Formation of A Novel Purine Metabolite through CYP3A4 Bioactivation and Glutathione Conjugation

Author(s): Julius L. Apuy, Cathie Xiang, Sarah Franc, Sayee G. Hegde, Robert Hubbard, Jingjing Zhao, Mehran F. Moghaddam

Journal Name: Drug Metabolism Letters

Volume 10 , Issue 2 , 2016

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Graphical Abstract:


Background: The study of novel sites of metabolism is important in understanding new mechanisms of biotransformation of a particular moiety by metabolic enzymes. This information is valuable in designing metabolically-stable compounds with drug-like properties. It may also provide insights into the existence of active and reactive metabolites.

Methods: We utilized small scale incubations to generate adequate amounts of the metabolite of interest. After purification, LC-MS/MS and Proton Nuclear Magnetic Resonance (1H-NMR) were utilized to unequivocally assign the novel site of glutathione conjugation on the purine ring system.

Results: A proposed novel site of glutathione conjugation was investigated on a diaminopurine-containing molecule. It was demonstrated that the formation of the glutathione conjugate at the C-6 position of the purine ring system was due to the bioactivation of the compound to a di-imine intermediate by CYP3A4, followed by the nucleophilic addition of glutathione.

Conclusion: S-glutathionylation at C-6 position of a purine was proven unequivocally. This previously unreported mechanism constitutes a novel biotransformation for purines.

Keywords: Biotransformation, CYP, drug discovery, glutathione, mass spectrometry, metabolite identification, metabolism, novel, NMR, purine, structural elucidation.

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Article Details

Year: 2016
Published on: 13 June, 2016
Page: [144 - 150]
Pages: 7
DOI: 10.2174/1872312810666160511150558

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