A drug discovery platform to identify compounds that inhibit EGFR triple mutants

Nat Chem Biol. 2020 May;16(5):577-586. doi: 10.1038/s41589-020-0484-2. Epub 2020 Feb 24.

Abstract

Receptor tyrosine kinases (RTKs) are transmembrane receptors of great clinical interest due to their role in disease. Historically, therapeutics targeting RTKs have been identified using in vitro kinase assays. Due to frequent development of drug resistance, however, there is a need to identify more diverse compounds that inhibit mutated but not wild-type RTKs. Here, we describe MaMTH-DS (mammalian membrane two-hybrid drug screening), a live-cell platform for high-throughput identification of small molecules targeting functional protein-protein interactions of RTKs. We applied MaMTH-DS to an oncogenic epidermal growth factor receptor (EGFR) mutant resistant to the latest generation of clinically approved tyrosine kinase inhibitors (TKIs). We identified four mutant-specific compounds, including two that would not have been detected by conventional in vitro kinase assays. One of these targets mutant EGFR via a new mechanism of action, distinct from classical TKI inhibition. Our results demonstrate how MaMTH-DS is a powerful complement to traditional drug screening approaches.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Non-Small-Cell Lung / drug therapy
  • Carcinoma, Non-Small-Cell Lung / genetics
  • Cell Line
  • Cell Line, Tumor
  • DNA Nucleotidyltransferases / genetics
  • Drug Discovery
  • Drug Resistance, Neoplasm / genetics
  • ErbB Receptors / antagonists & inhibitors
  • ErbB Receptors / genetics
  • Genes, Reporter
  • High-Throughput Screening Assays / methods*
  • Humans
  • Luciferases / genetics
  • Lung Neoplasms / drug therapy
  • Lung Neoplasms / genetics
  • Mutation
  • Phosphorylation / drug effects
  • Protein Kinase Inhibitors / pharmacology*
  • Reproducibility of Results
  • Small Molecule Libraries / pharmacology
  • Staurosporine / analogs & derivatives
  • Staurosporine / pharmacology

Substances

  • Protein Kinase Inhibitors
  • Small Molecule Libraries
  • Luciferases
  • EGFR protein, human
  • ErbB Receptors
  • DNA Nucleotidyltransferases
  • FLP recombinase
  • Staurosporine
  • midostaurin