α/β-Peptide Foldamers Targeting Intracellular Protein-Protein Interactions with Activity in Living Cells

J Am Chem Soc. 2015 Sep 9;137(35):11365-75. doi: 10.1021/jacs.5b05896. Epub 2015 Aug 28.

Abstract

Peptides can be developed as effective antagonists of protein-protein interactions, but conventional peptides (i.e., oligomers of l-α-amino acids) suffer from significant limitations in vivo. Short half-lives due to rapid proteolytic degradation and an inability to cross cell membranes often preclude biological applications of peptides. Oligomers that contain both α- and β-amino acid residues ("α/β-peptides") manifest decreased susceptibility to proteolytic degradation, and when properly designed these unnatural oligomers can mimic the protein-recognition properties of analogous "α-peptides". This report documents an extension of the α/β-peptide approach to target intracellular protein-protein interactions. Specifically, we have generated α/β-peptides based on a "stapled" Bim BH3 α-peptide, which contains a hydrocarbon cross-link to enhance α-helix stability. We show that a stapled α/β-peptide can structurally and functionally mimic the parent stapled α-peptide in its ability to enter certain types of cells and block protein-protein interactions associated with apoptotic signaling. However, the α/β-peptide is nearly 100-fold more resistant to proteolysis than is the parent stapled α-peptide. These results show that backbone modification, a strategy that has received relatively little attention in terms of peptide engineering for biomedical applications, can be combined with more commonly deployed peripheral modifications such as side chain cross-linking to produce synergistic benefits.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Apoptosis Regulatory Proteins / chemistry
  • Bcl-2-Like Protein 11
  • Cell Membrane Permeability
  • Cell Survival / drug effects
  • Cell-Penetrating Peptides / chemistry*
  • Cell-Penetrating Peptides / metabolism
  • Cell-Penetrating Peptides / pharmacology*
  • Cytochromes c / metabolism
  • HCT116 Cells
  • Humans
  • Intracellular Space / drug effects*
  • Intracellular Space / metabolism*
  • Membrane Proteins / chemistry
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Peptide Hydrolases / metabolism
  • Protein Binding / drug effects
  • Protein Folding*
  • Protein Stability
  • Protein Structure, Tertiary
  • Proteolysis
  • Proto-Oncogene Proteins / chemistry

Substances

  • Apoptosis Regulatory Proteins
  • BCL2L11 protein, human
  • Bcl-2-Like Protein 11
  • Bcl2l11 protein, mouse
  • Cell-Penetrating Peptides
  • Membrane Proteins
  • Proto-Oncogene Proteins
  • Cytochromes c
  • Peptide Hydrolases