.cp_wz table {border-top: твердое тело 1px #ccc; border-left: твердое тело 1px #ccc; } .cp_wz таблица td {border-right: 1px solid #ccc; нижняя граница: сплошной 1px #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 Молекулярный вес: \ n 394,36 JNJ-7706621 - пан-ингибитор CDK с наивысшей эффективностью в отношении CDK1 / 2 с IC50 9 нМ / 4 нМ и демонстрирующий> 6-кратную селективность в отношении CDK1 / 2, чем CDK3 / 4/6. Он также сильно ингибирует Aurora A / B и не проявляет активности в отношении Plk1 и Wee1. \ N Биологическая активность JNJ-7706621 также демонстрирует некоторое ингибирование VEGF-R2, FGF-R2 и GSK3β с IC50 154–254 нМ. JNJ-7706621 демонстрирует ингибирующее действие на панель типов раковых клеток человека, включая HeLa, HCT-116, SK-OV-3, PC3, DU145, A375, MDA-MB-231, MES-SA и MES-SA / Dx5. с IC50 112-514 нМ, независимо от p53, ретинобластомы или статуса P-гликопротеина. JNJ-7706621 в несколько раз менее эффективен при ингибировании роста нормальных типов клеток, включая MRC-5, HASMC, HUVEC и HMVEC, с IC50 3,67-5,42 мкМ. В клетках HeLa или U937 JNJ-7706621 (0,5–3 мкМ) задерживает выход из G1, задерживает клетки в G2-M, вызывает эндоредупликацию, активирует апоптоз и снижает образование колоний. В клеточной линии HeLa добавочная обработка увеличивающимися концентрациями JNJ-7706621 приводит к 16-кратной устойчивости, которая может быть опосредована ABCG2. В модели ксенотрансплантата мыши меланомы опухоли человека A375 JNJ-7706621 (100 или 125 мг / кг) вызывает регрессию опухоли. Протокол (только для справки) Анализ киназы: [1]
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In vitro kinase assay for CDK1 and Aurora kinases
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For CDK1 kinase activity, a method is developed using the CDK1/cyclin B complex purified from baculovirus to phosphorylate a biotinylated peptide substrate containing the consensus phosphorylation site for histone H1, which is phosphorylated in vivo by CDK1. Inhibition of CDK1 activity is measured by observing a reduced amount of 33P-γ-ATP incorporation into the immobilized substrate in streptavidin-coated 96-well scintillating microplates. CDK1 enzyme is diluted in 50 mM Tris-HCl (pH 8), 10 mM MgCl2, 0.1 mM Na3VO 4, 1 mM DTT, 1% DMSO, 0.25 μM peptide, 0.1 μCi per well 33P-γ-ATP, and 5 μM ATP in the presence or absence of various concentrations of JNJ-7706621 and incubated at 30 °C for 1 hour. The reaction is terminated by washing with PBS containing 100 mM EDTA and plates are counted in a scintillation counter. Linear regression analysis of the percent inhibition by JNJ-7706621 is used to determine IC50. The Aurora kinase assays are done with 10 μM ATP and a peptide containing a dual repeat of the kemptide phosphorylation motif.
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Клеточный анализ: [3]
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Cell lines
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HeLa, HCT-116, A375, SK-OV-3, MDA-MB-231, and PC-3 cells
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Concentrations
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1 nM - 10 μM, dissolved in DMSO
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Incubation Time
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48 hours
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Method
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The ability of JNJ-7706621 to inhibit the proliferation of cell growth is determined by measuring incorporation of 14C-labelled thymidine into newly synthesized DNA within the cells. Cells are trypsinized and counted and 3-8 × 103 cells are added to each well of a 96-well CytoStar tissue culture treated scintillating microplate in complete medium in a volume of 100 μL. Cells are incubated for 24 hours in complete medium at 37 °C in an atmosphere containing 5% CO2. Next, 1 μL of JNJ-7706621 is added to the wells of the plate. Cells are incubated for 24 more hours. Methyl 14C-thymidine 56 mCi/mmol is diluted in complete medium and 0.2 μCi/well is added to each well of the CytoStar plate in a volume of 20 μL. The plate is incubated for 24 hours at 37 °C in JNJ-7706621 plus 14C-thymidine. The contents of the plate are discarded and the plate is washed twice with 200 μL PBS. 200 μL of PBS is added to each well. The top of the plate is sealed with a transparent plate sealer and a white plate backing sealer is applied to the bottom of the plate. The degree of methyl 14C-thymidine incorporation is quantified on a Packard Top Count.
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Исследование на животных: [1]
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Animal Models
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Mouse xenograft model of A375 cells
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Formulation
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Dissolved in 0.5% methylcellulose containing 0.1% polysorbate 80 in sterile water.
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Dosages
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100 or 125 mg/kg
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Administration
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Orally or by intraperitoneal injection injection
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Solubility
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0.5% methylcellulose/0.2% Tween 80,
14 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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Конверсия различных модельных животных на основе BSA (значение на основе данных из проекта рекомендаций 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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Например, чтобы изменить дозу ресвератрола, используемую для мыши (22,4 мг / кг), на дозу, основанную на BSA для крысы, умножьте 22,4 мг / кг на коэффициент Km для мыши, а затем разделите на коэффициент Km для крыса. Этот расчет дает эквивалентную дозу ресвератрола для крыс 11,2 мг / кг.
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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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Химическая информация
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Molecular Weight (MW)
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394.36
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Formula
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C15H12F2N6O3S
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CAS No.
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443797-96-4
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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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79 mg/mL
(200.32 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% methylcellulose/0.2% Tween 80
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14 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-[[5-amino-1-(2,6-difluorobenzoyl)-1H-1,2,4-triazol-3-yl]amino]-benzenesulfonamide
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Калькулятор молярности Калькулятор разведения Калькулятор молекулярной массы
Группа Продуктов : Клеточный цикл > CDK ингибитор
