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CHIR-99021

Catalog No. T2310   CAS 252917-06-9
Synonyms: Laduviglusib, CT99021, CHIR-99021

CHIR-99021 (CT99021) is an activator of the Wnt/β-catenin signaling pathway and a GSK-3α/β inhibitor (IC50=10/6.7 nM) with selective and oral activity.CHIR-99021 induces cellular autophagy, which enhances self-renewal in mouse and human embryonic stem cells.

All products from TargetMol are for Research Use Only. Not for Human or Veterinary or Therapeutic Use.
CHIR-99021 Chemical Structure
CHIR-99021, CAS 252917-06-9
Pack Size Availability Price/USD Quantity
2 mg In stock $ 36.00
5 mg In stock $ 58.00
10 mg In stock $ 88.00
25 mg In stock $ 163.00
50 mg In stock $ 267.00
100 mg In stock $ 426.00
200 mg In stock $ 635.00
500 mg In stock $ 987.00
1 mL * 10 mM (in DMSO) In stock $ 64.00
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Purity: 98.91%
Purity: 98.63%
Purity: 98.51%
Purity: 98%
Purity: 98%
Purity: 98%
Purity: 97.94%
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Biological Description
Chemical Properties
Storage & Solubility Information
Description CHIR-99021 (CT99021) is an activator of the Wnt/β-catenin signaling pathway and a GSK-3α/β inhibitor (IC50=10/6.7 nM) with selective and oral activity.CHIR-99021 induces cellular autophagy, which enhances self-renewal in mouse and human embryonic stem cells.
Targets&IC50 GSK-3α:10 nM (cell free), GSK-3β:6.7 nM (cell free)
In vitro METHODS: Mouse stem cells ES-D3 were treated with CHIR-99021 (1-10 µM) for 72 h. Cell growth inhibition was detected using MTT.
RESULTS: CHIR-99021 dose-dependently inhibited ES-D3 cell growth with an IC50 of 4.9 µM.[1]
METHODS: Mouse embryonic stem cells J1 mESCs and mouse embryoma cells F9 mEC were treated with CHIR-99021 (3 μM) for 24 h. The expression levels of target proteins were detected by immunofluorescence.
RESULTS: After CHIR-99021 treatment, β-linker proteins were increased in the cytoplasm and nucleus of J1-mESCs and F9-mEC cells. [2]
METHODS: Human Tenon fibroblast HTFs were treated with CHIR-99021 (5 μM) for 48 h, and the expression levels of target proteins were detected by Western Blot.
RESULTS: The production of the active form of GSK-3β (p-GSK-3β (Y216)) was significantly reduced by CHIR-99021 treatment. [3]
In vivo METHODS: To test the antitumor activity in vivo, CHIR-99021 (37.5 mg/kg/twice daily on days 0-3, 6-10, 13-17, and 20) was orally administered and paclitaxel (10 mg/kg/one dose on day 0) was intraperitoneally injected into Balb/c nude mice harboring human non-small cell lung cancer tumor H1975.
RESULTS: CHIR-99021 and paclitaxel synergistically inhibited NSCLC tumor growth in vivo. [4]
METHODS: To investigate whether direct pharmacological inhibition of GSK-3 alters the positive potentiation of alcohol in mice, CHIR-99021 (1-10 mg/kg) was administered by single intraperitoneal injection to C57BL/6J mice with a history of alcohol or sucrose self-administration.
RESULTS: CHIR-99021 dose-dependently increased alcohol-enhanced responses with no effect on sucrose self-administration or locomotor activity. ChIR-99021 significantly decreased pGSK-3β expression in all brain regions tested, decreased PICK1 and increased total GluA2 expression only in NAcb. [5]
