AZD8330

• Catalog No.: S2134
• CAS Number: 869357-68-6
• M. F.: C16H17FIN3O4
• M. W.: 461.2267532
• Storage: -20°C

SYNONYMS

• IUPAC name: 2-[(2-fluoro-4-iodophenyl)amino]-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide
• IUPAC Traditional name: 2-[(2-fluoro-4-iodophenyl)amino]-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxopyridine-3-carboxamide
• Synonyms: ARRY-704; ARRY-424704; AZD-8330

DATABASE IDS

• CAS Number: 869357-68-6

PROPERTIES

• Target: MEK
• Salt Data: Free Base
• Storage Condition: -20°C

DETAILS

Description (English)

Research Area

• Description: Cancer

Biological Activity

• Description: AZD8330 (ARRY-424704) is a MEK 1/2 inhibitor with IC50 of 7 nM.
• Targets: MEK 1/2; ERK phosphorylation
• IC50: 7 nM; 0.4 nM ^[1]
• In Vitro: AZD8330 potently and strongly inhibits MEK 1/2. AZD8330 has no inhibitory activity against over 200 other kinases including at concentrations up to 10 μM. AZD8330 demonstrates sub-nanomolar potency in mechanistic (pERK) and low to sub-nanomolar potency in functional (proliferation) assays in MEK 1/2 inhibitor sensitive cell lines. ^[1]
• In Vivo: 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]
• Clinical Trials: A Phase I clinical trial for AZD8330 has been completed in the treatment of cancer.

Protocol

• Protocol: Kinase Assay ^[2]
• MEK1 enzymatic assays: NH2-terminal hexahistidine tagged, constitutively active MEK1 (S218D, S222D ΔR4F) 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 [γ- ^33P]phosphate from [γ-^33P]ATP onto ERK2. The assay is carried out in a 96-well polypropylene plate with an incubation mixture (100 μL) composed of 25 mM HEPES (pH 7.4), 10 mM MgCl2, 5 mM β-glycerolphosphate, 100 μM sodium orthovanadate, 5 mM DTT, 5 nM MEK1, 1 μM ERK2, and 0 to 80 nM AZD8330 (final concentration of 1% DMSO). The reactions are initiated by the addition of 10 μM ATP (with 0.5 μC k[γ-^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.
• Protocol: Cell Assay ^[1]
• Cell Lines: Malme-3M melanoma cells
• Concentrations: ~10 μM
• Incubation Time: 1 hour
• Methods: Malme-3M melanoma cells are plated in 96-wells and treated with various concentrations of AZD8330 for 1 hour at 37 °C. The cells are fixed, permeabilized, and incubated with an anti-phospho-ERK antibody and an anti-ERK 1/2 antibody. Plates are washed and fluorescently-labeled secondary antibodies are added. Plates are analyzed on a LICOR fluorescence imager. The pERK signal is normalized to the total ERK signal.
• Protocol: Animal Study ^[1]
• Animal Models: Female nude rats (NIH rnu/rnu) with Calu-6 cells, nude rats with SW620 cells
• Formulation: 0.5% HPMC-0.1% Tween
• Doses: 0.3 mg/kg, 1 mg/kg
• Administration: Oral administration

References

• References: [1] Wallace EM, et al. AACR Annual Meeting, 2009, Abst 3696.
• References: [2] Yeh TC, et al. Clin Cancer Res, 2007, 13(5), 1576-1583.

REFERENCES

• Wallace EM, et al. AACR Annual Meeting, 2009, Abst 3696.
• Yeh TC, et al. Clin Cancer Res, 2007, 13(5), 1576-1583.