.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; padding: 5px 0px 0px 5px;} \ n Peso molecular: \ n 479.4 SBE 13 HCl es un inhibidor de PLK1 potente y selectivo con IC50 de 200 pM, selectividad> 4000 veces superior a Aurora A quinasa, Plk2 y Plk3. \ n Actividad biológica SBE 13 disminuye la proliferación celular en varias líneas de células cancerosas y provoca una detención de G2 / M seguida de apoptosis. En las células primarias, SBE 13 no altera el ciclo celular y, por tanto, la proliferación de células primarias. SBE13 en combinación con Enzastaurin muestra una reducción sinérgica de la proliferación celular y una inducción de apoptosis mejorada en las células HCT116 (p53 - / -). Protocolo (solo como referencia) Ensayo de quinasa: [1]
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Kinase assays
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To assay Plk1 and Aurora A kinase activity, cells are lysed after 13 hrs release in the presence of SBE13 after double thymidine block, and kinases are immunoprecipitated from lysates using antibodies as described. In brief, for each immunoprecipitation 800 µg of total protein were incubated with 1.5 µg Plk1 antibody cocktail, 3 µg Plk2 antibody, 3 µg Plk3 antibody, or 5 µg Aurora A antibody, respectively, for 2 hrs at 4°C on a rotator. Immunoprecipitated protein is collected using Protein G Agarose beads. The Plk1, Plk2 and Plk3 immunoprecipitates are incubated with 1 µg casein and with 1 µCi of [γ32-P]ATP for 30 min at 37°C in kinase buffer. The Aurora A immunoprecipitates are incubated with 0.5 µl Histone and with 1 µCi of [γ32-P]ATP for 60 min at room temperature in kinase buffer. Products from the kinase assays are fractionated on 10% Bis-Tris-polyacrylamide gels, and the phosphorylated substrate is visualized by autoradiography after an exposure of 12 to 36 hrs. An equal amount of immunoprecipitates is subjected to western blot analysis to confirm equal loading of Plk1, Plk2, Plk3 or Aurora A protein in kinase reactions. Immunoprecipitated Plk1 after 13 hrs release in the presence of SBE13 is assayed after de-phosphorylation using λ protein phosphatase and compared to kinase activity of endogenous immunoprecipitated Plk1. Activity of Plk1 kinase with and wiiiuithout de-phosphorylation is compared and the ratio between de-phosphorylated and [normal" endogenous immunoprecipitated Plk1 kinase activity is calculated.
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Ensayo celular: sup> [1]
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Cell lines
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HeLa, A431, HCT-15, HT-29, MCF-7, U2OS, LN-229, SKW 6.4, PC-3, Det-562, SK-BR-3, SK-OV-3 and A549nb cells
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Concentrations
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100 μM
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Incubation Time
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72 hours
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Method
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Cells are treated with SBE13 one day after subculturing. Control cells are incubated with normal culture medium. Concentrations of SBE13 ranged from 1 nM–100 µM. The growth rate of 1 x 105 cells per 6-well is determined by counting cells at 24, 48 and 72 hours after treatment. Cell culture studies are performed in triplicate for each time point.
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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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479.4
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Formula
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C24H28Cl2N2O4
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CAS No.
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1052532-15-6
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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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N/A
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Chemical Name
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N-(4-((6-chloropyridin-3-yl)methoxy)-3-methoxybenzyl)-2-(3,4-dimethoxyphenyl)ethanamine hydrochloride
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Calculadora de molaridad Calculadora de dilución Calculadora de peso molecular
Grupos de Producto : Ciclo celular > Inhibidor de PLK
