.cp_wz table {border-top: 1px solid #ccc; border-left: 1px solid #ccc; } .cp_wz table td {border-right: 1px solid #ccc; borda inferior: 1px sólido #ccc; preenchimento: 5px 0px 0px 5px;} .cp_wz tabela th {border-right: 1px solid #ccc; border-bottom: 1px solid #ccc; preenchimento: 5px 0px 0px 5px;} \ n Peso molecular: 532,61 Oprozomibe (ONX 0912) é um inibidor oralmente biodisponível para a atividade CT-L do proteassoma 20S β5 / LMP7 com IC50 de 36 nM / 82 nM. Fase 1/2. \ N A atividade anti-MM de Oprozomib está associada à ativação de caspase-8, caspase-9, caspase-3 e PARP, bem como inibição da migração de células MM e angiogênese. Oprozomib demonstrou uma biodisponibilidade absoluta de até 39% em roedores e cães. É bem tolerado com administração oral repetida em doses resultando em> 80% de inibição do proteassoma na maioria dos tecidos e induziu uma resposta antitumoral em múltiplos xenoenxertos de tumor humano e modelos singênicos de camundongo. \ n
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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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Ensaio de células: [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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Estudo Animal: [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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Conversão de diferentes modelos de animais com base em BSA (valor com base em dados das diretrizes preliminares da 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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Por exemplo, para modificar a dose de resveratrol usada para um camundongo (22,4 mg / kg) para uma dose baseada na BSA para um rato, multiplique 22,4 mg / kg pelo fator Km para um camundongo e, em seguida, divida pelo fator Km para um rato. Este cálculo resulta em uma dose equivalente de rato para o resveratrol de 11,2 mg / kg.
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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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Informação Química
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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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Grupo de Produto : Ubiquitina > Inibidor de Proteassoma
