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In vitro and in vivo metabolite identification of a novel benzimidazole compound ZLN005 by liquid chromatography/tandem mass spectrometry
Author(s) -
Sun Wenchao,
Nguyen Khoa Dinh,
Fitch William L.,
Banister Samuel D.,
Tang Hongxiang,
Zhang Xiaolan,
Yu Lewis,
Engleman Edgar G.,
Rajadas Jayakumar
Publication year - 2018
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.8060
Subject(s) - chemistry , glucuronide , in vivo , metabolite , chromatography , tandem mass spectrometry , in vitro , mass spectrometry , metabolic pathway , biochemistry , metabolism , microbiology and biotechnology , biology
Rationale A novel benzimidazole compound ZLN005 was previously identified as a transcriptional activator of peroxisome proliferator‐activated receptor γ coactivator 1α (PGC‐1α) in certain metabolic tissues. Upregulation of PGC‐1α by ZLN005 has been shown to have a beneficial effect in a diabetic mouse model and in a coronary artery disease model in vitro . ZLN005 could also have therapeutic potential in neurodegenerative diseases involving down‐regulation of PGC‐1α. Given the phenotypic efficacy of ZLN005 in several animal models of human disease, its metabolic profile was investigated to guide the development of novel therapeutics using ZLN005 as the lead compound. Methods ZLN005 was incubated with both rat and human liver microsomes and S9 fractions to identify in vitro metabolites. Urine from rats dosed with ZLN005 was used to identify in vivo metabolites. Extracted metabolites were analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS) using a hybrid linear ion trap triple quadrupole mass spectrometer in full scan, enhanced product ion scan, neutral loss scan and precursor scan modes. Metabolites in plasma and brain of ZLN005‐treated rats were also profiled using multiple reaction monitoring. Results Identified in vitro transformations of ZLN005 include mono‐ and dihydroxylation, further oxidation to carboxylic acids, and mono‐ O ‐glucuronide and sulfate conjugation to hydroxy ZLN005 as well as glutathione conjugation. Identified in vivo metabolites are mainly glucuronide and sulfate conjugates of dihydroxyl, carboxyl, and hydroxy acid of the parent compound. The parent compound as well as several major phase I metabolites were found in rat plasma and brain. Conclusions Using both in vitro and in vivo methods, we elucidated the metabolic pathway of ZLN005. Phase I metabolites with hydroxylation and carboxylation, as well as phase II metabolites with glucuronide, sulfate and glutathione conjugation, were identified.