.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 500,33 CW069 - аллостерический и селективный ингибитор HSET моторного белка микротрубочек с IC50 75 мкМ, значительная селективность по сравнению с KSP. \ n Биологическая активность
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Description
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CW069 is an allosteric, and selective inhibitor of microtubule motor protein HSET with IC50 of 75 μM, significant selectivity over KSP.
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Targets
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HSET [1]
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IC50
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75 μM
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In vitro
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CW069 increases multipolar spindles in N1E-115 cells with supernumerary centrosomes without altering bipolar spindle morphology in normal human dermal fibroblast cells. CW069 inhibits growth in cancer N1E-115 cells with IC50 of 10 μM, but not in NHDF or primary human bone marrow cells.
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In vivo
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Features
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Allosteric, HSET-selective inhibitor.
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Протокол (только для справки) Анализ киназы: [1]
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In Vitro Enzymatic ATPase Assay
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The protocol is optimized for use with full-length, N-terminal, 6His-tagged HSET and KSP, and measured the MT-stimulated activity of the proteins. Inhibition of the Gsp synthetase activity of HSET/KSP is observed spectrophotometrically by coupling the hydrolysis of ATP to oxidation of NADH via pyruvate kinase/lactate dehydrogenase reactions. The assay is initiated by adding purified Gsp synthetase/amidase (12.8 nM) to an assay mixture containing the following components (final concentration): 6 nM protein, 0.07 mg/ml MTs (University Biologicals), 1.56 mM glutathione, 10 mM spermidine, 2 mM ATP, 2.7 mM MgCl2, 1 mM phospho(enol)-pyruvate, 0.2 mM NADH, 50 μg/ml lactate dehydrogenase, 100 μg/ml pyruvate kinase, and various concentrations of inhibitor all in 50 mM Na PIPES (pH 6.8) at 37°C. The ADP-Glo detection assay (Promega) is performed as described in the manufacturer`s instructions. All compound additions were performed using a multidrop BioMek Nxp. Plates were read using a Pherastar microplate reader.
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Клеточный анализ: [1]
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Cell lines
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N1E-115 cells, NHDF and primary human bone marrow cells.
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Concentrations
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~400 μM
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Incubation Time
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72 hours
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Method
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Cells are cultured in DMEM supplemented with 10% fetal calf serum (FCS) at 37°C and 5% CO2. All compounds used in the Sulforhodamine B colorimetric (SRB) assay are dissolved in DMSO and diluted in culture medium to a final concentration of 0.2% DMSO. For the SRB assay and live-cell imaging, cells are seeded in 96-well plates at a density of 2,500 cells per well. After 24 hr, the cells are treated with compound for 72 hr, with triplicate wells for each concentration. For the SRB assay, the cells are then fixed with trichloroacetic acid (TCA) and stained with SRB. Fluorescence is quantified using an Infinite 200 PRO plate-reader at a wavelength of 545 nm. Compound-treated wells are compared with solvent control wells and the concentration of compound that results in 50% of the solvent-control cell growth is designated as the IC50 concentration, calculated using Graphpad PRISM 6. At least three biological replicates are performed for each assay.
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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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500.33
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Formula
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C23H21IN2O3
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CAS No.
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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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N/A
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Solubility (25°C) *
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In vitro
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DMSO
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100 mg/mL
(199.86 mM)
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Water
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<1 mg/mL
(
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Ethanol
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4 mg/mL
(7.99 mM)
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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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(S)-2-(2-(benzylamino)-3-phenylpropanamido)-5-iodobenzoic acid
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Калькулятор молярности Калькулятор разведения Калькулятор молекулярной массы
Группа Продуктов : Цитоскелетная передача сигналов > Ингибитор, связанный с микротрубочками
