MEK2 (IC50 = 60 nM); MEK1 (IC50 = 70 nM)

U0126-EtOH 通过功能性拮抗 AP-1 转录活性来阻断参与炎症反应的多种细胞因子和金属蛋白酶的产生。 [1]通过降低 IL-2 mRNA 水平，U0126-EtOH 可减少响应抗原刺激或交联抗 CD3 加抗 CD28 抗体的 T 细胞增殖，而不影响 IL-2 引起的增殖。 [2]根据最近的一项研究，U0126-EtOH 抑制线粒体功能，在不借助 MEK 的情况下将去势抵抗性人类前列腺癌 C4-2 细胞转变为有氧糖酵解，并抵消白藜芦醇诱导的细胞凋亡。 [3]

U0126-EtOH 通过抑制细胞内 Raf/MEK/ERK 信号通路，抑制 2009 年大流行 IV H1N1 变种和高致病性禽流感病毒 (HPAIV) 在小鼠肺体内的传播，从而表现出抗病毒活性。 [4]通过在注射 A 的大鼠中激活核呼吸因子 1、线粒体转录因子 A 和过氧化物酶体增殖物激活受体 γ 共激活剂-1a，U0126-EtOH 表现出潜在的神经保护作用并改善莫里斯水迷宫 (MWM) 中的空间学习。 5]

在这些测定中，调整免疫沉淀的野生型 MEK 的量以产生与 10 nM 重组 MEK 相同数量的活性单位。 96 孔硝化纤维过滤器装置用于测量反应速度，如下详述。除非另有说明，所有反应均使用 10 nM 酶浓度、20 mM Hepes、10 mM MgCl2、5 mM β-巯基乙醇、0.1 mg/mL BSA、pH 7.4 和室温。向预混合的 MEK/ERK/抑制剂反应混合物中添加 [γ-33P]ATP 以启动反应。然后每 6 分钟取出 100 μL 等分试样，转移至含有 50 mM EDTA 的 96 孔硝酸纤维素膜板中以终止反应。将膜板拉出并用缓冲液真空清洗四次。然后将30μL Microscint-20闪烁液倒入孔中，使用Top Count闪烁计数器测量33P-磷酸化ERK的放射性。根据放射性与时间图的斜率，可以计算出速度。除非另有说明，ERK 和 ATP 浓度分别为 400 nM 和 40 μM。存在和不存在抑制剂时的初始反应速度分别称为 Vi 和 Vo，它们用于计算所有体外酶测定的抑制百分比。然后将数据绘制为抑制剂浓度与抑制百分比的函数关系，并通过使用非线性最小二乘回归将数据拟合到 Langmuir 等温线方程来计算 IC50。根据提供的信息，酶浓度不是基于活性位点滴定，而是基于最终测定体积中使用的蛋白质的分子量和质量。因此，酶活性位点的实际浓度可能与报告值不同。

A.E7 或 Th17 细胞与已用丝裂霉素 C 以及不同浓度的鸽子细胞色素 c、PR8 Ag 或 5 U/mL 人 rIL-2 处理的 B10.BR 或 BALB/c 脾细胞一起孵育。为了确定 MEK 抑制对 T 细胞增殖的直接影响，一些测定还包含 U0126 或无活性类似物 U0124。每个孔在培养开始两天后接受 1-μCi [3H]TdR 脉冲，并在第二天收获培养物。在不使用液体闪烁混合物的情况下，在 Packard Matrix 96 直接 β 计数器上测量 [3H]TdR 掺入 DNA 的情况。

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).
≤10 mM
Administered via aerosol.

[1]. Bioorg Med Chem Lett . 1998 Oct 20;8(20):2839-44.

[2]. J Immunol . 1998 May 1;160(9):4175-81.

[3]. Cancer Biol Ther . 2011 Dec 1;12(11):966-77.

[4]. Antiviral Res . 2011 Nov;92(2):195-203.

[5]. Behav Brain Res . 2012 Jun 15;232(1):165-73.

[6]. J Biol Chem . 1998 Jul 17;273(29):18623-32.

U0126.EtOH is an addition compound obtained by combining equimolar amounts of (2Z,3Z)-bis{amino[(2-aminophenyl)sulfanyl]methylidene}butanedinitrile (U0126) and ethanol. An inhibitor of mitogen-activated protein kinase that also exhibits anti-cancer properties. It has a role as an EC 2.7.11.24 (mitogen-activated protein kinase) inhibitor, an apoptosis inducer, an antineoplastic agent, an antioxidant, an osteogenesis regulator and a vasoconstrictor agent. It contains an U0126.

