Kinase assays

Purified kinase domains were incubated with AS602801 at two-fold dilutions over a concentration range of 100 uM to 0.006 uM or with vehicle (0.1% DMSO) in the presence of 10 uM ATP, 2.5 uCi of [γ-32P]ATP and substrate. Reactions were terminated by spotting onto nitrocellulose membranes; membrane was then washed four times to remove unbound radioactivity and dried. Transferred radioactivity was quantitated by phosphorimaging, and IC50 values were calculated by fitting the data to a sigmoidal dose-response.

Cell lines

PANC-1, A549, A2780 cell lines

Preparation method

The solubility of this compound in DMSO is >11.5 mg/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

7.5 μM, 3 days

Applications

AS602801 had selective cytotoxic activity against neoplastic cells and had anticancer effects. AS602801 treatment induced three serum-cultured human cancer cell lines (PANC-1, pancreatic cancer; A594, lung cancer; A2780, ovarian cancer) death and accordingly decreased the number of viable cells.

Animal models

BALB/cAJcl-nu/nu mice xenograft (PANC-1 CSLCs (primary tumors))

Dosage form

ip, 40 mg/kg daily for 10 consecutive days.

Application

Systemic AS602801 treatment reduced the proportion of tumor-initiating cells within the primary tumors (PANC-1 CSLCs).

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.

Kinases represent one of the most popular and promising target class in drug discovery. Success in finding new therapeutics will depend on the validation of the kinase chosen with respect to the disease of interest, and on the ability of chemists to design and synthesize inhibitors. AS602801 is a potent and selective JNK inhibitor with therapeutic potential inMS and fibrosis.

In vitro: AS602801 blocked T-lymphocyte proliferation and induced apoptosis. In RRMS CD4t and CD8t cells, AS602801 induced apoptosis genes and expression of surface markers, while in RRMS CD11bt cells it induced expression of innate immunity receptors and co-stimulatory molecules [1].

In vivo: AS602801, the best JNK inhibitors identified so far, were currently evaluated in preclinical studies, based on preliminary promising results in animal model of auto-immune diseases and neuronal apoptosis [1].

Clinical trial: Recently, a phase IIa proof of concept study of the pan-JNK inhibitor bentamapimod (PGL5001, AS602801) for the treatment of inflammatory endometriosis had been terminated.

Reference:
[1] Ferrandi C, Richard F, Tavano P, Hauben E, Barbié V, Gotteland JP, Greco B, Fortunato M, Mariani MF, Furlan R, Comi G, Martino G, Zaratin PF.  Characterization of immune cell subsets during the active phase of multiple sclerosis reveals disease and c-Jun N-terminal kinase pathway biomarkers. Mult Scler. 2011;17(1):43-56.