Discovery and validation of a novel class of necroptosis inhibitors targeting RIPK1
Author(s)
Type
Journal Article
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.
Date Issued
2025-07-18
Date Acceptance
2025-06-05
Citation
ACS Chemical Biology, 2025, 20 (7), pp.1527-1543
ISSN
1554-8929
Publisher
American Chemical Society (ACS)
Start Page
1527
End Page
1543
Journal / Book Title
ACS Chemical Biology
Volume
20
Issue
7
Copyright Statement
© 2025 The Authors. Published by American Chemical Society. This publication is licensed under CC-BY 4.0 (http://creativecommons.org/licenses/by/4.0/)
License URL
Identifier
10.1021/acschembio.5c00112
Publication Status
Published
Date Publish Online
2025-06-20