title: Staurosporine-derived inhibitors broaden the scope of analog-sensitive kinase technology.
abstract: Analog-sensitive (AS) kinase technology is a powerful approach for studying phospho-signaling pathways in diverse organisms and physiological processes. The key feature of this technique is that a kinase-of-interest can be mutated to sensitize it to inhibitor analogs that do not target wild-type (WT) kinases. In theory, this enables specific inhibition of any kinase in cells and in mouse models of human disease. Typically, these inhibitors are identified from a small library of molecules based on the pyrazolopyrimidine (PP) scaffold. However, we recently identified a subset of native human kinases, including the Ephrin A kinase family, that are sensitive to commonly used PP inhibitors. In an effort to develop a bioorthogonal AS-kinase inhibitor and to extend this technique to PP-sensitive kinases, we sought an alternative inhibitor scaffold. Here we report the structure-based design of synthetically tractable, potent, and extremely selective AS-kinase inhibitors based on the natural product staurosporine. We demonstrate that these molecules, termed staralogs, potently target AS kinases in cells, and we employ X-ray crystallography to elucidate their mechanism of efficacy. Finally, we demonstrate that staralogs target AS mutants of PP-sensitive kinases at concentrations where there is little to no inhibition of native human kinases. Thus, staralogs represent a new class of AS-kinase inhibitors and a core component of the chemical genetic tool kit for probing kinase-signaling pathways.
authors.affiliation: Howard Hughes Medical Institute and Department of Cellular & Molecular Pharmacology, University of California , San Francisco, California 94143, United States.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Lopez
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Choy
authors.forename: Ulf
authors.initials: U
authors.lastname: Peters
authors.forename: Martin L
authors.initials: ML
authors.lastname: Sos
authors.forename: David O
authors.initials: DO
authors.lastname: Morgan
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
mesh_terms: D000818:Animals; D001665:Binding Sites; D002227:Carbazoles / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D009154:Mutation; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D019311:Staurosporine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D002227:Carbazoles; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D019311:Staurosporine