| 名称 | TAK-632 |
| 描述 | TAK-632 is a potent pan-Raf inhibitor. |
| 细胞实验 | The cells are proliferated in appropriate medium (vender recommended) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and antibiotics, in tissue culture dishes placed in a humidified incubator maintained at 37°C in an atmosphere of 5% CO2 and 95% air. The anti-proliferative activity of compound is determined by treating cell lines with the compound for 3 days, followed by measurement of viable cell number in the Cell Titer-Glo assay. The cells are seeded in a 96-multiwell plate at 1500 to 4000 cells per well in medium containing FBS and cells allowed to sit down overnight. After 18–20 h, compounds at various concentrations by serial dilution are added to the cells and were cultured for 3 days in chamber. After the treatment culture, cellular proliferation is determined by a Cell Titer-Glo Luminescent Cell Viability Assay. In brief, 100 bL/well of Cell Titer-Glo Substrate solution is added to each well and the cells were cultured for an additional 10 minutes (approximately). The chemi-luminescence value is measured using a Luminescence Counter 1420 ARVO MX Light. Concentration response curves are generated by calculating the decrease in chemi-luminescence values in compound-treated samples relative to the vehicle (DMSO) treated controls. (Only for Reference) |
| 激酶实验 | Kinase Profile Assay: Assays for serine/threonine kinases using radio labeled [γ-33P] ATP are performed in 96 well plates. BRAF and c-RAF are expressed as N-terminal FLAG-tagged protein using a baculovirus expression system. The reaction conditions are optimized for each kinase: BRAF (25 ng/well of enzyme, 1 μg/well of GST-MEK1(K96R), 0.1 μCi/well of [γ-32P] ATP, room temperature, 20 min reaction); c-RAF (25 ng/well of enzyme, 1 μg/well of GST-MEK1 (K96R), 0.1 μCi/well of [γ-32P] ATP, room temperature, 20 min reaction). Enzyme reactions are performed in 25 mM HEPES, pH 7.5, 10 mM magnesium acetate, 1 mM dithiothreitol and 0.5 μM ATP containing optimized concentration of enzyme, substrate and radiolabeled ATP as described above in a total volume of 50 μL. Prior to the kinase reaction, compound and enzyme are incubated for 5 min at reaction temperature as described above. The kinase reactions are initiated by adding ATP. After the reaction period as described above, the reactions are terminated by the addition of 10% (final concentration) trichloroacetic acid. The [γ-33P] or [γ-32P]-phosphorylated proteins are filtered in GFC filter plates with a Cell Harvester and then the plates are washed out with 3% phosphoric acid. The plates are dried, followed by the addition of 40 μL of MicroScint0. The radioactivity is counted by a TopCount scintillation counter. |
| 体外活性 | 在携带NRAS突变型黑素瘤的SK-MEL-2异种移植小鼠模型中,口服 TAK-632（60 or 120 mg/kg）抑制 MAPK 信号通路,抑制肿瘤的发展. |
| 体内活性 | 在A375（GI50=66 nM）和HMVII系（和GI50=200 nM）中,TAK-632能够抑制细胞增殖。其中,在黑色素瘤A375细胞系(BRAFV600E)中,TAK-632抑制MEK（IC50=2 nM）和ERK磷酸化（IC50=16 nM）。在人类黑色素瘤HMVII细胞系(NRASQ61K/BRAFG469V)中,TAK-632抑制pMEK（IC50=49 nM）和pERK（IC50=50 nM）。 |
| 存储条件 | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | DMSO : 93 mg/mL (167.71 mM), Sonication is recommended.
Ethanol : 2 mg/mL (3.61 mM), Sonication is recommended.
10% DMSO+90% Corn Oil : 3.3 mg/mL (5.95 mM), Sonication is recommended.
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 关键字 | TAK-632 | TAK632 | TAK 632 | Raf kinases | Raf | PDGFRβ | Inhibitor | inhibit | FGFR3 | C-Raf | B-Raf | AuroraKinase | Aurora Kinase | Aurora B |
| 相关产品 | Amlexanox | Lenvatinib | Dabrafenib | Formononetin | Lenvatinib mesylate | Ferulic Acid | Regorafenib | Pazopanib | Sorafenib | Regorafenib monohydrate | Sunitinib | Nintedanib esylate |
| 相关库 | 抑制剂库 | 血管生成库 | 抗癌活性化合物库 | 经典已知活性库 | 已知活性化合物库 | 激酶抑制剂库 | 抗衰老化合物库 | 临床前化合物库 | 膜蛋白靶向化合物库 | 免疫/炎症分子化合物库 | 疼痛相关化合物库 | 酪氨酸激酶分子库 |