Cell lines

HEK293-ILR1 cells

Preparation method

Soluble in DMSO >25.4mg/mL. 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.

Reacting condition

0.1,0.3,1,3μM for 3hr

Applications

JNK-IN-8 was an extremely potent inhibitor of enzymatic and cellular JNK(c-Jun N-terminal kinase) that inhibited phosphorylation of c-Jun, the direct substrate of JNK kinase. The superior potency and selectivity of JNK-IN-8 in the HEK293 cells suggested that the compound would likely serve as very useful pharmacological probes of JNK-dependent cellular phenomena.

Animal models

Male KM mice (CL)(8-week-old)

Dosage form

3μg/μL, injection

Application

JNK-IN-8, a specific inhibitor of JNK pathway, could reduce the neuronal apoptosis significantly as compared to the DMSO group after brain injury in the mice.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

JNK-IN-8 is a specific JNK1/2/3 inhibitor with IC50 value of 4.67, 18.7, 0.98 nM respectively [1].

C-Jun N-terminal kinase (JNK) 1, 2 and 3 belong to the mitogen-activated protein kinase (MAPK) family, which are able to phosphorylate c-Jun on the Ser63 and Ser73 residue.They are responsive for stress stimuli, including cytokines and heat shock, and get involved in T cell differentiation and cell apoptosis process. JNK 1 and 2 are ubiquitous in all cell types but JNK 3 is only found in cells of brain, heart and testes tissues.

JNK-IN-8 is a JNK1/2/3 inhibitor with high specificity. When JNK-IN-8 was profiled with a panel of 400 kinases, it exhibited specific binding to JNK 1/2/3 but not to other kinases. Crystallization study also found that JNK-IN-8 forms covalent bonds with conserved cysteine residue of JNK 1/2/3, resulting in a conformational change of the activation loop that blocks the substrate binding, thereby inhibiting the activity of JNK 1/2/3 [1].

In Hela cells and A375 cells, pretreatment of cells with JNK-IN-8 resulted in the inhibition of c-Jun which is a direct phosphorylation substrate of JNK 1/2/3, confirming the inhibitory action of JNK-IN-8 on JNK 1/2/3. In HEK293-ILR1 cells following stimulation by anisomycin, the JNK-IN-8 was observed to inhibit c-Jun but not MSK1 and p38, and the inhibition was not reversible by removing JNK-IN-8 from culture medium. Additionally, JNK-IN-8 only exhibited on-pathway inhibition of JNK signaling pathway, which can be monitored by the phosphorylation of c-Jun [1].

Reference:
[1].  Zhang T et al., Discovery of potent and selective covalent inhibitors of JNK. Chemical Biology. 2012, 19(1):140-154.