.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: 488,74 PIK-75 é um inibidor de p110α com IC50 de 5,8 nM (200 vezes mais potente do que p110β), mutantes específicos de isoformas em Ser773 e também inibe potentemente DNA-PK com IC50 de 2 nM. \ n PIK-75 mostra a potência impressionante e seletividade de isoforma em p110α enquanto os valores de IC50 correspondentes são 1300 nM, 76 nM e 510 nM para outras isoformas de PI3K, p110β, -γ e -δ, respectivamente. Além disso, quando se liga a p110α purificado, PIK-75 é um inibidor não competitivo em relação ao ATP com Ki de 36 nM e competitivo em relação ao substrato PI com Ki de 2,3 nM. PIK-75 também mostra inibição potente de DNA-PK. PIK-75 (1 μM) reduz a sobrevivência celular diminuindo significativamente a atividade mitocondrial em células não estimuladas do músculo liso das vias aéreas (ASM), células ASM asmáticas e fibroblastos pulmonares. Enquanto em células ASM estimuladas por TGFβ, PIK75 apenas diminui a atividade mitocondrial em células asmáticas, sem efeitos em células não asmáticas. Um estudo recente mostra que PIK-75 (10 nM) inibe a expressão de mRNA de CD38 induzida por TNF-α e atenua significativamente a atividade de ADP-ribosil ciclase induzida por TNF-α em células de músculo liso humano das vias aéreas. \ n No modelo de tumor ErbB3WT, PIK-75 reduz a resposta quimiotática in vitro a HRGβ1 e reduz os níveis de pAkt em 40%. Além disso, PIK-75 reduz significativamente a motilidade das células tumorais e a invasão in vivo em tumores primários ErbB3WT. [5] Em camundongos machos CD1, PIK-75 leva a sérios prejuízos no teste de tolerância à insulina (ITT) e teste de tolerância à glicose (GTT), e um aumento na produção de glicose durante um teste de tolerância ao piruvato (PTT). Ensaio de cinase de protocolo (apenas para referência): [1]
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Inhibition Assays
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The PI3K inhibitor PIK-75 is dissolved at 10 mM in dimethyl sulfoxide and stored at −20°C until use. PI3K enzyme activity is determined in 50 μL of 20 mM HEPES, pH 7.5, and 5 mM MgCl2 containing 180 μM phosphatidyl inositol, with the reaction started by the addition of 100 μM ATP (containing 2.5 μCi of [γ-32P]ATP). After a 30-minute incubation at room temperature, the enzyme reaction is stopped by the addition of 50 μL of 1 M HCl. Phospholipids are then extracted with 100 μL of chloroform/methanol [1:1 (v/v)] and 250 μL of 2 M KCl followed by liquid scintillation counting. Inhibitors are diluted in 20% (v/v) dimethyl sulfoxide to generate a concentration versus inhibition of enzyme activity curve, which is then analyzed with the use of Prism version 5.00 for Windows to calculate the IC50. For kinetic analysis, a luminescent assay measuring ATP consumption is used. PI3K enzyme activity is determined in 50 μL of 20 mM HEPES, pH 7.5, and 5 mM MgCl2 with PI and ATP at various concentrations. After a 60-minute incubation at room temperature, the reaction is stopped by the addition of 50 μL of Kinase-Glo followed by a further 15-minute incubation. Luminescence is then read using a Fluostar plate reader. Results are analyzed using Prism.
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Ensaio de células: [3]
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Cell lines
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A2780, A2780/cp70, 2780AD, HCT116, HT29, WIL, CALU-3, MCF7, PC3 and HS852 cells
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Concentrations
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0-10 μM
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Incubation Time
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48 hours
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Method
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Mitochondrial activity is assessed after stimulation with TGFβ with or without inhibitors for 48 hours using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. Harvested washed cells are resuspended in DMEM-lO% FCS and aliquoted (500 μL) into 24-well cluster plates prior to serial dilution (1:2) in duplicates. To each well, 100 μL of an appropriate MTT concentration (dissolved in PBS and filtered through a 0.2 μm filter before use to remove any blue formazan product) is added immediately after diluting the cells, which are then incubated for 3.5 hours at 37 °C. The resulting blue formazan product is solubilized overnight (16 hours) at 37 °C by the addition of 500 μL of 10% sodium dodecyl sulfate (SDS) in 0.01 M HCl to each well. A sample (150 μL) from each duplicate well is transferred to a 96-well microplate, and the optical density determinedby automated spectrophotometry against a reagent blank (no cells). Absorbance is measured at a test wavelength of 570 nm and a reference wavelength of 690 nm. For each primary cell culture, results from three to six wells from each treatment are averaged, and data are expressed as absorbance 570 to 690 nm.
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Estudo Animal: [5]
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Animal Models
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MTLn3 cells are injected into the right fourth mammary fat pad from the head of female severe-combined immunodeficient/NCr mice.
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Formulation
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PIK-75 is dissolved in DMSO and then diluted in PBS.
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Dosages
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≤1 μM
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Administration
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Administered via i.p.
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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 \ n Informação Química
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Molecular Weight (MW)
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488.74
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Formula
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C16H14BrN5O4S.HCl
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CAS No.
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372196-77-5
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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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3 mg/mL
(6.13 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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(E)-N'-((6-bromoH-imidazo[1,2-a]pyridin-3-yl)methylene)-N,2-dimethyl-5-nitrobenzenesulfonohydrazide hydrochloride
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Calculadora de molaridade Calculadora de diluição Calculadora de peso molecular
Grupo de Produto : PI3K / Akt / mTOR > Inibidor PI3K