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In search of antiinflammatory drugs with a new mechanism of action, U0126 was found to functionally antagonize AP-1 transcriptional activity via noncompetitive inhibition of the dual specificity kinase MEK with an IC50 of 0.07 microM for MEK 1 and 0.06 microM for MEK 2. U0126 can undergo isomerization and cyclization reactions to form a variety of products, both chemically and in vivo, all of which exhibit less affinity for MEK and lower inhibition of AP-1 activity than parent, U0126.[1]

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Three mitogen-activated protein kinase pathways are up-regulated during the activation of T lymphocytes, the extracellular signal-regulated kinase (ERK), Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase pathways. To examine the effects of blocking the ERK pathway on T cell activation, we used the inhibitor U0126, which has been shown to specifically block mitogen-activated protein kinase/ERK kinase (MEK), the kinase upstream of ERK. This compound inhibited T cell proliferation in response to antigenic stimulation or cross-linked anti-CD3 plus anti-CD28 Abs, but had no effect on IL-2-induced proliferation. The block in T cell proliferation was mediated by down-regulating IL-2 mRNA levels. Blocking Ag-induced proliferation by inhibiting MEK did not induce anergy, unlike treatments that block entry into the cell cycle following antigenic stimulation. Surprisingly, induction of anergy in T cells exposed to TCR cross-linking in the absence of costimulation was also not affected by blocking MEK, unlike cyclosporin A treatment that blocks anergy induction. These results suggest that inhibition of MEK prevents T cell proliferation in the short term, but does not cause any long-term effects on either T cell activation or induction of anergy. These findings may help determine the viability of using mitogen-activated protein kinase inhibitors as immune suppressants.[2]

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Recent evidence has identified substantial overlap between metabolic and oncogenic biochemical pathways, suggesting novel approaches to cancer intervention. For example, cholesterol lowering statins and the antidiabetes medication metformin both act as chemopreventive agents in prostate and other cancers. The natural compound resveratrol has similar properties: increasing insulin sensitivity, suppressing adipogenesis, and inducing apoptotic death of cancer cells in vitro. However, in vivo tumor xenografts acquire resistance to resveratrol by an unknown mechanism, while mouse models of metabolic disorders respond more consistently to the compound. Here we demonstrate that castration-resistant human prostate cancer C4-2 cells are more sensitive to resveratrol-induced apoptosis than isogenic androgen-dependent LNCaP cells. The MEK inhibitor U0126 antagonized resveratrol-induced apoptosis in C4-2 cells, but this effect was not seen with other MEK inhibitors. U0126 was found to inhibit mitochondrial function and shift cells to aerobic glycolysis independently of MEK. Mitochondrial activity of U0126 arose through decomposition, producing both mitochondrial fluorescence and cyanide, a known inhibitor of complex IV. Applying U0126 mitochondrial inhibition to C4-2 cell apoptosis, we tested the possibility that glutamine supplementation of citric acid cycle intermediate α-ketoglutarate may be involved. Suppression of the conversion of glutamate to α-ketoglutarate antagonized resveratrol-induced death in C4-2 cells. A similar effect was also seen by reducing extracellular glutamine concentration in the culture medium, suggesting that resveratrol-induced death is dependent on glutamine metabolism, a process frequently dysregulated in cancer. Further work on resveratrol and metabolism in cancer is warranted to ascertain if the glutamine dependence has clinical implications.[3]

C18H16N6S2.C2H6O

426.56

426.129

C, 56.32; H, 5.20; N, 19.70; O, 3.75; S, 15.03

1173097-76-1

U0126;109511-58-2; 218601-62-8 (ZZ-isomer); 218601-64-0 (EE-isomer); 1173097-76-1 (EtOH)

16220066

White to off-white solid powder

612.5ºC at 760 mmHg

324.2ºC

5.684

222.49

5

9

5

29

612

0

S(/C(=C(\C#N)/C(/C#N)=C(\N([H])[H])/SC1=C([H])C([H])=C([H])C([H])=C1N([H])[H])/N([H])[H])C1=C([H])C([H])=C([H])C([H])=C1N([H])[H].O([H])C([H])([H])C([H])([H])[H]

CFQULUVMLGZVAF-OYJDLGDISA-N

InChI=1S/C18H16N6S2.C2H6O/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;1-2-3/h1-8H,21-24H2;3H,2H2,1H3/b17-11+,18-12+

(2Z,3Z)-2,3-bis[amino-(2-aminophenyl)sulfanylmethylidene]butanedinitrile;ethanol

U0126; U-0126-EtOH; U0126 EtOH; U 0126; U-0126; U0126-EtOH; U 0126-EtOH; U 0126 EtOH; U0126 Ethanol; U0126.EtOH; (2Z,3Z)-2,3-bis(amino(2-aminophenylthio)methylene)succinonitrile compound with ethanol (1:1); (2Z,3Z)-2,3-bis[amino-(2-aminophenyl)sulfanylmethylidene]butanedinitrile;ethanol; 2,3-Bis(amino((2-aminophenyl)thio)methylene)succinonitrile compound with ethanol (1:1); U0126 ethanolate; .U-0126 EtOH

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: ≥ 2.08 mg/mL (4.88 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 20.8 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.08 mg/mL (4.88 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.08 mg/mL (4.88 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 10% DMSO+50% PEG 300+ddH2O: 28mg/mL

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配方 5 中的溶解度: 5 mg/mL (11.72 mM) in 0.5% CMC-Na/saline water (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。