A high-throughput cell-based screening for L858R/T790M mutant epidermal growth factor receptor inhibitors

Anticancer Res. 2012 Jan;32(1):147-51.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Non-Small-Cell Lung / drug therapy*
  • Carcinoma, Non-Small-Cell Lung / genetics
  • Cell Line, Tumor
  • Drug Resistance, Neoplasm / drug effects
  • Drug Resistance, Neoplasm / genetics
  • Epidermal Growth Factor / pharmacology*
  • ErbB Receptors / antagonists & inhibitors*
  • ErbB Receptors / genetics*
  • Gefitinib
  • High-Throughput Screening Assays
  • Humans
  • Lung Neoplasms / drug therapy*
  • Lung Neoplasms / genetics
  • Mutation / drug effects*
  • Phosphorylation / drug effects
  • Protein Kinase Inhibitors / pharmacology
  • Quinazolines / pharmacology*
  • Signal Transduction / drug effects

Substances

  • Protein Kinase Inhibitors
  • Quinazolines
  • epidermal growth inhibitors
  • Epidermal Growth Factor
  • EGFR protein, human
  • ErbB Receptors
  • Gefitinib