U0126-EtOH 化学構造
分子量: 426.56

高品質保証

文献中の引用(57)

カスタマーフィードバック(11)

Quality Control & MSDS

製品説明

  • Compare MEK Inhibitors
    MEK製品生物活性の比較
  • 研究分野
  • U0126-EtOHのメカニズム

製品の説明

生物活性

製品説明 U0126-EtOH は化学的に合成されて非常に選択的な阻害剤で、MEK1 とMEK2 に作用すると、 IC50 がそれぞれ0.07 μM と 0.06 μMになる。
ターゲット

MEK1

MEK2

IC50

0.07 μM

0.06 μM [1]

In vitro試験 U0126-EtOH functionally antagonizes AP- 1 transcriptional activity and blocks the production of a variety of cytokines and metalloproteinases involved in the inflammatory response. [1] U0126-EtOH inhibits T cell proliferation in response to antigenic stimulation or cross-linked anti-CD3 plus anti-CD28 Abs without effect on IL-2-induced proliferation by down-regulating IL-2 mRNA levels. [2] A recent study shows that U0126-EtOH antagonizes resveratrol-induced apoptosis in castration-resistant human prostate cancer C4-2 cells, inhibits mitochondrial function and shifts cells to aerobic glycolysis independently of MEK. [3]
In vivo試験 U0126-EtOH, as the inhibitor of intracellular Raf/MEK/ERK signaling pathway, demonstrates antiviral activity by suppressing propagation of the 2009 pandemic IV H1N1 variant and highly pathogenic avian influenza viruses (HPAIV) in vivo in the mouse lung by inhibiting. [4] U0126-EtOH shows the potential neuroprotective effect and improving spatial learning in Morris water maze (MWM) by activating peroxisome proliferator-activated receptor gamma coactivator-1a, nuclear respiratory factor 1, and mitochondrial transcription factor A in Aβ-injected rats. [5]
臨床試験
特集 A chemically synthesized and highly selective inhibitor of both MEK1 and MEK2.

プロトコル (参考用のみ)

キナーゼアッセイ:

[1]

In Vitro Kinase Assays The amount of immunoprecipitated wild type MEK used in these assays is adjusted to give a similar amount of activity units as obtained with 10 nM recombinant MEK. Reaction velocities are measured using a 96-well nitrocellulose filter apparatus as described below. Unless otherwise noted, reactions are carried out at an enzyme concentration of 10 nM, in 20 mM Hepes, 10 mM MgCl2, 5 mM β-mercaptoethanol, 0.1 mg/mL BSA, pH 7.4, at room temperature. Reactions are initiated by the addition of [γ-33P]ATP into the premixed MEK/ERK/inhibitor reaction mixture, and an aliquot of 100 μL is taken every 6 minutes and transferred to the 96-well nitrocellulose membrane plate which has 50 mM EDTA to stop the reaction. The membrane plate is drawn and washed 4 times with buffer under vacuum. Wells are then filled with 30 μL of Microscint-20 scintillation fluid, and the radioactivity of 33P-phosphorylated ERK is counted with a Top Count scintillation counter. Velocities are obtained from the slopes of radioactivity versus time plots. Concentrations of ERK and ATP are 400 nM and 40 μM, respectively, unless otherwise indicated. For all of the in vitro enzyme assays, the percent inhibition is calculated 100 (1 −Vi/Vo) where Vi and Vo are the initial reaction velocities in the presence and absence of inhibitor, respectively. The data are then plotted as percent inhibition as a function of inhibitor concentration and fit, by nonlinear least squares regression, to the standard equation for a Langmuir isotherm to determine the IC50. As reported, enzyme concentrations are based upon molecular weights and mass of protein used in the final assay volume and not on active site titration. Thus, the actual enzyme active site concentration may differ from that reported.

細胞アッセイ:

[2]

細胞株 A.E7 or Th17 cells
濃度 0 to 10 μM
反応時間 48 hours
実験の流れ

A.E7 or Th17 cells are incubated with mitomycin C-treated B10.BR or BALB/c splenocytes plus varying concentrations of pigeon cytochrome c or PR8 Ag, or with 5 U/mL human rIL-2. In addition, some assays contains U0126 or an inactive analogue, U0124, to determine direct effects of MEK inhibition on T cell proliferation. Two days after culture initiation, each well is pulsed with 1 µCi of [3H]TdR and harvested the following day. The incorporation of [3H]TdR into DNA is quantitated on a Packard Matrix 96 direct beta counter without the use of liquid scintillation mixtures.

