Approach for targeting Ras with small molecules that activate SOS-mediated nucleotide exchange

Proc Natl Acad Sci U S A. 2014 Mar 4;111(9):3401-6. doi: 10.1073/pnas.1315798111. Epub 2014 Feb 18.

Abstract

Aberrant activation of the small GTPase Ras by oncogenic mutation or constitutively active upstream receptor tyrosine kinases results in the deregulation of cellular signals governing growth and survival in ∼30% of all human cancers. However, the discovery of potent inhibitors of Ras has been difficult to achieve. Here, we report the identification of small molecules that bind to a unique pocket on the Ras:Son of Sevenless (SOS):Ras complex, increase the rate of SOS-catalyzed nucleotide exchange in vitro, and modulate Ras signaling pathways in cells. X-ray crystallography of Ras:SOS:Ras in complex with these molecules reveals that the compounds bind in a hydrophobic pocket in the CDC25 domain of SOS adjacent to the Switch II region of Ras. The structure-activity relationships exhibited by these compounds can be rationalized on the basis of multiple X-ray cocrystal structures. Mutational analyses confirmed the functional relevance of this binding site and showed it to be essential for compound activity. These molecules increase Ras-GTP levels and disrupt MAPK and PI3K signaling in cells at low micromolar concentrations. These small molecules represent tools to study the acute activation of Ras and highlight a pocket on SOS that may be exploited to modulate Ras signaling.

Publication types

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

MeSH terms

  • Chromatography, Liquid
  • Chromatography, Thin Layer
  • Crystallography, X-Ray
  • Fluorescence Polarization
  • HeLa Cells
  • Humans
  • Indoles / metabolism*
  • Ligands
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Models, Molecular*
  • Molecular Structure
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / metabolism*
  • Piperidines / metabolism*
  • Protein Conformation*
  • Proto-Oncogene Proteins p21(ras) / antagonists & inhibitors*
  • Proto-Oncogene Proteins p21(ras) / chemistry
  • SOS1 Protein / chemistry
  • SOS1 Protein / metabolism*

Substances

  • Indoles
  • Ligands
  • Multiprotein Complexes
  • Piperidines
  • SOS1 Protein
  • Proto-Oncogene Proteins p21(ras)

Associated data

  • PDB/4NYI
  • PDB/4NYJ
  • PDB/4NYM