CHIR-99021 (CT99021) HCl

Catalog No.S2924

CHIR-99021 (CT99021) HCl is hydrochloride of CHIR-99021, which is a GSK-3α/β inhibitor with IC50 of 10 nM/6.7 nM in cell-free assays; ability to distinguish between GSK-3 and its closest homologs Cdc2 and ERK2.

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CHIR-99021 (CT99021) HCl Chemical Structure

CHIR-99021 (CT99021) HCl Chemical Structure
Molecular Weight: 501.8

Validation & Quality Control

Cited by 66 publications:

7 customer reviews :

Quality Control & MSDS

Related Compound Libraries

CHIR-99021 (CT99021) HCl is available in the following compound libraries:

GSK-3 Inhibitors with Unique Features

  • Selective GSK-3 Inhibitor

    TWS119 GSK-3β-selective, IC50 of 30 nM.

  • Most Potent GSK-3 Inhibitor

    CHIR-98014 GSK-3α, IC50=0.65 nM; GSK-3β, IC50=0.58 nM.

  • GSK-3 Inhibitor in Clinical Trial

    Tideglusib Phase II for Alzheimer's disease.

  • Newest GSK-3 Inhibitor

    AZD1080 Orally active, brain permeable GSK3 inhibitor, inhibits human GSK3α and GSK3β with Ki of 6.9 nM and 31 nM, respectively.

Product Information

  • Compare GSK-3 Inhibitors
    Compare GSK-3 Products
  • Research Area
  • Inhibition Profile

Product Description

Biological Activity

Description CHIR-99021 (CT99021) HCl is hydrochloride of CHIR-99021, which is a GSK-3α/β inhibitor with IC50 of 10 nM/6.7 nM in cell-free assays; ability to distinguish between GSK-3 and its closest homologs Cdc2 and ERK2.
Targets GSK-3β [1]
(Cell-free assay)
GSK-3α [1]
(Cell-free assay)
Tie-2 [1]
(Cell-free assay)
FLT1 [1]
(Cell-free assay)

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IC50 6.7 nM 10 nM >5 μM >5 μM
In vitro CHIR-99021 shows greater than 500-fold selectivity for GSK-3 versus its closest homologs CDC2 and ERK2, as well as other protein kinases. Furthermore, CHIR-99021 shows only weak binding to a panel of 22 pharmacologically relevant receptors and little inhibitory activity against a panel of 23 nonkinase enzymes. CHIR-99021 induces the activation of glycogen synthase (GS) in insulin receptor-expressing CHO-IR cells with EC50 of 0.763 μM. [1] In addition to simulating the actions of insulin, inhibition of GSK-3 by CHIR-99021 (3 μM) increases free cytosolic β-catenin by 1.9-fold, mimicking the canonical Wnt signaling pathway in 3T3-L1 preadipocytes. During any of the first 3 days of differentiation, CHIR-99021 treatment inhibits the preadipocyte differentiation with IC50 of 0.3 μM by blocking induction of CCAAT/enhancer-binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ). [2] Unlike lithium chloride and AR-A014418, CHIR-99021 treatment does not reduce the viability of INS-1E cells even at high concentrations. Instead, CHIR-99021 robustly increases the rate of proliferation of INS-1E cells in a dose-dependent manner, and significantly inhibits INS-E cell death induced by high glucose and high palmitate in a concentration-dependent manner. CHIR-99021 promotes primary beta cell replication in isolated rat islets at concentrations as low as 1 μM, with 2-3 fold increase of cell replication at 5 μM of CHIR-99021 treatment. [3]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
3T3-L1MmHjSpVv[3Srb36gRZN{[Xl?MoqyN{DPxE1?NEnGcVE1KGh?MmrZSG1UVw>?MkLrTY5pcWKrdIOgdJJm[WSrcH;jfZRmKGSrZn\ldoVvfGmjdHnvci=>MmT6NVIxPTV{MEC=
insulin receptor–expressing CHO-IRMX;GeY5kfGmxbjDBd5NigQ>?MXS1NFAhdk1?MUOzNEBucW5?M3TPR2ROW09?M3PHfGFkfGm4YYTld{BIdHmlb3flckBUgW62aHHz[UApT1NrMnjrNVI3ODZ2OUe=
H4IIENWDxd2JNTnWwY4Tpc44hSXO|YYm=MYGxNEDPxE1?NGLHc4ozPCCqNUC3SIlETE2VTx?=NEDON5BT\We3bHH0[ZMh|rJvY3H0[Y5qdiCjY4Tpeol1gSCjbnSgWGlTTSClb370ZYlvcW6pIIDyc41wfGW{IHHjeIl3cXS7M1nTe|E2OzVyMUm1
Oct4/Sox2/Klf4-infected MEFsM2HWUmZ2dmO2aX;uJGF{e2G7NEHo[4gyOCEQvF2=NXHRZnpVOjhiZB?=NUXmdYlOTE2VTx?=NUPxOIZlWHKxbX;0[ZMhVUWIczDy[ZBzd2e{YX3tbY5oKFS{YX7z[JVk\WRiYomgU4N1PCxiU3;4Nkwh[W6mIFvs[lQ>NWTVUpF7OTl6M{mwOVU>
Sca-1+ HPCMWfHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MW[yJO69VQ>?M2jCclYh\A>?M3fpT2ROW09?NV7SU3diW3mwZYLnbZN1cWOjbHz5JJBzd22xdHXzJHNk[S1zK9MgTHBEKHC{b3zp[oVz[XSrb36gZY5lKGOxbH;ufUBnd3KvYYTpc44hfG:pZYTo[ZIhf2m2aDDFS2Y>MWKyNVYzPDN2OB?=
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Endothelial cell(EC)M3L2O2Z2dmO2aX;uJGF{e2G7NHHDe44{KM7:TR?=MoD4SG1UVw>?MkHLVJJwdW:2ZYOg[Y5ld3SqZXzpZYwh[2WubDCoSWMqKGSrZn\ldoVvfGmjdHnvci=>MnHpNlIxPTJyME[=
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plateletsM{j1N2Z2dmO2aX;uJGF{e2G7NILEXZIyOCEQvF2=M2jsd|E2KG2rbh?=NG\ESIVFVVORMVPJcoNz\WG|ZYOgeIhzd22kaX6tcYVlcWG2ZXSgdIxifGWuZYSgZYdoemWpYYTpc44h[W6mIFHUVEB{\WO{ZYTpc44>NFPzcWgzOzJ|OUi3Oy=>
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... Click to View More Cell Line Experimental Data

