6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

• ChemBase ID: 72600
• Molecular Formular: C22H18Cl2N8
• Molecular Mass: 465.33792
• Monoisotopic Mass: 464.10314798
• SMILES: n1cc(c(nc1NCCNc1ccc(cn1)C#N)c1ccc(cc1Cl)Cl)c1[nH]cc(n1)C
• Canonical SMILES: N#Cc1ccc(nc1)NCCNc1ncc(c(n1)c1ccc(cc1Cl)Cl)c1[nH]cc(n1)C
• InChI: InChI=1S/C22H18Cl2N8/c1-13-10-29-21(31-13)17-12-30-22(32-20(17)16-4-3-15(23)8-18(16)24)27-7-6-26-19-5-2-14(9-25)11-28-19/h2-5,8,10-12H,6-7H2,1H3,(H,26,28)(H,29,31)(H,27,30,32)
• InChIKey: AQGNHMOJWBZFQQ-UHFFFAOYSA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: 6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile
• IUPAC Traditional name: 6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile
• Synonyms: CHIR-99021

Database IDs

• CAS Number: 252917-06-9
• PubChem SID: 162037525
• PubChem CID: 9956119

CALCULATED PROPERTIES

• Acid pKa: 13.594071
• H Acceptors: 7
• H Donor: 3
• LogD (pH = 5.5): 3.5425742
• LogD (pH = 7.4): 3.9773595
• Log P: 3.9875154
• Molar Refractivity: 138.0448 cm^3
• Polarizability: 48.580708 Å^3
• Polar Surface Area: 115.2 Å^2
• Rotatable Bonds: 7
• Lipinski's Rule of Five: true

PROPERTIES

Safety Information

• Storage Condition: -20°C

Pharmacology Properties

• Target: GSK-3

Product Information

• Salt Data: Free Base

DETAILS

Selleck Chemicals - S1263

Research Area

• Description: Cancer

Protocol

• Protocol: Kinase Assay ^[1]
• Cell-free kinase assays: Polypropylene 96-well plates are filled with 300 μL/well buffer (50 mM tris HCl, 10 mM MgCl₂, 1 mM EGTA, 1 mM dithiothreitol, 25 mM β-glycerophosphate, 1 mM NaF, 0.01% BSA, pH 7.5) containing 27 nM GSK-3α or 29 nM GSK-3β, and 0.5 μM biotin-CREB peptide substrate. Various concentrations of CHIR-99021 are added in 3.5 μL of DMSO, followed by 50 μL of ATP stock to yield a final concentration of 1 μM ATP in all cell-free assays. After incubation, triplicate 100-μL aliquots are transferred to Combiplate eight plates containing 100 μL/well 50 μM ATP and 20 mM EDTA. After 1 hour, the wells are rinsed five times with PBS, filled with 200 μL of scintillation fluid, sealed, left 30 minutes, and counted in a scintillation counter. All steps are performed at room temperature.
• Protocol: Cell Assay ^[3]
• Cell Lines: INS-1E
• Concentrations: Dissolved in DMSO, final concentrations ~ 20 μM
• Incubation Time: 1, or 4 days
• Methods: Cells are maintained for 24 hours in starvation medium (culture medium with only 5 mM glucose, 1% fetal calf serum). Then cells are exposed to various concentrations of CHIR-99021 for 1, or 4 days. Cell number is measured by staining of cellular DNA with CyQuant dye, which becomes fluorescent when bound to DNA. Fluorescence is measured after 30 minutes of incubation using the FLUOstar Optima reader. Cell replication is determined by BrdUrd incorporation. BrdUrd labeling solution is added to the medium for the last 4 hours before the cells are fixed using FixDenat solution and incubated with monoclonal anti-BrdUrd-POD antibodies. After substrate solution is added to each well, the light emission is measured in a microplate luminometer using the Analyst HT detection system.
• Protocol: Animal Study ^[1]
• Animal Models: Female db/db mice or male ZDF rats with type 2 diabetes
• Formulation: Formulated as solutions in 20 mM citrate-buffered 15% Captisol or as fine suspensions in 0.5% carboxymethylcellulose.
• Doses: ~48 mg/kg
• Administration: Orally

References

• References: [1] Ring DB, et al. Diabetes, 2003, 52(3), 588-595.
• References: [2] Bennett CN, et al. J Biol Chem, 2002, 277(34), 30998-31004.
• References: [3] Mussmann R, et al. J Biol Chem, 2007, 282(16), 12030-12037.
• References: [4] Trowbridge JJ, et al. Nat Med, 2006, 12(1), 89-98.

REFERENCES

• Ring DB, et al. Diabetes, 2003, 52(3), 588-595.
• Bennett CN, et al. J Biol Chem, 2002, 277(34), 30998-31004.
• Mussmann R, et al. J Biol Chem, 2007, 282(16), 12030-12037.
• Trowbridge JJ, et al. Nat Med, 2006, 12(1), 89-98.