A Discovery Strategy for Selective Inhibitors of c-Src in Complex with the Focal Adhesion Kinase SH3/SH2-binding Region

Chem Biol Drug Des. 2015 Aug;86(2):144-55. doi: 10.1111/cbdd.12473. Epub 2014 Nov 28.


The c-Src tyrosine kinase co-operates with the focal adhesion kinase to regulate cell adhesion and motility. Focal adhesion kinase engages the regulatory SH3 and SH2 domains of c-Src, resulting in localized kinase activation that contributes to tumor cell metastasis. Using assay conditions where c-Src kinase activity required binding to a tyrosine phosphopeptide based on the focal adhesion kinase SH3-SH2 docking sequence, we screened a kinase-biased library for selective inhibitors of the Src/focal adhesion kinase peptide complex versus c-Src alone. This approach identified an aminopyrimidinyl carbamate compound, WH-4-124-2, with nanomolar inhibitory potency and fivefold selectivity for c-Src when bound to the phospho-focal adhesion kinase peptide. Molecular docking studies indicate that WH-4-124-2 may preferentially inhibit the 'DFG-out' conformation of the kinase active site. These findings suggest that interaction of c-Src with focal adhesion kinase induces a unique kinase domain conformation amenable to selective inhibition.

Keywords: SH2 domain; SH3 domain; Src kinase; cancer drug discovery; focal adhesion kinase; kinase inhibitors.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • CSK Tyrosine-Protein Kinase
  • Crystallography, X-Ray
  • Drug Evaluation, Preclinical / methods
  • Focal Adhesion Kinase 1 / antagonists & inhibitors*
  • Focal Adhesion Kinase 1 / chemistry
  • Focal Adhesion Kinase 1 / metabolism
  • Humans
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Binding
  • Protein Kinase Inhibitors / chemistry
  • Protein Kinase Inhibitors / pharmacology*
  • Protein Structure, Tertiary
  • src Homology Domains
  • src-Family Kinases / antagonists & inhibitors*
  • src-Family Kinases / chemistry
  • src-Family Kinases / metabolism


  • Protein Kinase Inhibitors
  • CSK Tyrosine-Protein Kinase
  • Focal Adhesion Kinase 1
  • PTK2 protein, human
  • src-Family Kinases
  • CSK protein, human