In vivo Oral administration of CHIR-99021 at 30 mg/kg enhances glucose metabolism in a rodent model of type 2 diabetes, with a maximal plasma glucose reduction of nearly 150 mg/dl 3-4 hours after administration, while plasma insulin remains at or below control levels. Oral administration of CHIR-99021 at 16 or 48 mg/kg 1 hour before oral glucose challenges in ZDF rats significantly improves glucose tolerance with 14% and 33% reduction in plasma glucose at 16 mg/kg and 48 mg/kg, respectively, and the higher dose of CHIR-99021 also reduces hyperglycemia before the oral glucose challenge. [1] Administration of CHIR-99021 significantly augments hematopoietic repopulation in recipient mice transplanted with mouse or human hematopoietic stem cells (HSCs), suggesting that GSK-3 is a specific modulator of HSC activity. [4]
Features

Protocol(Only for Reference)

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.

Cell Assay:

[3]

Cell lines INS-1E
Concentrations Dissolved in DMSO, final concentrations ~ 20 μM
Incubation Time 1, or 4 days
Method

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.

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.
Dosages ~48 mg/kg
Administration Orally

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Ring DB, et al. Diabetes, 2003, 52(3), 588-595.

[2] Bennett CN, et al. J Biol Chem, 2002, 277(34), 30998-31004.

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Chemical Information

Download CHIR-99021 (CT99021) HCl SDF
Molecular Weight (MW) 501.8
Formula

C22H18Cl2N8.HCl

CAS No. 252917-06-9 (free base)
Storage 3 years -20℃powder
2 years -80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 93 mg/mL (185.33 mM)
Ethanol 2 mg/mL (3.98 mM)
Water <1 mg/mL
In vivo 15% Captisol 30 mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name 6-(2-(4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)pyrimidin-2-ylamino)ethylamino)nicotinonitrile hydrochloride

Frequently Asked Questions

  • Question 1
    How about the half-life of this compound? If it is necessary for daily treatment when the compound is used in long-term experiment?

    Answer: Please change the culture medium each day to add the new comound in it if the duration is more than 72h.

  • Question 2
    What is the difference between S2924 and S1263? Which one is better for differentiation?

    Answer: S1263 is the free base format of Chir-99021, while S2924 is the HCl salt format. They have the same biological activity but are slightly different in solubility.

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.

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

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