Temsirolimus (CCI-779)

Pfizer辉瑞授权 目录号:S1044 别名: NSC 683864 中文名称:替西罗莫司

仅限科研使用

Temsirolimus (CCI-779, NSC 683864)是一种特定的mTOR抑制剂,在无细胞试验中IC50为1.76 μM。Temsirolimus 可诱导自噬和凋亡。

Temsirolimus (CCI-779) Chemical Structure

CAS: 162635-04-3

规格 价格 库存 购买数量
10mM (1mL in DMSO) RMB 1087.98 现货
RMB 953.84 现货
RMB 3016.49 现货
RMB 7122.99 现货
大包装 有超大折扣
大剂量询单有大折扣!

今日订购,明日送达 全国免运费 分装免费 分装成小规格产品(单支最小为1mg)
如1支10mg产品可分装为10支1mg产品

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客户使用Selleck生产的Temsirolimus (CCI-779)发表文献83篇:

产品安全说明书

mTOR抑制剂选择性比较

相关mTOR产品

生物活性

产品描述 Temsirolimus (CCI-779, NSC 683864)是一种特定的mTOR抑制剂,在无细胞试验中IC50为1.76 μM。Temsirolimus 可诱导自噬和凋亡。
靶点
mTOR [1]
(Cell-free assay)
1.76 μM
体外研究

Temsirolimus抑制核糖体蛋白S6磷酸化,作用于PTEN阳性DU145细胞比作用于PTEN阴性PC-3细胞更有效,且抑制细胞生长,和细胞的克隆生存,这种作用存在浓度依赖性。[5] Temsirolimus按100 ng/mL剂量处理原代人淋巴细胞性白血病(ALL) 细胞,显著抑制增殖,且诱导凋亡。[6]在 FKBP12存在时, Temsirolimus 有效抑制mTOR 激酶活性,IC50为1.76 μM, 与Rapamycin(IC50为1.74 μM)类似。Temsirolimus按纳摩尔浓度(10 nM 到<5 μM) 处理,通过FKBP12依赖机制,具有适度的选择性抗增殖活性,在低微摩尔浓度 (5-15 μM)时,可以完全抑制多种肿瘤细胞增殖,涉及FKBP12非依赖性的 mTOR信号阻断。Temsirolimus 按微摩尔而不是纳摩尔浓度 (20 μM)处理,显著降低全部蛋白质合成,也显著降低多聚核糖体的分解, 伴随着翻译延伸因子 eEF2和翻译起始因子eIF2A的磷酸化快速提高。[1]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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Assay
Methods Test Index PMID
Western blot p-mTOR / mTOR / p-S6 / S6 / p-4E-BP1 / 4E-BP1 p-rS6 / rS6 / p-AKT /AKT 23291985 22039466
Immunofluorescence NRF2 22848625
Growth inhibition assay Cell growth (HGC-3 cells) Cell viability 30866995 22039466
体内研究 Temsirolimus 按20 mg/kg 剂量腹腔注射,每周5天,显著延迟DAOY 移植瘤生长,与对照组相比,处理一周,延迟160%,处理两周,延迟240%。Temsirolimus按 100 mg/kg高剂量单独给药腹腔注射,一周内,诱导肿瘤体积衰退37%。 Temsirolimus 处理2两周,也延迟抗Rapamycin的U251移植瘤生长,延迟148%。[2]Temsirolimus 作用于患Huntington疾病的小鼠模型,抑制mTOR,提高四种不同行为任务的表现,且降低聚合形成。[3] Temsirolimus处理8226, OPM-2,和 U266移植瘤的皮下生长,诱导显著的抗癌反应,这种作用存在剂量依赖性,作用于8226和 OPM-2移植瘤,ED50分别为20 mg/kg 和2 mg/kg,与抑制增殖和血管生长,诱导凋亡,和肿瘤细胞尺寸降低相关。 [4] 每天按10 mg/kg剂量处理携带人ALL的NOD/SCID移植瘤模型,降低外周血膨胀和脾肿大。[6]

推荐的实验操作(此推荐来自于公开的文献所以Selleck并不保证其有效性)

激酶实验:[1]
  • 体外mTOR 催化活性检测:

