Pelitinib (EKB-569)

Cell experiment:

For experiments using cells in culture, A431 cells or 3T3/c-erbB-2 cells over-expressing c-erbB2 are are treated with various concentrations of EKB-569 for 2.75 h before co-incubation with 100 ng/mL EGF (A431 cells) or no growth factor (3T3/c-erbB-2 cells) for 0.25 h. Cells are ished twice with cold phosphate-buffered saline (PBS) before adding to lysis buffer for 20 min on ice, before immunoprecipitation and SDS–PAGE-immunoblotting[1].

Animal experiment:

Mice: For in vivo experiments, athymic nu/nu female mice are implanted subcutaneously with 5×106 A431 tumor cells. When tumors reach a mass of 200-300 mg, animals are treated with a single dose of 10 mg/kg EKB-569 in pH 2.0 water per gavage. Tumors from control and drug-treated animals are excised and minced into 1-mm pieces for anlysis[1].

References:

[1]. Torrance CJ, et al. Combinatorial chemoprevention of intestinal neoplasia. Nat Med. 2000 Sep;6(9):1024-8.
[2]. Nunes M, et al. Phosphorylation of extracellular signal-regulated kinase 1 and 2, protein kinase B, and signal transducer and activator of transcription 3 are differently inhibited by an epidermal growth factor receptor inhibitor, EKB-569, in tumor cells and normal human keratinocytes. Mol Cancer Ther. 2004 Jan;3(1):21-7.