.cp_wz table {border-top: 1px solid #ccc; border-left: 1px solid #ccc; } .cp_wz table td {border-right: 1px solid #ccc; borda inferior: 1px sólido #ccc; preenchimento: 5px 0px 0px 5px;} .cp_wz tabela th {border-right: 1px solid #ccc; border-bottom: 1px solid #ccc; preenchimento: 5px 0px 0px 5px;} \ n Peso molecular: 337,85 AT7867 é um potente inibidor competitivo de ATP de Akt1 / 2/3 e p70S6K / PKA com IC50 de 32 nM / 17 nM / 47 nM e 85 nM / 20 nM, respectivamente; pouca atividade fora da família de AGC quinase. \ n AT7867 também inibe AGC quinases estruturalmente relacionadas p70S6K e PKA com IC50 de 20 nM e 85 nM, respectivamente. AT7867 mostra atividade competitiva de ATP para Akt2 com Ki de 18 nM. AT7867 exibe antiproliferação em linhas celulares com mutações PTEN ou PIK3CA e mostra grande potente para MES-SA, MDA-MB-468, MCF-7, HCT116 e HT29 com IC50 de 0,94 μM, 2,26 μM, 1,86 μM, 1,76 μM e 3,04 μM , respectivamente. AT7867 também suprime o crescimento celular das células U87MG, PC-3 e DU145 com IC50 de 8,22 μM, 10,37 μM e 11,86 μM, respectivamente. AT7867 suprime a atividade Akt inibindo a fosforilação de GSK-3β em células tumorais humanas com IC50 de 2-4 μM. AT7867 também induz a fosforilação dos seguintes substratos diretos de Akt, incluindo fatores de transcrição pró-apoptóticos FKHR (FoxO1a), FKHRL1 (FoxO3a) e o alvo a jusante S6RP em células U87MG. AT7867 mostra biodisponibilidade de 44% em camundongos por via po. AT7867 pode aumentar a PARP clivada em xenoenxertos MES-SA a 20 mg / kg ip ou 90 mg / kg po. AT7867 inibe significativamente o crescimento do tumor em xenoenxertos MES-SA ou xenoenxertos U87MG com T / C de 0,37 e 0,51, respectivamente. \ n
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In vitro kinase assays
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Kinase assays for Akt2, PKA, p70S6K, and CDK2/cyclin A are all carried out in a radiometric filter binding format. Assay reactions are set up in the presence of AT7867. For Akt2, the Akt2 enzyme and 25 μM Aktide-2T peptide (HARKRERTYSFGHHA) are incubated in 20 mM MOPS (pH 7.2), 25 mM β-glycerophosphate, 5 mM EDTA, 15 mM MgCl2, 1 mM sodium orthovanadate, 1 mM DTT, 10 μg/mL bovine serum albumin, and 30 μM ATP (1.16 Ci/mmol) for 4 hours. For PKA, the PKA enzyme and 50 μMpeptide (GRTGRRNSI) are incubated in 2 mM MOPS (pH 7.2), 25 mM β-glycerophosphate, 5 mM EDTA, 15 mM MgCl2, 1 mM orthovanadate, 1 mM DTT, and 40 μM ATP (0.88 Ci/mmol) for 20 minutes. For p70S6K, the p70S6K enzyme and 25 μMpeptide substrate (AKRRRLSSLRA) are incubated in 10 mM MOPS (pH 7), 0.2 mM EDTA, 1 mM MgCl2, 0.01% β-mercaptoethanol, 0.1 mg/mL bovine serum albumin, 0.001% Brij-35, 0.5% glycerol, and 15 μM ATP (2.3 Ci/mmol) for 60 min. For CDK2, the CDK2/cyclin A enzyme and 0.12 μg/mL histone H1 are incubated in 20 mM MOPS (pH 7.2), 25 mM β-glycerophosphate, 5 mM EDTA, 15 mM MgCl2, 1 mM sodium orthovanadate, 1 mM DTT, 0.1 mg/mL bovine serum albumin, and 45 μM ATP (0.78 Ci/mmol) for 4 hours. Assay reactions are stopped by adding an excess of orthophosphoric acid, and the stopped reaction mixture is then transferred to Millipore MAPH filter plates and filtered. The plates are then washed, scintillant is added, and radioactivity is measured by scintillation counting on a Packard TopCount. IC50 values are calculated from replicate curves using GraphPad Prism software. Akt1 and Akt3 enzyme assays are carried out.
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Ensaio de células: [1]
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Cell lines
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MES-SA, MDA-MB-468, MCF-7, HCT116, HT29, U87MG, PC-3 and DU145 cells
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Concentrations
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0-100 μM , dissolved to a 10 mM stock in DMSO
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Incubation Time
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72 hours
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Method
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Cells are plated in 96-well microplates at 5 × 103 per well in medium supplemented with 10% fetal bovine serum and grown for 24 hours before treatment with AT7867. AT7867 or vehicle control is added to the cells for 72 hours. Following this, Alamar Blue solution is added. The IC50 value for AT7868 is calculated in GraphPad Prism using nonlinear regression analysis and a sigmoidal dose-response (variable slope) equation.
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Estudo Animal: [1]
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Animal Models
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Human MES-SA uterine sarcoma cells or U87MG human glioblastoma cells are injected s.c. in the right flank of BALB/c mice.
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Formulation
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Formulated in a vehicle containing 10% DMSO, 20% water, and 70% hydroxypropyl-β-cyclodextrin (25% aqueous, w/v)
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Dosages
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20 mg/kg (i.p.) or 90 mg/kg (p.o.) once every 3 days
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Administration
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Administered via i.p. or p.o.
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Solubility
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15% Captisol, pH 9,
10 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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Conversão de diferentes modelos de animais com base em BSA (valor com base em dados das diretrizes preliminares da 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 exemplo, para modificar a dose de resveratrol usada para um camundongo (22,4 mg / kg) para uma dose baseada na BSA para um rato, multiplique 22,4 mg / kg pelo fator Km para um camundongo e, em seguida, divida pelo fator Km para um rato. Este cálculo resulta em uma dose equivalente de rato para o 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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\ n Informação Química
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Molecular Weight (MW)
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337.85
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Formula
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C20H20ClN3
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CAS No.
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857531-00-1
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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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68 mg/mL
(201.27 mM)
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Water
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<1 mg/mL
(
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Ethanol
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5 mg/mL
(14.79 mM)
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In vivo
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15% Captisol, pH 9
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10 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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4-(4-(1H-pyrazol-4-yl)phenyl)-4-(4-chlorophenyl)piperidine
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Calculadora de molaridade Calculadora de diluição Calculadora de peso molecular
Grupo de Produto : PI3K / Akt / mTOR > Inibidor de Akt
