Non-targeted metabolomics-guided sildenafil metabolism study in human liver microsomes

•We revisited sildenafil metabolism in human liver microsomes using a metabolomics approach.•12 metabolites were identified in HLMs, and elucidated by MS2 fragmentation using HR-MS system.•7 Novel metabolites of sildenafil were identified in human liver microsomes.•The global metabolic picture of si...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2018-01, Vol.1072, p.86-93
Main Authors: Kim, Ju-Hyun, Jo, Jun Hyun, Seo, Kyung-ah, Hwang, Hayoung, Lee, Hye Suk, Lee, Sangkyu
Format: Article
Language:English
Subjects:
Cytochrome P450
Human liver microsomes
LC–MS/MS
Metabolite
Metabolomics
Sildenafil
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Summary:•We revisited sildenafil metabolism in human liver microsomes using a metabolomics approach.•12 metabolites were identified in HLMs, and elucidated by MS2 fragmentation using HR-MS system.•7 Novel metabolites of sildenafil were identified in human liver microsomes.•The global metabolic picture of sildenafil were performed by HR-MS and multivariate data analysis. Metabolomics combined with high-resolution mass spectrometry (HR-MS) and multivariate data analysis has broad applications in the study of xenobiotic metabolism. Although information about xenobiotic metabolism is essential to understand toxic mechanisms, pharmacokinetic parameters and excretion pathways, it is limited to predict all generated metabolites in biological fluids. Here, we revisited sildenafil metabolism in human liver microsomes using a metabolomics approach to achieve a global picture of sildenafil phase 1 metabolism. Finally, 12 phase 1 metabolites were identified in human liver microsomes; M1-M5 were previously known metabolites. The chemical structures of the novel metabolites were elucidated by MS2 fragmentation using an HR-MS system as follows: M6, reduced sildenafil; M7, N,N-deethylation and mono-oxidation; M8, demethanamine, N,N-deethylation and mono-hydroxylation; M9, demethanamine and N,N-deethylation; M10 and M11, mono-oxidation in the piperazine ring after N-demethylation; and M12, mono-oxidation. All metabolites, except M1, were formed by CYP3A4 and CYP3A5. In conclusion, we successfully updated the metabolic pathway of sildenafil in human liver, including 7 novel metabolites using metabolomics combined with HR-MS and multivariate data analysis.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2017.11.009