title: Quantitative determination of zebularine (NSC 309132), a DNA methyltransferase inhibitor, and three metabolites in murine plasma by high-performance liquid chromatography coupled with on-line radioactivity detection.
abstract: The metabolism of zebularine (NSC 309132), a novel agent that inhibits DNA methyltransferases, is still uncharacterized. To examine the in vivo metabolism of zebularine, an analytical method was developed and validated (based on FDA guidelines) to quantitate 2-[(14)C]-zebularine and its major metabolites in murine plasma. Zebularine and its metabolites uridine, uracil and dihydrouracil were baseline-separated based on hydrophilic interaction chromatography by using an amino column. The assay was accurate and precise in the concentration ranges of 5.0-100 microg/mL for zebularine, 2.5-50 microg/mL for uridine, 1.0-10 microg/mL for uracil and 0.5-5.0 microg/mL for dihydrouracil. This assay is being used to quantitate zebularine and its metabolites in ongoing pharmacokinetic studies of zebularine.
authors.affiliation: Molecular Therapeutics/Drug Discovery Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA. beumerjh@upmc.edu
authors.forename: Jan H
authors.initials: JH
authors.lastname: Beumer
authors.forename: Erin
authors.initials: E
authors.lastname: Joseph
authors.forename: Merrill J
authors.initials: MJ
authors.lastname: Egorin
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Covey
authors.forename: Julie L
authors.initials: JL
authors.lastname: Eiseman
mesh_terms: D000818:Animals; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003562:Cytidine / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors*; D004355:Drug Stability; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009862:Online Systems; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D013056:Spectrophotometry, Ultraviolet; D014498:Uracil / Q000031:analogs & derivatives / Q000097:blood; D014529:Uridine / Q000097:blood
chemical_list: D002250:Carbon Radioisotopes; C007419:dihydrouracil; D014498:Uracil; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside; D015254:DNA Modification Methylases; D014529:Uridine