.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 431.20 Pimasertib (AS-703026) es un inhibidor alostérico no competitivo de ATP altamente selectivo y potente de MEK1 / 2 con IC50 de 5 nM-2 μM en células MM líneas. Fase 2. \ n Actividad biológica AS703026 es un inhibidor de MEK1 / 2 novedoso, selectivo y biodisponible por vía oral que se une al sitio alostérico distintivo de MEK y, por lo tanto, exhibe una exquisita selectividad de quinasa. AS703026 inhibe el crecimiento y la supervivencia de las células de mieloma múltiple (MM) humano, incluidas U266 e INA-6, con IC50 de 5 nM y 11 nM, respectivamente. Tal efecto inhibidor por AS703026 está mediado por la detención del ciclo celular G0-G1 y está acompañado por una expresión reducida del oncogén MAF. AS703026 además induce la apoptosis a través de la escisión de la caspasa-3 y PARP en las células MM, tanto en presencia como en ausencia de células estromales de la médula ósea (BMSC). AS703026 puede ser una terapia eficaz en el cáncer colorrectal causado por la mutación K-Ras. AS703026 (10 μM) inhibe eficazmente la vía de ERK, la proliferación y la transformación en células de cáncer colorrectal humano DLD-1 que llevan un alelo mutante de K-Ras (D-MUT). AS703026 (15 y 30 mg / kg) inhibe significativamente el crecimiento tumoral en un modelo de xenoinjerto de plasmacitoma humano de células H929 MM. Esto puede correlacionarse con pERK1 / 2 regulado negativamente, escisión de PARP inducida y microvasos disminuidos. AS703026 (10 mg / kg) inhibe el crecimiento tumoral y disminuye notablemente el nivel de p-ERK en un modelo de ratón xenoinjerto de tumor colorrectal humano con mutación de K-Ras (D-MUT). Protocolo (solo como referencia) Ensayo de quinasa: [3]
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MEK1 enzyme assays
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AS703026 is dissolved in 2.5% DMSO. Activated diphosphorylated MEK (pp-MEK) assays contained 40 μM 33P-γATP (AppKm 8.5 μM), 0.5 nM human-activated MEK1 or MEK2, 1 μM kinase-dead ERK2 (AppKm 0.73 μM). All assays are done in buffer containing 20 mM HEPES (pH 7.2), 5 mM 2-mercaptoethanol, 0.15 mg/mL BSA, and 10 mM MgCl2. The final concentration of 33P- ATP is 0.02 μCi/μL for all the assays. pp-MEK kinase reactions are stopped after 40 min by transferring 30 μL of reaction mixture to Durapore 0.45-μm filters plates containing 12.5% TCA. Filters are dried and read with liquid scintilant on a TopCount. Concentration response data are analyzed for IC50. 0.2 nM recombinant human MEK1 or MEK2 is preincubated with vehicle or with AS703026 for 40 minutes in reaction buffer to determine IC50 of initially unphosphorylated MEK (u-MEK). Phosphorylation/activation is initiated by the addition of a final concentration of 20 nM final B-RafV600E and 30 μM final ATP for 10 min. B-Raf activity is then quenched by addition of the B-Raf inhibitor SB590885 (final concentration 100 nM), and MEK kinase activity is assayed by the addition of 1 μM KD-ERK2 and 0.02 μCi/μL 33P-ATP in reaction buffer. The kinase reactions are stopped after 90 min by transferring 30μL of reaction mixture to a Durapore filter plate, and read as above.
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Ensayo celular: [1]
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Cell lines
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U266 and INA-6 cells
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Concentrations
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2 nM - 20 μM (stock: 10 mM in DMSO)
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Incubation Time
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48 hours
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Method
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Cytotoxicity assays for AS703026 are assessed by measuring both [3H]thymidine incorporation and MTT dye absorbance. Cells (1 × 104 per well) are cultured in 96-well plates for 3 days. For the [3H]thymidine incorporation assay, cells are pulsed with 18.5 kBq/well [3H]thymidine for 6 hours, harvested onto glass fibre filters, and counted in a β-scintillation counter.
Cell cycle analysis is assessed by propidium iodide (PI) staining using flow cytometry. AS703026-induced apoptosis is determined by annexin-V/PI staining and flow cytometric data analysis.
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Estudio con animales: [1]
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Animal Models
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H929 MM xenografts are established in CB17 (SCID) mice
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Formulation
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10 mg/mL, dissolved in 0.5% carboxymethylcellulose / 0.25% Tween20
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Dosages
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15 or 30 mg/kg
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Administration
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Oral gavage twice daily
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Solubility
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0.5% CMC/0.25% Tween 80,
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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431.20
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Formula
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C15H15FIN3O3
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CAS No.
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1236699-92-5
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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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Solubility (25°C) *
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In vitro
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DMSO
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86 mg/mL
(199.44 mM)
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Water
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<1 mg/mL
(
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Ethanol
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<1 mg/mL
(
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In vivo
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0.5% CMC/0.25% Tween 80
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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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(S)-N-(2,3-dihydroxypropyl)-3-(2-fluoro-4-iodophenylamino)isonicotinamide
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Calculadora de molaridad Calculadora de dilución Calculadora de peso molecular
Grupos de Producto : MAPK > Inhibidor de MEK
