title: A comprehensive understanding of thioTEPA metabolism in the mouse using UPLC-ESI-QTOFMS-based metabolomics.
abstract: ThioTEPA, an alkylating agent with anti-tumor activity, has been used as an effective anticancer drug since the 1950s. However, a complete understanding of how its alkylating activity relates to clinical efficacy has not been achieved, the total urinary excretion of thioTEPA and its metabolites is not resolved, and the mechanism of formation of the potentially toxic metabolites S-carboxymethylcysteine (SCMC) and thiodiglycolic acid (TDGA) remains unclear. In this study, the metabolism of thioTEPA in a mouse model was comprehensively investigated using ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOFMS) based-metabolomics. The nine metabolites identified in mouse urine suggest that thioTEPA underwent ring-opening, N-dechloroethylation, and conjugation reactions in vivo. SCMC and TDGA, two downstream thioTEPA metabolites, were produced from thioTEPA from two novel metabolites 1,2,3-trichloroTEPA (VII) and dechloroethyltrichloroTEPA (VIII). SCMC and TDGA excretion were increased about 4-fold and 2-fold, respectively, in urine following the thioTEPA treatment. The main mouse metabolites of thioTEPA in vivo were TEPA (II), monochloroTEPA (III) and thioTEPA-mercapturate (IV). In addition, five thioTEPA metabolites were detected in serum and all shared similar disposition. Although thioTEPA has a unique chemical structure which is not maintained in the majority of its metabolites, metabolomic analysis of its biotransformation greatly contributed to the investigation of thioTEPA metabolism in vivo, and provides useful information to understand comprehensively the pharmacological activity and potential toxicity of thioTEPA in the clinic.
authors.affiliation: Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20852, USA. lif3@mail.nih.gov
authors.forename: Fei
authors.initials: F
authors.lastname: Li
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Patterson
authors.forename: Constance C
authors.initials: CC
authors.lastname: Höfer
authors.forename: Kristopher W
authors.initials: KW
authors.lastname: Krausz
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Gonzalez
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Idle
mesh_terms: D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000097:blood / Q000378:metabolism* / Q000652:urine; D002233:Carbocysteine / Q000097:blood / Q000378:metabolism* / Q000652:urine; D002853:Chromatography, Liquid; D008297:Male; D055432:Metabolomics / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D015999:Multivariate Analysis; D021241:Spectrometry, Mass, Electrospray Ionization; D013864:Thioglycolates / Q000097:blood / Q000378:metabolism* / Q000652:urine; D013852:Thiotepa / Q000097:blood / Q000378:metabolism* / Q000652:urine
chemical_list: D018906:Antineoplastic Agents, Alkylating; D013864:Thioglycolates; C010518:thiodiacetic acid; D002233:Carbocysteine; D013852:Thiotepa