Discovery and Validation of a Novel Class of Necroptosis Inhibitors Targeting RIPK1

  • ACS Chem Biol. 2025 Jul 18;20(7):1527-1543. doi: 10.1021/acschembio.5c00112.
Lior Soday  1 ,  Chotima Seripracharat  1 ,  Janine L Gray  1 ,  André F S Luz  2 ,  Ryan T Howard  1 ,  Ravi Singh  1 ,  Thomas J Burden  1 ,  Erika Bernardini  1 ,  Miguel Mateus-Pinheiro  2 ,  Jens Petersen  3 ,  Anders Gunnarsson  3 ,  Jenny Gunnarsson  3 ,  Anna Aagaard  3 ,  Tove Sjögren  3 ,  Sarah Maslen  4 ,  Edward J Bartlett  1 ,  Abigail F Iles  1 ,  David M Smith  5 ,  James S Scott  5 ,  Mark Skehel  4 ,  Andrew M Davis  5 ,  Ana S Ressurreição  2 ,  Rui Moreira  2 ,  Cecília M P Rodrigues  2 ,  Avinash R Shenoy  6 ,  Edward W Tate  1  4
Affiliations
  • 1. Department of Chemistry, Molecular Sciences Research Hub, Imperial College London, London W12 0BZ, U.K.
  • 2. Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Lisbon 1649-004, Portugal.
  • 3. Discovery Sciences, R&D Gothenburg, AstraZeneca, Pepparedsleden 1, SE-431 83 Mölndal, Sweden.
  • 4. The Francis Crick Institute, London NW1 1AT, U.K.
  • 5. Hit Discovery, Discovery Sciences, R&D, AstraZeneca, Cambridge CB2 0AA, U.K.
  • 6. Department of Infectious Diseases, Imperial College London, Flowers Building, South Kensington Campus, London SW7 2AZ, U.K.
PMID: 40539767 DOI: 10.1021/acschembio.5c00112
Abstract

Necroptosis is a form of programmed cell death that, when dysregulated, is associated with Cancer and inflammatory and neurodegenerative diseases. Here, starting from hits identified from a phenotypic high-throughput screen for inhibitors of Necroptosis, we synthesized a library of compounds containing a 7-phenylquinoline motif and validated their anti-necroptotic activity in a novel live-cell assay. Based on these data, we designed an optimized photoaffinity probe for target engagement studies and through biochemical and cell-based assays established receptor-interacting kinase 1 (RIPK1) as the cellular target, with inhibition of Necroptosis arising from the prevention of RIPK1 autophosphorylation and activation. X-ray crystallography and mass spectrometry revealed that these compounds bind at the hinge region of the active conformation of RIPK1, establishing them as type I kinase inhibitors. In addition, we demonstrated in vitro synergy with type III kinase inhibitors, such as necrostatin-1 and found that lead compounds protected mice against acute inflammation in Necroptosis models in vivo. Overall, we present a novel pharmacophore for inhibition of human RIPK1, a key protein involved in Necroptosis, and provide a photoaffinity probe to explore RIPK1 target engagement in cells.

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