META060 inhibits multiple kinases in the NF-kappaB pathway and suppresses LPS--mediated inflammation in vitro and ex vivo

• Published in: Inflammation research Vol. 58; no. 5; pp. 229 - 234
• Main Authors: Desai, A, Konda, V R, Darland, G, Austin, M, Prabhu, K S, Bland, J S, Carroll, B J, Tripp, M L
• Format: Journal Article
• Language: English
• Published: Switzerland 01.05.2009
• Subjects: Animals; Anti-Inflammatory Agents - chemistry; Anti-Inflammatory Agents - metabolism; Anti-Inflammatory Agents - pharmacology; Cell Line; Cyclooxygenase 2 - metabolism; Cyclopentanes - chemistry; Cyclopentanes - metabolism; Cyclopentanes - pharmacology; Dinoprostone - metabolism; Humans; Inflammation - chemically induced; Inflammation - metabolism; Interleukin-6 - metabolism; Lipopolysaccharides - immunology; Macrophages - cytology; Macrophages - drug effects; Macrophages - metabolism; Mice; Molecular Structure; NF-kappa B - genetics; NF-kappa B - metabolism; Nitric Oxide - metabolism; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - metabolism; Protein Kinase Inhibitors - pharmacology; Signal Transduction - drug effects; Signal Transduction - physiology; Tumor Necrosis Factor-alpha - metabolism
• ISSN: 1420-908X
• DOI: 10.1007/s00011-008-8162-y

We investigated whether a novel candidate META060 targeted the inflammatory signal transduction without affecting constitutive COX-2 enzymatic activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. We also investigated its bioavailability in humans and its anti-inflammatory effect ex vivo. We measured prostaglandin E(2), nitric oxide, TNFalpha and IL-6 by ELISA, COX-2 protein by Western blot, NF-kappaB nuclear binding by electrophoretic mobility shift assays, and NF-kappaB activation by luciferase assay. Kinase inhibitions were measured by cell-free assays. Bioavailability was tested in 4 human subjects consuming 940 mg META060. LPS-activated TNFalpha and IL-6 were measured in peripheral blood mononuclear cells (PBMC) isolated from 1 subject up to 6 hours post administration. META060 dose-dependently inhibited prostaglandin E(2) and nitric oxide formation, COX-2 abundance, and NF-kappaB activation. In cell-free assays, META060 inhibited multiple kinases in the NF-kappaB signaling pathway, including BTK, PI3K, and GSK3. META060 was detected in the plasma of the subjects; isolated PBMC were resistant to LPS-stimulated TNFalpha and IL-6 production. Without inhibiting COX-2 enzyme, META060 reduces the inflammation by inhibiting multiple kinases involved in NF-kappaB pathway, and may have potential as a safe anti-inflammatory therapeutic.