Abstract
Panobinostat (LBH589) is a novel pan-deacetylase inhibitor that is currently being evaluated in phase III clinical trials for treatment of Hodgkin's lymphoma and multiple myeloma. Under catalysis of recombinant human CYP3A4 and CYP2D6 coexpressed with human cytochrome P450 reductase in Escherichia coli JM109, five metabolites of panobinostat were produced via whole-cell biotransformation. The structures of the metabolites were elucidated with the spectroscopic methods mass spectrometry (MS) and NMR and revealed an oxidative cyclization of the ethyl-amino group to the methylindole moiety. The MS2 spectrum of the cyclized metabolite showed a base peak, where the closed ring is reopened and that, taken as sole base for structure proposals, would have lead to wrong conclusions. The metabolites were substantially less potent deacetylase inhibitors than the parent compound.
Footnotes
Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
ABBREVIATIONS:
- BMS-690514
- (3R,4R)-4-amino-1-((4-((3-methoxyphenyl)-amino)pyrrolo[2,1-f][1,2,4] triazin-5-yl)methyl)-3-piperidinol
- HDAC
- histone deacetylase
- ACN
- acetonitrile
- CID
- collision-induced dissociation
- COSY
- correlation spectroscopy
- P450
- cytochrome P450
- HMBC
- heteronuclear multiple bond correlation
- HR-MS
- high-resolution mass spectrometry
- HSQC
- heteronuclear single quantum coherence
- LB
- Luria-Bertani
- OD600
- optical density at 600 nm
- rh
- recombinant human
- TXI
- triple resonance, heteronuclei inverse detected
- MS/MS
- tandem mass spectrometry
- DMSO
- dimethyl sulfoxide
- LC-MS
- liquid chromatography-mass spectrometry
- 2D
- two-dimensional
- HPLC
- high-performance liquid chromatography
- HMBC
- heteronuclear multiple-bond correlation spectroscopy
- HEK
- human embryonic kidney
- PSE buffer
- phosphate - sucrose - EDTA buffer.
- Received October 31, 2011.
- Accepted February 16, 2012.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
DMD articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|