AZD8330

461, 23

Female nude rats (NIH rnu/rnu) with Calu-6 cells, nude rats with SW620 cells

A Phase I clinical trial for AZD8330 has been completed in the treatment of cancer.

0.3 mg/kg, 1 mg/kg

7 nM, 7 nM, 7 nM, 7 nM, 7 nM, 7 nM

In a Calu-6 rat xenograft pharmacokinetic/pharmacodynamic (PK/PD) model a single, 1.25 mg/kg oral dose of AZD8330 inhibits ERK phosphorylation by > 90% for between 4 and 8 hours. Doses as low as 0.4 mg/kg once daily are sufficient for > 80% tumor growth inhibition in the Calu-6 nude rat xenograft model. In the Calu-6 model, AZD8330 inhibits tumor growth in a dose-dependent fashion, at 0.3 mg/kg and 1.0 mg/kg once daily. [1]

[2], MEK1 enzymatic assays, NH2-terminal hexahistidine tagged, constitutively active MEK1 (S218D, S222D deltaR4F) is expressed in baculovirus-infected Hi5 insect cells and purified by immobilized metal affinity chromatography, ion exchange, and gel filtration. The activity of MEK1 is assessed by measuring the incorporation of [gamma- 33P]phosphate from [gamma-33P]ATP onto ERK2. The assay is carried out in a 96-well polypropylene plate with an incubation mixture (100 uL) composed of 25 mM HEPES (pH 7.4), 10 mM MgCl2, 5 mM beta-glycerolphosphate, 100 uM sodium orthovanadate, 5 mM DTT, 5 nM MEK1, 1 uM ERK2, and 0 to 80 nM AZD8330 (final concentration of 1% DMSO). The reactions are initiated by the addition of 10 uM ATP (with 0.5 uC k[gamma-33P]ATP/well) and incubated at room temperature for 45 min. An equal volume of 25% trichloracetic acid is added to stop the reaction and precipitate the proteins. Precipitated proteins are trapped onto glass fiber B filter plates, excess labeled ATP is washed off with 0.5% phosphoric acid, and radioactivity is counted in a liquid scintillation counter. ATP dependence is determined by varying the amount of ATP in the reaction mixture. The data are globally fitted.

DMSO 92 mg/mL, Water <1 mg/mL, Ethanol 92 mg/mL

2-(2-fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-1, 5-dimethyl-6-oxo-1, 6-dihydropyridine-3-carboxamide