Inhibitory effect of T-614 on tumor necrosis factor-alpha induced cytokine production and nuclear factor-kappaB activation in cultured human synovial cells.
OBJECTIVE
To investigate the mechanism of the immunosuppressive effect of T-614 [N-(3-formylamino-4-oxo-6-phenoxy-4H-chromen-7-yl)methanesulfonamide], a new antirheumatic drug whose clinical efficacy has been determined for the treatment of patients with rheumatoid arthritis (RA).


METHODS
RA synovial fibroblast-like cells were cultured with tumor necrosis factor-alpha (TNF-alpha, 10 ng/ml) in the presence or absence of T-614. After incubation, cytokine production was measured by ELISA. Expression of interleukin 6 (IL-6) and IL-8 mRNA was examined by real-time quantitative reverse transcriptase-polymerase chain reaction analysis and TNF-alpha induced nuclear factor-kappaB (NF-kappaB) activation was observed using immunostaining with an antibody against NF-kappaB p65.


RESULTS
T-614 suppressed TNF-alpha induced production of IL-6, IL-8, and monocyte chemoattractant protein 1, and also reduced the accumulation of IL-6 and IL-8 mRNA in a concentration dependent manner. T-614 interfered with the TNF-alpha induced translocation of NF-kappaB to the nucleus from the cytoplasm.


CONCLUSIONS
Inhibition of NF-kappaB activation and transcription of proinflammatory cytokines by T-614 contributes to its clinical antirheumatic effect.
Kohno
M
M
First Department of Internal Medicine, Kyoto Prefectural University of Medicine, Japan.
Aikawa
Y
Y
Tsubouchi
Y
Y
Hashiramoto
A
A
Yamada
R
R
Kawahito
Y
Y
Inoue
K
K
Kusaka
Y
Y
Kondo
M
M
Sano
H
H
D018501:Antirheumatic Agents / Q000494:pharmacology*; D001578:Benzopyrans / Q000494:pharmacology*; D002135:Calcium-Binding Proteins; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007124:Immunoenzyme Techniques; D015850:Interleukin-6 / Q000096:biosynthesis / Q000235:genetics; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics; D007378:Interleukins / Q000096:biosynthesis* / Q000235:genetics; D008562:Membrane Glycoproteins / Q000378:metabolism; D016328:NF-kappa B / Q000096:biosynthesis*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013449:Sulfonamides / Q000494:pharmacology*; D050863:Synaptotagmin I; D050857:Synaptotagmins; D013583:Synovial Membrane / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D018501:Antirheumatic Agents; D001578:Benzopyrans; D002135:Calcium-Binding Proteins; D015850:Interleukin-6; D016209:Interleukin-8; D007378:Interleukins; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D013449:Sulfonamides; D050863:Synaptotagmin I; D014409:Tumor Necrosis Factor-alpha; C077313:T 614; D050857:Synaptotagmins