2. Academic Validation
  3. Discovery and Validation of a Novel Class of Necroptosis Inhibitors Targeting RIPK1

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

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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