| Size | Price | Stock | Qty |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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Purity: ≥98%
U0126 (also known as U0126-EtOH) is a novel potent, highly selective and non-ATP competitive inhibitor of MEK1/2 with IC50 of 0.07 μM/0.06 μM in cell-free assays.Through noncompetitive inhibition of the dual specificity kinase MEK1/2, it was discovered that U0126 functionally inhibits AP-1 transcriptional activity. Compared to PD9805, it exhibits a 100-fold higher affinity for ΔN3-S218E/S222D MEK. The anti-inflammatory drug U0126-EtOH was eliminated because it inhibited AP-1 transcription with an IC50 value of 1μM and did not interact with GREs. With an IC50 of 0.07 microM for MEK 1 and 0.06 microM for MEK 2, U0126-EtOH treatment significantly reduced the cell damage brought on by oxidative glutamate toxicity in HT22 cells. It also remarkably blocked the phosphorylation of ERK1/2. Chemically and in vivo, U0126 can go through isomerization and cyclization reactions to produce a number of products, all of which have lower MEK affinities and lower inhibitions of AP-1 activity than the parent, U0126.
| Targets |
MEK2 (IC50 = 60 nM); MEK1 (IC50 = 70 nM)
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| ln Vitro |
Treatment with U0126 effectively lowers progeny virus titers of all examined strains in A549 cells. While nM concentrations of U0126 are effective at reducing H1N1v and H5N1 (MB1),μM concentrations of U0126 are needed to lower H5N1 (GSB) and H7N7 virus titers. The EC50 values for U0126-EtOH against H1N1v are 1.2±0.4 μM in A549 cells and 74.7±1.0 μM in MDCKII cells[2]. Rat hepatocarcinoma cells (FAO) stimulated by fetal calf serum (FCS) exhibit a significant proportion in S phase (32.62%), whereas U0126 significantly reduces the proportion of cells in S phase (9.92%) and increases the proportion of cells in G0-G1 phase and to a lesser extent in G2/M[3].
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| ln Vivo |
Mice are administered U0126-EtOH (U0126; i.p., 10.5 mg/kg) every day. The tumor sizes throughout the control experiment are either constant or slightly increasing. On the other hand, engraftment and early tumor growth are markedly reduced in all U0126-EtOH experiments. Additionally, 9 days after injection and thereafter, the volume of tumors treated with U0126-EtOH is reduced by 60–70%[3]. Rats are given transient middle cerebral artery occlusion (tMCAO) for 120 minutes, and at 0 and 24 hours after reperfusion, they are given U0126-EtOH (U0126; i.p., 30 mg/kg). After receiving U0126-EtOH therapy, the vasoconstriction to S6c is significantly decreased[4].
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| Enzyme Assay |
In these assays, the amount of immunoprecipitated wild type MEK is adjusted to produce an equivalent number of activity units to that of 10 nM recombinant MEK. A 96-well nitrocellulose filter apparatus is used to measure reaction velocities, as detailed below. A 10 nM enzyme concentration, 20 mM Hepes, 10 mM MgCl2, 5 mM β-mercaptoethanol, 0.1 mg/mL BSA, pH 7.4, and room temperature are used for all reactions unless otherwise specified. The pre-mixed MEK/ERK/inhibitor reaction mixture is added [γ-33P]ATP to start the reactions. An aliquot of 100 μL is then taken every 6 minutes and transferred to the 96-well nitrocellulose membrane plate containing 50 mM EDTA to stop the reaction. The membrane plate is drawn and vacuum-washed four times with buffer. After that, 30 L of Microscint-20 scintillation fluid is poured into the wells, and a Top Count scintillation counter is used to measure the radioactivity of 33P-phosphorylated ERK. From the slopes of radioactivity versus time plots, speeds are calculated. Unless otherwise stated, ERK and ATP concentrations are 400 nM and 40 μM, respectively. The initial reaction velocities in the presence and absence of the inhibitor, respectively, are called Vi and Vo, and they are used to calculate the percent inhibition for all in vitro enzyme assays. The IC50 is then calculated by fitting the data using nonlinear least squares regression to the standard equation for a Langmuir isotherm. The data are then plotted as a function of inhibitor concentration as a function of percent inhibition. As stated, rather than active site titration, enzyme concentrations are determined by the molecular weights and protein mass used in the final assay volume. The actual concentration of the enzyme active site may therefore vary from the reported value.
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| Cell Assay |
A.E7 or Th17 cells are incubated with B10.BR or BALB/c splenocytes that have been treated with mitomycin C, along with varying concentrations of pigeon cytochrome c, PR8 Ag, or 5 U/mL human rIL-2. In order to ascertain the direct effects of MEK inhibition on T cell proliferation, some assays also contain U0126 or an inactive analog, U0124. Each well receives a 1µCi [3H]TdR pulse two days after culture initiation, and the following day, the cultures are harvested. Without the use of liquid scintillation mixtures, the incorporation of [3H]TdR into DNA is measured on a Packard Matrix 96 direct beta counter.
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| Animal Protocol |
Athymic female nude mice (SWISS, nu/nu)[3].
10.5 mg/kg. Intraperitoneal injection daily. |
| References |
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| Molecular Formula |
C18H16N6S2
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| Molecular Weight |
380.49
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| Exact Mass |
380.09
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| Elemental Analysis |
C, 56.82; H, 4.24; N, 22.09; S, 16.85
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| CAS # |
109511-58-2
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| Related CAS # |
U0126-EtOH;1173097-76-1
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| Appearance |
White to off-white solid powder
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| SMILES |
C1=CC=C(C(=C1)N)S/C(=C(/C(=C(/SC2=CC=CC=C2N)\N)/C#N)\C#N)/N
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| InChi Key |
DVEXZJFMOKTQEZ-JYFOCSDGSA-N
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| InChi Code |
InChI=1S/C18H16N6S2/c19-9-11(17(23)25-15-7-3-1-5-13(15)21)12(10-20)18(24)26-16-8-4-2-6-14(16)22/h1-8H,21-24H2/b17-11+,18-12+
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| Chemical Name |
(2Z,3Z)-2,3-bis[amino-(2-aminophenyl)sulfanylmethylidene]butanedinitrile
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.57 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.57 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.57 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 10% DMSO+50% PEG 300+ddH2O: 28mg/mL |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6282 mL | 13.1409 mL | 26.2819 mL | |
| 5 mM | 0.5256 mL | 2.6282 mL | 5.2564 mL | |
| 10 mM | 0.2628 mL | 1.3141 mL | 2.6282 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.
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