| 中文名称: | b-AP15 | ||||
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| 英文名称: | b-AP15 | ||||
| 别名: | (3E,5E)-3,5-二[(4-硝基苯基)亚甲基]-1-(1-氧代-2-丙烯-1-基)-4-哌啶酮;b-AP15 抑制剂 NSC 687852 | ||||
| CAS No: | 1009817-63-3 | 分子式: | C22H17N3O6 | 分子量: | 419.39 |
| CAS No: | 1009817-63-3 | ||||
| 分子式: | C22H17N3O6 | ||||
| 分子量: | 419.39 | ||||
| EINEC: | 687852 | ||||
| EINEC: | 687852 | ||||
![9-氯-5,6,7,8-四氢-N-[3-[甲基(苄基)氨基]丙基]-2-吖啶甲酰胺](http:///www.jm-bio.com/content/xtheme/beijingjinming/images/noimage.jpg)
基本信息
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产品编号： |
A11276 |
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产品名称： |
b-AP15 |
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CAS: |
储存条件 |
粉末 |
-20℃ |
四年 |
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分子式： |
溶于液体 |
-80℃ |
两年 |
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分子量 |
419.39 |
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化学名： |
(3E,5E)-3,5-bis[(4-nitrophenyl)methylidene]-1-prop-2-enoylpiperidin-4-one |
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Solubility (25°C)： |
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体外:
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DMSO |
48mg/mL warmed with 50ºC water bath(114.45mM) |
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Ethanol |
Insoluble |
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Water |
Insoluble |
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体内(现配现用): |
1.请依序添加每种溶剂：10% DMSO→40% PEG300→5% Tween-80→45% saline，Solubility: 2.08mg/mL (4.96mM); Suspended solution; Need ultrasonic |
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此⽅案可获得2.08mg/mL(4.96mM)的均匀悬浊液，悬浊液可⽤于⼝服和腹腔注射。以1mL⼯作液为例，取100μL20.8mg/mL的澄清DMSO储备液加到400μLPEG300中，混合均匀；向上述体系中加⼊50μLTween-80，混合均匀；然后继续加⼊450μL⽣理盐⽔定容⾄1mL。 |
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＜1mg/ml表示微溶或不溶。 |
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普西唐提供的所有化合物浓度为内部测试所得，实际溶液度可能与公布值有所偏差，属于正常的批间细微差异现象。 |
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请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 |
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制备储备液
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浓度
溶液体积 质量 |
1mg |
5mg |
10mg |
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1mM |
2.3844mL |
11.9221mL |
23.8442mL |
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5mM |
0.4769mL |
2.3844mL |
4.7688mL |
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10mM |
0.2384mL |
1.1922mL |
2.3844mL |
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50mM |
0.0477mL |
0.2384mL |
0.4769mL |
生物活性
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产品描述 |
特异性抑制去泛素化酶 (deubiquitinating) UCHL5 和 Usp14活性。 |
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靶点 |
USP14 |
UCHL5 2.1μM |
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体外研究 |
