Catalytic in vivo protein knockdown by small-molecule PROTACs

Nat Chem Biol. 2015 Aug;11(8):611-7. doi: 10.1038/nchembio.1858. Epub 2015 Jun 10.

Abstract

The current predominant therapeutic paradigm is based on maximizing drug-receptor occupancy to achieve clinical benefit. This strategy, however, generally requires excessive drug concentrations to ensure sufficient occupancy, often leading to adverse side effects. Here, we describe major improvements to the proteolysis targeting chimeras (PROTACs) method, a chemical knockdown strategy in which a heterobifunctional molecule recruits a specific protein target to an E3 ubiquitin ligase, resulting in the target's ubiquitination and degradation. These compounds behave catalytically in their ability to induce the ubiquitination of super-stoichiometric quantities of proteins, providing efficacy that is not limited by equilibrium occupancy. We present two PROTACs that are capable of specifically reducing protein levels by >90% at nanomolar concentrations. In addition, mouse studies indicate that they provide broad tissue distribution and knockdown of the targeted protein in tumor xenografts. Together, these data demonstrate a protein knockdown system combining many of the favorable properties of small-molecule agents with the potent protein knockdown of RNAi and CRISPR.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Binding Sites
  • Biocatalysis
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology
  • Female
  • Humans
  • MCF-7 Cells
  • Mice
  • Models, Molecular
  • Molecular Targeted Therapy
  • Neoplasm Proteins / antagonists & inhibitors*
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Neoplasm Transplantation
  • Proteasome Endopeptidase Complex / metabolism
  • Protein Binding
  • Proteolysis
  • Receptor-Interacting Protein Serine-Threonine Kinase 2 / antagonists & inhibitors*
  • Receptor-Interacting Protein Serine-Threonine Kinase 2 / genetics
  • Receptor-Interacting Protein Serine-Threonine Kinase 2 / metabolism
  • Receptors, Estrogen / antagonists & inhibitors*
  • Receptors, Estrogen / genetics
  • Receptors, Estrogen / metabolism
  • Small Molecule Libraries / pharmacology*
  • Ubiquitin / genetics
  • Ubiquitin / metabolism
  • Ubiquitination
  • Von Hippel-Lindau Tumor Suppressor Protein / genetics
  • Von Hippel-Lindau Tumor Suppressor Protein / metabolism

Substances

  • Antineoplastic Agents
  • ERRalpha estrogen-related receptor
  • Neoplasm Proteins
  • Receptors, Estrogen
  • Small Molecule Libraries
  • Ubiquitin
  • Von Hippel-Lindau Tumor Suppressor Protein
  • RIPK2 protein, human
  • Receptor-Interacting Protein Serine-Threonine Kinase 2
  • Proteasome Endopeptidase Complex
  • VHL protein, human

Associated data

  • PubChem-Substance/251963825
  • PubChem-Substance/251963826
  • PubChem-Substance/251963827
  • PubChem-Substance/251963828
  • PubChem-Substance/251963829
  • PubChem-Substance/251963830
  • PubChem-Substance/251963831
  • PubChem-Substance/251963832
  • PubChem-Substance/251963833
  • PubChem-Substance/251963834
  • PubChem-Substance/251963835
  • PubChem-Substance/251963836
  • PubChem-Substance/251963837
  • PubChem-Substance/251963838
  • PubChem-Substance/251963839