spectrum, 2000, 35(11): 1360-1361.
[16] 高凯, 余伟, 杨静, 等.大鼠肝微粒体CYP3A1 2 和CYP2C9 10 参与甘草次酸羟化代谢[J].中国临床药理学与治疗学, 2007, 12(11):1255-1260.