Cell-Permeable Peptide Inhibitors of the p53-hDM2 Interaction via Foldamer Helix Mimicry and Bis-Thioether Stapling

J Med Chem. 2025 Jan 9;68(1):236-246. doi: 10.1021/acs.jmedchem.4c01762. Epub 2024 Dec 24.

Abstract

Combining helical foldamers with α-peptides can produce α-helix mimetics with a reduced peptide character and enhanced resistance to proteolysis. Previously, we engineered a hybrid peptide-oligourea sequence replicating the N-terminal α-helical domain of p53 to achieve high affinity binding to hDM2. Here, we further advance this strategy by combining the foldamer approach with side chain cross-linking to create more constrained cell-permeable inhibitors capable of effectively engaging the target within cells. Starting from the crystal structure of the foldamer-hDM2 complex, we identified specific sites suitable for stapling, and generated a small library of macrocyclic foldamer-peptide hybrids. The most promising binders were subsequently optimized for cellular uptake and tested in a cellular assay. We observed that the introduction of a short segment of positively charged residues at the N-terminus of the sequence led to inhibitors that exhibited cytotoxic activity independently of p53. In contrast, neutral acetylated peptide-foldamer macrocycles demonstrated activity in a p53-dependent manner.

MeSH terms

  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / metabolism
  • Antineoplastic Agents / pharmacology
  • Cell Line, Tumor
  • Cell-Penetrating Peptides / chemistry
  • Cell-Penetrating Peptides / metabolism
  • Cell-Penetrating Peptides / pharmacology
  • Humans
  • Models, Molecular
  • Peptides / chemistry
  • Peptides / metabolism
  • Peptides / pharmacology
  • Protein Binding
  • Proto-Oncogene Proteins c-mdm2* / antagonists & inhibitors
  • Proto-Oncogene Proteins c-mdm2* / chemistry
  • Proto-Oncogene Proteins c-mdm2* / metabolism
  • Structure-Activity Relationship
  • Tumor Suppressor Protein p53* / antagonists & inhibitors
  • Tumor Suppressor Protein p53* / metabolism

Substances

  • Tumor Suppressor Protein p53
  • Proto-Oncogene Proteins c-mdm2
  • MDM2 protein, human
  • Cell-Penetrating Peptides
  • Peptides
  • Antineoplastic Agents