Osimertinib azd9291

Parental PC9 cells were kindly provided by F. Hoffman-La Roche Ltd (Basel, Switzerland) with the authorization of Dr. Mayumi Ono (Kyushu University, Fukuoka, Japan). Parental 11–18 cells were kindly provided by Dr. Mayumi Ono. All cell culture materials were obtained from Biological Industries (Kibbutz Beit Haemek, Israel) or Gibco Life Technologies (Carlsbad, CA, USA). Cells were cultured in a humidified atmosphere with 5% CO₂ at 37 °C in RPMI1640 + 10% fetal bovine serum (FBS), 50 µg/mL penicillin-streptomycin and 2 mM l-Glutamine. EGFR inhibitors (erlotinib, gefitinib, afatinib, dacomitinib, and osimertinib (AZD9291)) and the Akt inhibitor AZD5363 were purchased from Selleck Chemicals (Houston, TX, USA). The Akt inhibitor GSK2141795 (Uprosertib) was purchased from Medchemexpress LLC (Monmouth Junction, NJ, USA). All drugs were dissolved in DMSO, aliquoted and kept at −20 °C.

H1299 and HEK 293T cells were grown in DMEM (Gibco, NY, USA) containing 10% FBS supplemented with penicillin (100 U/mL)/streptomycin (100 μg/mL). Human NSCLC cells H292, PC-9 or H1975 were grown in RPMI-1640 containing 10% FBS supplemented with penicillin (100 U/mL)/streptomycin (100 μg/mL) (Gibco, NY, USA). H1299 and H292 cells contain wild-type EGFR, H1975 or PC-9 cells bear EGFR^L858R/T790M or exon 19 deletion mutation, respectively. Cells were cultured at 37 °C in a humidified 5% CO₂ incubator.
MG132 (S2619), erlotinib (S1023), gefitinib (S1025), ganetespib (STA-9090) (S1159), nebivolol HCl (S1549), flibanserin (S3716), raltegravir (S2005), and osimertinib (AZD9291) (S7297) were purchased from Selleck Chemicals (Houston, USA). BC-1215 (SML1049), chloroquine diphosphate salt (C6628), and cycloheximide (C7698) were purchase from Sigma-Aldrich (St. Louis, USA). Recombinant Human EGF (236-EG) was purchase from R&D systems (Minnesota, USA).