CHIR-99021

SKU: orb1305728

Description

CHIR-99021 is a potent, selective, and orally active GSK-3α/β inhibitor that activates Wnt/β-catenin signaling. It is widely used in stem cell research to promote self-renewal of embryonic stem cells by inducing autophagy, and has applications in studies of development, metabolism, and regeneration in vitro and in vivo.

Research Area

Cell Biology, Signal Transduction, Stem Cell & Developmental Biology

Key Properties

• CAS Number: 252917-06-9
• MW: 465.34
• Purity: 99.29% (May vary between batches)
• Formula: C₂₂H₁₈Cl₂N₈
• SMILES: Cc1c[nH]c(n1)-c1cnc(NCCNc2ccc(cn2)C#N)nc1-c1ccc(Cl)cc1Cl
• Target: Autophagy,Wnt/beta-catenin,GSK-3
• Solubility: DMSO: 105 mg/mL (225.64 mM), Sonication is recommended.

Bioactivity

• Target IC50:
  GSK-3α:10 nM (cell free)|GSK-3β:6.7 nM (cell free)|CHO cells:0.13 μM (EC50)|HepG2 cells:1.5 μM (EC50)|CDC2:8800 nM
• 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. 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.
• 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. 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. 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.
• 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 .
• 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 .

Storage & Handling

• Storage: store at low temperature | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

Beta catenin, beta-catenin, betacatenin, CHIR 99021, CHIR99021, CHIR-99021, CT99021, CT-99021, CT 99021, Autophagy, bcatenin, Laduviglusib, Inhibitor, Glycogen synthase kinase 3, Glycogen synthase kinase-3, GSK-3β, GSK-3α, GSK3, GSK-3, inhibit, βcatenin, β-catenin, Wnt, Wnt/β-catenin, Wnt/b-catenin, Wnt/betacatenin

Compound Identifiers

PubChem CID

9956119