000K  utf8
1100  $c2025
1500  eng
2050  urn:nbn:de:gbv:8:3-2025-00293-4
2051  10.1021/acschembio.4c00812
3000  Miletić, Nebojša
3010  Abdul Azeez, Kamal Rayees
3010  Adhikari, Bikash
3010  Berner, Nicola
3010  Elson, Lewis
3010  Gehrtz, Paul
3010  Hartung, Ingo V
3010  Hoffmann, Lasse
3010  Hoffmann, Marina E
3010  Holzmann, Hanna
3010  Knapp, Stefan
3010  Kuster, Bernhard
3010  Mosler, Thorsten
3010  Müller, Juliane
3010  Müller, Susanne
3010  Němec, Václav
3010  Rathore, Rajeshwari
3010  Schlesiger, Sarah
3010  Schwalm, Martin P
3010  Sivashanmugam, Saran Aswathaman
3010  Weckesser, Janik
3010  Wilhelm, Stephanie
3010  Wolf, Elmar
3010  Đikić, Ivan
4000  Workflow for E3 Ligase Ligand Validation for PROTAC Development$hAmerican Chemical Society (ACS)  [Miletić, Nebojša]
4030  $nAmerican Chemical Society (ACS)
4209  Proteolysis targeting chimeras (PROTACs) have gained considerable attention as a new modality in drug discovery. The development of PROTACs has been mainly focused on using CRBN (Cereblon) and VHL (Von Hippel-Lindau ligase) E3 ligase ligands. However, the considerable size of the human E3 ligase family, newly developed E3 ligase ligands, and the favorable druggability of some E3 ligase families hold the promise that novel degraders with unique pharmacological properties will be designed in the future using this large E3 ligase space. Here, we developed a workflow aiming to improve and streamline the evaluation of E3 ligase ligand efficiency for PROTAC development and the assessment of the corresponding "degradable" target space using broad-spectrum kinase inhibitors and the well-established VHL ligand VH032 as a validation system. Our study revealed VH032 linker attachment points that are highly efficient for kinase degradation as well as some of the pitfalls when using protein degradation as a readout. For instance, cytotoxicity was identified as a major mechanism leading to PROTAC- and VHL-independent kinase degradation. The combination of E3 ligase ligand negative controls, competition by kinase parent compounds, and neddylation and proteasome inhibitors was essential to distinguish between VHL-dependent and -independent kinase degradation events. We share here the findings and limitations of our study and hope that this study will provide guidance for future evaluations of new E3 ligase ligand systems for degrader development.
4950  https://doi.org/10.1021/acschembio.4c00812$xR$3Volltext$534
4950  https://nbn-resolving.org/urn:nbn:de:gbv:8:3-2025-00293-4$xR$3Volltext$534
4961  https://macau.uni-kiel.de/receive/macau_mods_00005818
5051  540
5550  Adaptor Proteins, Signal Transducing
5550  Drug Discovery
5550  Humans
5550  Ligands
5550  Protein Kinase Inhibitors
5550  Proteolysis
5550  Ubiquitin-Protein Ligases
5550  Von Hippel-Lindau Tumor Suppressor Protein
5550  Workflow