Size | Price | Stock | Qty |
---|---|---|---|
5mg |
|
||
10mg |
|
||
25mg |
|
||
50mg |
|
||
100mg |
|
||
250mg |
|
||
Other Sizes |
|
Purity: ≥98%
TPCA-1 (also known as TPCA1; GW-683965; GW683965) is novel, potent, and selective inhibitor of IKK-2 with potential anti-inflammatory activity. In a cell-free assay, it inhibits IKK-2 with an IC50 of 17.9 nM and shows 22-fold selectivity for IKK-1 over IKK-1. An excellent in vivo anti-inflammatory effect of TPCA-1 is seen in a murine collagen-induced arthritis model.
Targets |
STAT3 ; NF-κB; IKK2 (IC50 = 17.9 nM)
|
---|---|
ln Vitro |
TPCA-1 has an IC50 of 17.9 nM in a time-resolved fluorescence resonance energy transfer assay to inhibit the activity of human IKK-2. Additionally, it has been shown that TPCA-1 competes with ATP. Additionally, TPCA-1 has IC50 values against IKK-1 and JNK3 of 400 nM and 3600 nM, respectively. TPCA-1 exhibits concentration-dependent inhibition of TNF-α, IL-6, and IL-8 production, with IC50 values of 170, 290, and 320 nM, respectively. [1] NFκB-dependent IL8 gene expression, TNF-induced RelA (p65) nuclear translocation, and glioma cell proliferation are all inhibited by TPCA-1. Importantly, TPCA-1 completely blocks IFN-induced gene expression of MX1 and GBP1, while only slightly affecting the expression of ISG15. [2]
|
ln Vivo |
Murine collagen-induced arthritis (CIA) is less severe when TPCA-1 is administered prophylactically at doses of 3, 10, or 20 mg/kg, intravenously, every day. The effects of the anti-rheumatic drug etanercept when given prophylactically at 4 mg/kg, i.p., every other day are comparable to the effects of the significantly reduced disease severity and delayed disease onset caused by the administration of TPCA-1 at 10 mg/kg, i.p., b.i.d. In the paw tissue of TPCA-1 and etanercept-treated mice, nuclear localization of p65, as well as levels of IL-1beta, IL-6, TNF-alpha, and interferon-gamma, are markedly decreased. In addition, administration of TPCA-1 in vivo significantly reduces collagen-induced T cell proliferation ex vivo.
|
Enzyme Assay |
A time-resolved fluorescence resonance energy transfer assay is used to determine the activity of recombinant human IKK-2 (residues 1-756) expressed in baculovirus as an N-terminally GST-tagged fusion protein. In a nutshell, 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 the substance or dimethyl sulfoxide (DMSO) vehicle (3% final). In a total volume of 30 L, GST-IB substrate (25 nM final) and ATP (1 μM final) are added to start the reaction. After 30 minutes of incubation at room temperature, the reaction is stopped by adding 15 μL of 50 mM EDTA. The reaction is further incubated for 60 min at room temperature with the addition of 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. Using a Packard Discovery plate reader, the amount of phosphorylation of GST-IκBα is calculated as the ratio of a specific 665-nm energy transfer signal to a reference 620-nm europium signal.
|
Cell Assay |
Ten microliters of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) from stock solution (10 mg/mL) are added to each well of 96-well plates containing glioma cells, and the mixture is then incubated at 37 °C for 2-4 h. Plating is carried out at 37 °C for 4 hours in a humid environment after adding 100 μL of 10% sodium dodecyl sulfate (SDS) in 0.01 N HCL to solubilize the oxidized MTT. At 570 nm, a plate reader reads plates.
|
Animal Protocol |
Murine collagen-induced arthritis
3, 10, or 20 mg/kg Administered via i.p. or b.i.d. |
References |
Molecular Formula |
C12H10FN3O2S
|
---|---|
Molecular Weight |
279.29
|
Exact Mass |
279.290
|
Elemental Analysis |
C, 51.61; H, 3.61; F, 6.80; N, 15.05; O, 11.46; S, 11.48
|
CAS # |
507475-17-4
|
Appearance |
Solid powder
|
SMILES |
C1=CC(=CC=C1C2=CC(=C(S2)NC(=O)N)C(=O)N)F
|
InChi Key |
SAYGKHKXGCPTLX-UHFFFAOYSA-N
|
InChi Code |
InChI=1S/C12H10FN3O2S/c13-7-3-1-6(2-4-7)9-5-8(10(14)17)11(19-9)16-12(15)18/h1-5H,(H2,14,17)(H3,15,16,18)
|
Chemical Name |
2-(carbamoylamino)-5-(4-fluorophenyl)thiophene-3-carboxamide
|
Synonyms |
GW683965; TPCA-1; GW-683965; TPCA1; TPCA 1; GW 683965
|
HS Tariff Code |
2934.99.9001
|
Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
Solubility (In Vitro) |
|
|||
---|---|---|---|---|
Solubility (In Vivo) |
|
Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 3.5805 mL | 17.9025 mL | 35.8051 mL | |
5 mM | 0.7161 mL | 3.5805 mL | 7.1610 mL | |
10 mM | 0.3581 mL | 1.7903 mL | 3.5805 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration: mg/mL;
Method for preparing DMSO stock solution: mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation::Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.