.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 438,3 P276-00 é um novo inibidor de CDK1, CDK4 e CDK9 com IC50 de 79 nM, 63 nM e 20 nM, respectivamente. Fase 2/3. \ N A atividade biológica P276-00 mostra seletividade de 40 vezes para Cdk4-D1, em comparação com Cdk2-E [1]. Mostra efeitos antiproliferativos potentes contra várias linhas de células de câncer humano, incluindo HCT-116, U2OS, H-460, HL-60, HT-29, SiHa, MCF-7, Colo-205, SW-480, PC-3, Caco2 , T-24 com um IC50 variando de 300 a 800 nmol / L, e é considerado altamente seletivo para células cancerosas em comparação com células de fibroblastos normais. P276-00 pode regular negativamente a ciclina D1 e Cdk4 de uma maneira competitiva com ATP e diminuir a fosforilação de pRb Ser780 específica de Cdk4. O P276-00 também induz a apoptose por meio da atuação da atividade da caspase-3 celular e da formação da escada de DNA. P276-00, administrado ip a 50 mg / kg por dia durante 20 tratamentos pode induzir significativamente a inibição do crescimento do câncer de cólon murino (CA-51). No entanto, no modelo de carcinoma de pulmão murino (pulmão de Lewis), uma dose aumentada de 60 mg / kg (30 mg / kg duas vezes ao dia) administrada em dias alternados ip para 7 tratamentos mostra inibição significativa no crescimento. E também inibe o crescimento de xenoenxerto de carcinoma de cólon humano HCT-116 e xenoenxerto de carcinoma de pulmão de células não pequenas humano H-460. Estudos de eficácia mostram que a dose máxima tolerada é de 78 mg / kg / d. Ensaio de cinase de protocolo (apenas para referência): [1]
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Cdk4-D1/Cdk2-E enzyme assay
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The Cdk4-D1/Cdk2-E enzyme assay is run in 96-well format using Millipore Multiscreen filtration plates. All assay steps are done in a single filter plate. The filtration wells are prewetted with 100 μL of kinase buffer [50 mmol/L HEPES (pH, 7.5), 10 mmol/L MgCl2, 1 mmol/L EGTA], and then the solution is removed by vacuum. With filter plate on vacuum manifold, 50 μL GST-Rb bound to GSH-Sepharose beads in kinase buffer (0.5 μg GST-Rb/50 μL) is added to each well, and vacuum is applied to the filter plate. About 25 μL of a reaction mix containing ATP (cold + hot) and 4× phosphatase inhibitor mix (40 μmol/L unlabeled ATP, 10 μCi/mL γ32P-ATP, 40 mmol/L h-glycerophosphate, 4 mmol/L DTT, 0.4 mmol/L NaF, 0.4 mmol/L sodium orthovanadate)
diluted in kinase buffer is added to each well. The test compound (4×final concentration in kinase buffer) or kinase buffer alone (control) is then added in an additional 25 μL volume. To each well, 50 μL (100 ng) of human Cdk4-D1/Cdk2-E enzyme in kinase buffer is added to initiate the reaction, which is allowed to continue for 30 min at 30°C.
When the reaction is completed, vacuum is applied again, and the plate is washed with the TNEN buffer [20 mmol/L Tris (pH, 8.0), 100 mmol/L NaCl, 1 mmol/L EDTA, 0.5% nonidet-P40] thrice; the filter plate is air-dried and is placed in a Multiscreen adapter plate. Packard Microscint-O cocktail (30 μL) is added, and the plate is covered with a Top-Seal A film. Quantitation of 32P-GST-Rb in 96-well filter plates is carried out by Top Count scintillation counter. All compounds are tested initially at 1 μmol/L concentration. Compounds showing more than or equal to 50% inhibition are further profiled for IC50 determination.
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Ensaio de células: [1]
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Cell lines
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HCT-116, U2OS, H-460, HL-60, HT-29, SiHa, MCF-7, Colo-205, SW-480, PC-3, Caco2, T-24
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Concentrations
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~5.0 μM
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Incubation Time
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48 h
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Method
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The cells are seeded at a density of 3,000-5,000 cells per well, depending on cell type in 180 μL of culture medium in 96-well plate and incubated overnight to allow the cells to adhere. Varying concentrations of compounds are added to the wells and incubated for 48 h at 37°C. 3H-thymidine (0.25 μCi) is added to each well, and incorporation of the radiolabel is allowed to proceed for
5 to 7 h. Following this incubation, cells are harvested onto GF/B unifilter plates using a Packard Filtermate Universal harvester, and the plates are counted in a Packard Top Count 96-well liquid scintillation counter.
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Estudo Animal: [2]
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Animal Models
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H-460 xenograft
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Formulation
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water
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Dosages
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50 mg/kg once daily or 30 mg/kg twice daily
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Administration
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i.p.
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Solubility
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30% propylene glycol, 5% Tween 80, 65% D5W,
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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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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438.3
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Formula
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C21H20ClNO5.HCl
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CAS No.
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920113-03-7
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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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88 mg/mL
(200.77 mM)
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Water
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88 mg/mL
(200.77 mM)
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Ethanol
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7 mg/mL
(15.97 mM)
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In vivo
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30% propylene glycol, 5% Tween 80, 65% D5W
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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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4H-1-Benzopyran-4-one, 2-(2-chlorophenyl)-5,7-dihydroxy-8-[(2R,3S)-2-(hydroxymethyl)-1-methyl-3-pyrrolidinyl]-, hydrochloride (1:1)
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Calculadora de molaridade Calculadora de diluição Calculadora de peso molecular
Grupo de Produto : Ciclo de célula > Inibidor CDK
