.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 409,23 HA14-1 - это непептидный лиганд поверхностного кармана Bcl-2 с IC50 ~ 9 мкМ. \ n Биологическая активность HA14-1 представляет собой небольшую молекулу и непептидный лиганд поверхностного кармана Bcl-2 с IC50 приблизительно 9 мкМ. HA14-1 вызывает апоптоз клеток HL-60 дозозависимым образом через активацию каспазы. НА14-1 проявляет цитотоксическое действие на линии В-клеток фолликулярной лимфомы HF1A3, HF4.9 и HF28RA с LC50 4,5 мкМ, 12,6 мкМ и 8,1 мкМ соответственно. HA14-1 индуцирует апоптоз клеток HF1A3, HF4.9 и HF28RA через каспазу и АФК. Обработка HA14-1 (400 нМ) не оказывала значительного влияния на рост опухолей глиобластомы у иммунодефицитных мышей. Но НА14-1 (400 нМ) усиливает действие повреждающего ДНК агента этопозида (2,5 мг / кг) на рост глиобластомы in vivo. Протокол (только для справки) Анализ киназы: [1]
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Affinity determination
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The binding affinity of organic compounds to Bcl-2 protein in vitro is determined by a competitive binding assay based on fluorescence polarization. For this assay, 5-carboxyfluorecein is coupled to the N terminus of a peptide, GQVGRQLAIIGDDINR, derived from the BH3 domain of Bak (Flu-BakBH3), which has been shown to bind to the surface pocket of the Bcl-xL protein with high-affinity. According to our molecular modeling studies and binding measurement using fluorescence polarization, the Flu-BakBH3 peptide binds the surface pocket of Bcl-2 with a similar affinity. Bcl-2 used in this assay is a recombinant GST-fused soluble protein. Flu-BakBH3 and Bcl-2 protein are mixed in the presence or absence of organic compounds under standard buffer conditions and are incubated for 30 min. The binding of Flu-BakBH3 to Bcl-2 protein is measured on a LS-50 luminescence spectrometer equipped with polarizers using a dual path length quartz cell (500 μl). The fluorophore is excited with vertical polarized light at 480 nm (excitation slit width 15 nm), and the polarization value of the emitted light is observed through vertical and horizontal polarizers at 530 nm (emission slit width 15 nm). The binding affinity of each compound for Bcl-2 protein is assessed by determining the ability of different concentrations of the compound to inhibit Flu-BakBH3 binding to Bcl-2.
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Клеточный анализ: [2]
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Cell lines
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HF1A3, HF4.9 and HF28RA cells
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Concentrations
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~25 μM
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Incubation Time
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20 h
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Method
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The cytotoxic effects of HA14-1 against different FL cell lines are determined by the MTT assay. Briefly, the cells (5000/well) are incubated in triplicate in 96-well plate in the presence or absence of HA14-1 for 20 h at 37 °C. Thereafter, the MTT solution is added to each well. After 4 h incubation at 37 °C, the optical density (OD) is measured by means of 96-well plate reader, with the extraction buffer as a blank. The following formula is used: percentage cell viability = (OD of the experiment samples/OD of the control) × 100. Sigmoidal dose-response curves are fitted to the mean cell viability plotted against log HA14-1 dose and lethal concentration 50% (LC50) values are calculated from the resulting curves using Prism 4.0 softw
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Исследование на животных: [3]
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Animal Models
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Female Swiss nude mice bearing BeGBM xenografts.
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Formulation
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Free RPMI 1640-50% DMSO.
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Dosages
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400 nM
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Administration
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Inject at the site of cell injection.
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Solubility
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1% DMSO/30% polyethylene glycol/1% Tween 80,
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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409.23
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Formula
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C17H17BrN2O5
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CAS No.
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65673-63-4
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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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82 mg/mL
(200.37 mM)
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Water
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<1 mg/mL
(
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Ethanol
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82 mg/mL
(200.37 mM)
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In vivo
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1% DMSO/30% polyethylene glycol/1% Tween 80
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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)-ethyl 2-amino-6-bromo-4-((R)-1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate
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Калькулятор молярности Калькулятор разведения Калькулятор молекулярной массы
Группа Продуктов : Апоптоз > Bcl-2 ингибитор
