Molecular Weight (MW)

377.41

Formula

C22H20FN3O2

CAS No.

1129669-05-1

Storage

-20℃ for 3 years in powder form

-80℃ for 2 years in solvent

Solubility (In vitro)

DMSO: 75 mg/mL (198.7 mM)

Water: <1 mg/mL

Ethanol: 10 mg/mL (26.5 mM)

SMILES

FC(C=C1)=CC=C1CCNC(C2=O)=C(C(N2C)=O)C(C3=C(C=CC=C3)N4)=C4C

Synonyms

IM-12; IM 12; IM12; 3-(4-Fluorophenylethylamino)-1-methyl-4-(2-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione

In Vitro

In vitro activity: In hNPCs, IM-12 inhibits GSK-3β activity with IC50 of 3.8 μM and subsequently increases β-catenin concentration significantly. Through the activation of canonical Wnt signalling pathway, IM-12 promotes the neuronal differentiation of human neural progenitor cells.

Kinase Assay: IC50 of new synthesised compound IM-12 to GSK-3β is determined by a luminometric GSK-3β activity assay. Briefly, compounds are tested in different concentrations diluted in assay buffer containing final concentrations of: 4 mM MOPS pH 7.2; 0.4 mM EDTA; 1 mM EGTA; 2.5 mM β-glycerophosphate; 4 mM MgCl2; 40 μM BSA; 0.05 mM DTT. Four microlitres of diluted compounds are added to 25 μM pGS-2 peptide substrate, 20 ng recombinant GSK-3β and 1 μM ATP to a total assay volume of 40 μl. The enzymatic reaction is stopped after 30 min of incubation at 30 °C by adding 40 μl KinaseGlo. The luminometric signal is allowed to stabilise for 10 min and the measured with a Glomax(R) 96 Microplate Luminometer.

Cell Assay: To measure viable cells, 50-100 μL of cell suspension is analyzed using CASY technology with the appropriate program. ReNcell VM cells are seeded at a defined cell number and proliferated for 24 h. Then the medium is changed to proliferation medium with added substances at indicated concentrations. The cell number was determined every 24 h. Cells were exposed to the added drugs during the whole experiment, whereas the media is changed every 24 h.

In Vivo

Animal model

Formulation & Dosage

References

Bioorg Med Chem. 2010 Sep 15;18(18):6785-95.