U0126-EtOH

Biological Activity

U0126-EtOH is a highly selective inhibitor of both MEK1 and MEK2, a type of MAPK/ERK kinase. U0126 inhibited anchorage-independent growth of Ki-ras-transformed rat fibroblasts by simultaneously blocking both extracellular signal-regulated kinase (ERK) and mammalian target of rapamycin (mTOR)-p70(S6K) pathways. U0126 inhibits MEK-1 and MEK-2 with little or no effect on the activities of PKC, Abl, Raf, MEKK, ERK, JNK, MKK-3, MKK-4/SEK, MKK-6, Cdk2 or Cdk4. U0126 selectively repressed anchorage-independent growth of MDA-MB231 and HBC4 cells, two lines with constitutively activated ERK. In addition, i.v. administration of U0126 (100-200 mg/kg), a specific inhibitor of MEK (MAPK/ERK kinase), protects the hippocampus against forebrain ischemia. Moreover, treatment with U0126 at 3 h after ischemia significantly reduces infarct volume after transient (3 h) focal cerebral ischemia in mice. 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.

Product Citations

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  Ecotoxicol Environ Saf. 2021 Sep 1;220:112341.

  Cadmium induces apoptosis of human granulosa cell line KGN via mitochondrial dysfunction-mediated pathways
  U0126-EtOH purchased from AbMole

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  Ecotoxicol Environ Saf. 2021 Jan 15;208:111429.

  Bisphenol A induces apoptosis through GPER-dependent activation of the ROS/Ca 2+-ASK1-JNK pathway in human granulosa cell line KGN
  U0126-EtOH purchased from AbMole

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  Environ Pollut. 2021 Feb 1;270:116051.

  Bisphenol AF induces apoptosis via estrogen receptor beta (ERβ) and ROS-ASK1-JNK MAPK pathway in human granulosa cell line KGN
  U0126-EtOH purchased from AbMole

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  Front Immunol. 2020 Oct 14;11:571959.

  Threonyl-tRNA Synthetase Promotes T Helper Type 1 Cell Responses by Inducing Dendritic Cell Maturation and IL-12 Production via an NF-κB Pathway
  U0126-EtOH purchased from AbMole

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  Mar Drυgs. 2020 Aug 17;18(8):429.

  Comparison of Physicochemical Characteristics and Macrophage Immunostimulatory Activities of Polysaccharides from Chlamys farreri
  U0126-EtOH purchased from AbMole

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  Oncology. 2020 Jul 20.

  5-Fluorouracil enhances the chemosensitivity of gastric cancer to TRAIL via inhibition of the MAPK pathway
  U0126-EtOH purchased from AbMole

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  Int J Oncol. 2020 Jul;57(1):197-212.

  G6PD Facilitates Clear Cell Renal Cell Carcinoma Invasion by Enhancing MMP2 Expression Through ROS‑MAPK Axis Pathway
  U0126-EtOH purchased from AbMole

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  Stem Cell Res Ther. 2019 Dec 10;10(1):378.

  Magnesium-enriched microenvironment promotes odontogenic differentiation in human dental pulp stem cells by activating ERK/BMP2/Smads signaling.
  U0126-EtOH purchased from AbMole

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  Stem Cell Res Ther. 2019 Jan 31;10(1):48.

  MagT1 regulated the odontogenic differentiation of BMMSCs induced byTGC-CM via ERK signaling pathway.
  U0126-EtOH purchased from AbMole

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  J Immunol. 2018 Oct 1.

  Lysyl–Transfer RNA Synthetase Induces the Maturation of Dendritic Cells through MAPK and NF-κB Pathways, Strongly Contributing to Enhanced Th1 Cell Responses.
  U0126-EtOH purchased from AbMole

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  BMC Genomics. 2017 Sep 11;18(1):714.

  The dynamic landscape of gene regulation during Bombyx mori oogenesis
  U0126-EtOH purchased from AbMole

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  Int Immunopharmacol. 2016 Nov;40:508-516.

