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

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, TNF alpha and IL-6 by ELISA, COX-2 protein by Western blot, NF-kappa B nuclear binding by electrophoretic mobility shift assays, and NF-kappa B 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 TNF alpha 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-kappa B activation. In cell-free assays, META060 inhibited multiple kinases in the NF-kappa B signaling pathway, including BTK, PI3K, and GSK3. META060 was detected in the plasma of the subjects; isolated PBMC were resistant to LPS-stimulated TNF alpha and IL-6 production. Without inhibiting COX-2 enzyme, META060 reduces the inflammation by inhibiting multiple kinases involved in NF-kappa B pathway, and may have potential as a safe anti-inflammatory therapeutic.