    Flag标记的野生型人 mTOR (Flag-mTOR) DNA 结构短暂转染进HEK293 细胞。48小时后,进行蛋白提取和Flag-mTOR纯化。在体外,在不同浓度Temsirolimus存在而没 FKBP12 时,在96孔板上进行纯化的Flag-mTOR激酶实验,通过解离增强镧系荧光免疫分析(DELFIA),使用His6-S6K1 作为底物来测量。在激酶实验 buffer (10 mM Hepes pH 7.4, 50 mM NaCl, 50 mM β-甘油磷酸,10 mM MnCl2, 0.5 mM DTT, 0.25 μM微囊藻素 LR, 和 100 μg/mL BSA)上第一次对酶进行稀释。每孔中,12 μL 稀释的酶与0.5 μL Temsirolimus混合。加入含ATP 和His6-S6K 的12.5 μL 激酶buffer开始激酶反应,终体积为25 μL,含800 ng/mL FLAG-mTOR, 100 μM ATP, 和 1.25 μM His6-S6K。反应板在室温下温和震荡温育2小时,加入 25 μL 终止液(20 mM Hepes pH 7.4, 20 mM EDTA, 和 20 mM EGTA)终止反应。磷酸化的(Thr-389) His6-S6K 的DELFIA 检测在室温下使用单克隆抗P(T389)-p70S6K抗体进行,抗体使用铕-N1-ITC (Eu) (每种抗体10.4 Eu)标记。45 μL终止的激酶反应混合物转移到含55 μL PBS的MaxiSorp板上。His6-S6K 粘附2小时后,吸除每孔,使用PBS冲洗。加入含40 ng/mL Eu-P(T389)-S6K 抗体的 100 μL DELFIA buffer。轻轻搅拌,再使抗体结合持续进行1小时。吸除每孔,然后使用含0.05% Tween-20(PBST)的PBS冲洗4次。每孔加入100 μL of DELFIA增强溶液,然后在 PerkinElmer Victor 模板酶标仪上读数。

细胞实验:[1]
  • Cell lines: A549, H157, H460, H446, HCT116, HT29, SW480, DLD1, Caco2, LNCap, DU145, MDA468, MDA231, HEK293, 和PC3-MM2
  • Concentrations: 溶于DMSO,终浓度为~20 μM
  • Incubation Time: 72小时
  • Method: 使用不同浓度Temsirolimus处理细胞72小时。处理后,通过MTS染料转换,使用CellTiter AQ 检测试剂盒测定存活细胞密度。
动物实验:[2]
  • Animal Models: 皮下注射DAOY, 或U251细胞的雌性无胸腺裸鼠
  • Dosages: 20 mg/kg
  • Administration: 每天注射,每周5次

溶解度(25°C)

体外

体内

从左到右依次将纯溶剂加入产品,现配现用(数据来自Selleck实验检测而非文献):
2% DMSO+30% PEG 300 +2% Tween 80+ddH2O

7mg/mL

* 溶解度检测是由Selleck技术部门检测的,可能会和文献中提供的溶解度有所差异,这是由于生产工艺和批次不同产生的正常现象。请按照顺序依次加入各个纯溶剂。

化学数据

分子量 1030.29
化学式

C56H87NO16

CAS号 162635-04-3
储存条件 3年 -20°C 粉状
2年 -80°C 溶于溶剂

动物体内配方计算器(澄清溶液)

第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量)

mg/kg g μL

第二步:请输入动物体内配方组成(配方适用于不溶于水的药物;不同批次药物配方比例不同,请联系Selleck为您提供正确的澄清溶液配方)

% DMSO % % Tween 80 % ddH2O
%DMSO %

计算结果:

工作液浓度: mg/ml;

DMSO母液配制方法: mg 药物溶于μL DMSO溶液(母液浓度mg/mL,:如该浓度超过该批次药物DMSO溶解度,请先联系Selleck);

体内配方配制方法:μL DMSO母液,加入μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入μL ddH2O,混匀澄清。

体内配方配制方法:μL DMSO母液,加入μL Corn oil,混匀澄清。

注意:1. 首先保证母液是澄清的;
2.一定要按照顺序依次将溶剂加入,进行下一步操作之前必须保证上一步操作得到的是澄清的溶液,可采用涡旋、超声或水浴加热等物理方法助溶。

摩尔浓度计算器

质量 浓度 体积 分子量

临床试验信息

NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT01653067 Unknown status Drug: Rituximab Temsirolimus DHAP intravenous Diffuse Large B-Cell Lymphoma Mathias Witzens-Harig|Johannes Gutenberg University Mainz|Technische Universität München|Ludwig-Maximilians - University of Munich|University Hospital Ulm|University Hospital Erlangen|Charite University Berlin Germany|University Hospital Freiburg|Johann Wolfgang Goethe University Hospital|University Hospital Heidelberg September 2012 Phase 2
NCT02093598 Completed Drug: Temsirolimus Carcinoma Endometrioid|mTOR Protein MedSIR May 2012 Phase 2

(data from https://clinicaltrials.gov, updated on 2022-01-17)

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