.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: \ n 361,48 PHA-793887 é um novo e potente inibidor de CDK2, CDK5 e CDK7 com IC50 de 8 nM, 5 nM e 10 nM. É 6 vezes mais seletivo para CDK2, 5 e 7 do que CDK1, 4 e 9. Fase 1. \ n Atividade biológica PHA-793887 tem baixa atividade contra CDK1, CDK4, CDK9 e GSK3β com IC50 de 60 nM , 62 nM, 138 nM e 79 nM, respectivamente. PHA-793887 inibe a proliferação celular de muitas linhas de células tumorais, incluindo A2780, HCT-116, COLO-205, C-433, DU-145, A375, PC3, MCF-7 e BX-PC3, com IC50 de 88 nM- 3,4 μM. PHA-793887 (1 μM) mostra uma diminuição na fase S, um aumento subsequente da fase G1 e um leve acúmulo da fase G2 / M nas células A2780. PHA-793887 (3 μM) aumenta significativamente a fase G2 / M e reduz a síntese de DNA. PHA-793887 é citotóxico para linhas celulares leucêmicas, incluindo K562, KU812, KCL22 e TOM1, com IC50 de 0,3–7 μM, mas não é citotóxico para células mononucleares de sangue periférico não estimuladas normais ou células-tronco hematopoiéticas CD34 +. Em ensaios de colônia, PHA-793887 mostra atividade muito alta contra linhas de células de leucemia com IC50 inferior a 0,1 μM. PHA-793887 induz a parada do ciclo celular, inibe Rb e fosforilação da nucleofosmina e modula a expressão de ciclina E e cdc6 a 0,2-1 μM e induz apoptose a 5 μM. PHA-793887 (10-30 mg / kg) mostra boa eficácia em modelos de xenoenxerto de carcinoma pancreático A2780 humano, HCT-116 de cólon e BX-PC3 pancreático. PHA-793887 (20 mg / kg) é eficaz em modelos de xenoenxerto de células K562 e HL60, modelo disseminado de leucemia primária e um modelo de ALL-2 disseminado de alta carga derivado de um paciente com leucemia linfóide aguda positiva Filadélfia recidivante. Ensaio de cinase de protocolo (apenas para referência): [3]
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CDK Kinase Assay
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The biochemical activity of compounds is determined by incubation with specific enzymes and substrates, followed by quantitation of the phosphorylated product. PHA-793887 (1.5 nM–10 μM) is incubated for 30−90 min at room temperature in the presence of ATP/33P-γ-ATP mix, substrate, and the specific enzyme (0.7−100 nM) in a final volume of 30 μL of kinase buffer, using 96 U bottom plates. After incubation, the reaction is stopped and the phosphorylated substrate is separated from nonincorporated radioactive ATP using SPA beads, Dowex resin, or Multiscreen phosphocellulose filter as follows: (1) For SPA Assays. The reaction is stopped by the addition of 100 μL of PBS + 32 mM EDTA + 0.1% Triton X-100 + 500 μM ATP, containing 1 mg of streptavidin-coated SPA beads. After 20 min of incubation for substrate capture, 100 μL of the reaction mixture is transferred into Optiplate 96-well plates containing 100 μL of 5 M CsCl, left to stand for 4 hours to allow stratification of beads to the top of the plate, and counted using TopCount to measure substrate-incorporated phosphate. (2) For Dowex Resin Assay. An amount of 150 μL of resin/formate, pH 3.00, is added to stop the reaction and capture unreacted 33P-γ-ATP, separating it from the phosphorylated substrate in solution. After 60 min of rest, 50 μL of supernatant is transferred to Optiplate 96-well plates. After the additon of 150 μL of Microscint 40, the radioactivity is counted in the TopCount. (3) For Multiscreen Assay. The reaction is stopped with the addition of 10 μL of EDTA (150 mM). An amount of 100 μL is transferred to a MultiScreen plate to allow substrate binding to phosphocellulose filter. Plates are then washed three times with 100 μL of H2PO4 (75 mM) filtered by a MultiScreen filtration system, and dried. After the additon of 100 μL of Microscint 0, radioactivity is counted in the TopCount.
IC50 values are obtained by nonlinear regression analysis.
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Ensaio de células: [1]
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Cell lines
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A2780 cells
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Concentrations
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0.1 nM-1 μM, dissolved 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 seeded into 96- or 384-wells plates at final concentration ranging from 1 × 104 to 3 × 104 per cm2. After 24 hours, cells are treated using serial dilution of PHA-793887. At 72 hours after the treatment, the amount of cells are evaluated using the CellTiter-Glo assay. IC50 values are calculated using a sygmoidal fitt
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Estudo Animal: [1]
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Animal Models
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Mouse xenograft models of human ovarian A2780, colon HCT-116 and pancreatic BX-PC3 carcinoma
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Formulation
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Dissolved in 5% dextrose solution
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Dosages
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10, 20, and 30 mg/kg
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Administration
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Intravenous injection once daily
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Solubility
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30% propylene glycol, 5% Tween 80, 65% D5W,
15 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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Informação Química
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Molecular Weight (MW)
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361.48
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Formula
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C19H31N5O2
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CAS No.
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718630-59-2
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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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72 mg/mL
(199.18 mM)
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Water
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<1 mg/mL
(
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Ethanol
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72 mg/mL
(199.18 mM)
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In vivo
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30% propylene glycol, 5% Tween 80, 65% D5W
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15 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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N-(6,6-dimethyl-5-(1-methylpiperidine-4-carbonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-3-methylbutanamide
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Calculadora de molaridade Calculadora de diluição Calculadora de peso molecular
Grupo de Produto : Ciclo de célula > Inibidor CDK
