.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 553,52 SB743921 - ингибитор белка веретена кинезина (KSP) с Ki 0,1 нМ, почти не имеет сродства к MKLP1, Kin2, Kif1A, Kif15, KHC, Kif4 и CENP- Э. Фаза 1/2. \ N Биологическая активность. Ki SB 743921 для KSP человека и мыши составляет 0,1 нМ и 0,12 нМ, соответственно, тогда как Ki SB 743921 для других кинезинов, включая MKLP1, Kin2, составляет более 70 мкМ. SB 743921 блокирует сборку функционального митотического веретена, тем самым вызывая остановку клеточного цикла в митозе и последующую гибель клеток. SB-743921 обладает большей эффективностью по сравнению с испинезибом как в биохимических, так и в клеточных анализах. SB-743921 обладает большей эффективностью in vivo против лейкемии P388. SB-743921 обладает значительной эффективностью в широком спектре моделей опухолей, отличных от таксанов. SB-743921 обладает активностью против продвинутых ксенотрансплантатов опухоли человека Colo205 (полная регрессия), MCF-7, SK-MES, H69, OVCAR-3 (полная и частичная регрессия) и HT-29, MX-1, MDA- MB-231, A2780 (задержка роста опухоли). SB-743921 не вызывает невропатию, часто связанную с тубулиновыми агентами. Протокол (только для справки) Анализ киназы: [2]
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Biochemistry assay
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The motor domains of KSP (amino acids 1–360) is expressed as in Escherichia coli BL21(DE3) as COOH-terminal 6-his fusion proteins. Bacterial pellets are lysed in a microfluidizer with a lysis buffer [50 mM Tris-HCl; 50 mM KCl, 10 mM imidazole, 2 mM MgCl2, 8 mM β-mercaptoethanol, 0.1 mM ATP (pH 7.4)], and proteins are purified using Ni-NTA agarose affinity chromatography, with an elution buffer consisting of 50 mM PIPES, 10% sucrose, 300 mM imidazole, 50 mM KCl, 2 mM MgCl2, mM β-mercaptoethanol, and 0.1 mM ATP (pH 6.8). Steady-state measurements of ATPase activity are performed with a pyruvate kinase–lactate dehydrogenase detection system that coupled the appearance of ADP with oxidation of NADH. Absorbance changes are monitored at 340 nm. All biochemical experiments are performed in PEM25 buffer [25 mM Pipes/KOH (pH 6.8), 2 mM MgCl2, 1 mM EGTA] supplemented with 10 μM SB 743921 for experiments involving microtubules. Rates of ADP release are measured in a stopped-flow apparatus; the decrease in fluorescence of MANT-ATP is monitored. Rates of Pi release are measured in a stopped-flow apparatus, using bacterial phosphate binding protein modified with 7-diethylamino-3-((((2 maleimidyl)ethyl)amino)carbonyl)coumarin (MDCC) dye. Ki estimates of KSP inhibitors are extracted from the dose–response curves, with explicit correction for enzyme concentration. Tubulin polymerization by measuring changes in absorbance at 340 nm is monitored. The assay is performed in 100-μL volumes in 96-well half-area microtiter plates, using a microplate reader with the incubation temperature set at 37 °C.
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Клеточный анализ: [2]
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Cell lines
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HeLa cells
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Concentrations
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~1 μM
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Incubation Time
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24 hours
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Method
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All cells including HeLa cells are cultured in 10% FCS in RPMI 1640 in 5% CO2. We assessed 48-hour growth inhibition by serial dilution of SB 743921 relative to DMSO-treated cells in 96-well microtiter plates, using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium. Cell growth is represented as the ratio of absorbance of treated cells to DMSO control, plotted by concentration and fitted to a four-parameter curve. Concentrations at which cellular growth is inhibited by 50% are extrapolated from the curve fit. The DNA content of HeLa cells cultured in the presence or absence of 1 μM SB 743921 for 24 hours is assessed by propidium iodide staining and flow cytometry. Immunofluorescence images are collected of HeLa cells treated for 24 hours with 1 μM SB 743921, fixed with 2% formaldehyde, permeabilized, and stained with DM1-α, anti-γ-tubulin, and 1 μg/mL 4′,6-diamidino-2-phenylindole, and with Alexa 488 secondary goat antirabbit IgG and Rhodamine-X goat antimouse IgG. Images are collected with a DeltaVision Restoration Microscopy System at a magnification of ×600. Z stacks (0.2 μm) are collected, and out of focus information is removed by constrained iterative deconvolution. Z stacks are then compressed into to a single image plane.
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Исследование на животных: [1]
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Animal Models
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Female BDF1 mice with P388 lymphocytic leukemia cells
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Formulation
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2% dimethylacetamide + 2% Cremophor EL + 96% acidified water [pH 5.0]
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Dosages
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7.5 mg/kg- 30 mg/kg
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Administration
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Administered via i.p.
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Solubility
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Saline pH5.0,
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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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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553.52
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Formula
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C31H33N2O3.HCl
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CAS No.
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940929-33-9
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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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111 mg/mL
(200.53 mM)
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Water
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22 mg/mL
(39.74 mM)
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Ethanol
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111 mg/mL
(200.53 mM)
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In vivo
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Saline pH5.0
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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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(R)-N-(3-aminopropyl)-N-(1-(3-benzyl-7-chloro-4-oxo-4H-chromen-2-yl)-2-methylpropyl)-4-methylbenzamide hydrochloride
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Калькулятор молярности Калькулятор разведения Калькулятор молекулярной массы
Группа Продуктов : Цитоскелетная передача сигналов > Ингибитор кинезина
