title: Inflammatory Signaling by NOD-RIPK2 Is Inhibited by Clinically Relevant Type II Kinase Inhibitors.
abstract: RIPK2 mediates pro-inflammatory signaling from the bacterial sensors NOD1 and NOD2, and is an emerging therapeutic target in autoimmune and inflammatory diseases. We observed that cellular RIPK2 can be potently inhibited by type II inhibitors that displace the kinase activation segment, whereas ATP-competitive type I inhibition was only poorly effective. The most potent RIPK2 inhibitors were the US Food and Drug Administration-approved drugs ponatinib and regorafenib. Their mechanism of action was independent of NOD2 interaction and involved loss of downstream kinase activation as evidenced by lack of RIPK2 autophosphorylation. Notably, these molecules also blocked RIPK2 ubiquitination and, consequently, inflammatory nuclear factor κB signaling. In monocytes, the inhibitors selectively blocked NOD-dependent tumor necrosis factor production without affecting lipopolysaccharide-dependent pathways. We also determined the first crystal structure of RIPK2 bound to ponatinib, and identified an allosteric site for inhibitor development. These results highlight the potential for type II inhibitors to treat indications of RIPK2 activation as well as inflammation-associated cancers.
authors: Structural Genomics Consortium, University of Oxford, Old Road Campus, Roosevelt Drive, Oxford OX3 7DQ, UK.
authors: Peter
authors: P
authors: Canning
authors: Department of Developmental, Molecular & Chemical Biology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA.
authors: Qui
authors: Q
authors: Ruan
authors: Translational Gastroenterology Unit, Nuffield Department of Medicine and Department of Pediatrics, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DU, UK.
authors: Tobias
authors: T
authors: Schwerd
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7DQ, UK.
authors: Matous
authors: M
authors: Hrdinka
authors: Department of Developmental, Molecular & Chemical Biology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA.
authors: Jenny L
authors: JL
authors: Maki
authors: Medical Scientist Training Program and Program in Neuroscience, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, MA 02111, USA.
authors: Danish
authors: D
authors: Saleh
authors: Department of Chemistry, University of Houston, Houston, TX 77204, USA.
authors: Chalada
authors: C
authors: Suebsuwong
authors: Center for Neurologic Diseases, Department of Neurology, Brigham & Women's Hospital and Harvard Medical School, Cambridge, MA 02139, USA.
authors: Soumya
authors: S
authors: Ray
authors: Structural Genomics Consortium, University of Oxford, Old Road Campus, Roosevelt Drive, Oxford OX3 7DQ, UK; Target Discovery Institute, University of Oxford, NDM Research Building, Roosevelt Drive, Oxford OX3 7LD, UK.
authors: Paul E
authors: PE
authors: Brennan
authors: Department of Pharmacological and Pharmaceutical Sciences, University of Houston, Houston, TX 77204, USA.
authors: Gregory D
authors: GD
authors: Cuny
authors: Translational Gastroenterology Unit, Nuffield Department of Medicine and Department of Pediatrics, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DU, UK.
authors: Holm H
authors: HH
authors: Uhlig
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7DQ, UK.
authors: Mads
authors: M
authors: Gyrd-Hansen
authors: Department of Developmental, Molecular & Chemical Biology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA. Electronic address: alexei.degterev@tufts.edu.
authors: Alexei
authors: A
authors: Degterev
authors: Structural Genomics Consortium, University of Oxford, Old Road Campus, Roosevelt Drive, Oxford OX3 7DQ, UK. Electronic address: alex.bullock@sgc.ox.ac.uk.
authors: Alex N
authors: AN
authors: Bullock
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D006801:Humans; D007093:Imidazoles / Q000737:chemistry / Q000494:pharmacology; D007249:Inflammation / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D053474:Nod1 Signaling Adaptor Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism; D053473:Nod2 Signaling Adaptor Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011724:Pyridazines / Q000737:chemistry / Q000494:pharmacology; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D061987:Sf9 Cells; D015398:Signal Transduction / Q000187:drug effects; D054875:Ubiquitination / Q000187:drug effects
chemical_list: D007093:Imidazoles; C119067:NOD1 protein, human; C429018:NOD2 protein, human; D053474:Nod1 Signaling Adaptor Protein; D053473:Nod2 Signaling Adaptor Protein; D047428:Protein Kinase Inhibitors; D011724:Pyridazines; C545373:ponatinib; C506253:RIPK2 protein, human; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2