Fragment‐Based Drug Discovery of Novel High‐affinity, Selective, and Anti‐inflammatory Inhibitors of the Keap1‐Nrf2 Protein‐Protein Interaction

Abstract Activating the cytoprotective response of nuclear factor erythroid 2‐related factor 2 (Nrf2) can reduce oxidative stress and inflammation. A promising strategy is to inhibit the protein‐protein interaction between Kelch‐like ECH‐associated protein 1 (Keap1) and Nrf2 using noncovalent compounds that target the Keap1 Kelch domain. These compounds may be more specific than covalent Keap1‐reacting Nrf2 activators. However, the development of drug‐like noncovalent Keap1‐Nrf2 inhibitors faces challenges due to the size and polarity of the Kelch binding pocket. Here, we present a new series of noncovalent Keap1‐Nrf2 inhibitors developed from a weak fragment hit identified by crystallographic screening. A two‐step growing strategy and optimization guided by several X‐ray cocrystal structures led to compounds with low nanomolar affinities and complete selectivity for Keap1 in a panel of homologous Kelch domains. In cells, compounds 24 and 28 potently activated the expression of Nrf2‐controlled genes and showed anti‐inflammatory effects by downregulating NLRP3 inflammasome and STING signalling activation. RNA sequencing revealed activation of cytoprotective pathways and a different profile from typical covalent Nrf2 activators. This work highlights the potential of fragment‐based drug discovery for challenging targets like Keap1 and introduces novel Keap1‐Nrf2 inhibitors as chemical probes and drug leads.

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Journal
Angewandte Chemie
Published
2025-08-21
DOI
https://doi.org/10.1002/ange.202508121
Primary Topic
Genomics, phytochemicals, and oxidative stress
Type
article
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0.00

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article

Fragment‐Based Drug Discovery of Novel High‐affinity, Selective, and Anti‐inflammatory Inhibitors of the Keap1‐Nrf2 Protein‐Protein Interaction

Michael Gajhede, David Olagnier, Charlotte E. Manning, Mahya Dezfouli et al.
1 links
Angewandte Chemie
Genomics, phytochemicals, and oxidative stress
article

Fragment‐Based Drug Discovery of Novel High‐affinity, Selective, and Anti‐inflammatory Inhibitors of the Keap1‐Nrf2 Protein‐Protein Interaction

Michael Gajhede, David Olagnier, Charlotte E. Manning, Mahya Dezfouli, Dilip Narayanan, Chun‐Yu Lin, Camilla B. Chan, William Richardson, David J. Blake, Leandro Silva da Costa, Zhuoyao Chen, Helena Käck, Jaslin L. Lindsay, Anders Bach, Alex N. Bullock, Cecilie Poulsen, Marilia Barreca, Jingyi Wang, Jie Zang, Xiangrong Chen, Dimitra Vlissari, Thomas S. Mortensen, Kendall G. Wichman, Cherisse A. Charley
article en

Abstract

Abstract Activating the cytoprotective response of nuclear factor erythroid 2‐related factor 2 (Nrf2) can reduce oxidative stress and inflammation. A promising strategy is to inhibit the protein‐protein interaction between Kelch‐like ECH‐associated protein 1 (Keap1) and Nrf2 using noncovalent compounds that target the Keap1 Kelch domain. These compounds may be more specific than covalent Keap1‐reacting Nrf2 activators. However, the development of drug‐like noncovalent Keap1‐Nrf2 inhibitors faces challenges due to the size and polarity of the Kelch binding pocket. Here, we present a new series of noncovalent Keap1‐Nrf2 inhibitors developed from a weak fragment hit identified by crystallographic screening. A two‐step growing strategy and optimization guided by several X‐ray cocrystal structures led to compounds with low nanomolar affinities and complete selectivity for Keap1 in a panel of homologous Kelch domains. In cells, compounds 24 and 28 potently activated the expression of Nrf2‐controlled genes and showed anti‐inflammatory effects by downregulating NLRP3 inflammasome and STING signalling activation. RNA sequencing revealed activation of cytoprotective pathways and a different profile from typical covalent Nrf2 activators. This work highlights the potential of fragment‐based drug discovery for challenging targets like Keap1 and introduces novel Keap1‐Nrf2 inhibitors as chemical probes and drug leads.

Angewandte ChemieVol. 137(39)
University of Copenhagen (DK), Fort Lewis College (US), Aarhus University (DK), Novo Nordisk Foundation (DK), Nuffield Orthopaedic Centre (GB), University of Oxford (GB), AstraZeneca (Finland) (FI), Oxford BioMedica (United Kingdom) (GB), AstraZeneca (Germany) (DE), University of Palermo (IT)
McGill University, Hørslev-Fonden, University of Dundee, Diamond Light Source, European Federation of Pharmaceutical Industries and Associations, European Commission, VINNOVA, Svenska Forskningsrådet Formas, Lundbeckfonden, Novo Nordisk, Vetenskapsrådet, China Scholarship Council, Eva og Henry Frænkels Mindefond, Th. Maigaards Eftf. Fru Lily Benthine Lunds Fond af 1.6. 1978, Novo Nordisk Fonden, Hartmann Fonden, Fabrikant Einar Willumsens Mindelegat, H. Lundbeck A/S, National Institutes of Health, Horizon 2020, NIH Office of the Director, National Institute of General Medical Sciences, Common Fund
Openalex Percentile: Top 81%
Genomics, phytochemicals, and oxidative stress
DOIOpen Access PDFOpenAlex
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