JNK-IN-7 (Synonyms: JNK-IN-7, c-Jun N-terminal Kinase Inhibitor VII) |
Catalog No.GC15028 |
A non-selective JNK inhibitor
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 1408064-71-0
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Kinase experiment [1]: | |
Cell-Based assays for c-Jun phosphorylation |
The cell-based kinase assays for c-Jun phosphorylation were carried out by using the LanthaScreen c-Jun (1 ~ 79) HeLa cell lines which stably expressed GFP-c-Jun 1 ~ 79 and GFP-ATF2 19 ~ 106, respectively. Phosphorylation was determined by measuring the TR-FRET between a terbium-labeled phospho-c-Jun specific antibody and GFP. The cells were plated in white tissue culture treated 384 well plates at a density of 10,000 cells per well in 32 μL assay medium (Opti-MEM, supplemented with 0.5% charcoal/dextran-treated FBS, 100 U/mL Penicillin, 100 μg/mL Streptomycin, 0.1 mM nonessential amino acids, 1 mM sodium pyruvate, 25 mM HEPES [pH 7.3] and lacking phenol red). After overnight incubation, cells were pretreated for 90 mins with JNK-IN-7 (at indicated concentration) diluted in 4 μL assay buffer followed by 30 mins of stimulation with 5 ng/mL of TNF-α in 4 μL assay buffer (final assay volume was 40 μL). The medium was then removed by aspiration and the cells were lysed by adding 20 μL of lysis buffer (20 mM Tris-HCl [pH 7.6], 5 mM EDTA, 1% Nonidet P-40 substitute, 5 mM NaF, 150 mM NaCl and 1:100 protease and phosphatase inhibitor mix, P8340 and P2850, respectively). The lysis buffer included 2 nM of the terbium-labeled anti-c-Jun (pSer73) detection antibodies. After allowing the assay to equilibrate for 60 mins at room temperature, TR-FRET emission ratios were determined on a BMG Pherastar fluorescence plate reader using the following parameters: excitation at 340 nm, emission 520 and 490 nm; 100 ms lag time; 200 μs integration time; emission ratio = Em 520/Em 490. All data were analyzed and plotted using GraphPad Prism 4. |
Cell experiment [2]: | |
Cell lines |
Human IL-1R cells and RAW264.7 macrophages |
Preparation method |
Soluble in DMSO. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
Reaction Conditions |
0.1, 1 and 10 mM; 1 hr |
Applications |
In human IL-1R cells, JNK-IN-7 inhibited IL-1β-stimulated phosphorylation of c-Jun and the activation of Pellino 1. In Pam3CSK4-stimulated RAW macrophages, JNK-IN-7 also inhibited the phosphorylation of c-Jun. |
References: [1]. Zhang T, Inesta-Vaquera F, Niepel M, et al. Discovery of potent and selective covalent inhibitors of JNK. Chem Biol, 2012, 19(1): 140-154. [2]. Goh ET, Arthur JS, Cheung PC, et al. Identification of the protein kinases that activate the E3 ubiquitin ligase Pellino 1 in the innate immune system. Biochem J, 2012, 441(1): 339-346. |
JNK-IN-7 is a selective JNK inhibitor with IC50 values of 1.54 nM, 1.99 nM, 0.75 nM to JNK1, JNK2, JNK3, respectively. It also inhibits phosphorylation of c-Jun, which is a direct substrate of JNK kinase.
JNK-IN-8, an analog of JNK-IN-7 with an extra flag methyl, dramatically improved in selectivity and eliminated binding to IRAK1, PIK3C3, PIP4K2C and PIP5K3, comparing to JNK-IN-7. JNK-IN-7 and JNK-IN-8 require Cys116 for JNK2 inhibition. JNK-IN-7 can indeed inhibit IRAK-1 dependent E3 ligase activity of pellino, which plays an role in the Toll receptor signaling pathway in cells at relative high compound concentrations (1–10 mM).
The IRAK1 inhibitor JNK-IN-7 inhibited the IL-1-stimulated activation of Pellino 1 in IL-1R cells, but not the Pam3CSK4-stimulated activation of Pellino 1 in RAW264.7 macrophages. JNK-IN-7 also suppressed the phosphorylation of c-Jun in Pam3CSK4-stimulated RAW macrophages, but in contrast with IL-1R cells it did not affect the activation of Pellino 1.
References:
[1]. Zhang T, Inesta-Vaquera F, Niepel M et al. Discovery of potent and selective covalent inhibitors of JNK. Chem Biol. 2012 Jan 27;19(1):140-54.
[2]. Goh ET, Arthur JS, Cheung PC et al. Identification of the protein kinases that activate the E3 ubiquitin ligase Pellino 1 in the innate immune system. Biochem J. 2012 Jan 1;441(1):339-46.
Cas No. | 1408064-71-0 | SDF | |
Synonyms | JNK-IN-7, c-Jun N-terminal Kinase Inhibitor VII | ||
Chemical Name | 3-[[(E)-4-(dimethylamino)but-2-enoyl]amino]-N-[4-[(4-pyridin-3-ylpyrimidin-2-yl)amino]phenyl]benzamid | ||
Canonical SMILES | CN(C)CC=CC(=O)NC1=CC=CC(=C1)C(=O)NC2=CC=C(C=C2)NC3=NC=CC(=N3)C4=CN=CC=C4 | ||
Formula | C28H27N7O2 | M.Wt | 493.56 |
Solubility | ≥ 24.7 mg/mL in DMSO | Storage | Store at -20°C |
General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. |
Prepare stock solution | |||
1 mg | 5 mg | 10 mg | |
1 mM | 2.0261 mL | 10.1305 mL | 20.261 mL |
5 mM | 0.4052 mL | 2.0261 mL | 4.0522 mL |
10 mM | 0.2026 mL | 1.013 mL | 2.0261 mL |
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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
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