.cp_wz tabla {borde superior: 1px sólido #ccc; borde izquierdo: 1px sólido #ccc; } .cp_wz table td {borde derecho: 1px sólido #ccc; borde inferior: 1px sólido #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 Peso molecular: \ n 501.5 PND-1186 (VS-4718) es un inhibidor de FAK reversible y selectivo con IC50 de 1,5 nM. Fase 1. \ n Actividad biológica In vitro, PND-1186 inhibe la motilidad del carcinoma de mama 4T1, promueve la apoptosis de 4T1 en condiciones de suspensión y reduce el número y el tamaño de las colonias de agar blando 4T1. En las células HEY y OVCAR8, VS-4718 promueve la detención del ciclo celular G0-G1 seguida de la muerte celular. En ratones que portan tumores 4T1, PND-1186 (100 mg / kg sc) inhibe el crecimiento tumoral subcutáneo 4T1 por inducción de apoptosis. En ratones que portan tumores ID8, PND-1186 (0,5 mg / ml por vía oral) también causa inhibición del crecimiento tumoral de carcinoma de ovario. Protocolo (solo como referencia) Ensayo de quinasa: [1]
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In vitro kinase activity
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GST-FAK in vitro kinase activity is measured and compared to His-tagged FAK 411–686 using the K-LISA screening kit and poly(Glu:Tyr) (4:1) copolymer as a substrate immobilized on microtiter plates. IC50 values are determined with various concentrations of test compounds in a buffer containing 50 µM ATP and 10 mM MnCl2, 50 mM HEPES (pH 7.5), 25 mM NaCl, 0.01% BSA, and 0.1 mM Na orthovanadate for 5 min at room temperature. Serial diluted compounds are tested in triplicate. Substrate phosphorylation is measured using horseradish peroxidase-conjugated anti-pTyr antibodies with spetrophometic color quantitation. IC50 values are determined using the Hill-Slope Model. Kinase selectivity profiling is performed by using the KinaseProfiler service.
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Ensayo celular: [1]
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Cell lines
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Murine ID8 ovarian carcinoma cells
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Concentrations
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~1 μM
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Incubation Time
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6 days
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Method
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For soft agar assays, 48-well plates are coated with a 1:4 mix of 2% agar (EM Science) in 0.2 mL growth media (bottom layer). 5?04 cells are plated per well (in triplicate) in a mixture of 0.3% agar in 0.2 mL growth media (top layer). After agar solidification, 0.2 mL growth media is added containing DMSO or PND-1186 (final concentration for 0.6 mL). In separate experiments, PND-1186 is added after 4 days. After 10 days, colonies are imaged in phase contrast, enumerated by counting 9 fields (3 fields per well), and total area determined using Image J. For all analyses, experimental points are performed in triplicate and repeated at least two times.
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Estudio con animales: [1]
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Animal Models
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Mice bearing ID8 tumors or 4T1 tumors
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Formulation
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polyethylene glycol 400 (PEG400) in PBS (1:1) for s.c.; 5% sucrose for p.o.
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Dosages
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100 mg/kg every 12 hours for s.c.; 0.5 mg/mL for p.o.
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Administration
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s.c. or p.o.
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Conversión de diferentes modelos de animales basados en BSA (valor basado en datos del Borrador de Directrices de la 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 ejemplo, para modificar la dosis de resveratrol utilizada para un ratón (22,4 mg / kg) a una dosis basada en el BSA para una rata, multiplique 22,4 mg / kg por el factor Km para un ratón y luego divida por el factor Km para una rata. Este cálculo da como resultado una dosis equivalente para ratas de 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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Información química
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Molecular Weight (MW)
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501.5
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Formula
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C25H26F3N5O3
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CAS No.
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1061353-68-1
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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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SR-2156
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Chemical Name
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2-(2-(2-methoxy-4-morpholinophenylamino)-5-(trifluoromethyl)pyridin-4-ylamino)-N-methylbenzamide
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Calculadora de molaridad Calculadora de dilución Calculadora de peso molecular
Grupos de Producto : Angiogénesis > Inhibidor de FAK
