.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: 532,61 Опрозомиб (ONX 0912) представляет собой пероральный биодоступный ингибитор активности CT-L 20S протеасомы β5 / LMP7 с IC50 36 нМ / 82 нМ. Фаза 1/2. \ N Активность опрозомиба против ММ связана с активацией каспазы-8, каспазы-9, каспазы-3 и PARP, а также с ингибированием миграции клеток ММ и ангиогенеза. Абсолютная биодоступность опрозомиба у грызунов и собак достигает 39%. Он хорошо переносится при повторном пероральном введении в дозах, приводящих к ингибированию протеасом> 80% в большинстве тканей и вызывая противоопухолевый ответ на множественных ксенотрансплантатах опухоли человека и сингенных моделях мышей. \ п
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ELISA-based active site binding assay
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Samples (lysed cells or tissue homogenates) are treated for 1 h at room temperature with the biotinylated active site probe PR-584 (5-15 μM). Samples are denatured by addition of SDS (0.9% final) and heating to 100 °C for 5 min. The denatured samples are transferred to a 96-well or 384-well filter plat, mixed with streptavidin-sepharose beads (2.5-5 μL packed beads/well), and incubated for 1 h at room temperature on a plate shaker. The beads are washed 5 times with 100-200 μL /well of ELISA buffer (PBS, 1% bovine serum albumin, 0.1% Tween-20) by vacuum filtration. The beads are incubated overnight at 4 °C on a plate shaker with the following antibodies recognizing the six catalytic subunits diluted into ELISA buffer: β5, β1, and β2 diluted 1:3000, LMP7 and LMP2 diluted 1:5000, and MECL-1 diluted 1:1000. The beads are washed 5 times with 100-200 μL /well of ELISA buffer and incubated with HRP-conjugated secondary antibody diluted 1:5000 in ELISA buffer and incubated 2 h at room temperature on a plate shaker. The beads are washed 5 times with 100-200 μL /well of ELISA buffer and developed for chemiluminsecence signal using the supersignal ELISA pico substrate following the manufacturer's instructions. Luminescence is measured on a plate reader and converted to ng of proteasome or μg/ml of lysate by comparison with 20S proteasome or untreated cell lysate standard curves. For proteasome inhibitor studies, active site probe binding values are expressed as the percent of binding relative to DMSO treated cells.
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Клеточный анализ: [2]
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Cell lines
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MM.1S, MM.1R, RPMI-8226, KMS12, INA-6, OPM-2, Dox-40
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Concentrations
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~1 μM
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Incubation Time
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48 h
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Method
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MTT assay
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Исследование на животных: [1]
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Animal Models
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Non-Hodgkin`s lymphoma cell line RL xenograft, colorectal tumor cell
line CT-26 xenograft
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Formulation
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10% (v/v) EtOH and 10% (v/v) PS80 in citrate buffer (pH 3.5)
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Dosages
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30 mg/kg, twice weekly on days 1 and 2
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Administration
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p.o.
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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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532.61
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Formula
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C25H32N4O7S
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CAS No.
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935888-69-0
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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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100 mg/mL
(187.75 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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Chemical Name
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L-Serinamide, O-methyl-N-[(2-methyl-5-thiazolyl)carbonyl]-L-seryl-O-methyl-N-[(1S)-2-[(2R)-2-methyl-2-oxiranyl]-2-oxo-1-(phenylmethyl)ethyl]-
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Stock Solution (1ml DMSO)
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1mM
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10mM
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20mM
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30mM
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Mass(mg)
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0.53261
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5.3261
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10.6522
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15.9783
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Группа Продуктов : Убиквитин > Ингибитор протеасомы
