.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; relleno: 5px 0px 0px 5px;} \ n Peso molecular: \ n 517.06 Ispinesib (SB-715992, CK0238273) es un inhibidor potente, específico y reversible de la proteína del huso de kinesina (KSP) con una aplicación de Ki de 1,7 nM, sin inhibición de CENP -E, RabK6, MCAK, MKLP1, KHC o Kif1A. Fase 2. \ n Actividad biológica Ispinesib es un inhibidor potente, alostérico, reversible y específico de KSP, que cambia la propiedad de unión de KSP a los microtúbulos y altera su movimiento al inhibir la liberación de ADP sin alterar la liberación del complejo KSP-ADP de el microtúbulo. Ispinesib muestra una potente actividad citotóxica en un panel de líneas de células tumorales, que incluyen Colo205, Colo201, HT-29, M5076, Madison-109 y MX-1, con IC50 de 1,2 nM a 9,5 nM. En las células de cáncer de próstata PC-3, Ispinesib (15 nM y 30 nM) bloquea la proliferación celular e induce la apoptosis regulando los niveles de expresión de genes que controlan la apoptosis, la proliferación celular, el ciclo celular y la señalización celular, como EGFR, p27, p15 e IL-11. En un panel de 53 líneas de células mamarias, Ispinesib (7,4 nM-600 nM) demuestra una amplia actividad inhibidora. En las células BT-474 y MDA-MB-468, Ispinesib (150 nM) induce apoptosis, como lo revela una mayor proporción de células apoptóticas, menor nivel antiapoptótico de Bcl-XL y mayores niveles proapoptóticos de Bax y Bid. Ispinesib (4.5 mg / kg-15 mg / kg) exhibe efectos inhibidores contra las células Colo205, Colo201, HT-29, pero no contra las células MX-1, en modelos de xenoinjerto de ratón. SB-715992 (6 mg / kg – 10 mg / kg) también inhibe los tumores sólidos murinos, incluido el carcinoma pulmonar Madison 109, el sarcoma M5076 y las leucemias L1210 y P388. En modelos de xenoinjerto de ratones de células de cáncer de mama MCF-7, HCC1954, MDA-MB-468 y KPL4, Ispinesib (8 mg / kg-10 mg / kg) inhibe el crecimiento tumoral. Protocolo (solo como referencia) Ensayo de quinasa: [1]
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Steady-State Kinetic Analysis of Human KSP ATPase Activity and Inhibition by Ispinesib
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Kinesin specificity analysis is carried out using a pyruvate kinase-lactate dehydrogenase detection system that couples the production of ADP to oxidation of NADH. Absorbance changes are monitored at 340 nm. Steady-state studies using nanomolar concentrations of KSP are performed using a sensitive fluorescence-based assay utilizing a pyruvate kinase, pyruvate oxidase, and horseradish peroxidase (HRP) coupled detection system that couples the generation of ADP to oxidation of Amplex Red to fluorescent resorufin. Generation of resorufin is monitored by fluorescence (λexcitation = 520 nm and λemission = 580 nm). Steady-state biochemical experiments are performed in PEM25 buffer [25 mM Pipes-K+ (pH 6.8), 2 mM MgCl2, 1 mM EGTA] supplemented with 10 µM paclitaxel for experiments involving microtubules. The IC50 for steady-state inhibition is determined at 500 µM ATP, 5 µM Microtubules, and 1 nM KSP in PEM25 buffer. Ki app (apparent inhibitor dissociation constant) values of Ispinesib are extracted from the dose-response curves, with explicit correction for enzyme concentration by using the Morrison equation.
Inhibitor modality (e.g., competitive, noncompetitive, uncompetitive, or mixed) under steady-state conditions is determined by measuring the effect of inhibitor concentration on initial velocity as a function of substrate concentrations. Data are fit using equations in GraFit to velocity equations for the various modes of inhibition.
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Ensayo celular: [4]
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Cell lines
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Breast cancer cells, including MCF-7, HCC1954, MDA-MB-468, and KPL4
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Concentrations
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0.085 nM–33 µM
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Incubation Time
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72 hours
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Method
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Cells are plated in log phase of growth in 96-well plates and treated with Ispinesib for 72 hours. Then, cell growth is measured using CellTiter-Glo, and luminescence is detected using BioTek FLx800. Data are analyzed and the IC50 value, defined as the drug concentration that results in 50% growth inhibition relative to control, is calculated.
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Estudio con animales: [4]
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Animal Models
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Nude (nu/nu) mice models of MCF7, KPL4, and HCC1954 cells; severe combined immunodeficient (SCID) mice model of MDA-MB-468 cells;
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Formulation
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Dissolved in 10% ethanol, 10% cremophor, and 80% D5W (dextrose 5%)
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Dosages
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10 mg/kg for nude mice or 8 mg/kg for SCID mice
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Administration
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Intraperitoneal injection on a q4d× schedule (3 doses, every 4 day
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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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517.06
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Formula
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C30H33ClN4O2
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CAS No.
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336113-53-2
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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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CK0238273
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Solubility (25°C) *
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In vitro
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DMSO
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103 mg/mL
(199.2 mM)
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Water
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<1 mg/mL
(
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Ethanol
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103 mg/mL
(199.2 mM)
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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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(R)-N-(3-aminopropyl)-N-(1-(3-benzyl-7-chloro-4-oxo-3,4-dihydroquinazolin-2-yl)-2-methylpropyl)-4-methylbenzamide
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Calculadora de molaridad Calculadora de dilución Calculadora de peso molecular
Grupos de Producto : Señalización citoesquelética > Inhibidor de kinesina
