PF-2341066

• Catalog No.: S1068
• CAS Number: 877399-52-5
• M. F.: C21H22Cl2FN5O
• M. W.: 450.3366832
• Storage: -20°C

SYNONYMS

• IUPAC name: 3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5-[1-(piperidin-4-yl)-1H-pyrazol-4-yl]pyridin-2-amine
• IUPAC Traditional name: crizotinib
• Synonyms: Crizotinib; PF-02341066

DATABASE IDS

• CAS Number: 877399-52-5

PROPERTIES

• Target: c-Met
• Target: ALK
• Salt Data: Free Base
• Solubility: DMSO
• Storage Condition: -20°C

DETAILS

Description (English)

Research Area

• Description: Cancer

Protocol

• Protocol: Kinase Assay ^[1]
• Biochemical kinase assays: c-Met catalytic activity is quantitated using a continuous-coupled spectrophotometric assay in which the time-dependent production of ADP by c-Met is determined by analysis of the rate of consumption of NADH. NADH consumption is measured by a decrease in absorbance at 340 nm by spectrophotometry at designated time points. To determine K_i values, PF-2341066 is introduced into test wells at various concentrations in the presence of assay reagents and incubated for 10 minutes at 37 °C. The assay is initiated by the addition of the c-Met enzyme.
• Protocol: Cell Assay ^[1]
• Cell Lines: GTL-16 gastric carcinoma cells and T47D breast carcinoma cells
• Concentrations: 0-256 nM
• Incubation Time: 1 hour
• Methods: Cells including GTL-16 gastric carcinoma cells and T47D breast carcinoma cells are seeded in 96-well plates in media supplemented with 10% fetal bovine serum (FBS) and transferred to serum-free media [with 0.04% bovine serum albumin (BSA)] after 24 hours. In experiments investigating ligand-dependent RTK phosphorylation, corresponding growth factors are added for up to 20 minutes. After incubation of cells with PF-2341066 for 1 hour and/or appropriate ligands for the designated times, cells are washed once with HBSS supplemented with 1 mM Na₃VO₄, and protein lysates are generated from cells. Subsequently, phosphorylation of selected protein kinases is assessed by a sandwich ELISA method using specific capture antibodies used to coat 96-well plates and a detection antibody specific for phosphorylated tyrosine residues. Antibody-coated plates are (a) incubated in the presence of protein lysates at 4 °C overnight; (b) washed seven times in 1% Tween 20 in PBS; (c) incubated in a horseradish peroxidase–conjugated anti–total-phosphotyrosine (PY-20) antibody (1:500) for 30 min; (d) washed seven times again; (e) incubated in 3,3^′,5,5^′-tetramethyl benzidine peroxidase substrate to initiate a colorimetric reaction that is stopped by adding 0.09 N H₂SO₄; and (f) measured for absorbance in 450 nm using a spectrophotometer.
• Protocol: Animal Study ^[1]
• Animal Models: Female or male nu/nu mice bearing NCI-H441,or DLD-1, or MDA-MB-231
• Doses: 12.5 mg/kg/day, 25 mg/kg/day, and 50 mg/kg/day
• Administration: Administered via p.o.

References

• References: [1] Zou HY, et al. Cancer Res. 2007, 67(9), 4408-4417.
• References: [2] Christensen JG, et al. Mol Cancer Ther. 2007, 6(12 Pt 1), 3314-3322.
• References: [3] Sampson ER, et al. J Bone Miner Res. 2011, 26(6), 1283-1294.
• References: [4] Cullinane C, et al. J Nucl Med. 2011, 52(8), 1261-1267.
• References: [5] Gong HC, et al. Int J Proteomics. 2011, 2011, 215496.

REFERENCES

• Zou HY, et al. Cancer Res. 2007, 67(9), 4408-4417.
• Christensen JG, et al. Mol Cancer Ther. 2007, 6(12 Pt 1), 3314-3322.
• Sampson ER, et al. J Bone Miner Res. 2011, 26(6), 1283-1294.
• Cullinane C, et al. J Nucl Med. 2011, 52(8), 1261-1267.
• Gong HC, et al. Int J Proteomics. 2011, 2011, 215496.