  Costunolide, a sesquiterpene lactone, inhibits the differentiation of pro-inflammatory CD4+ T cells through the modulation of mitogen-activated protein kinases.
  U0126-EtOH purchased from AbMole

Customer Product Validations & Biological Datas

	Source	J Immunol (2018 Oct). Figure 3. U0126 (AbMole BioScience)
	Method	Western blots
	Cell Lines	DCs
	Concentrations	10 μM
	Incubation Time	30 min
	Results	The addition of NF-kB- and JNK-specific inhibitors signifi-cantly attenuated the KRS-induced IL-12 production from DCs, whereas p38 kinase and ERK inhibitors only slightly affected the IL-12 production from KRS-treated DCs.

	Source	BMC Genomics (2017). Additional file 7. U0126 (Abmole Bioscience, USA)
	Method	qRT-PCR
	Cell Lines	pupa abdomen
	Concentrations
	Incubation Time	48 h
	Results	We performed injection experiments involving several inhibitors of the insulin signaling pathway. It was observed that the development of follicles was clearly delayed and the follicles showed abnormalities (Additional file 7), and the expressions of marker genes, like Cyp18a1 and early chorion gene, were up-regulated at choriogenic stages.

	Source	International Immunopharmacology(2016).Figure 4. U0126 (AbMole BioScience, Houston, TX)
	Method	CD4+ T cell isolation and T cell polarization in vitro
	Cell Lines	CD4+ T cells
	Concentrations	0.5–2.0 μM
	Incubation Time
	Results	When CD4+ T cells were treated with costunolide, U0126, or SB203580, the expression of CD69 upon TCR stimulation was significantly down-regulated (Fig. 4B). Th1 and Th2 cell generation was suppressed when ERK activation was inhibited by U0126 in a similar fashion to that of costunolide, albeit the development of Th17 cells was unchanged in the presence of U0126 (Fig. 4C).

Protocol (for reference only)

Cell Experiment
Cell lines	EHMES-10 cells and MSTO211H cells
Preparation method	Cell proliferation assay. The cell proliferation assay reagent WST-1 (4-[3-(4-lodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]- 1,3-benzene disulfonate) (Roche Diagnostics GmbH, Mannheim, Germany) was used to assess the effect of U0126 or LY294002 on cell growth. MPM cells (1x104 cells/well) were plated in 96-well plates (Nunc, Roskilde, Denmark) and were exposed to various concentrations of test agents dissolved in DMSO. Controls received DMSO vehicle at a concentration equal to that of drug treated cells. After drug treatment for 72 h, 10 µl of WST-1 reagent were added to each well. Absorbance was measured at 450 nm with a reference wavelength at 690 nm by an E max precision microplate reader (Molecular Devices, Tokyo, Japan).
Concentrations	0~200µM
Incubation time	72 h

Animal Experiment
Animal models	Orthotopic implantation model
Formulation	DMSO + PBS
Dosages	20, 30 and 40 mg/kg twice a week
Administration	intraperitoneally

Chemical Information

Molecular Weight	426.56
Formula	C18H16N6S2.C2H6O
CAS Number	1173097-76-1
Solubility (25°C)	DMSO ≥ 50 mg/mL (warmed)
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Priscila B Rosa, et al. Purinergic Signal. The antidepressant-like effect of guanosine is dependent on GSK-3β inhibition and activation of MAPK/ERK and Nrf2/heme oxygenase-1 signaling pathways

[2] Satoshi Deyama, et al. Int J Neuropsychopharmacol. Resolvin D1 and D2 Reverse Lipopolysaccharide-Induced Depression-Like Behaviors Through the mTORC1 Signaling Pathway

[3] Ana Lucia Bertarello Z, et al. Pharmacol Biochem Behav. Involvement of PKA, CaMKII, PKC, MAPK/ERK and PI3K in the acute antidepressant-like effect of ferulic acid in the tail suspension test

[4] Namura S, et al. Proc Natl Acad Sci U S A. Intravenous administration of MEK inhibitor U0126 affords brain protection against forebrain ischemia and focal cerebral ischemia.

[5] Duncia JV, et al. Bioorg Med Chem Lett. MEK inhibitors: the chemistry and biological activity of U0126, its analogs, and cyclization products.