b-AP15抑制两种19S调节颗粒相关的去泛素化酶，泛素C-末端水解酶5（UCHL5）和泛素特异性肽酶14（USP14），导致多聚泛素的积累。b-AP15导致UbG76V-YFP受体累积， IC50为0.8μM, 这种作用存在剂量依赖性，说明损伤的蛋白酶体降解受损。b-AP15 (1μM)作用于人类结肠癌HCT-116细胞，导致多聚泛素化蛋白快速累积。b-AP15(2.2μM)作用于HCT-116细胞，提高细胞周期蛋白依赖性激酶CDKN1A和CDKNIB，及肿瘤抑制基因TP53的数量，这种作用存在剂量依赖性，但不改变鸟氨酸脱羧酶1（ODC1）的数量。b-AP15(1μM)作用于HCT-116细胞，导致细胞周期停滞在G2/M期，相应地，导致细胞周期抑制剂累积。b-AP15处理提高亚二倍体细胞的数量，并相应地增加，细胞凋亡标记的数量，包括活化的caspase-3，caspase裂解的多聚ADP核糖聚合酶（PARP）和细胞角蛋白-18（CK18）。与永生化上皮细胞（hTERT-RPE1）或外周血单核细胞相比，b-AP15对HCT-116细胞毒性更大。b-AP15抑制去泛素活性，使用各种底物，包括Ub-AMC, Ub-GFP22, 泛素化p53-结合蛋白同源物(HDM2), 及K48-和K63-连接的泛素四聚体链。b-AP15是UPS抑制剂，通过诱导cathepsin-D依赖性的溶酶体凋亡途径而诱导细胞死亡。b-AP15引出特征性的UPS缺陷，包括泛素偶联物，细胞周期抑制剂，如p21，p27，和肿瘤抑制基因p53的积累。b-AP15抑制半胱氨酸DUBs,的去泛素化酶活性，USP14稍微比UCHL5敏感。b-AP15作用于过量表达抗凋亡Bcl-2蛋白的细胞和缺乏p53基因的细胞，诱导细胞凋亡。b-AP15 (1μM) 抑制ATP诱导的IL-1β从LPS诱导的腹腔巨噬细胞中释放。b-AP15(1μM)作用于THP-1细胞，降低Nigericin处理诱导的细胞死亡水平。b-AP15(1μM)处理LPS诱导的THP-1细胞，显著降低Nigericin处理后形成的ASC斑点数量。 |
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体内研究 |
b-AP15 (5mg/kg)处理携带鳞状细胞癌移植瘤的重症联合免疫缺陷（SCID）小鼠，具有显著的抗肿瘤活性。b-AP15(5mg/kg)处理携带HCT-116结肠癌移植瘤的小鼠，显著延缓肿瘤发病。 |
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特征 |
b-AP15不是一般的去泛素化酶抑制剂，对重组和胞浆非蛋白酶体半胱氨酸去泛素化酶具有最低抑制效果。 |
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推荐实验方法(仅供参考)
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Kinase Assay |
For deubiquitinase inhibition assays, 19S regulatory particle (5nM), 26S (5nM) UCH-L1 (5nM), UCH-L3 (0.3nM), USP2CD (5nM) USP7CD (5nM) USP8CD (5nM) or BAP1 (5nM) is incubated with DMSO or b-AP15 and monitored the cleavage of ubiquitin-AMC (1,000nM) using a Wallac VICTOR Multilabel counter or a Tecan Infinite M1000 equipped with 380nM excitation and 460nM emission filters |
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Cell Assay |
Cell viability is monitored by either the fluorometric microculture cytotoxicity assay or the MTT assay. For the MTT assay, cells are seeded into 96-well flat-bottomed plates overnight and exposed to drugs, using DMSO as the control. At the end of incubations, 10 µl of a stock solution of 5mg/mL MTT is added into each well, and the plates are incubated 4 hours at 37°C. Formazan crystals are dissolved with 100 µL 10% SDS/10 mM HCl solution overnight at 37°C. Absorbance is measured using an enzyme-linked immunosorbent assay (ELISA) plate reader at 590nM |
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Animal Administration |
Mice For the squamous carcinoma model, 1×106 FaDu cells are subcutaneously injected into the right rear flank of female SCID mice. Tumor growth is measured by the formula length×width2×0.44. When tumors have grown to a size of approximately 200 mm3 (defined as day 0), mice are randomized to receive either vehicle (n=10) or b-AP15 (n=15) at 5mg per kg of body weight by daily subcutaneous injection. For the colon carcinoma model, we subcutaneously injected 2.5×106 HCT-116 colon carcinoma cells overexpressing Bcl2 into the right flank of female nude mice. We treated mice with 5mg of b-AP15 per kg of body weight by intraperitoneal injection. For the lung carcinoma model, we subcutaneously injected 2×105 LLC cells into the right rear flank of female C57/B6 mice. When tumors had grown to a size of approximately 50 mm3 (defined as day 0), we randomized mice to receive either vehicle (n=4) or b-AP15 (n=4) at 5mg per kg of body weight intraperitoneally, with a treatment cycle consisting of 2 d of treatment followed by 2 d of rest (2 d on, 2 d off) for 2 weeks. |
本计算器可帮助您计算出特定溶液中溶质的质量、溶液浓度和体积之间的关系，公式为：
质量 (g) = 浓度 (mol/L) x 体积 (L) x 分子量 (g/mol)
摩尔浓度计算公式
用本工具协助配置特定浓度的溶液，使用的计算公式为：
开始浓度 x 开始体积 = 最终浓度 x 最终体积
稀释公式
稀释公式一般简略地表示为：C1V1 = C2V2 ( 输入 输出 )
