首页Cytoskeletal SignalingAktMK-2206 dihydrochloride
MK-2206 dihydrochloride
编号 T1952     别名: MK-2206 2HCl
CAS 1032350-13-2     分子式 C25H23Cl2N5O     分子量 480.39
靶点: Akt1; Akt2; Akt3;
MK-2206 is a highly specific inhibitor of Akt1/2/3 (IC50: 8/12/65 nM in cell-free assays) and no effect on the activities of 250 other protein kinases.
纯度:99.69%
规格 库存 单价 数量
2 mg 上海现货 290.00
5 mg 上海现货 586.00
10 mg 上海现货 878.00
25 mg 上海现货 1489.00
50 mg 上海现货 2062.00
100 mg 上海现货 3502.00
200 mg 上海现货 6310.00
1 mL * 10 mM (in DMSO) 上海现货 643.00
大包装特惠,请咨询
大包装特惠,请咨询

生物活性

产品描述

MK-2206 is a highly specific inhibitor of Akt1/2/3 (IC50: 8/12/65 nM in cell-free assays) and no effect on the activities of 250 other protein kinases.

靶点活性

Akt1,8 nM (cell free)

Akt2,12 nM (cell free)

Akt3,65 nM (cell free)

实验溶液

15% Captisol: 17 mg/mL

体外活性

MK-2206 by itself causes growth inhibition of the eight cell lines with an IC50 that ranges between 3.4 and 28.6 μmol/L. MK-2206 alone more potently inhibited the cell growth of Ras wild-type (WT) cell lines (A431, HCC827, and NCI-H292; IC50s of 5.5, 4.3, and 5.2 μmol/L, respectively) as compared with Ras-mutant cell lines (NCI-H358, NCI-H23, NCI-H1299, and Calu-6; IC50s of 13.5, 14.1, 27.0, and 28.6 μmol/L, respectively), with the exception of NCI-H460, which has a PIK3CA E545K mutation (IC50, 3.4 μmol/L) [2]. MK-2206 inhibited the four NPC cell line growths and reduced the sizes of the colonies in a dose-dependent manner. At 72 and 96 hours, the half maximal inhibitory concentration (IC50) values of MK-2206 in CNE-1, CNE-2, and HONE-1 cell lines were 3-5 μM, whereas, in SUNE-1, IC50 was less than 1 μM, and MK-2206 induced cell cycle arrest at the G1 phase [3].

体内活性

MK-2206 (120 mg/kg) was orally administered 2 hours after erlotinib (50 mg/kg), and tumors were isolated 14 hours after erlotinib administration. MK-2206 alone moderately inhibited phospho-Akt (53.1 ± 6.2%) and only slightly inhibited phospho-Erk (27.7 ± 5.9%). Erlotinib alone only moderately inhibited phospho-Erk, whereas it only slightly, if at all, inhibited phosphor-Akt in the tumor. Cotreatment potentiated the inhibition of phospho-Akt (79.9 ± 3.1%) and phosphor-Erk (53.5 ± 5.8%) [2]. Compared with the control group (30% Captisol diluent), both MK-2206 doses (480 mg/kg once a week and 240 mg/kg three times a week) can inhibit the growth of human CNE-2 xenografts in nude mice. In the two MK-2206 groups, the tumor weights were much lighter than the control group. Temporal body weight reduction was observed after receiving the MK-2206 treatment [3].

激酶实验

Akt kinases assay: Akt kinases are assayed by a GSK-derived biotinylated peptide substrate. The extent of peptide phosphorylation is determined by Homogeneous Time Resolved Fluorescence (HTRF) using a lanthanide chelate (Lance)-coupled monoclonal antibody specific for the phosphopeptide in combination with a streptavidin-linked allophycocyanin (SA-APC) fluorophore which will bind to the biotin moiety on the peptide. When the Lance and APC are in proximity, a non-radiative energy transfer takes place from the Lance to the APC, followed by emission of light from APC at 655 nm. Working Solution: 100X protease inhibitor cocktail (PIC): 1mg/mL benzamidine, 0.5 mg/mL pepstatin, 0.5 mg/mL leupeptin, 0.5 mg/mL aprotinin; 10X assay buffer: 500 mM HEPES, pH7.5, 1% PEG, 16.6 mM EDTA, 1 mM EGTA, 1% BSA, 20 mM 9-glycerol phosphate; Quench buffer 50 mM HEPES pH 7.3, 16.6 mM EDTA, 0.1% BSA, 0.1% Triton X-100, 0.17 nM labeled monoclonal antibody, 0.0067 mg/mL SA-APC; ATP/MgCl2 working solution: 1X Assay buffer, 1 mM DTT, 1X PIC, 5% glycerol, active Akt; Peptide working solution: 1X Assay buffer, 1 mM DTT, 1X PIC, 5% glycerol, 2 TM GSK biotinylated peptide. The reaction is assembled by adding 16 µL of ATP/MgCl2 working solution to the appropriate wells. MK-2206 or vehicle (1.0 µL) is added followed by 10 µL of peptide working solution. The reaction is started by adding 13 μL of the enzyme working solution and mixing. The reaction is allowed to proceed for 50 min and then stopped by the addition of 60 µL HTRF quench buffer. The stopped reactions are incubated at room temperature for at least 30 min and then read in the instrument.

