A Novel, Highly Selective, Tight Binding IκB Kinase-2 (IKK-2) Inhibitor: A Tool to Correlate IKK-2 Activity to the Fate and Functions of the Components of the Nuclear Factor-κB Pathway in Arthritis-Relevant Cells and Animal Models
| Autor: | Julia A. Guzova, Gabriel Mbalaviele, Alexander F. Shaffer, Sheri L. Bonar, John F. Schindler, David C. Thompson, Nandini Kishore, Catherine S. Tripp, Cynthia D. Sommers, Yiding Hu, Sumathy Mathialagan, Po Chang Chiang, Michele A. Melton, Lori Christine |
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| Rok vydání: | 2009 |
| Předmět: |
Lipopolysaccharides
Blotting Western Arthritis Electrophoretic Mobility Shift Assay Enzyme-Linked Immunosorbent Assay Inflammation IκB kinase Biology p38 Mitogen-Activated Protein Kinases environment and public health Tandem Mass Spectrometry In vivo Synovial Fluid medicine Animals Electrophoretic mobility shift assay Enzyme Inhibitors Cells Cultured Chromatography High Pressure Liquid Pharmacology NF-kappa B Transcription Factor RelA Streptococcus Fibroblasts medicine.disease Arthritis Experimental Recombinant Proteins I-kappa B Kinase Rats Cell biology enzymes and coenzymes (carbohydrates) Biochemistry Rats Inbred Lew Molecular Medicine Experimental pathology Phosphorylation Female medicine.symptom Tomography X-Ray Computed Ex vivo Signal Transduction |
| Zdroj: | Journal of Pharmacology and Experimental Therapeutics. 329:14-25 |
| ISSN: | 1521-0103 0022-3565 |
| DOI: | 10.1124/jpet.108.143800 |
| Popis: | Nuclear factor (NF)-kappaB activation has been clearly linked to the pathogenesis of multiple inflammatory diseases including arthritis. The central role that IkappaB kinase-2 (IKK-2) plays in regulating NF-kappaB signaling in response to inflammatory stimuli has made this enzyme an attractive target for therapeutic intervention. Although diverse chemical classes of IKK-2 inhibitors have been identified, the binding kinetics of these inhibitors has limited the scope of their applications. In addition, safety assessments of IKK-2 inhibitors based on a comprehensive understanding of the pharmacokinetic/pharmacodynamic relationships have yet to be reported. Here, we describe a novel, potent, and highly selective IKK-2 inhibitor, PHA-408 [8-(5-chloro-2-(4-methylpiperazin-1-yl)isonicotinamido)-1-(4-fluorophenyl)-4,5-dihydro-1H-benzo[g]indazole-3-carboxamide]. PHA-408 is an ATP-competitive inhibitor, which binds IKK-2 tightly with a relatively slow off rate. In arthritis-relevant cells and animal models, PHA-408 suppresses inflammation-induced cellular events, including IkappaBalpha phosphorylation and degradation, p65 phosphorylation and DNA binding activity, the expression of inflammatory mediators, and joint pathology. PHA-408 was efficacious in a chronic model of arthritis with no adverse effects at maximally efficacious doses. Stemming from its ability to bind tightly to IKK-2, as a novelty, we demonstrated that PHA-408-mediated inhibition of IKK-2 activity correlated very well with its ability to modulate the fate of IKK-2 substrates and downstream transcriptional events. We ultimately directly linked IKK-2 activity ex vivo and in vivo to markers of inflammation with the inhibitor plasma concentrations. Thus, PHA-408 represents a powerful tool to further gain insight into the mechanisms by which IKK-2 regulates NF-kappaB signaling and validates IKK-2 as a therapeutic target. |
| Databáze: | OpenAIRE |
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