Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, U.K.
European Screening Centre, University of Dundee, Biocity, Bo'Ness Road, Motherwell ML1 5UH, U.K.
Biochem J. 2020 Feb 28;477(4):801-814. doi: 10.1042/BCJ20190846.
Autophagy is a critical cellular homeostatic mechanism, the dysfunction of which has been linked to a wide variety of disease states. It is regulated through the activity of specific kinases, in particular Unc-51 like autophagy activating kinase 1 (ULK1) and Phosphatidylinositol 3-kinase vacuolar protein sorting 34 (VPS34), which have both been suggested as potential targets for drug development. To identify new chemical compounds that might provide useful chemical tools or act as starting points for drug development, we screened each protein against the Published Kinase Inhibitor Set (PKIS), a library of known kinase inhibitors. In vitro screening and analysis of the published selectivity profiles of the hits informed the selection of three relatively potent ATP-competitive inhibitors against each target that presented the least number of off-target kinases in common. Cellular assays confirmed potent inhibition of autophagy in response to two of the ULK1 inhibitors and all three of the VPS34 inhibitors. These compounds represent not only a new resource for the study of autophagy but also potential chemical starting points for the validation or invalidation of these two centrally important autophagy kinases in disease models.
自噬是一种重要的细胞内稳态机制，其功能障碍与多种疾病状态有关。它通过特定激酶的活性来调节，特别是自噬激活激酶 1 (ULK1) 和磷脂酰肌醇 3-激酶液泡蛋白分选 34 (VPS34)，它们都被认为是药物开发的潜在靶点。为了鉴定可能提供有用化学工具或作为药物开发起点的新化学化合物，我们针对已发表的激酶抑制剂集 (PKIS) 筛选了每种蛋白质，这是一个已知激酶抑制剂的文库。对命中物的已发表选择性概况进行的体外筛选和分析为针对每个靶标选择三种相对有效的 ATP 竞争性抑制剂提供了信息，这些抑制剂共同具有最少数量的非靶标激酶。细胞测定法证实了两种 ULK1 抑制剂和三种 VPS34 抑制剂均能有效抑制自噬。这些化合物不仅是自噬研究的新资源，也是在疾病模型中验证或否定这两种重要的自噬激酶的潜在化学起点。
