CHIR-99021

SKU: orb1305728

Description

CHIR-99021

Research Area

Product Categories/Products/Small Molecules,Cytoskeletal Signaling|||Autophagy|||PI3K/Akt/mTOR signaling|||Stem Cells

Key Properties

• CAS Number: 252917-06-9
• MW: 465.34
• Purity: ≥95%
• Formula: C₂₂H₁₈Cl₂N₈
• SMILES: Cc1c[nH]c(n1)-c1cnc(NCCNc2ccc(cn2)C#N)nc1-c1ccc(Cl)cc1Cl

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: -20°C
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Compound Identifiers

PubChem CID

9956119