Abstract
UTL-5b is an anti-inflammatory and anti-arthritic small-molecule tumor necrosis factor-alpha inhibitor and a structural analogue of the anti-arthritic drug, leflunomide. Leflunomide is known to be metabolized to teriflunomide, but the metabolites of UTL-5b have not been reported. The objective of this study was to investigate whether UTL-5b has a similar metabolic behavior as leflunomide. Preliminary studies showed that when exposed to microsomes in vitro with or without NADPH, UTL-5b disappeared within 30 min. To further investigate the microsomal metabolism, liquid chromatography-ultraviolet (LC-UV) and LC/tandem mass spectrometry (LC–MS/MS) were employed to, respectively, monitor the microsomal metabolites and identify the structure of the metabolites using LC-full scan MS and LC combined with multiple-ion monitoring MS. Fragmentation determination was analyzed by two types of scans: product ion scans and precursor ion scan. The in vitro microsomal treatment of UTL-5b resulted in two major metabolites: 5-methylisoxazole-3-carboxylic acid and 2-chloroaniline. Thus, the in vitro metabolic behavior of UTL-5b appears to be different from that of leflunomide in that the isoxazole ring is cleaved.
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Abbreviations
- DHODH:
-
Dihydroorotate dehydrogenase
- ESI:
-
Electrospray ionization
- HPLC:
-
High performance liquid chromatography
- LC:
-
Liquid chromatography
- LC–MS/MS:
-
Liquid chromatography/tandem mass spectrometry
- MIM:
-
Multiple ion monitoring
- MS:
-
Mass spectrometry/mass spectrometer
- NMR:
-
Nuclear magnetic resonance
- Rt:
-
Retention time
- SIR:
-
Single ion recording
- TIC:
-
Total ion current
- TNF-α:
-
Tumor necrosis factor-alpha
- UV:
-
Ultraviolet
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Acknowledgments
This work was supported by SBIR grant 1R43 AR052193 from the National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Department of Heath and Human Services, USA.
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Shaw, J., Shay, B., Jiang, J. et al. Metabolism studies of a small-molecule tumor necrosis factor-alpha (TNF-α) inhibitor, UTL-5b (GBL-5b). Eur J Drug Metab Pharmacokinet 37, 83–89 (2012). https://doi.org/10.1007/s13318-011-0072-7
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DOI: https://doi.org/10.1007/s13318-011-0072-7