Home PI3K/Akt/mTOR GSK-3 inhibitor AZD2858

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AZD2858 GSK-3 inhibitor

AZD2858 is a selective GSK-3 inhibitor with an IC50 of 68 nM, activating Wnt signaling, increases bone mass in rats.

AZD2858 GSK-3 inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 453.52

Purity & Quality Control

Batch: S725301 DMSO]7 mg/mL]false]Water]Insoluble]false]Ethanol]Insoluble]false Purity: 99.56%
99.56

Related Products

Signaling Pathway

GSK-3 Signaling Pathways

GSK-3 Signaling Pathway

Mechanism of Action

Targets
GSK-3 [1]
68 nM

In vitro
In vitro AZD2858 is a selective GSK-3 inhibitor with an IC50 of 68 nM, inhibits tau phosphorylation at the S396 site, activates Wnt signaling pathway. This compound treatment (1 μM, 12 h) on primary isolated human osteoblast-like cells results in a 3-fold increase of β-catenin levels. [1] It causes β-catenin stabilisation in human and rat mesenchymal stem cells, stimulates hADSC commitment towards osteoblasts and osteogenic mineralisation in vitro. [2]
Kinase Assay Tau phosphorylation assay
NIH-3T3 cells expressing 4-repeat Tau are used to assess functional activity of AZD2858 in vitro. The cells are grown in DMEM media and 2 mM L-glut, and 10% HiFCS, and plated at a concentration of 6×105 cells/well in 6-well plates. In each experiment, this compound is dosed in triplicates at a concentration of 1, 10, 100, 500, 1000, 2000 and 10,000 nM. Cells are treated for 4 h prior to cell lysis using 100 μL ice cold lysis buffer (0.5% NP-40, 10 mM Tris, pH 7.2, 150 mM NaCl, 2 mM EDTA). A suspension is made with addition of protease and phosphatase inhibitors: 50 mM NaF, 0.2 mM NaVO4 and Cocktail Protease inhibitors. The solution is then snap frozen at − 80 °C for at least 1 h, before thawing on ice and lysate clarification by centrifugation, followed by Western blot according to standard protocols. After blocking, the blots are exposed to the primary antibody, Phospho-Ser396-tau (1:1000) over night, washed and incubated with the secondary antibody (donkey anti-rabbit, 1:5000), followed by a final wash. For re-probing, the primary antibody Tau5 (1:200) and the secondary horseradish peroxidase linked antibody (sheep anti-mouse, 1:10000) are used. All blots are developed using ECL Western blot detection reagents, Kodak X-ray films, quantified using densitometric analysis, and the ratio of S396 tau to total tau (tau5) is calculated.
Cell Research Cell lines Human adipose derived stem cells and rat MSCs
Concentrations ~20 mM
Incubation Time 24 h
Method Human adipose derived stem cells and rat MSCs (isolated from bone marrow of Sprague Dawley rats at less than 8 weeks after gestation) are cultured in a basal media of DMEM containing 5% FBS and 2 mM GlutaMax. Cells are seeded in basal media into 96-well plates (3–5000 cells/well) for 18 h before treatment with AZD2858 (0.3 nM to 20 mM). After 24 h, β-catenin stabilisation is measured.

In Vivo
In vivo In rats, oral AZD2858 treatment causes a dose-dependent increase in trabecular bone mass compared to control after a two-week treatment with a maximum effect at a dose of 20 mg/kg once daily (total BMC: 172% of control). A small but significant effect is also seen at cortical sites (total BMC: 111% of control). [1] This compound (30 μmol/kg) on rats daily for up to 3 weeks shows an increase in both mineral density (of 28% at 2 weeks and 38% at 3 weeks) and mineral content (of 81% at 2 weeks and 93% at 3 weeks) in the calluses. This treatment makes the fractures heals more rapidly, with a bony callus without an obvious endochondral component. [3] It produces time-dependent changes in serum bone turnover biomarkers and increases bone mass over 28 days exposure in rats. After 7 days, this compound increases the bone formation biomarker P1NP, and reduces the resorption biomarker TRAcP-5b, indicating increased bone anabolism and reduced resorption in rats. [2]
Animal Research Animal Models rat
Dosages 30 μmol/kg/day
Administration orally

References
  • https://pubmed.ncbi.nlm.nih.gov/22142634/
  • https://pubmed.ncbi.nlm.nih.gov/23872097/
  • https://pubmed.ncbi.nlm.nih.gov/23337038/

Chemical Information

Molecular Weight 453.52 Formula

C21H23N7O3S
CAS No. 486424-20-8 SDF Download SDF
Synonyms N/A
Smiles CN1CCN(CC1)S(=O)(=O)C2=CC=C(C=C2)C3=CN=C(C(=N3)C(=O)NC4=CN=CC=C4)N

Storage and Stability

Storage (From the date of receipt)

In vitro
Batch:

DMSO : 7 mg/mL ( (15.43 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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In vivo
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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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