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

IF 3.5 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
ACS Chemical Biology Pub Date : 2025-07-18 Epub Date: 2025-06-20 DOI:10.1021/acschembio.5c00112
Lior Soday, Chotima Seripracharat, Janine L Gray, André F S Luz, Ryan T Howard, Ravi Singh, Thomas J Burden, Erika Bernardini, Miguel Mateus-Pinheiro, Jens Petersen, Anders Gunnarsson, Jenny Gunnarsson, Anna Aagaard, Tove Sjögren, Sarah Maslen, Edward J Bartlett, Abigail F Iles, David M Smith, James S Scott, Mark Skehel, Andrew M Davis, Ana S Ressurreição, Rui Moreira, Cecília M P Rodrigues, Avinash R Shenoy, Edward W Tate
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引用次数: 0

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.

一类新的靶向RIPK1的坏死性下垂抑制剂的发现和验证。
坏死性坏死是一种程序性细胞死亡的形式,当失调时,与癌症、炎症和神经退行性疾病有关。在这里,我们从从表型高通量筛选中鉴定出的坏死性坏死抑制剂开始,合成了一个含有7-苯基喹啉基序的化合物库,并在一种新的活细胞试验中验证了它们的抗坏死性坏死活性。基于这些数据,我们设计了一种优化的光亲和探针用于靶标接合研究,并通过生化和细胞实验建立了受体相互作用激酶1 (RIPK1)作为细胞靶标,通过阻止RIPK1的自磷酸化和激活来抑制坏死坏死。x射线晶体学和质谱分析显示,这些化合物结合在RIPK1活性构象的铰链区域,确定它们是I型激酶抑制剂。此外,我们在体外证明了与III型激酶抑制剂(如necrostatin-1)的协同作用,并发现铅化合物可以保护小鼠免受体内坏死性坏死模型的急性炎症。总之,我们提出了一种新的抑制人类RIPK1的药效团,RIPK1是参与坏死坏死的关键蛋白,并提供了一种光亲和探针来探索细胞中RIPK1靶点的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ACS Chemical Biology
ACS Chemical Biology 生物-生化与分子生物学
CiteScore
7.50
自引率
5.00%
发文量
353
审稿时长
3.3 months
期刊介绍: ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology. The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies. We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.
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