GZD824 Димезилат 1421783-64-3

.cp_wz table {border-top: твердое тело 1px #ccc; border-left: твердое тело 1px #ccc; } .cp_wz таблица td {border-right: 1px solid #ccc; нижняя граница: сплошной 1px #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 Молекулярный вес: \ n 724,77 GZD824 Димезилат - новый пероральный биодоступный ингибитор Bcr-Abl для Bcr-Abl (WT) и Bcr-Abl (T315I) с IC50 0,34 нМ и 0,68 нМ, соответственно. \ n Биологическая активность GZD824 сильно подавляет рост клеток Ba / F3, экспрессирующих Bcr-Abl дикого типа, с IC50 1,0 нМ. В высокой степени согласуясь с биохимическим ингибированием киназ и сильным сродством связывания с белками, GZD824 также сильно ингибирует пролиферацию клеток Ba / F3, экспрессирующих мутант Bcr-AblT315I и 14 других мутантов Bcr-Abl, имеющих отношение к устойчивости. Кроме того, GZD824 эффективно подавляет активацию Bcr-Abl, а также нижестоящих Crk1 и STAT5 в зависимости от концентрации в клетках K562 CML. GZD824 в дозах 5 и 10 мг / кг / день дозозависимо ингибирует рост опухоли в ксенотрансплантате опухоли K562 и модели ксенотрансплантата Ku812 без смертности или значительной потери тела. Кроме того, GZD824 (20 мг / кг / день) подавляет рост опухоли у мышей, несущих аллогенные клетки Ba / F3, экспрессирующие Bcr-AblWT и Bcr-AblT315I. Протокол (только для справки) Анализ киназы: [1]

FRET-Based Z′-Lyte Assay Detecting Peptide Substrate Phosphorylation

The kinases ABL1, ABL1(E255K), ABL1 (G250E), ABL1(T315I), and ABL1(Y253F) are P3049, PV3864, PV3865, PV3866, and PV3863 are full-length human recombinant protein expressed in insect cells and histidine-tagged. Inhibition activities of inhibitors against Abl1 and its mutants are performed in 384-well plates using the FRET-based Z′-Lyte assay system. Briefly, the kinase substrate is diluted into 5 μL of kinase reaction buffer; and the kinase is added. Compounds (10 nL) with indicated concentrations are then delivered to the reaction by using Echo liquid handler, and the mixture is incubated for 30 min at room temperature. Then 5 μL of 2X ATP solution is added to initiate the reaction, and the mixture is further incubated for 2 h at room temperature. The resulting reactions contains 10 μM (for wild-type Abl1, and mutants Y253F, Q252H, M351T, and H396P) or 5 μM (for mutants E255K, G250E, T315I) of ATP, 2 μM of Tyr2 Peptide substrate in 50 mM HEPES (PH 7.5), 0.01% BRIJ-35, 10 mM MgCl2, 1 mM EGTA, 0.0247 μg/mL Abl1, and inhibitors as appropriate. Then, 5 μL of development reagent is added, and the mixture is incubated for 2 h at room temperature before 5 μL of stop solution is added. Fluorescence signal ratio of 445 nm (Coumarin)/520 nm (fluorescin) is examined on an EnVision Multilabel Reader. The data are analyzed using Graphpad Prism5. The data are the mean value of three experiments.

Клеточный анализ: [1]

Cell lines	Ba/F3 cells expressing wildtype and mutant Bcr-Abl
Concentrations	0-50 μM
Incubation Time	72 hours
Method	Cells in the logarithmic phase are plated in 96-well culture dishes. Twenty-four hours later, cells are treated with the corresponding compounds or vehicle control at the indicated concentration for 72 h. CCK-8 is added into the 96-well plates (10 μL/well) and incubated with the cells for 3 h. OD450 and OD650 are determined by a microplate reader. Absorbance rate (A) for each well is calculated as OD450 – OD650. The cell viability rate for each well is calculated as V% = (As – Ac)/(Ab – Ac) × 100%, and the data were further analyzed using Graphpad Prism5. The data are the mean value of the three experiments. As is the absorbance rate of the test compound well, Ac is the absorbance rate of the well without either cell or test compound, and Ab is the absorbance rate of the well with cell and vehicle control.

Исследование на животных: [1]

Animal Models	Mice xenograft or allograft tumor models using K562 and transformed Ba/F3 cells expressing native Bcr-Abl or Bcr-Abl mutants.
Formulation	1% DMSO, 22.5% Cremophor, 7.5% ethanol, and 69% normal saline
Dosages	~20 mg/kg
Administration	oral gavage
Solubility	Saline, 20 mg/mL
* 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.

Конверсия различных модельных животных на основе BSA (значение на основе данных из проекта рекомендаций FDA)

Species	Baboon	Dog	Monkey	Rabbit	Guinea pig	Rat	Hamster	Mouse
Weight (kg)	12	10	3	1.8	0.4	0.15	0.08	0.02
Body Surface Area (m2)	0.6	0.5	0.24	0.15	0.05	0.025	0.02	0.007
Km factor	20	20	12	12	8	6	5	3

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

Например, чтобы изменить дозу ресвератрола, используемую для мыши (22,4 мг / кг), на дозу, основанную на BSA для крысы, умножьте 22,4 мг / кг на коэффициент Km для мыши, а затем разделите на коэффициент Km для крыса. Этот расчет дает эквивалентную дозу ресвератрола для крыс 11,2 мг / кг.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×	mouse Km(3)	= 11.2 mg/kg
	rat Km(6)

\ n Химическая информация

Molecular Weight (MW)	724.77
Formula	C29H27F3N6O.2CH4O3S
CAS No.	1421783-64-3

Storage	3 years -20℃Powder
	6 months-80℃in solvent (DMSO, water, etc.)
Synonyms

Solubility (25°C) *	In vitro	DMSO	100 mg/mL (137.97 mM)
		Water	100 mg/mL (137.97 mM)
		Ethanol	<1 mg/mL (
	In vivo	Saline	20 mg/mL
* <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.

Chemical Name	Benzamide, 4-methyl-N-[4-[(4-methyl-1-piperazinyl)methyl]-3-(trifluoromethyl)phenyl]-3-[2-(1H-pyrazolo[3,4-b]pyridin-5-yl)ethynyl]-, methanesulfonate (1:2)

Калькулятор молярности Калькулятор разведения Калькулятор молекулярной массы

Группа Продуктов : Ангиогенез > Bcr-Abl ингибитор