Kinase Assay Kinases were purified from SF9 cells through the use of their His or Glu tag. Glu-tagged proteins were purified as described, and His-tagged proteins were purified according to the manufacturer's instructions. Kinase assays were performed in 96-well plates with appropriate peptide substrates in a 300-μl reaction buffer (variations on 50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 1 mM EGTA, 1 mM dithiothreitol, 25 mMβ-glycerophosphate, 1 mM NaF, and 0.01% bovine serum albumin). Peptides had Km values from 1 to 100 μM. CHIR 99021 or CHIR GSKIA was added in 3.5 μl of Me2SO, followed by ATP to a final concentration of 1 μM. After incubation, triplicate 100-μl aliquots were transferred to Combiplate 8 plates containing 100 μl/well of 50 μM ATP and 20 mM EDTA. After 1 hour, the wells were rinsed five times with phosphate-buffered saline, filled with 200 μl of scintillation fluid, sealed, and counted in a scintillation counter 30 min later. All of the steps were at room temperature. The percentage of inhibition was calculated as 100 × (inhibitor ? no enzyme control)/(Me2SO control ? no enzyme control) [4].
Cell Research The Wnt/beta-catenin reporter assay was performed with the M50 Super 8× TOPFlash and M51 Super 8× FOPFlash vector containing the firefly luciferase gene under the control of TCF/LEF binding sites or mutated bindings sites. 12,500 cells were seeded overnight on gelatine-coated 96-well plates in LIF-containing ES cell medium. On the next day, the cells were transfected using Lipofectamine with one of the aforementioned vectors plus pGL4.75 [hRluc/CMV] encoding the renilla luciferase reporter gene hRluc as a transfection control. Six hours after transfection the medium was changed to medium devoid of LIF, with reduced serum, and supplemented with 5 μM CHIR-99021. The Dual-Luciferase? reporter assay system was employed 48 and 72 h after the medium change to follow the luminescence reaction using a GloMax?-multi detection system [4].
Animal Research Blood was obtained by shallow tail snipping at lidocaine-anesthetized tips. Blood glucose was measured directly or heparinized plasma was collected for measurement of glucose or insulin. Animals were pre-bled and randomized to vehicle control or GSK-3 inhibitor treatment groups. For glucose tolerance tests (GTTs), animals fasted throughout the procedure with food removal early in the morning, 3 h before the first prebleed (db/db mice), or the previous night, 16 h before the bleed (ZDF rats). When the time course of plasma glucose and insulin changes in fasting ZDF rats was measured, food was removed ~16 h before test agent administration. The glucose challenges in the GTT were 1.35 g/kg i.p. (ipGTT) or 2 g/kg via oral gavage (oGTT). CHIR-99021 were formulated as solutions in 20 mmol/l citrate-buffered 15% Captisol or as fine suspensions in 0.5% carboxymethylcellulose [1].
Synonyms Laduviglusib, CT99021, CHIR-99021
Molecular Weight 465.34
Formula C22H18Cl2N8
CAS No. 252917-06-9

