(2Z,3Z)-bis({amino[(2-aminophenyl)sulfanyl]methylidene})butanedinitrile; ethanol

• ChemBase ID: 72510
• Molecular Formular: C20H22N6OS2
• Molecular Mass: 426.55828
• Monoisotopic Mass: 426.12965135
• SMILES: c1ccc(c(c1)S/C(=C(/C(=C(/Sc1c(cccc1)N)\N)/C#N)\C#N)/N)N.CCO
• Canonical SMILES: N#C/C(=C(\Sc1ccccc1N)/N)/C(=C(/Sc1ccccc1N)\N)/C#N.CCO
• InChI: 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+
• InChIKey: CFQULUVMLGZVAF-OYJDLGDISA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: (2Z,3Z)-bis({amino[(2-aminophenyl)sulfanyl]methylidene})butanedinitrile; ethanol; bis({amino[(2-aminophenyl)sulfanyl]methylidene})butanedinitrile; ethanol
• IUPAC Traditional name: (2Z,3Z)-bis({amino[(2-aminophenyl)sulfanyl]methylidene})butanedinitrile; ethyl alcohol; bis({amino[(2-aminophenyl)sulfanyl]methylidene})butanedinitrile; ethyl alcohol
• Synonyms: U0126-EtOH; 1,4-Diamino-2,3-dicyano-1,4-bis(o-aminophenylmercapto)butadiene monoethanolate; U0126 monoethanolate

Database IDs

• CAS Number: 1173097-76-1
• PubChem SID: 24278770; 162037435
• PubChem CID: 16220066

CALCULATED PROPERTIES

• H Acceptors: 6
• H Donor: 4
• LogD (pH = 5.5): 1.9509677
• LogD (pH = 7.4): 1.9532725
• Log P: 1.9533019
• Molar Refractivity: 130.886 cm^3
• Polarizability: 40.824818 Å^3
• Polar Surface Area: 151.66 Å^2
• Rotatable Bonds: 5
• Lipinski's Rule of Five: true

PROPERTIES

Physical Property

• Solubility: DMSO; DMSO: >10 mg/mL (Stock solutions in DMSO should be used within a month. Acidic solutions exhibit weaker inhibition)
• Apperance: white to off-white powder

Safety Information

• Storage Condition: -20°C; protect from light
• German water hazard class: 3
• Personal Protective Equipment: Eyeshields, Gloves, type N95 (US), type P1 (EN143) respirator filter
• Storage Temperature: 2-8°C

Pharmacology Properties

• Target: MEK
• Gene Information: human ... MAP2K1(5604), MAP2K2(5605)

Product Information

• Purity: ≥98% (HPLC)
• Salt Data: Ethanolate
• Empirical Formula (Hill Notation): C18H16N6S2 · C2H5OH

DETAILS

Selleck Chemicals - S1102

Research Area

• Description: Cancer

Biological Activity

• Description: U0126-EtOH is a highly selective inhibitor of MEK1 and MEK2 with IC50 of 0.07 μM and 0.06 μM, respectively.
• Targets:

  MEK1

  ;

  MEK2

• IC50:

  0.07 μM

  ;

  0.06 μM ^[1]

• In Vitro: 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: 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]
• Features: U0126-EtOH is a chemically synthesized and highly selective inhibitor of both MEK1 and MEK2.

Protocol

• Protocol: Kinase Assay ^[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.
• Protocol: Cell Assay ^[2]
• Cell Lines: A.E7 or Th17 cells
• Concentrations: 0 to 10 μM
• Incubation Time: 48 hours
• Methods:

  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.

• Protocol: Animal Study ^[4]
• Animal Models: 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).
• Formulation: U0126-EtOH is dissolved in 10% DMSO, 30% of Cremophor EL and 60% PBS.
• Doses: ≤10 mM
• Administration: Administered via aerosol.

References

• References: [1] Duncia JV, et al. Bioorg Med Chem Lett. 1998, 8(20), 2839-2844.
• References: [2] DeSilva DR, et al. J Immunol. 1998, 160(9), 4175-4181.
• References: [3] Freeman MR, et al. Cancer Biol Ther. 2011, 12(11), 966-977.
• References: [4] Droebner K, et al. Antiviral Res. 2011, 92(2), 195-203.
• References: [5] Ashabi G, et al. Behav Brain Res. 2012, http://dx.doi.org/10.1016/j.bbr.2012.04.006.

Sigma Aldrich - U120

Caution
photosensitive
Biochem/physiol Actions
U0126, 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio] butadiene, is a highly selective inhibitor of both MEK1 and MEK2. U0126 was found to functionally antagonize AP-1 transcriptional activity via noncompetitive inhibition of the dual specificity kinase MEK with IC50 of 72 nM for MEK1 and 58 nM for MEK2. 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. The effects of U0126 on the growth of eight human breast cancer cell lines shown that U0126 selectively repressed anchorage-independent growth of MDA-MB231 and HBC4 cells, two lines with constitutively activated ERK. Loss of contact with substratum triggers apoptosis in many normal cell types, a phenomenon termed anoikis. U0126 sensitized MDA-MB231 and HBC4 to anoikis, i.e., upon treatment with U0126, cells deprived of anchorage entered apoptosis.

REFERENCES

• Duncia JV, et al. Bioorg Med Chem Lett. 1998, 8(20), 2839-2844.
• DeSilva DR, et al. J Immunol. 1998, 160(9), 4175-4181.
• Freeman MR, et al. Cancer Biol Ther. 2011, 12(11), 966-977.
• Droebner K, et al. Antiviral Res. 2011, 92(2), 195-203.
• Ashabi G, et al. Behav Brain Res. 2012, http://dx.doi.org/10.1016/j.bbr.2012.04.006.