An Exploration of Chemical Properties Required for Cooperative Stabilization of the 14-3-3 Interaction with NF-κB-Utilizing a Reversible Covalent Tethering Approach

J Med Chem. 2021 Jun 24;64(12):8423-8436. doi: 10.1021/acs.jmedchem.1c00401. Epub 2021 Jun 2.


Protein-protein modulation has emerged as a proven approach to drug discovery. While significant progress has been gained in developing protein-protein interaction (PPI) inhibitors, the orthogonal approach of PPI stabilization lacks established methodologies for drug design. Here, we report the systematic ″bottom-up″ development of a reversible covalent PPI stabilizer. An imine bond was employed to anchor the stabilizer at the interface of the 14-3-3/p65 complex, leading to a molecular glue that elicited an 81-fold increase in complex stabilization. Utilizing protein crystallography and biophysical assays, we deconvoluted how chemical properties of a stabilizer translate to structural changes in the ternary 14-3-3/p65/molecular glue complex. Furthermore, we explore how this leads to high cooperativity and increased stability of the complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 14-3-3 Proteins / metabolism*
  • Benzaldehydes / chemistry*
  • Drug Design
  • Escherichia coli
  • Escherichia coli Proteins / metabolism*
  • Molecular Structure
  • Protein Binding / drug effects*
  • Small Molecule Libraries / chemistry*
  • Structure-Activity Relationship
  • Transcription Factor RelA / metabolism*


  • 14-3-3 Proteins
  • Benzaldehydes
  • Escherichia coli Proteins
  • Small Molecule Libraries
  • Transcription Factor RelA