ASTRAZENECA: Octan-1-ol/water (pH7.4) distribution coefficent measured by a shake flask method described in J. Biomol. Screen. 2011, 16, 348-355. Experimental range -1.5 to 4.5
ASTRAZENECA: Solubility in pH7.4 buffer using solid starting material using the method described in J. Assoc. Lab. Autom. 2011, 16, 276-284. Experimental range 0.10 to 1500 uM
ASTRAZENECA: Intrinsic clearance measured in rat hepatocytes following incubation at 37C. Experimental range <3 to >150 microL/min/1E6 cells. Rapid Commun. Mass Spectrom. 2010, 24, 1730-1736
ASTRAZENECA: Intrinsic clearance measured in human hepatocytes following incubation at 37C. Experimental range <3 to >150 microL/min/1E6 cells. Rapid Commun. Mass Spectrom. 2010, 24, 1730-1736.
Biochemical Kinase Assay : Recombinant FGFR1 (2.5 nM), or FGFR4 (12 nM) was prepared as a mixture with substrate KKKSPGEYVNIEFG (SEQ ID NO:1) (20 μM, FGFR1 substrate); Poly [E,Y]4:1 (0.2 mg/ml, FGFR2,3,4 substrate)] in kinase reaction buffer (20 mM HEPES-HCl, pH 7.5, 10 mM MgCl2, 2 mM MnCl2, 1 mM EGTA, 0.02% Brij35, 0.1 mM Na3VO4, 0.02 mg/ml BSA, 2 mM DTT, and 1% DMSO). Compound was added to the enzyme/substrate mixture using acoustic technology and pre-incubated for 0, 15, or 60 minutes at room temperature. After compound pre-incubation, 33P-γ-ATP was added at a final concentration of 10 μM to initiate kinase reactions. Reactions were incubated for 120 minutes at room temperature.
Biochemical Kinase Assay : Recombinant FGFR1 (2.5 nM), or FGFR4 (12 nM) was prepared as a mixture with substrate KKKSPGEYVNIEFG (SEQ ID NO:1) (20 μM, FGFR1 substrate); Poly [E,Y]4:1 (0.2 mg/ml, FGFR2,3,4 substrate)] in kinase reaction buffer (20 mM HEPES-HCl, pH 7.5, 10 mM MgCl2, 2 mM MnCl2, 1 mM EGTA, 0.02% Brij35, 0.1 mM Na3VO4, 0.02 mg/ml BSA, 2 mM DTT, and 1% DMSO). Compound was added to the enzyme/substrate mixture using acoustic technology and pre-incubated for 0, 15, or 60 minutes at room temperature. After compound pre-incubation, 33P-γ-ATP was added at a final concentration of 10 μM to initiate kinase reactions. Reactions were incubated for 120 minutes at room temperature.
Inhibition of recombinant GST fused FGFR3 (unknown origin) using poly(EY) 4:1 as substrate in presence of [gamma-32P]ATP after 10 mins by scintillation counting method
In vivo inhibition of FGFR2 in rat assessed as reduction in bFGF-induced CCL2 production at 10 mg/kg, po measured at 5 hrs post dose relative to control
In vivo inhibition of FGFR2 in rat assessed as reduction in bFGF-induced CCL2 production at 10 mg/kg, po measured at 12 hrs post dose relative to control
Inhibition of SMO V404M mutant in gefitinib resistant human HCC827 cells assessed as decrease in GLI1 activity at 2 uM by GLI1 luciferase reporter assay relative to control
Inhibition of SMO V404M mutant in gefitinib resistant human HCC827 cells assessed as decrease in GLI1 activity at 60 uM by GLI1 luciferase reporter assay relative to control
Inhibition of SMO V404M mutant in gefitinib resistant human HCC827 cells assessed as decrease in GLI1 activity at 2 to 60 uM in presence of SMO agonist SAG by GLI1 luciferase reporter assay