Home PI3K/Akt/mTOR GSK-3 inhibitor AR-A014418

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AR-A014418 GSK-3 inhibitor

Cat.No.S7435

AR-A014418 (GSK-3β Inhibitor VIII) is an ATP-competitive, and selective GSK3β inhibitor with IC50 and Ki of 104 nM and 38 nM in cell-free assays, without significant inhibition on 26 other kinases tested.
AR-A014418 GSK-3 inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 308.31

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Quality Control

Batch: Purity: 99.76%
99.76

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human BxPC3 cells Growth inhibition assay 72 h Growth inhibition of human BxPC3 cells after 72 hrs by MTS assay, IC50=14 μM
human HUPT3 cells Growth inhibition assay 72 h Growth inhibition of human HUPT3 cells after 72 hrs by MTS assay, IC50=22 μM
human MIAPaCa2 cells Growth inhibition assay 72 h Growth inhibition of human MIAPaCa2 cells after 72 hrs by MTS assay, IC50=29 μM
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 61 mg/mL (197.85 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

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Mass Concentration Volume Molecular Weight
Dilution Calculator Molecular Weight Calculator

In vivo
Batch:

In vivo Formulation Calculator (Clear solution)

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
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Chemical Information, Storage & Stability

Molecular Weight 308.31 Formula

C12H12N4O4S

 Storage (From the date of receipt)
CAS No. 487021-52-3 Download SDF Storage of Stock Solutions

Synonyms GSK-3β Inhibitor VIII Smiles COC1=CC=C(C=C1)CNC(=O)NC2=NC=C(S2)[N+](=O)[O-]

Mechanism of Action

Features
Cell-permeable GSK3-selective inhibitor.
Targets/IC50/Ki
GSK-3β
(Cell-free assay)
38 nM(Ki)
GSK-3β
(Cell-free assay)
38 nM(Ki)
In vitro
AR-A014418 inhibits tau phosphorylation at a GSK3-specific site (Ser-396) in 3T3 fibroblasts expressing human four-repeat tau protein with IC50 of 2.7 μM, and protects cultured N2A cells from death induced by blocking PI3K/PKB pathway. In hippocampal slices, this compound inhibits neurodegeneration mediated by beta-amyloid peptide. While in NGP and SH-5Y-SY cells, this chemical reduces neuroendocrine markers and suppresses neuroblastoma cell growth.
Kinase Assay
GSK3 Scintillation Proximity Assay
The competition experiments are carried out in duplicate with 10 concentrations of the inhibitor in clear-bottomed microtiter plates. The biotinylated peptide substrate, biotin-AAEELDSRAGS(PO3H2)PQL, is added at a final concentration of 2 μM in an assay buffer containing 6 milliunits of recombinant human GSK3 (equal mix of both α and β), 12 mM MOPS, pH 7.0, 0.3 mM EDTA, 0.01% β-mercaptoethanol, 0.004% Brij 35, 0.5% glycerol, and 0.5 μg of bovine serum albumin/25 μl and preincubated for 10-15 min. The reaction is initiated by the addition of 0.04 μCi of [γ-33P]ATP and unlabeled ATP in 50 mM Mg(Ac)2 to a final concentration of 1 μM ATP and assay volume of 25 μl. Blank controls without peptide substrate are used. After incubation for 20 min at room temperature, each reaction is terminated by the addition of 25 μl of stop solution containing 5 mM EDTA, 50 μM ATP, 0.1% Triton X-100, and 0.25 mg of streptavidin-coated SPA beads corresponding to ∼35 pmol of binding capacity. After 6 h the radioactivity is determined in a liquid scintillation counter. Inhibition curves are analyzed by non-linear regression using GraphPad Prism.
In vivo
In ALS mouse model with the G93A mutant human SOD1, AR-A014418 (0-4 mg/kg, i.p.) delays the onset of symptoms, improves motor activity, slows down disease progression, and postpons the endpoint of the disease. In addition, this compound produces inhibition effect on acetic acid- and formalin-induced nociception in mice by modulating NMDA and metabotropic receptor signaling as well as TNF-α and IL-1β transmission in the spinal cord.
References
  • [4] https://pubmed.ncbi.nlm.nih.gov/20705523/

Applications

Methods Biomarkers Images PMID
Western blot β-catenin β-catenin / GSK3α / GSK3β / Notch1 TAK1 / TAB1 / TAB2 / p-p65 / p65
S7435-WB1
26292722
Growth inhibition assay Cell viability
S7435-viability1
26292722

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