title: A "Click Chemistry Platform" for the Rapid Synthesis of Bispecific Molecules for Inducing Protein Degradation.
abstract: Proteolysis targeting chimeras (PROTACs) are bispecific molecules containing a target protein binder and an ubiquitin ligase binder connected by a linker. By recruiting an ubiquitin ligase to a target protein, PROTACs promote ubiquitination and proteasomal degradation of the target protein. The generation of effective PROTACs depends on the nature of the protein/ligase ligand pair, linkage site, linker length, and linker composition, all of which have been difficult to address in a systematic way. Herein, we describe a "click chemistry" approach for the synthesis of PROTACs. We demonstrate the utility of this approach with the bromodomain and extraterminal domain-4 (BRD4) ligand JQ-1 (3) and ligase binders targeting cereblon (CRBN) and Von Hippel-Lindau (VHL) proteins. An AlphaScreen proximity assay was used to determine the ability of PROTACs to form the ternary ligase-PROTAC-target protein complex and a MSD assay to measure cellular degradation of the target protein promoted by PROTACs.
authors.forename: Ryan P
authors.identifier: 0000-0003-1413-5208
authors.initials: RP
authors.lastname: Wurz
authors.forename: Ken
authors.initials: K
authors.lastname: Dellamaggiore
authors.affiliation: Department of Therapeutic Discovery-Discovery Technologies, Amgen Inc. , 1120 Veterans Boulevard, South San Francisco, California 94080, United States.
authors.forename: Hannah
authors.initials: H
authors.lastname: Dou
authors.affiliation: Department of Therapeutic Discovery-Discovery Technologies, Amgen Inc. , 1120 Veterans Boulevard, South San Francisco, California 94080, United States.
authors.forename: Noelle
authors.initials: N
authors.lastname: Javier
authors.affiliation: Department of Therapeutic Discovery-Discovery Technologies, Amgen Inc. , 1120 Veterans Boulevard, South San Francisco, California 94080, United States.
authors.forename: Mei-Chu
authors.initials: MC
authors.lastname: Lo
authors.forename: John D
authors.initials: JD
authors.lastname: McCarter
authors.forename: Dane
authors.initials: D
authors.lastname: Mohl
authors.forename: Christine
authors.initials: C
authors.lastname: Sastri
authors.forename: J Russell
authors.initials: JR
authors.lastname: Lipford
authors.forename: Victor J
authors.initials: VJ
authors.lastname: Cee
mesh_terms: D006801:Humans; D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D057930:Click Chemistry / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D008024:Ligands; D009687:Nuclear Proteins / Q000378:metabolism; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000378:metabolism; D000092468:Bromodomain Containing Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D008024:Ligands; D009687:Nuclear Proteins; D010447:Peptide Hydrolases; D010455:Peptides; D011994:Recombinant Proteins; D014157:Transcription Factors; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C441289:BRD4 protein, human; D000092468:Bromodomain Containing Proteins; C502543:CRBN protein, human; C497446:VHL protein, human