Cell lines

Human oral squamous carcinoma cell lines, OSC-19 and HSC-3-M3.

Preparation Method

Cells were grown in DMEM supplemented with 10% FBS and penicillin (50 units / mL) / streptomycin (50 μg / mL) in a humidified atmosphere (5% CO₂) at 37℃. T-5224 was dissolved in DMSO and diluted in culture medium to the target concentration for each experiment. For oral gavage administration, T-5224 was dissolved in polyvinylpyrrolidone (PVP) solution according to the manufacturer’s recommended procedure.

Reaction Conditions

Cells were incubated in T-5224 (0–80 μM), and cell counts were carried out at 24, 48, and 72 h after treatment.

Applications

T-5224 treatment could inhibit the invasion activity of highly metastatic tumor cell line HSC-3-M3. Cell migration was potently inhibited by T-5224 in both cell lines in a dose-dependent manner. Migration activity was almost completely inhibited in the medium containing 80 μM T-5224.

Animal models

Female 5–7-week-old BALB/ c mice

Preparation Method

HSC-3-M3 (1 x 10⁵cells) were suspended in 50 μL Hank’s Balanced Salts Solution and injected in the flank of the tongue at day 0. T-5224 was diluted in PVP solution, and T-5224 was given orally to the mice of the treatment group every day from day 1 for 4 weeks.

Dosage form

150 mg/ kg

Applications

T-5224 showed significant inhibitory effects against lymph node metastasis in the animal model of HNSCC. The rate of positive metastasis was significantly lower by T-5224 treatment. It is possible that higher doses (150 mg/kg) might be required for cells in which activity of AP-1 is intensively and constitutively activated by genetic mutations or substantial inflammation.

• Int J Biol Sci (2020)

T-5224 is a non-peptidic small molecule AP-1 inhibitor that specifically inhibits the binding of AP-1 to the AP-1 binding site of the promoter region and was originally developed as an anti-inflammatory drug for the treatment of rheumatoid arthritis (RA) suppressing inflammatory cytokines and MMPs without any side-effects.^[1]

In vitro study indicated that transcriptional inhibition of the expression of MMPs by T-5224 is not limited to tumor cells only; it also occurs in the ECM of the surrounding tissue, and consequently, inhibits tumor cells from infiltrating the surrounding tissue. In addition, T-5224 potently inhibits the essential steps of metastasis, infiltration through the basement membrane barrier and migration into the ECM, by transcriptionally suppressing the expression of MMP-2 and -9.^[1]

In vivo experiments demonstrated that T-5224 blocked serum TNF-α, HMGB-1, BUN, and creatinine concentrations, reducing mortality of LPS-induced AKI. These findings suggest that T-5224 may be protective against lethal LPS-induced AKI. T-5224 attenuated LPS-induced liver injury in mice. T-5224 may have beneficial effects in sepsis-induced organ dysfunctions.^[3]

References:
[1]. Daisuke K. et al. Selective activator protein-1 inhibitor T-5224 prevents lymph node metastasis in an oral cancer model. Cancer Sci. 2016 May; 107 (5) 666–673.
[2]. Mari I. et al. T-5224, a selective inhibitor of c-Fos/activator protein-1, improves survival by inhibiting serum high mobility group box-1 in lethal lipopolysaccharide-induced acute kidney injury model. Journal of Intensive Care (2015) 3:49.