.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: 333,36 GSK1059615 - двойной ингибитор PI3Kα / β / δ / γ (обратимый) и mTOR с IC50 0,4 нМ / 0,6 нМ / 2 нМ / 5 нМ и 12 нМ, соответственно. Фаза 1. \ n Производное пиридинилхинолина GSK1059615 представляет собой новый, АТФ-конкурентный и обратимый ингибитор семейства PI3K класса I. GSK1059615 ингибирует передачу сигналов PI3K, вызывает остановку G1 и апоптоз, особенно в клетках опухоли молочной железы. Данные, обобщенные в другом обзоре, также показывают, что GSK1059615 ингибирует PI3Kα, β, γ и δ с Ki 0,42 нМ, 0,6 нМ, 0,47 нМ и 1,7 нМ, соответственно. Другой отчет показывает, что GSK1059615 ингибирует PI3Kα с IC50 2 нМ. В раковых клетках T47D и BT474 GSK1059615 ингибирует фосфорилирование Akt по S473 с IC50 40 нМ. В моделях клеток рака молочной железы BT474 или HCC1954 на мышах с ксенотрансплантатом GSK1059615 (25 мг / кг) эффективно подавляет рост опухоли. Новый, АТФ-конкурентный и обратимый ингибитор как PI3Kα, β, δ и γ, так и mTOR. \ n Протокол (только для справки) Анализ киназы: [4]
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HTRF In vitro Profiling Assays for PI3K Inhibition
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The measurement of the GSK1059615-dependent inhibition of the PI3Ks is accessed using a HTRF based PI3K profiling assay kit. 400 pM enzyme is used in PI3Kα and δ assays, 200 pM in PI3Kβ assays, and 1 nM in PI3Kγ assay. In addition, the PI3Kα, β, and δ assays are run with 150 mM NaCl and 100 μM ATP, while the PI3Kγ assay is run with no NaCl and 15 μM ATP. All reactions are run at 10 μM PIP2. GSK1059615 is serially diluted (3-fold in DMSO), and 50 nL is transferred to a 384-well low-volume assay plate. PI3K Reaction Buffer is prepared by diluting the stock 1:4 with de-ionized water. Freshly prepared DTT is added at a final concentration of 5 mM on the day of use. Enzyme addition and GSK1059615 pre-incubation are initiated by the addition of 2.5 µL of PI3K in reaction buffer. Plates are incubated at room temperature for 15 min. Reactions are initiated by addition of 2.5 µL of 2× substrate solution (PIP2 and ATP in 1× reaction buffer). Plates are incubated at room temperature for one hour. Reactions are quenched by the addition of 2.5 µL of stop solution. The quenched reactions are then processed to detect product formation by adding 2.5 µL of Detection Solution. Following a 1-hour incubation in the dark, the HTRF signal is measured on the Envision plate reader set for 330nm excitation and dual emission detection at 620nm (Eu) and 665nm (APC). The IC50 value is then obtained.
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Клеточный анализ: [4]
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Cell lines
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T47D and BT474 cells
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Concentrations
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0–1 μM, serially diluted (3-fold) in DMSO
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Incubation Time
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30 min
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Method
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Cells are plate at a density of 1 × 104 cells per well in clear flat-bottomed 96-well plates and incubated overnight. Then, GSK1059615 is added and the plates are incubated for 30 min. At the end of incubation, media is aspirated from the plates, and the plate is wash once with cold PBS. 80 μL MSD Lysis buffer is added into each well and the plates are incubated on a shaker at 4 °C for at least 30 min. For Akt duplex assay, plates are washed with 200 μL/well wash buffer for 4 times and tapped on paper towel to blot. Then, 60 μL lysates is added to each well and the plates are incubated on shaker at room temperature for 1 hour. After another 4 times washing, antibody is added (25 μL per well) and the plates are incubated on shaker for 1 hour and washed again. Finally, Read Buffer is added (150 μL per well) and the plates are read immediately. IC50 values are then obtained.
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Исследование на животных: [3]
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Animal Models
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Xenograft mice models of BT474 or HCC1954 cells
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Formulation
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Dosages
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25 mg/kg
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Administration
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Oral gavage
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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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333.36
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Formula
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C18H11N3O2S
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CAS No.
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958852-01-2
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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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2 mg/mL
(5.99 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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* <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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(Z)-5-((4-(pyridin-4-yl)quinolin-6-yl)methylene)thiazolidine-2,4-dione
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Калькулятор молярности Калькулятор разведения Калькулятор молекулярной массы
Группа Продуктов : PI3K / Akt / mTOR > Ингибитор PI3K