细胞实验

Cells were seeded at a density of 2 to 3 × 103 per well in 96-well plates. Twenty-four hours after plating, varying concentrations of the drug, either as a single agent or in combination, were added to the wells. Cell proliferation was determined by using the CellTiter-Glo assay at 72 or 96 hours after dosing. The nature of the drug interaction was evaluated by using the combination index (CI) according to the method of Chou and Talalay. A commercial software package was obtained from Calcusyn. In combination with docetaxel, we tested three treatment sequences: (a) MK-2206 followed by docetaxel—cells were exposed to MK-2206 for 24 hours, and then after washout of MK-2206, cells were treated with docetaxel for an additional 72 hours; (b) docetaxel followed by MK-2206—cells were exposed to docetaxel for 24 hours, and then after washout of docetaxel, cells were treated with MK-2206 for an additional 72 hours; and (c) concurrent treatment—cells were exposed to both MK-2206 and docetaxel for 72 hours [2].

细胞系: A431, HCC827, NCI-H292, NCI-H358, NCI-H23, NCI-H1299, Calu-6 and NCI-H460 cells

动物实验

When the mean tumor size reached 0.13 cm3 for the SK-OV-3 or 0.2 cm3 for the NCI-H292, HCC70, PC-3, and NCI-H460 models, the mice were randomized into control and treatment groups with approximately equivalent ranges of tumor volume between groups (n = 5 animals per group). The following vehicles were used to dose the compounds: 30% Captisol (Cydex) for MK-2206; 0.5% methylcellulose + 0.1% Tween 80 for erlotinib; distilled water for lapatinib; 0.73% ethanol in saline for docetaxel; and saline for carboplatin and gemcitabine. The control group received vehicle only. Tumor volume was measured with calipers twice a week. Animal body weight and physical signs were monitored during the experiments. Tumor volume was calculated, taking length to be the longest diameter across the tumor and width to be the perpendicular diameter, by using the following formula: (length × width)2 × 0.5. Relative tumor volume was assessed by dividing the tumor volume on different observation days with the starting tumor volume. Statistical significance was evaluated by using the two-way repeated ANOVA test followed by Dunnett's test or an unpaired t-test [2].

动物模型:SK-OV-3, NCI-H292, HCC70, PC-3, and NCI-H460 models in CD1-nude mice

化学信息

分子量

480.39

分子式

C25H23Cl2N5O

CAS

1032350-13-2

溶解度

DMSO: 12 mg/mL (25 mM)

Ethanol: <1 mg/mL

( < 1 mg/ml refers to the product slightly soluble or insoluble )

储存条件

store at -80°C

备注

For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.

配制溶液

1 mg 5 mg 10 mg
1 mM 2.082 ml 10.408 ml 20.816 ml
5 mM 0.416 ml 2.082 ml 4.163 ml
10 mM 0.208 ml 1.041 ml 2.082 ml
50 mM 0.042 ml 0.208 ml 0.416 ml
技术支持
在订购、运输、储存和使用我们的产品的任何阶段,您遇到的任何问题,均可以通过拨打我们的热线电话400-998-7375,或者邮箱 info@tsbiochem.com,直接联系到我们。我们会在24小时内尽快联系您。
订购方式
联系陶素
电话:400-821-2233
传真:021-34692979
邮箱:info@tsbiochem.com


  • 关注公众号
    - 超值特惠
    - 客户福利
    - 业界动态
    - 实用攻略
陶素的所有产品和服务仅用于科学研究,我们不为任何其他用途提供产品和服务(也不为任何个人提供产品和服务)。