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.
Jacob I Stuckey  1 ,  Nico R Cantone  1 ,  Alexandre Côté  1 ,  Shilpi Arora  1 ,  Valerie Vivat  1 ,  Ashwin Ramakrishnan  1 ,  Jennifer A Mertz  1 ,  Avinash Khanna  1 ,  Jehrod Brenneman  1 ,  Victor S Gehling  1 ,  Ludivine Moine  1 ,  Robert J Sims 3rd  1 ,  James E Audia  1 ,  Patrick Trojer  1 ,  Julian R Levell  1 ,  Richard T Cummings  2
Affiliations
  • 1. Constellation Pharmaceuticals, Cambridge, Massachusetts, USA.
  • 2. Constellation Pharmaceuticals, Cambridge, Massachusetts, USA. Electronic address: [email protected].
PMID: 33524394 DOI: 10.1016/j.jbc.2021.100349
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.
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