Unprotected peptide macrocyclization and stapling via a fluorine-thiol displacement reaction

Nat Commun. 2022 Jan 17;13(1):350. doi: 10.1038/s41467-022-27995-5.

Abstract

We report the discovery of a facile peptide macrocyclization and stapling strategy based on a fluorine thiol displacement reaction (FTDR), which renders a class of peptide analogues with enhanced stability, affinity, cellular uptake, and inhibition of cancer cells. This approach enabled selective modification of the orthogonal fluoroacetamide side chains in unprotected peptides in the presence of intrinsic cysteines. The identified benzenedimethanethiol linker greatly promoted the alpha helicity of a variety of peptide substrates, as corroborated by molecular dynamics simulations. The cellular uptake of benzenedimethanethiol stapled peptides appeared to be universally enhanced compared to the classic ring-closing metathesis (RCM) stapled peptides. Pilot mechanism studies suggested that the uptake of FTDR-stapled peptides may involve multiple endocytosis pathways in a distinct pattern in comparison to peptides stapled by RCM. Consistent with the improved cell permeability, the FTDR-stapled lead Axin and p53 peptide analogues demonstrated enhanced inhibition of cancer cells over the RCM-stapled analogues and the unstapled peptides.

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

  • Amino Acid Sequence
  • Axin Protein / chemistry
  • Cell Membrane Permeability
  • Cell-Penetrating Peptides / chemistry
  • Cross-Linking Reagents / chemistry
  • Cyclization
  • Fluorine / chemistry*
  • HEK293 Cells
  • Humans
  • Macrocyclic Compounds / chemistry*
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Dynamics Simulation
  • Peptides / chemistry*
  • Sulfhydryl Compounds / chemistry*
  • Thermodynamics
  • Tumor Suppressor Protein p53 / chemistry

Substances

  • Axin Protein
  • Cell-Penetrating Peptides
  • Cross-Linking Reagents
  • Macrocyclic Compounds
  • Peptides
  • Sulfhydryl Compounds
  • Tumor Suppressor Protein p53
  • Fluorine