化合物COH000 T5199

Product Introduction
Bioactivity


英文名: COH000

描述: 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。

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).

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).

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.

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。

存储条件: Powder: -20°C for 3 years | In solvent: -80°C for 1 year

溶解度: H2O : < 0.1 mg/mL (insoluble)
DMSO : 35.71 mg/mL (85.13 mM)


关键字: Ubiquitin conjugating enzyme | E1 activating enzyme | COH000 | E1/E2/E3 Enzyme | E2 conjugating enzyme | Ubiquitin ligase | Ubiquitin activating enzyme | COH-000 | COH 000 | Inhibitor | E3 ligating enzyme | inhibit

相关产品: 2-D08 | Cbl-b-IN-6 | Cbl-b-IN-8 | Mezigdomide | Skp2 Inhibitor C1 | Siremadlin | Cbl-b-IN-9 | RO8994 | Pevonedistat | BI8626

相关库: HIF-1 Signaling Pathway Compound Library | Bioactive Compounds Library Max | Ubiquitination Compound Library | Inhibitor Library | Target-Focused Phenotypic Screening Library | NO PAINS Compound Library | Covalent Inhibitor Library | Bioactive Compound Library

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注：该产品未在中华人民共和国食品药品监督管理部门申请医疗器械注册和备案，不可用于临床诊断或治疗等相关用途