Identification of phenylbutyrate-generated metabolites in Huntington disease patients using parallel liquid chromatography/electrochemical array/mass spectrometry and off-line tandem mass spectrometry.
Oral sodium phenylbutyrate (SPB) is currently under investigation as a histone deacetylation (HDAC) inhibitor in Huntington disease (HD). Ongoing studies indicate that symptoms related to HD genetic abnormalities decrease with SPB therapy. In a recently reported safety and tolerability study of SPB in HD, we analyzed overall chromatographic patterns from a method that employs gradient liquid chromatography with series electrochemical array, ultraviolet (UV), and fluorescence (LCECA/UV/F) for measuring SPB and its metabolite phenylacetate (PA). We found that plasma and urine from SPB-treated patients yielded individual-specific patterns of approximately 20 metabolites that may provide a means for the selection of subjects for extended trials of SPB. The structural identification of these metabolites is of critical importance because their characterization will facilitate understanding the mechanisms of drug action and possible side effects. We have now developed an iterative process with LCECA, parallel LCECA/LCMS, and high-performance tandem MS for metabolite characterization. Here we report the details of this method and its use for identification of 10 plasma and urinary metabolites in treated subjects, including indole species in urine that are not themselves metabolites of SPB. Thus, this approach contributes to understanding metabolic pathways that differ among HD patients being treated with SPB.
Ebbel
Erika N
EN
Center for Biomedical Mass Spectrometry and Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
Leymarie
Nancy
N
Schiavo
Susan
S
Sharma
Swati
S
Gevorkian
Sona
S
Hersch
Steven
S
Matson
Wayne R
WR
Costello
Catherine E
CE
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056572:Histone Deacetylase Inhibitors / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D006655:Histone Deacetylases / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006816:Huntington Disease / Q000188:drug therapy / Q000378:metabolism*; D010654:Phenylbutyrates / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D056572:Histone Deacetylase Inhibitors; D010654:Phenylbutyrates; D006655:Histone Deacetylases