title: Cell-based assays in combination with ultra-high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry for screening bioactive capilliposide C metabolites generated by rat intestinal microflora.
abstract: Many plant-derived glycosides are used as medications. It is common that these glycosides show poor intestinal absorption but their metabolites generated by intestinal microflora demonstrate strong bioactivity. Hence, it is crucial to develop a method for the identification and characterization of the metabolites, and consequently reveal the pathway in which the glycosides are processed in gut. In this study, cell-based assays in combination with ultra-high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS/MS) were developed for rapid discovery and evaluation of the metabolites of a glycoside compound, capilliposide C (LC-C). 92.7% of LC-C was biotransformed by rat intestinal microflora after 36-h incubation at 37°C. Human cancer cell lines HepG2, PC-3 and A549 was treated with metabolites pool, respectively, which was followed by cell viability assays and characterization of metabolites using UHPLC-QTOF-MS/MS. As a result, significant cytotoxicity was observed for the metabolites pool, from which six metabolites were identified. Based on the metabolites identified, deglycosylation and esterolysis were proposed as the major metabolic pathways of LC-C in rat intestinal microflora. In addition, M4, an esterolysis product of LC-C, was obtained and evaluated for its bioactivity in vitro. As a result, M4 exhibited a reduction in cell viability in HepG2 with an IC50 value of 17.46±1.55μg/mL.
authors.affiliation: School of Pharmacy, Weifang Medical University, 7166 Baotong West Street, Weifang, Shandong 261053, China; Tongji School of Pharmacy, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
authors.forename: Zhongzhe
authors.initials: Z
authors.lastname: Cheng
authors.affiliation: Tongji School of Pharmacy, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
authors.forename: Meilin
authors.initials: M
authors.lastname: Huang
authors.affiliation: Tongji School of Pharmacy, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
authors.forename: Guiying
authors.initials: G
authors.lastname: Chen
authors.affiliation: Tongji School of Pharmacy, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
authors.forename: Guangjie
authors.initials: G
authors.lastname: Yang
authors.affiliation: Tongji School of Pharmacy, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Tongji School of Pharmacy, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
authors.forename: Chang
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Pharmacy, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, 13Hangkong Road, Wuhan, Hubei 430030, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Medical Research Center, Humanwell Healthcare (Group) Co., Ltd., 666 Gao xin Road, Wuhan, Hubei 430075, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China.
authors.forename: Yulin
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, Zhejiang, China. Electronic address: zhanglin@zju.edu.cn.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Tongji School of Pharmacy, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China. Electronic address: jianghongliang@hust.edu.cn.
authors.forename: Hongliang
authors.initials: H
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D001711:Biotransformation; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002853:Chromatography, Liquid / Q000379:methods*; D000069196:Gastrointestinal Microbiome / Q000502:physiology; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000382:microbiology*; D015394:Molecular Structure; D051381:Rats; D012503:Saponins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014315:Triterpenes / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D012503:Saponins; D014315:Triterpenes; C511098:capilliposide C
keywords: UHPLC-QTOF-MS/MS; bioactive metabolites; capilliposide C; glycosides; intestinal microflora