A high-throughput cell-based screening for L858R/T790M mutant epidermal growth factor receptor inhibitors.
A high-throughput 32D(L858R/T790M) cell-based assay to identify inhibitors of the L858R/T790M mutant epidermal growth factor receptor (EGFR) pathway was established. After screening, ten hits from among 60,000 compounds in our in-house compound library were initially identified. In the secondary assays, one hit, 1-[2-(decyloxy)-2-oxoethyl]-3-methyl-2-[(4-methylphenoxy) methyl]-1H-benzimidazol-3-ium, was confirmed to directly inhibit the kinase activity of recombinant L858R/T790M EGFR and the phosphorylation of EGFR-L858R/T790M in gefitinib-resistant H1975 cells. Thus, this high-throughput assay system may be useful for identifying novel inhibitors which suppress mutant EGFR-T790M signalling and for overcoming T790M-mediated acquired resistance for future anticancer drug discovery.
Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, No.35, Keyan Road, Zhunan Town, Miaoli County, 35053, Taiwan, ROC.
Wen-Hsing
WH
Lin
Jen-Shin
JS
Song
Tzu-Wen
TW
Lien
Chun-Yu
CY
Chang
Szu-Huei
SH
Wu
Yu-Wen
YW
Huang
Teng-Yuan
TY
Chang
Ming-Yu
MY
Fang
Kuei-Jung
KJ
Yen
Chun-Hwa
CH
Chen
Chang-Ying
CY
Chu
Hsing-Pang
HP
Hsieh
Yi-Rong
YR
Chen
Yu-Sheng
YS
Chao
John T-A
JT
Hsu
D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000235:genetics; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000187:drug effects / Q000235:genetics; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000235:genetics*; D000077156:Gefitinib; D057166:High-Throughput Screening Assays; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000235:genetics; D009154:Mutation / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects
D047428:Protein Kinase Inhibitors; D011799:Quinazolines; C017867:epidermal growth inhibitors; D004815:Epidermal Growth Factor; C512478:EGFR protein, human; D066246:ErbB Receptors; D000077156:Gefitinib