Biochemical characterization of EphA2 antagonists with improved physico-chemical properties by cell-based assays and surface plasmon resonance analysis

Biochem Pharmacol. 2016 Jan 1:99:18-30. doi: 10.1016/j.bcp.2015.10.006. Epub 2015 Oct 14.

Abstract

Amino acid conjugates of lithocholic acid (LCA) have been recently described as effective disruptors of the EphA2-ephrin-A1 interaction able to inhibit EphA2 phosphorylation in intact cells and thus able to block prometastatic responses such as cellular retraction and angiogenesis. However, these LCA-based compounds were significantly more potent at disrupting the EphA2-ephrin-A1 interaction than at blocking phenotype responses in cells, which might reflect an unclear mechanism of action or a metabolic issue responsible for a reduction of the compound concentration at the cell's surface. Through the synthesis of new compounds and their examination by a combination of cell-based assays and real-time interaction analysis by surface plasmon resonance, we showed at molecular level that l-tryptophan conjugates of lithocholic acid disrupt EphA2-ephrin-A1 interaction by targeting the EphA 2 receptor and that the presence of a polar group in position 3 of steroid scaffold is a key factor to increase the effective concentration of the compounds in cancer cell lines.

Keywords: Antagonist; Bile acid; Eph kinases; Ephrin; PPI.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Chemical Phenomena
  • Humans
  • Lithocholic Acid / analogs & derivatives
  • Lithocholic Acid / chemistry
  • Lithocholic Acid / metabolism
  • Lithocholic Acid / pharmacology
  • Molecular Docking Simulation / methods
  • Protein Kinase Inhibitors / chemistry*
  • Protein Kinase Inhibitors / metabolism*
  • Protein Kinase Inhibitors / pharmacology
  • Protein Structure, Secondary
  • Receptor, EphA2 / antagonists & inhibitors*
  • Receptor, EphA2 / metabolism*
  • Structure-Activity Relationship
  • Surface Plasmon Resonance / methods*
  • Tryptophan / analogs & derivatives
  • Tryptophan / chemistry
  • Tryptophan / metabolism
  • Tryptophan / pharmacology

Substances

  • Protein Kinase Inhibitors
  • UniPR129
  • Lithocholic Acid
  • Tryptophan
  • Receptor, EphA2