T-5224 C-Fos/AP-1 inhibitor

Inhibitory activities

The DNA binding activity of transcription factors was measured using the TransAM kits (Active Motif). Nuclear extracts containing factors such as c-Fos/AP-1, c-Jun/AP-1, ATF-2, C/EBPa, MyoD, Sp-1 or NF-kB/p65 and various concentrations of T-5224 were added in the multi-well plates precoated with respective consensus double-stranded (ds)DNA oligomers. After incubation for 1 h, the transcription factor bound to its respective consensus dsDNA sequences was detected by using antibodies reactive against the respective transcription factors according to the manufacturer’s protocol.

Cell lines

Human synovial SW982 cells; human chondrocyte SW1353 cells

Preparation method

Dissolved in DMSO. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

0.1, 1, 10, 100, 1000 μM.

Applications

In human synovial SW982 cells stimulated by IL-1β, T-5224 inhibits the production of MMP-1, MMP-3, IL-6 and TNFα. In human chondrocyte SW1353 cells stimulated by IL-1β, T-5224 inhibits the production of MMP-3 and MMP-13 with IC50 value of 10 μM.

Animal models

Male DBA/1J mice with type II collagen–induced arthritis (CIA).

Dosage form

0.3, 1, 3, 30 mg/kg, once daily from day 21, before the onset of arthritis; administered orally.

Applications

T-5224 significantly inhibits the development of arthritis by 64% and 91% on day 50 at 3 and 30 mg/kg, respectively. T-5224 inhibits joint destruction and completely protects the joint from destruction at 30 mg/kg. At 30 mg/kg dose, T-5224 significantly inhibits MMP-2, MMP-3, MMP-9, MMP-13, IL-1β, IL-6 and TNFα.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Aikawa Y, Morimoto K, Yamamoto T, et al. Treatment of arthritis with a selective inhibitor of c-Fos/activator protein-1. Nat Biotechnol, 2008, 26(7): 817-823.