.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: 470,52 ETP-46464 é um inibidor potente e seletivo de ATR com IC50 de 25 nM. \ n ETP-46464 mostra atividade quase total contra ATR a 100 nM. ETP-46464 inibe a atividade de PI3Kα, mTOR e DNA-PKcs com IC50s de 170, 0,6 e 36 nM. O ETP-46464 é capaz de promover a quebra de garfos de replicação paralisados. A exposição ao ETP-46464 leva à geração de quantidades substanciais de danos ao DNA em células em replicação. 1 μM ETP-46464 anula o ponto de verificação G2 / M induzido por radiação ionizante e leva à presença de micronúcleos ou núcleos completamente fragmentados nas células. ETP-46464 é particularmente tóxico para células deficientes em p53, o que é exacerbado por condições geradoras de estresse replicativo, como a superexpressão de ciclina E. \ n Inibidor seletivo de ATR e particularmente tóxico para células cancerosas deficientes em p53. Freqüentemente usado como uma sonda farmacológica devido às suas propriedades farmacológicas ideais. \ n
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ATR kinase activity assay
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For the in vitro kinase assay flag-tagged ATR kinase is immunoprecipitated from 293T cells using Anti-flag M2 affinity gel. As a substrate, p53/1-101 peptide fused to GST is purified from IPTG stimulated BL21 bacteria by conventional GST purification methods. Briefly, pBJF-flag-ATR plasmids (wild type and kinase dead versions) is transfected into 293T cells. 48 hours after transfection, cells are lysed in TGN buffer (50 mM Tris (pH 7.5), 50 mM glycerophosphate, 150 mM NaCl, 10% glycerol, 1% tween-20 and protease inhibitors cocktail) for 15 minutes on ice and, after centrifugation, the supernatant is pre-cleared for 1h with unspecific rabbit IgG and protein A on a rotator at 4 degrees. Next, the supernatant is incubated with M2-slurry for 2 h at 4 degrees. For a confluent p100 plate of 293T cells, 400 μL of buffer, 10 μg of IgG, 30 μL of protein A slurry, and 25 μL of M2 beads are used. Anti flag M2 beads are then washed 5 times in ice cold TNG, 3 times in ice cold LiCl buffer (100mM Tris pH 7.5, 0.5 M LiCl), and 5 times in ice cold Kinase buffer (10 mM Hepes (pH 7.5), 50 mM NaCl, 10 mM MgCl2, 10 mM Mn2, 1 mM DTT). After the last wash, IPs are splitted in the proper number of tubes for the assay (approximately one confluent 293T p100 is used per condition). For the kinase reaction, 100 μM `cold` ATP is added to the buffer. Only fresh IPs are used. Prior to every experiment a mix is prepared containing 2 μg of GST-p53 and 10 μ Ci of 32P-γATP per condition. Inhibitor are first added to the beads. Last, 25μL of the mix is added to each tube. Reactions are performed for 20 minutes at 30 degrees and stopped by adding 15 microliters of 4 × NuPAGE SDS buffer and boiling for 5 minutes. Beads are pelleted and supernatants are resolved by SDS-PAGE on a polyacrylamide gel. Radioactive gels are slightly washed and directly exposed to Phospor-Imager screens for 2h to 16h. Radioactivity is then detected using a STORM scanner. Finally, gels are stained with coomasie to check for protein levels.
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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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\ n Informação Química
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Molecular Weight (MW)
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470.52
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Formula
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C30H22N4O2
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CAS No.
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1345675-02-6
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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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6 mg/mL
(12.75 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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* <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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Benzeneacetonitrile, α,α-dimethyl-4-[2-oxo-9-(3-quinolinyl)-2H-[1,3]oxazino[5,4-c]quinolin-1(4H)-yl]-
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Calculadora de molaridade Calculadora de diluição Calculadora de peso molecular
Grupo de Produto : PI3K / Akt / mTOR > Inibidor ATM / ATR
