Powder: -20°C for 3 years | In solvent: -80°C for 1 year
COH000 是不可逆的泛素样激活酶1 共价变构抑制剂,体外测得对 SUMO 化修饰的IC50=0.2 μM。
规格 | 价格/CNY | 货期 | 数量 | |
---|---|---|---|---|
1 mg | ¥ 1,860 | 现货 | ||
2 mg | ¥ 2,810 | 现货 | ||
5 mg | ¥ 4,850 | 现货 | ||
10 mg | ¥ 6,590 | 现货 | ||
25 mg | ¥ 9,880 | 现货 | ||
50 mg | ¥ 13,300 | 现货 | ||
100 mg | ¥ 17,900 | 现货 | ||
1 mL * 10 mM (in DMSO) | ¥ 4,930 | 现货 |
产品描述 | COH000 is a covalent and irreversible inhibitor of small ubiquitin-like modifier (SUMO)-activating enzyme and inhibited SUMOylation (IC50: ~ 0.2 μM in vitro). |
靶点活性 | SUMO activating enzyme:0.2 μM (cell free) |
体外活性 | COH000 inhibited SUMOylation with an average IC50 of approximately 0.2 μM in vitro but did not inhibit ubiquitylation in a Ubc13-mediated poly-ubiquitylation assay tested under the same condition at concentrations up to 100 μM. COH000 treatment induced apoptosis in HCT-116 cells. The level of apoptosis was reduced when SAE2 was overexpressed by transfection and was increased when SAE2 was knocked down in HCT116 cells. |
体内活性 | 当肿瘤变得可触及时,携带肿瘤的Es1e/SCID小鼠接受了COH000或对照剂的治疗,为期14天。COH000显著抑制了肿瘤生长,并且显著降低了肿瘤组织中SAE2的水平。 |
激酶实验 | Briefly, SAE, Ubc9, GST-SUMO and His6-RanGap-1 proteins were expressed and purified as described previously.Assay buffer contained 50 mMTris-HCl pH 7.4, 0.3 mM DTT, 10 mM MgCl2, and 0.005% Tween-20. The assays were conducted using 1536-well, white plates. 2 μL of Mixture 1, containing 12.5 nM SAE and 100 nM His6-RanGap-1 in the assay buffer was mixed with 70 nl of 2 mM compounds dissolved in DMSO. Then 2 μL of Mixture 2, containing 20 mM ATP, 12.5 nM E2 and 30 nM GST-SUMO in assay buffer, was added. After incubation for 90 min at room temperature, 1 μl of pre-mixed Ni acceptor beads and Glutathione Donor beads, at 10 μg/ml each, was added. After incubation for 60 min at room temperature, readings were made on a BMG LabtechPheraStar in an AlphaScreen mode (Ex: 680 nm; Em: 570 nm). |
细胞实验 | Cell proliferation was measured using a CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS-based) after COH000 or its analog (54 or 55) treatment at the indicated concentrations and the time points. Briefly, cells were incubated with 20 mL of CellTiter 96 AQueous reagentafter the treatment and incubated at 37C until color development. Absorbance measurements were performed using a SpectraMax M5 reader. For assays measuring anti-proliferation effects, all values were normalized to the vehicle treatment. All values are represented graphically as mean ± standard deviation (STDEV) from three independent samples (n=3). |
动物实验 | Mice were housed in a controlled environment (12-h light/12-h dark cycle) with access to waterand fed a standard diet generally from 6 to 8 weeks of age. Body weight was measured weekly and food intake was monitored. For colorectal cancer xenograft with COH000 treatment, HCT116 cells were s.c. injected into a plasma esterase-deficient SCID mouse strain (Es1e/SCID), male, 8-10 weeks. When the tumors became palpable, COH000 was administered subcutaneously peritumoral injection once a day at a dose of 10 mg per kilogram of body weight. Mice in the control group received equal volumes of vehicle (5% DMSO, 30% solutol in PBS). Tumor volume was measured until the endpoint was reached. Mice were euthanized using CO2 inhalation and tumors were excised. |
分子量 | 419.47 |
分子式 | C25H25NO5 |
CAS No. | 1534358-79-6 |
Powder: -20°C for 3 years | In solvent: -80°C for 1 year
H2O: < 0.1 mg/mL (insoluble)
DMSO: 35.71 mg/mL (85.13 mM), Sonication is recommended.
可选溶剂 | 浓度 体积 质量 | 1 mg | 5 mg | 10 mg | 25 mg |
DMSO | 1 mM | 2.384 mL | 11.9198 mL | 23.8396 mL | 59.599 mL |
5 mM | 0.4768 mL | 2.384 mL | 4.7679 mL | 11.9198 mL | |
10 mM | 0.2384 mL | 1.192 mL | 2.384 mL | 5.9599 mL | |
20 mM | 0.1192 mL | 0.596 mL | 1.192 mL | 2.98 mL | |
50 mM | 0.0477 mL | 0.2384 mL | 0.4768 mL | 1.192 mL |
对于不同动物的给药剂量换算,您也可以参考 更多...
请在以下方框中输入您的动物实验信息后点击计算,可以得到母液配置方法和体内配方的制备方法: 比如您的给药剂量是10 mg/kg,每只动物体重20 g,给药体积100 μL,一共给药动物10 只,您使用的配方为5% DMSO+30% PEG300+5% Tween 80+60% ddH2O。那么您的工作液浓度为2 mg/mL。
母液配置方法:2 mg 药物溶于 50 μL DMSO (母液浓度为 40 mg/mL), 如您需要配置的浓度超过该产品的溶解度,请先与我们联系。
体内配方的制备方法:取 50 μL DMSO 主液,加入 300 μL PEG300, 混匀澄清,再加 50 μL Tween 80,混匀澄清,再加 600 μL ddH2O, 混匀澄清。
您可能有的问题的答案可以在抑制剂处理说明中找到,包括如何准备库存溶液,如何存储产品,以及基于细胞的分析和动物实验需要特别注意的问题。
COH000 1534358-79-6 Ubiquitination E1/E2/E3 Enzyme Ubiquitin conjugating enzyme E1 activating enzyme E2 conjugating enzyme Ubiquitin ligase Ubiquitin activating enzyme COH-000 COH 000 Inhibitor E3 ligating enzyme inhibit inhibitor