動物実験:

[4]

動物モデル Female C57Bl/6 mice infected by Mouse-adapted highly pathogenic avian influenza A/FPV/Bratislava/79 (H7N7; FPV) virus and swine origin human influenza A virus (SOIV) A/Regensburg/D6/2009 (H1N1v; RB1).
製剤 U0126-EtOH is dissolved in 10% DMSO, 30% of Cremophor EL and 60% PBS.
投薬量 ≤10 mM
投与方法 Administered via aerosol.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDogMonkeyBaboon
Weight (kg)0.020.151.80.40.0810312
Body Surface Area (m2)0.0070.0250.150.050.020.50.240.6
Km factor361285201220
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

参考

化学情報

Download U0126-EtOH SDF
分子量 426.56
化学式

C18H16N6S2.C2H6O

CAS No. 1173097-76-1
保管 2年-20℃
6月-80℃in solvent
別名 N/A
溶解度 (25°C) * In vitro DMSO 85 mg/mL warming (199.26 mM)
<1 mg/mL (<1 mM)
エタノール <1 mg/mL (<1 mM)
In vivo 10% DMSO+50% PEG 300+ddH2O 28mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
化学名 2,3-bis(amino(2-aminophenylthio)methylene)succinonitrile,ethanol

カスタマーフィードバック (11)


Click to enlarge
Rating
Source J Natl Cancer Inst, 2012, 104(21), 1673-9. U0126-EtOH purchased from Selleck
Method Western Blot
Cell Lines primary human melanocytes
Concentrations 1 μM
Incubation Time 18 h
Results In BRAFV600E melanoma cells, the highly selective BRAFV600E inhibitor GDC-0879 and three selective MEK inhibitors [PD184352/CI-1040, U0126, PD98059] did not suppress c-Jun levels, although they effectively reduced phospho-ERK levels.

Click to enlarge
Rating
Source Proc Natl Acad Sci U S A, 2014, 111(15):E1528-37. U0126-EtOH purchased from Selleck
Method Western blot
Cell Lines J-Lat cells
Concentrations 10/20/40 uM
Incubation Time 24 h
Results J-Lat cells were stimulated with U0126 and TPA as escribed. Samples were analyzed by Western blot to detect phosphorylated MAPK p42/p44. TPA induced robust MAPK phosphorylation, which was repressed by U0126 in a dose-dependent manner.

Click to enlarge
Rating
Source Biomaterials, 2011, 32, 7524-7531. U0126-EtOH purchased from Selleck
Method Cell migration tracking and quantification
Cell Lines Stromal cells
Concentrations 0-1 nM
Incubation Time
Results Partial ERK inhibition with 0.3 nM of U0126 (data not shown) significantly increased MSC persistence time by 30% from 17 min to 22 min(Fig. B) and increased MFP by 50%, from 16 m m to 24 m m (Fig. A). Speed was decreased mildly, by <10% (Fig. C)

Click to enlarge
Rating
Source Cell Death Differ, 2012, 19, 1908-16. U0126-EtOH purchased from Selleck
Method Real-time RT-PCR/ Determination of apoptosis/Immunoblot analysis
Cell Lines MCF10A cells
Concentrations 10 μM
Incubation Time 15 h
Results Inhibition of the ERK1/2 pathway by the MAPK/ERK kinase 1 (MEK1) inhibitor U0126 blocked FLIPL downregulation induced by EGF re-addition to EGF-deprived cells.

Click to enlarge
Rating
Source J Invest Dermatol, 2012, 132, 2780-90. U0126-EtOH purchased from Selleck
Method Western blot
Cell Lines UACC62 cells
Concentrations 1 μM
Incubation Time 6 h
Results Induced p-Erk1/2 was associated with hyperphosphorylation of its target p90 ribosomal S6 kinase 1, which could significantly be suppressed by the Mek inhibitor, U0126.

Click to enlarge
Rating
Source J Immunol, 2010, 184, 7219-7228. U0126-EtOH purchased from Selleck
Method Western blotting
Cell Lines Il6ra-/-mice
Concentrations 0.4 μg/μL
Incubation Time 0-10 d
Results Stat3 phosphorylation was reduced to a similar extent in Il6-/-, Il6ra-/-, and Il6-/-/Il6ra-/-mice (Fig. A) compared with WT mice. phosphorylation of the MAPK ERK1/2 was undetectable in Il6-/-mice yet increased in Il6ra-/-relative to WT mice 1 d after wounding (Fig. A). Topical application of U0126 reduced ERK phosphorylation in wounds of both WT and Il6ra-/- mice compared with vehicle-treated controls (Fig. B). The difference in wound contraction rates was significantly delayed in Il6ra-/-mice treated with U0126 compared with vehicle control-treated mice but did not reach significance in WT mice (Fig. C).

Click to enlarge
Rating
Source Lung Cancer, 2011, 73, 274-82. U0126-EtOH purchased from Selleck
Method Cell viability assay
Cell Lines A549 cells/ H322 cells
Concentrations 2.5 μM
Incubation Time 72 h
Results When 2.5 μM U0126 was added to the gemcitabine/AG1478 combination in A549 (Fig. A), reduction in cytotoxicity was observed for 0.5 μM AG1478 but this effect became less apparent with increasing AG1478 concentration. This is represented as increased CI (greater antagonism) at low concentrations of AG1478 (Fig. B). Similarly in H322 (Fig. C), addition of U0126 attenuated cytotoxicity at 0.5 μM and 2.5 μM AG1478 but this effect was lost at higher AG1478 concentrations. This was seen as reduced synergism on the CI curve (Fig. D).

