Molecular Weight (MW)

273.08

Formula

C9H9BrN2O3

CAS No.

188011-69-0

Storage

-20℃ for 3 years in powder form

-80℃ for 2 years in solvent

Solubility (In vitro)

DMSO: 54 mg/mL (197.7 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

SMILES

BrC1=CN=C(NC(CCC(O)=O)=O)C=C1

Synonyms

4-((5-bromopyridin-2-yl)amino)-4-oxobutanoic acid; Abrasin;

InChi Key: XFYYQDHEDOXWGA-UHFFFAOYSA-N

InChi Code: InChI=1S/C9H9BrN2O3/c10-6-1-2-7(11-5-6)12-8(13)3-4-9(14)15/h1-2,5H,3-4H2,(H,14,15)(H,11,12,13)

SMILES Code: BrC1=CN=C(NC(CCC(O)=O)=O)C=C1

In Vitro

In vitro activity: Bikinin directly targets BIN2 of Arabidopsis GSK-3s and activates brassinosteroid-dependent gene expression. Bikinin also inhibits RD26 expression, and inhibits the induction of GUS activity by 150 mM NaCl in the RD26pro:GUS line.

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

Bikinin strongly inhibits abscisic acid signaling outputs, and inhibits T180 phosphorylation of SnRK2.3 and its kinase activity in vivo.

Animal model

Formulation & Dosage

References

Mol Cells. 2014 Nov;37(11):795-803; Proc Natl Acad Sci U S A. 2014 Jul 1;111(26):9651-6.