TPCA-1 是一种选择性IKK-2抑制剂，IC50值为 17.9 nM。它也是反式激活、DNA结合以及STAT3磷酸化的一种抑制剂。

TPCA-1 is an effective and specific IKK-2 inhibitor (IC50: 17.9 nM); shows >22-fold selectivity over IKK-1 and >550-fold selectivity over others kinases and enzymes.

TPCA-1可显著减少体内胶原诱导的T细胞增殖.TPCA-1（3/10/20 mg/kg,i.p.,b.i.d.）预防性给药,使小鼠体内胶原诱导性关节炎的严重程度呈剂量依赖性降低.TPCA-1（20 mg/kg,i.p.,b.i.d.）可明显降低CIA的严重程度,与依那西普（12.5 mg/kg,i.p.）每隔一天给药的效果相同.TPCA-1（10 mg/kg,i.p.,b.i.d.）使疾病严重程度明显降低,并使疾病发作的延迟,这与依那西普（4 mg/kg,i.p.）每隔一天预防性给药的效果相当.

TPCA-1对TNF-α（IC50：170 nM）,IL-6（IC50：290 nM）和IL-8（IC50：320 nM）的产生具有浓度依赖性的抑制作用。 TPCA-1对胶质瘤细胞增殖、TNF诱导的RelA (p65)核转运及NFκB依赖性IL8基因表达均有抑制作用。TPCA-1对IFN诱导的基因表达有抑制效果,并完全抑制MX1和GBP1基因表达,但对ISG15表达的作用很小。此外,TPCA-1对IKK-1（IC50：400 nM）和JNK3（IC50：3600 nM）也有一定抑制效果。

IKK-2 Assay: Recombinant human IKK-2 (residues 1-756) is expressed in baculovirus as an N-terminal GST-tagged fusion protein, and its activity is assessed using a time-resolved fluorescence resonance energy transfer assay. In brief, IKK-2 (5 nM final) diluted in assay buffer (50 mM HEPES, 10 mM MgCl2, 1 mM CHAPS, pH 7.4, with 1 mM DTT and 0.01% w/v BSA) is added to wells containing various concentrations of compound or dimethyl sulfoxide (DMSO) vehicle (3% final). The reaction is initiated by the addition of GST-IκBα substrate (25 nM final)/ATP (1 μM final), in a total volume of 30 μL. The reaction is incubated for 30 min at room temperature, then terminated by the addition of 15 μL of 50 mM EDTA. Detection reagent (15 μL) in buffer (100 mM HEPES, pH 7.4, 150 mM NaCl, and 0.1% w/v BSA) containing antiphosphoserine- IκBα-32/36 monoclonal antibody 12C2, labeled with W-1024 europium chelate, and an allophycocyanin-labeled anti-GST antibody is added, and the reaction is further incubated for 60 min at room temperature. The degree of phosphorylation of GST- IκBαis measured as a ratio of specific 665-nm energy transfer signal to reference europium 620-nm signal, using a Packard Discovery plate reader.

Ten microliters of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) from stock solution (10?mg/mL) is added to each well of 96-well plates containing glioma cells and incubated at 37 °C for 2–4?h. Oxidized MTT is solubilized by adding 100?μL of 10% sodium dodecyl sulfate (SDS) in 0.01?N HCL, and plates are incubated at 37 °C for 4?h in a humidified chamber. Plates are read at 570?nm on a plate reader.(Only for Reference)

507475-17-4

C12H10FN3O2S

279.29

TPCA1;GW683965;IKK2 Inhibitor IV

Ethanol:2.8 mg/mL (10 mM)
DMSO:27.9 mg/mL (100 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years