title: Determination of GDC-0449, a small-molecule inhibitor of the Hedgehog signaling pathway, in human plasma by solid phase extraction-liquid chromatographic-tandem mass spectrometry.
abstract: To support clinical development, a solid phase extraction (SPE) liquid chromatographic-tandem mass spectrometry (LC-MS/MS) method for the determination of GDC-0449 concentrations in human plasma has been developed and validated. Samples (200 microl) were extracted using an Oasis MCX 10 mg 96-well SPE plate and the resulting extracts were analyzed using reverse-phase chromatography coupled with a turbo-ionspray interface. The method was validated over calibration curve range 5-5000 ng/mL. Quadratic regression and 1/x(2) weighing were used. Within-run relative standard deviation (%RSD) was within 10.1% and accuracy ranged from 88.6% to 109.0% of nominal. Between-run %RSD was within 8.6% and accuracy ranged from 92.4% to 105.3% of nominal. Extraction recovery of GDC-0449 was between 88.3% and 91.2% as assessed using quality control sample concentrations. GDC-0449 was stable in plasma for 315 days when stored at -70 degrees C and stable in reconstituted sample extracts for 117 h when stored at room temperature. Quantitative matrix effect/ion suppression experiment was performed and no significant matrix ion suppression was observed. This assay allows for the determination of GDC-0449 plasma concentrations over a sufficient time period to determine pharmacokinetic parameters at relevant clinical doses.
authors.affiliation: Genentech Inc., Drug Metabolism and Pharmacokinetics, MS 412A, 1 DNA Way, South San Francisco, CA 94080, United States. ding.xiao@gene.com
authors.forename: X
authors.initials: X
authors.lastname: Ding
authors.forename: B
authors.initials: B
authors.lastname: Chou
authors.forename: R A
authors.initials: RA
authors.lastname: Graham
authors.forename: S
authors.initials: S
authors.lastname: Cheeti
authors.forename: S
authors.initials: S
authors.lastname: Percey
authors.forename: L C
authors.initials: LC
authors.lastname: Matassa
authors.forename: S A
authors.initials: SA
authors.lastname: Reuschel
authors.forename: M
authors.initials: M
authors.lastname: Meng
authors.forename: S
authors.initials: S
authors.lastname: Liu
authors.forename: T
authors.initials: T
authors.lastname: Voelker
authors.forename: B L
authors.initials: BL
authors.lastname: Lum
authors.forename: P J
authors.initials: PJ
authors.lastname: Rudewicz
authors.forename: C E C A
authors.initials: CE
authors.lastname: Hop
mesh_terms: D000813:Anilides / Q000097:blood* / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D006801:Humans; D011725:Pyridines / Q000097:blood* / Q000493:pharmacokinetics; D015398:Signal Transduction; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000813:Anilides; D053823:Hedgehog Proteins; C538724:HhAntag691; D011725:Pyridines