Identification and characterization of second-generation EZH2 inhibitors with extended residence times and improved biological activity

J Biol Chem. 2021 Jan-Jun:296:100349. doi: 10.1016/j.jbc.2021.100349. Epub 2021 Jan 30.

Abstract

The histone methyltransferase EZH2 has been the target of numerous small-molecule inhibitor discovery efforts over the last 10+ years. Emerging clinical data have provided early evidence for single agent activity with acceptable safety profiles for first-generation inhibitors. We have developed kinetic methodologies for studying EZH2-inhibitor-binding kinetics that have allowed us to identify a unique structural modification that results in significant increases in the drug-target residence times of all EZH2 inhibitor scaffolds we have studied. The unexpected residence time enhancement bestowed by this modification has enabled us to create a series of second-generation EZH2 inhibitors with sub-pM binding affinities. We provide both biophysical evidence validating this sub-pM potency and biological evidence demonstrating the utility and relevance of such high-affinity interactions with EZH2.

Keywords: EZH2; TR-FRET-binding kinetics; femtomolar; residence time; small-molecule kinetics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Regulation / drug effects
  • Animals
  • Drug Discovery
  • Enhancer of Zeste Homolog 2 Protein / antagonists & inhibitors*
  • Enhancer of Zeste Homolog 2 Protein / metabolism
  • Enzyme Inhibitors / chemistry*
  • Enzyme Inhibitors / pharmacology*
  • Female
  • HeLa Cells
  • Humans
  • Mice
  • Mice, SCID
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology

Substances

  • Enzyme Inhibitors
  • Small Molecule Libraries
  • EZH2 protein, human
  • Enhancer of Zeste Homolog 2 Protein