Title:Identification of <i>In Vivo</i> Metabolites of a Potential Anti-tumor Drug Candidate AMAC, in Rat Plasma, Urine and Feces Samples Using UHPLC/QTOF /MS/MS
VOLUME: 17 ISSUE: 4
Author(s):Caixia Dou, Minghai Tang, Yuanyuan Xia, Linyu Yang, Xiang Qiu, Yong Li, Haoyu Ye* and Li Wan*
Affiliation:School of pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, Sichuan Province, Lab of Natural Product Drugs, Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu, Sichuan Province, School of pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, Sichuan Province, Lab of Natural Product Drugs, Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu, Sichuan Province, School of pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, Sichuan Province, Lab of Natural Product Drugs, Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu, Sichuan Province, Lab of Natural Product Drugs, Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu, Sichuan Province, School of pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, Sichuan Province
Keywords:AMAC, UHPLC/Q-TOF-MS/MS, in vivo metabolites, metabolic pathways, structural confirmation, cytoskeletal.
Abstract:
Background: Drugs based on natural products targeting the microtubule system remain an
important component in cancer therapy. Compound 10, 4-((3-amino-4-methoxyphenyl) amino)-2Hcoumarin,
derived from coumarin, showed excellent anti-proliferative activity through directly binding
to the colchicine-binding site in β-tubulin, suggesting that it could be a perfect drug candidate for antitumor
drug research and development. Identification and structural characterization of metabolites is a
critical step of both drug discovery and development research.
Objective: Compound 10, 4-((3-amino-4-methoxyphenyl) amino)-2H-coumarin, derived from coumarin.
Method: In this study, an efficient and sensitive method using Ultra High-Performance Liquid Chromatography
couple with Quadrupole Time of Flight tandem Mass Spectrometry (UHPLC/QTOF/
MS/MS) was successfully established and applied to identify the in vivo metabolites in plasma,
urine and feces samples of rats after intravenous administration of Compound 10 with a single dose of
10 mg/kg.
Result: A total of eight metabolites (including two phase I and six phase II metabolites) had been detected
or tentatively identified in plasma, urine and feces, indicating the prominent metabolic pathways
were glucuronidation, demethylation and hydroxylation. In addition, in order to understand the structure
of metabolites more accurately, synthesis strategy was used to confirm the metabolite M3.
Conclusion: The present study provides important information on the metabolism of Compound 10 in
vivo for the first time, which would be helpful for understanding the potential metabolic processes of
Compound 10 and paving the way for pharmacology and toxicology research.