Click to enlarge
Rating
Source 2010, Dr. Zhang of Tianjin Medical University. U0126-EtOH purchased from Selleck
Method Western blotting
Cell Lines Breast cancer cells
Concentrations 0-10 nM
Incubation Time 24 h
Results U0126 treatment resulted in a reduction of Erk phosphorylation in Breast cancer cells.

Click to enlarge
Rating
Source Biochim Biophys Acta, 2013, 1832(12):1998-2008. U0126-EtOH purchased from Selleck
Method Immunohistochemistry staining
Cell Lines C57BL/6 male mice
Concentrations 10 mg/kg
Incubation Time 24 h
Results In the sham-operated kidneys, Na,K-ATPase was localized basolaterally in a stripe pattern in proximal tubular epithelial cells. In the ischemic injured kidneys, Na,K-ATPase was much fainter and distributed over the entire cell membrane of proximal tubular epithelial cells, which was treated by U0126,compared with vehicle.

Click to enlarge
Rating
Source Biochim Biophys Acta, 2013, 1832(12):1998-2008. U0126-EtOH purchased from Selleck
Method Immunohistochemistry
Cell Lines C57BL/6 male mice
Concentrations 10 mg/kg
Incubation Time 24 h
Results Nine days after ischemia, tubular atrophy, dilation, and congestion of tubules were significantly greater in the U0126-treated mouse kidneys than in the vehicle-treated mouse kidneys. The number of interstitial cells also was greater in the U0126-treated mouse kidneys when compared with that in the vehicle-treated mouse kidneys. Consistent with the morphological evaluation, tubular damage scores were significantly higher in the U0126-treated mouse kidneys than in the vehicle-treated mouse kidneys.

Click to enlarge
Rating
Source U0126-EtOH purchased from Selleck
Method Western Blot
Cell Lines HeLa cells
Concentrations 0/30/300/3000 nM
Incubation Time 24 h
Results

文献中の引用 (57)

技術サポート&よくある質問(FAQ)

ストックの作り方、阻害剤の保管する方法、細胞実験や動物実験に注意すべきな点を全部含めており、製品を取扱う時よくあった質問に対して取扱説明書でお答えいたします。

電話番号: +1-832-582-8158 Ext:3月曜日〜金曜日 9:00 AM–5:00 PM (米国中部標準時)

他の質問がある場合は、お気軽くお問合せください。

* 必須

Related MEK 阻害剤

  • Cobimetinib (GDC-0973, RG7420)

    Cobimetinib (GDC-0973, RG7420) is a potent and highly selective MEK1 inhibitor with IC50 of 4.2 nM. Phase 3.

  • BI-847325

    BI-847325 is an orally bioavailable, and selective dual MEK/Aurora kinase inhibitor. Phase 1.

  • GDC-0623

    GDC-0623 is a potent and ATP-uncompetitive MEK1 inhibitor with Ki of 0.13 nM. Phase 1.

  • GDC-0994

    GDC-0994 is a potent, orally available ERK1/2 inhibitor. Phase 1.

  • Trametinib (GSK1120212)

    Trametinib (GSK1120212)は、非常に特定で有力なMEK1MEK2 阻害剤で、IC50 がそれぞれ 0.92 nM と 1.8 nMです。

    Features:More potent than PD0325901 or AZD6244.

  • PD0325901

    PD0325901は選択的です、そして、非ATP競争的有糸分裂促進物質はMEK活動のinhibitonのためにプロテインキナーゼ・キナーゼ(MEKまたはMAPKK)MEK阻害剤を起動させました。IC50 が 0.33 nMになる。

  • Selumetinib (AZD6244)

    Selumetinib (AZD6244) は有効的にMEK1を抑制して、IC50 が14 nMになる.

    Features:First MEK inhibitor being tested in Phase II clinical trials.

  • PD98059

    PD98059は、非競争的MEK</bです> 阻害剤で、IC50 が2 μMです.

    Features:Does not inhibit c-Raf phosphorylated MEK1.

  • PD184352 (CI-1040)

    PD184352 (CI-1040)は、ATP非競争MEK1/2阻害剤で、IC50 が 17 nMです。

    Features:First MEK inhibitor to begin clinical development.

最近チェックしたアイテム

Tags: U0126-EtOHを買う | U0126-EtOH供給者 | U0126-EtOHを購入する | U0126-EtOH費用 | U0126-EtOH生産者 | オーダーU0126-EtOH | U0126-EtOH代理店
×
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID