.cp_wz tabla {borde superior: 1px sólido #ccc; borde izquierdo: 1px sólido #ccc; } .cp_wz table td {borde derecho: 1px sólido #ccc; borde inferior: 1px sólido #ccc; padding: 5px 0px 0px 5px;} .cp_wz table th {border-right: 1px solid #ccc; border-bottom: 1px solid #ccc; relleno: 5px 0px 0px 5px;} \ n Peso molecular: 356.17 BIO (6-bromoindirrubina-3'-oxima) es un inhibidor específico de GSK-3 con IC50 de 5 nM para GSK-3α / β, muestra> 16 veces selectividad sobre CDK5, también un inhibidor de pan-JAK. \ n BIO (6-bromoindirrubina-3'-oxima) es un inhibidor específico de la glucógeno sintasa quinasa-3 (GSK-3), con IC50 de 5 nM para GSK-3α / β, muestra una selectividad> 16 veces mayor que la de CDK5. BIO interactúa dentro del bolsillo de unión de ATP de estas quinasas, reduce la fosforilación de β-catenina en un sitio específico de GSK-3 en modelos celulares, imita de cerca la señalización de Wnt en embriones de Xenopus. En células madre embrionarias humanas y de ratón, BIO mantiene el fenotipo indiferenciado y mantiene la expresión de los factores de transcripción específicos del estado pluripotente Oct-3/4, Rex-1 y Nanog. La activación de Wnt mediada por BIO es funcionalmente reversible, ya que la retirada del compuesto conduce a programas normales de diferenciación múltiple en células madre embrionarias de ratón y humano. BIO promueve la proliferación en cardiomiocitos de mamíferos. 6BIO también es un inhibidor de pan-JAK, con valores de IC50 de 0.03, 1.5, 8.0, 0.5 μM para TYK2, JAK1, JAK2 y JAK3. BIO inhibe selectivamente la fosforilación de STAT3 e induce la apoptosis de las células de melanoma humano. BIO suprime el crecimiento del tumor de melanoma en un modelo de xenoinjerto de ratón. El primer agente farmacológico que se ha demostrado que mantiene la autorrenovación en células madre embrionarias humanas y de ratón.
orte
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Kinase assay
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Kinase activities are assayed in Buffer A or C at 30°C, at a final ATP concentration of 15 μM. Blank values are subtracted and activities calculated as pmoles of phosphate incorporated during a 10 min incubation. Controls are performed with appropriate dilutions of dimethylsulfoxide. In a few cases phosphorylation of the substrate is assessed by autoradiography after SDS-PAGE. GSK-3α/β is purified from porcine brain by affinity chromatography on immobilized axin. It is assayed, following a 1/100 dilution in 1 mg BSA/ml 10 mM DTT, with 5 μl 40 μM GS-1 peptide, a specific GSK-3 substrate, (YRRAAVPPSPSLSRHSSPHQSpEDEEE), in buffer A, in the presence of 15 μM [γ-32P] ATP (3,000 Ci/mmol; 1 mCi/ml) in a final volume of 30 μl. After 30 min incubation at 30°C, 25 μl aliquots of supernatant are spotted onto 2.5 × 3 cm pieces of Whatman P81 phosphocellulose paper, and 20 seconds later, the filters are washed five times (for at least 5 min each time) in a solution of 10 ml phosphoric acid/liter of water. The wet filters are counted in the presence of 1 ml ACS scintillation fluid.
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Ensayo celular: [1]
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Cell lines
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COS1, Hepa or SH-SY5Y cells
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Concentrations
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~10 μM
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Incubation Time
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12 or 24 h
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Method
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COS1, Hepa (wild-type, CEM/LM AhR deficient and ELB1 ARNT deficient), or SH-SY5Y cells are grown in 6 cm culture dishes in Dulbecco's Modified Medium (DMEM) containing 10% fetal bovine serum. For treatment, IO (5 μM), BIO (5 or 10 μM), MeBIO (5 or 50 μM), LiCl (20 or 40 mM), or mock solution (DMSO, 0.5% final concentration) is added to medium when cell density reaches ∼70% confluence. After 12 (SH-SY5Y) or 24 hours, the cells, while still in plate, are lysed with lysis buffer (1% SDS, 1 mM sodium orthovanadate, 10 mM Tris [pH 7.4]). The lysate is passed several times through a 26G needle, centrifuged at 10,000 × g for 5 min, and adjusted to equal protein concentration. About 8 μg of each sample is loaded for immunoblotting. Enhanced chemiluminescence is used for detection. The following primary antibodies are used: mouse anti-β-catenin CT (Upstate Biotechnolgies, Clone 7D8, recognizes total β-catenin), mouse anti-phospho-β-catenin (Upstate Biotechnologies, Clone 8E7, recognizes dephosphorylated β-catenin), mouse anti-GSK-3 β, mouse anti-GSK-3 phosphoTyr216, rabbit anti-AhR (Aryl hydrocarbon receptor), and rabbit anti-actin.
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Estudio con animales: [4]
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Animal Models
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mouse
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Formulation
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freshly prepared in 30% Solutol (Basf) at a concentration of 10 mg/mL.
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Dosages
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50 mg/kg
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Administration
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Oral gavage
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Solubility
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30% PEG400/0.5% Tween80/5% propylene glycol,
30 mg/mL
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* 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.
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Conversión de diferentes modelos de animales basados en BSA (valor basado en datos del Borrador de Directrices de la FDA)
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Species
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Baboon
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Dog
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Monkey
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Rabbit
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Guinea pig
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Rat
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Hamster
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Mouse
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Weight (kg)
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12
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10
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3
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1.8
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0.4
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0.15
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0.08
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0.02
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Body Surface Area (m2)
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0.6
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0.5
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0.24
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0.15
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0.05
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0.025
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0.02
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0.007
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Km factor
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20
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20
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12
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12
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8
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6
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5
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3
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Animal A (mg/kg) = Animal B (mg/kg) multiplied by
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Animal B Km
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Animal A Km
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Por ejemplo, para modificar la dosis de resveratrol utilizada para un ratón (22,4 mg / kg) a una dosis basada en el BSA para una rata, multiplique 22,4 mg / kg por el factor Km para un ratón y luego divida por el factor Km para una rata. Este cálculo da como resultado una dosis equivalente para ratas de resveratrol de 11,2 mg / kg.
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Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×
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mouse Km(3)
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= 11.2 mg/kg
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rat Km(6)
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Información química
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Molecular Weight (MW)
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356.17
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Formula
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C16H10BrN3O2
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CAS No.
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667463-62-9
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Storage
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3 years -20℃Powder
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6 months-80℃in solvent (DMSO, water, etc.)
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Synonyms
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GSK-3 Inhibitor IX, 6-bromoindirubin-3-oxime
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Solubility (25°C) *
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In vitro
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DMSO
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71 mg/mL
(199.34 mM)
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Water
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<1 mg/mL
(
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Ethanol
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21 mg/mL
(58.96 mM)
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In vivo
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30% PEG400/0.5% Tween80/5% propylene glycol
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30 mg/mL
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* <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.
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Chemical Name
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2H-Indol-2-one, 6-bromo-3-[(3E)-1,3-dihydro-3-(hydroxyimino)-2H-indol-2-ylidene]-1,3-dihydro-, (3Z)-
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Calculadora de molaridad Calculadora de dilución Calculadora de peso molecular
Grupos de Producto : PI3K / Akt / mTOR > Inhibidor GSK-3
