CHIR-99021

SKU: orb1305728

Description

CHIR-99021 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.

Research Area

Cell Biology, Signal Transduction, Stem Cell & Developmental Biology

Key Properties

• CAS Number: 252917-06-9
• MW: 465.34
• Purity: 99.29%
• 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 tes 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-s All 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 alter 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, an the expression levels of target proteins were detected by Western Blot. Results the production o 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 wit the M50 Super × TOPFlash and M51 Super × FOPFlash vector containin the firefly luciferase gene unde 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. O the Next day the cells were transfected using Lipofectamine with one o the aforementioned vectors plus pGL4.75 [hRluc/CMV] encodin the renilla luciferase Reporter gene hRluc as a transfection control. Six hours after transfectio 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 afte the medium change to follo 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 andomized to Vehicle control or GSK-3 inhibitor treatment groups. For glucose tolerance tests (GTTs), animals fasted throughou the procedure with food removal early in the morning, 3 h befor the first prebleed (db/db mice), o the previous night, 16 h befor the bleed (ZDF rats). Whe 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.
• 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