Storage

Powder: -20°C for 3 years | In solvent: -80°C for 1 year

Solubility Information

DMSO: 9.3 mg/mL (20 mM)

TargetMolReferences and Literature

1. Naujok O, et al. Cytotoxicity and activation of the Wnt/beta-catenin pathway in mouse embryonic stem cells treated with four GSK3 inhibitors. BMC Res Notes. 2014 Apr 29;7:273. 2. Wu Y, et al. GSK3 inhibitors CHIR99021 and 6-bromoindirubin-3'-oxime inhibit microRNA maturation in mouse embryonic stem cells. Sci Rep. 2015 Mar 2;5:8666. 3. Lee SY, et al. The Effect of CHIR 99021, a Glycogen Synthase Kinase-3β Inhibitor, on Transforming Growth Factor β-Induced Tenon Fibrosis. Invest Ophthalmol Vis Sci. 2021 Dec 1;62(15):25. 4. O'Flaherty L, et al. Tumor growth suppression using a combination of taxol-based therapy and GSK3 inhibition in non-small cell lung cancer. PLoS One. 2019 Apr 10;14(4):e0214610. 5. Faccidomo S, et al. Pharmacological inhibition of glycogen synthase kinase 3 increases operant alcohol self-administration in a manner associated with altered pGSK-3β, protein interacting with C kinase and GluA2 protein expression in the reward pathway of male C57BL/6J mice. Behav Pharmacol. 2020 Feb;31(1):15-26. 6. Gong-Bo Fu, Wei-Jian Huang, Min Zeng, Xu Zhou, Hong-Ping Wu, Chang-Cheng Liu, Han Wu, Jun Weng, Hong-Dan Zhang, Yong-Chao Cai, Charles Ashton, Min Ding, Dan Tang, Bao-Hua Zhang, Yi Gao, Wei-Feng Yu, Bo Zhai, Zhi-Ying He, Hong-Yang Wang, and He-Xin Yan . Expansion and differentiation of human hepatocyte-derived liver progenitor-like cells and their use for the study of hepatotropic pathogens [J]. Cell Research. 2019 Jan;29(1):8-22.

TargetMolCitations

1. Lin Y Y, Yao R, Zhuang J, et al.PACT inhibits the replication of SARS‐CoV‐2 through the blockage of GSK‐3β‐N‐nsp3 cascade.Journal of Medical Virology.2023, 95(6): e28832. 2. Yu Y, Li X, Jiao R, et al.H3K27me3-H3K4me1 transition at bivalent promoters instructs lineage specification in development.Cell & Bioscience.2023, 13(1): 1-20. 3. Wu M, Zhang X, Zhang W, et al.Paracrine secretion of IL8 by breast cancer stem cells promotes therapeutic resistance and metastasis of the bulk tumor cells.Cell Communication and Signaling.2023, 21(1): 1-17. 4. Han L, Song B, Zhang P, et al.PC3T: a signature-driven predictor of chemical compounds for cellular transition.Communications Biology.2023, 6(1): 989. 5. Yang Y, Xiao L, Xue Y, et al.ZBTB7A regulates primed‐to‐naïve transition of pluripotent stem cells via recognition of the PNT‐associated sequence by Zinc Fingers 1–2 and recognition of γ‐globin− 200 gene element by Zinc Fingers 1–4.The FEBS Journal.2023 6. Yang Z, Liu H, Song R, et al. Reduced MAGI3 level by HPV18E6 contributes to Wnt/β‐catenin signaling activation and cervical cancer progression. FEBS Open bio. 2021, 11(11): 3051. 7. He W, Zhu X, Xin A, et al. Long-term maintenance of human endometrial epithelial stem cells and their therapeutic effects on intrauterine adhesion. Cell & Bioscience. 2022, 12(1): 1-20. 8. Fu G B, Huang W J, Zeng M, et al. Expansion and differentiation of human hepatocyte-derived liver progenitor-like cells and their use for the study of hepatotropic pathogens. Cell Research. 2019, 29(1): 8-22 9. Ma X, Lu Y, Zhou Z, Human expandable pancreatic progenitor–derived β cells ameliorate diabetes. Science Advances. 2022, 8(8): eabk1826. 10. Wang W, Ren S, Lu Y, et al. Inhibition of Syk promotes chemical reprogramming of fibroblasts via metabolic rewiring and H2S production. The EMBO Journal. 2021 Jun 1;40(11):e106771. doi: 10.15252/embj.2020106771. Epub 2021 Apr 28.
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This product is contained In the following compound libraries:
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Keywords

CHIR-99021 252917-06-9 Autophagy Cytoskeletal Signaling PI3K/Akt/mTOR signaling Stem Cells Wnt/beta-catenin GSK-3 Glycogen synthase kinase 3 CHIR99021 β-catenin Laduviglusib inhibit CT99021 Beta catenin CT-99021 CHIR 99021 Wnt Inhibitor Glycogen synthase kinase-3 CT 99021 inhibitor

 

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