JAK2-targeted anti-inflammatory effect of a resveratrol derivative 2,4-dihydroxy-N-(4-hydroxyphenyl)benzamide

Abstract

Chemical derivatization of resveratrol has been widely conducted in an effort to overcome its chemical instability and therapeutic potential. In the present study, we examined the anti-inflammatory effects of resveratrol derivatives containing an amide functionality using in vitro macrophage models that were stimulated by Toll-like receptor (TLR) ligands, and using several animal inflammatory disease models. Of the resveratrol derivatives tested, compound 8 (2,4-dihydroxy-N-(4-hydroxyphenyl)benzamide) most strongly inhibited the production of nitric oxide (NO), tumor necrosis factor (TNF)-α, and prostaglandin E2 (PGE2), as well as the mRNA expression of inducible NO synthase (iNOS), TNF-α, and cyclooxygenase (COX)-2 in lipopolysaccharide (LPS)-activated RAW264.7 cells, differentiated U937 cells, and peritoneal macrophages. The inhibitory activity of compound 8 was apparently mediated by suppressing the phosphorylation of signal transducer and activator of transcription (STAT)-1, STAT-3, STAT-5, and interferon regulatory factor (IRF)-3. The direct target of compound 8 was revealed to be Janus kinase 2 (JAK2) but not TANK-binding kinase (TBK) 1 using the direct kinase assay and analyses of complex formation with these molecules. Additionally, upstream kinase of TBK1 seems to be also inhibited by compound 8. This compound also strongly ameliorated mouse inflammatory symptoms seen in arachidonic acid-induced ear edema, dextran sodium sulfate (DSS)-treated colitis, EtOH/HCl-induced gastritis, collagen type II-triggered arthritis, and acetic acid-induced writhing. Therefore, of the resveratrol derivatives that we tested, compound 8 was determined to have the strongest anti-inflammatory activities in vitro and in vivo and may potentially be developed for use as a novel anti-inflammatory drug.

Keywords: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (a tetrazole); AP-1; Amide functionality; Anti-inflammatory effects; COX; DTT; EIA; ELISA; ERK; IRF-3; Inflammatory mediators; JAK2; JNK; Janus kinase 2; LPS; MAPK; MTT; NF-κB; NO; Na CMC; PG; PI3K; RT-PCR; Resveratrol derivatives; TLR; TNF-α; Toll-like receptors; activator protein-1; and STAT-1; c-Jun N-terminal kinase; cyclooxygenase; dithiothreitol; enzyme immunoassay; enzyme linked immunosorbent assay; extracellular signal-related kinase; iNOS; inducible NO synthase; interferon regulatory factor-3; lipopolysaccharide; mitogen activated protein kinase; nitric oxide; nuclear factor-κB; phosphoinositide 3-kinase; prostaglandin; reverse transcriptase-polymerase chain reaction; signal transducer and activator of transcription 1; sodium carboxyl methylcellulose; tumor necrosis factor α.