PMID- 36473314 OWN - NLM STAT- MEDLINE DCOM- 20230117 LR - 20230220 IS - 2213-2317 (Electronic) IS - 2213-2317 (Linking) VI - 59 DP - 2023 Feb TI - Design and characterization of a heterobifunctional degrader of KEAP1. PG - 102552 LID - S2213-2317(22)00324-X [pii] LID - 10.1016/j.redox.2022.102552 [doi] LID - 102552 AB - The Kelch-like ECH-associated protein 1 (KEAP1) - nuclear factor erythroid 2-related factor 2 (NRF2) signaling pathway senses reactive oxygen species and regulates cellular oxidative stress. Inhibiting KEAP1 to activate the NRF2 antioxidant response has been proposed as a promising strategy to treat chronic diseases caused by oxidative stress. Here, we developed a proteolysis targeting chimera (PROTAC) that depletes KEAP1 from cells through the ubiquitin-proteasome pathway. A previously developed KEAP1 inhibitor and thalidomide were incorporated in the heterobifunctional design of the PROTAC as ligands for KEAP1 and CRBN recruitment, respectively. Optimization of the chemical composition and linker length resulted in PROTAC 14 which exhibited potent KEAP1 degradation with low nanomolar DC(50) in HEK293T (11 nM) and BEAS-2B (<1 nM) cell lines. Furthermore, PROTAC 14 increased the expression of NRF2 regulated antioxidant proteins and prevented cell death induced by reactive oxygen species. Together, these results established a blueprint for further development of KEAP1-targeted heterobifunctional degraders and will facilitate the study of the biological consequences of KEAP1 removal from cells. This approach represents an alternative therapeutic strategy to existing treatments for diseases caused by oxidative stress. CI - Copyright (c) 2022 The Authors. Published by Elsevier B.V. All rights reserved. FAU - Chen, Hao AU - Chen H AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Nguyen, Nghi H AU - Nguyen NH AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Magtoto, Charlene M AU - Magtoto CM AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Cobbold, Simon A AU - Cobbold SA AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Bidgood, Grace M AU - Bidgood GM AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Meza Guzman, Lizeth G AU - Meza Guzman LG AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Richardson, Lachlan W AU - Richardson LW AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Corbin, Jason AU - Corbin J AD - Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Au, Amanda E AU - Au AE AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Lechtenberg, Bernhard C AU - Lechtenberg BC AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Feltham, Rebecca AU - Feltham R AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Sutherland, Kate D AU - Sutherland KD AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Grohmann, Christoph AU - Grohmann C AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Nicholson, Sandra E AU - Nicholson SE AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. FAU - Sleebs, Brad E AU - Sleebs BE AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Australia; Department of Medical Biology, The University of Melbourne, Parkville, 3010, Australia. Electronic address: sleebs@wehi.edu.au. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20221126 PL - Netherlands TA - Redox Biol JT - Redox biology JID - 101605639 RN - 0 (Reactive Oxygen Species) RN - 0 (Antioxidants) RN - 0 (Kelch-Like ECH-Associated Protein 1) RN - 0 (NF-E2-Related Factor 2) RN - 0 (KEAP1 protein, human) SB - IM MH - Humans MH - Reactive Oxygen Species/metabolism MH - *Antioxidants/pharmacology/metabolism MH - Kelch-Like ECH-Associated Protein 1/metabolism MH - *NF-E2-Related Factor 2/metabolism MH - HEK293 Cells MH - Oxidative Stress PMC - PMC9720105 OTO - NOTNLM OT - Antioxidant OT - KEAP1-NRF2 pathway OT - Oxidative stress OT - PROTAC OT - ROS COIS- Declaration of competing interest All authors have approved the final version of the manuscript and declare no conflict of interest related to this work. EDAT- 2022/12/07 06:00 MHDA- 2023/01/18 06:00 PMCR- 2022/11/26 CRDT- 2022/12/06 18:16 PHST- 2022/09/28 00:00 [received] PHST- 2022/11/20 00:00 [revised] PHST- 2022/11/22 00:00 [accepted] PHST- 2022/12/07 06:00 [pubmed] PHST- 2023/01/18 06:00 [medline] PHST- 2022/12/06 18:16 [entrez] PHST- 2022/11/26 00:00 [pmc-release] AID - S2213-2317(22)00324-X [pii] AID - 102552 [pii] AID - 10.1016/j.redox.2022.102552 [doi] PST - ppublish SO - Redox Biol. 2023 Feb;59:102552. doi: 10.1016/j.redox.2022.102552. Epub 2022 Nov 26.