title: Rapid and sensitive LC/MS/MS analysis of the novel tyrosine kinase inhibitor ZD6474 in mouse plasma and tissues.
abstract: ZD6474 (N-(4-Bromo-2-fluorophenyl)-6-methoxy-7-[(1-methylpiperidin-4-yl)methoxy] quinazolin-4-amine) is a tyrosine kinase inhibitor with anti-angiogenic and anti-tumor activity that is currently undergoing human trials for cancer treatment. Pharmacokinetic studies in animal models are an important component in clinical development of this agent to relate pre-clinical models to patient treatment. A liquid chromatography tandem mass spectrometry method was developed for the determination of ZD6474 levels in mouse plasma and tissues. Plasma (0.05 mL) and tissue homogenates (0.1 mL of 10 mg/mL) were extracted under alkaline conditions with ethyl acetate:pentane (1:1, v/v) after addition of the internal standard (trazodone, 2-[3-[4-(3-chlorophenyl)-1-piperazinyl]propyl]-1,2,4-triazolo[4,3-a]pyridine-3(2H)-one). Separation was achieved on a C18, 50 mm x 2 mm column with quantitation by internal standard reference and multiple reaction monitoring of the ion transitions m/z 475-->112 (ZD6474) and m/z 372-->176 (trazodone). The calibration curve was linear from a range spanning 20-20,000 ng/mL in plasma and 10-320 ng/mg in tissue homogenates. Mean recoveries from plasma and tissue homogenates were 88 and 90%, respectively. The accuracy in plasma was 88% at the lower limit of quantitation (20 ng/mL with a 50 microL plasma sample) with high precision (R.S.D.%<10%). Assay performance in liver and other tissue homogenates is also reported. The assay was applied to a pharmacokinetic study in mice to determine dosing schedules that would approximate therapeutic ZD6474 levels determined in humans.
authors.affiliation: School of Pharmacy, Department of Pharmaceutical Sciences, University of Colorado Health Sciences Center, 4200 East 9th Ave., Denver, CO 80262, USA.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Zirrolli
authors.forename: Erica L
authors.initials: EL
authors.lastname: Bradshaw
authors.forename: Michael E
authors.initials: ME
authors.lastname: Long
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Gustafson
mesh_terms: D000085:Acetates / Q000032:analysis; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002138:Calibration; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002845:Chromatography / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D007477:Ions; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D008956:Models, Chemical; D009369:Neoplasms / Q000188:drug therapy*; D010420:Pentanes / Q000032:analysis; D010880:Piperidines / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011799:Quinazolines / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014018:Tissue Distribution; D014196:Trazodone / Q000032:analysis
chemical_list: D000085:Acetates; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D007477:Ions; D010420:Pentanes; D010880:Piperidines; D011799:Quinazolines; C033353:pentane; C007650:ethyl acetate; D011505:Protein-Tyrosine Kinases; D014196:Trazodone; C452423:vandetanib