Torin 1 是一种有效的 mTOR 抑制剂，IC₅₀ 为 3 nM。Torin 1 抑制 mTORC1/2 复合物，IC₅₀ 值在 2 和 10 nM 之间。

基本信息

产品描述

Torin 1是一种有效的mTORC1/2抑制剂，在无细胞试验中IC50为2 nM/10 nM；作用于mTOR比作用于PI3K选择性高1000倍。

靶点（IC50 & Targe）

DNA-PK,6.34nM

mTOR,4.32nM

mTORC1,2nM

mTORC2,10nM

p110γ,171nM

体外研究

Torin1在2和10 nM浓度下分别抑制mTORC1 和mTORC2底物的磷酸化。此外，Torin1对mTOR比对PI3K (EC50 = 1800 nM)的选择性高1000倍，比对450种其它蛋白激酶的结合选择性高100倍。[1] [2] Torin1通过耐rapamycin机制引起细胞周期阻滞，并且其不依赖于mTORC2。Torin1比rapamycin更完全地干扰mTORC1依赖表现型。Cap依赖性翻译需要mTORC1耐Rapamycin的功能。[1]在近期的一项研究中，据报道Torin1通过活化人内分泌细胞系BON 中MEK/ERK/c-Jun通路，能够增加神经降压素分泌和基因表达。[3]

体内研究

Torin1在20 mg/kg剂量下，在U87MG异种移植模型中是有效的，并且在肿瘤和外周组织中对mTOR下游效应蛋白表现出良好的药效学抑制作用。[2]

激酶实验

mTORC1和 mTORC2体外激酶试验:

为产生可溶性mTORC1，HEK-293T细胞系，稳定表达N端FLAG标记的Raptor，使用水泡性口炎病毒G-假型病毒MSCV逆转录酶病毒产生。对于mTORC2，产生相似的HeLa细胞，能够稳定表达N端FLAG-标记的Protor-1。两个复合物通过在50 mm HEPES，pH 7.4，10 mm 焦磷酸钠，10 mm β-甘油磷酸钠，100 mm NaCl，2 mm EDTA，0.3% CHAPS中裂解细胞纯化。细胞在4 °C下裂解30分钟，不溶解部分通过以13,000 rpm微量离心10分钟移除。上清液与FLAG-M2单克隆抗体-琼脂糖培育1小时，然后用裂解缓冲液洗涤3次，用包含终浓度0.5 M NaCl的裂解缓冲液洗涤1次。纯化的mTORC1用100 μg/mL 3×FLAG多肽在50 mm HEPES，pH 7.4，100 mm NaCl中洗脱。将洗出液等分收集，储存在-80 °C。纯化底物S6K1 和 Akt1。激酶试验在30 °C，由激酶缓冲液(25 mm HEPES，pH 7.4，50 mm KCl，10 mm MgCl2，500 μm ATP)和150 ng 灭活的S6K1或Akt1底物组成的20 μL终体积混合物中进行。反应通过加入80 μL样品缓冲液停止，并沸煮5分钟。随后，样品通过SDS-PAGE和免疫印迹法分析。

细胞实验

Cell lines: MEFs

Concentrations: ～250 nM

Incubation Time: 4天

Method: 细胞活性通过CellTiter-Glo发光法细胞活性测定评估。在第0天，细胞以500细胞每孔接种到96孔板，并生长过夜。第1天，细胞用适当的化合物处理，随后在第3-5天分析。对于分析，板在室温下培育60分钟，将50 μL CellTiter-Glo试剂加入每孔中，板在定轨摇床上混合12分钟。发光在标准平板光度计上定量。

(Only for Reference)

动物实验

Animal Models: U87MG 异种移植物模型

Formulation: Torin1粉末首先以25 mg/mL在100% N-甲基-2-吡咯烷酮中溶解，随后以1:4 与无菌 50% PEG400稀释到5 mg/mL的终浓度

Dosages: 20 mg/kg

Administration: 腹腔注射给药，每天一次

(Only for Reference)

参考文献

[1] Thoreen CC, et al, J Biol Chem, 2009, 284(12), 8023-8032.

[2] Liu Q, et al, J Med Chem, 2010, 53(19), 7146-7155.

[3] Li J, et al, Am J Physiol Cell Physiol, 2011, 301(1), C213-C226.

[4] Dowling RJ, et al, Science, 2010, 328(5982), 1172-1176.

产品说明

一般描述

A cell-permeable pyridinonequinoline derivative that acts as a highly potent, ATP-competitive inhibibitor against mTOR and DNA-PK (IC₅₀ = 4.32 and 6.34 nM, respectively, in cell-free kinase assays), inhibiting class I (IC₅₀ =171, 250, and 564 nM, respectively, against P110γ, P110α/P85α, and P110δ/P85α, respectively), class II (IC₅₀ =176 and 564 nM, respectively, against C2α, and C2β, respectively), and class III (IC₅₀ = 533 nM against hVPS34) PI 3-K only at much higher concentrations. Effectively inhibits mTORC1-mediated S6K1 Thr389 phosphorylation in MEF cultures (IC₅₀ = 2 nM) in vitro, as well as mTORC2-mediated Akt Ser473 and mTORC1-dependent S6 Ser235/236 phosphorylations in murine lung and liver in vivo (up to 6 h after single i.p. dose at 20 mg/kg). Despite its poor in vivo stability (T_1/2 = 4.52 h in mice; 10 mg/kg i.p.), Torin1 is reported to completely suppress U87MG-derived tumor expansion in mice when administered via daily i.p. at a high dosage of 200 mg/kg. Also reported to effectively inhibit cellular mTORC1 functionalities known to be rapamycin-resistant (Cat. Nos. 553210, 553211, and 553212).
A cell-permeable pyridinonequinoline derivative that acts as a highly potent, ATP-competitive inhibibitor against mTOR and DNA-PK (IC₅₀ = 4.32 and 6.34 nM, respectively), while inhibiting PI 3-K only at much higher concentrations (IC₅₀ =171 to 533 nM). Effectively inhibits mTORC1-mediated S6K1 phosphorylation in MEF (IC₅₀ = 2 nM) in vitro, as well as mTORC2-mediated Akt and mTORC1-dependent S6 phosphorylations in murine lung and liver in vivo (up to 6 h post single 20 mg/kg i.p. dose). Despite its poor in vivo stability (T_1/2 = 4.52 h in mice; 10 mg/kg i.p.), Torin1 is reported to completely suppress U87MG-derived tumor expansion in mice when administered via daily i.p. at a high dosage of 200 mg/kg. Also reported to effectively inhibit cellular mTORC1 functionalities known to be rapamycin-resistant (Cat. Nos. 553210, 553211, and 553212).

包装

10 mg in Glass bottle
Packaged under inert gas

生化/生理作用

Cell permeable: yes
Primary Target
MTOR
Reversible: yes

警告

Toxicity: Standard Handling (A)

重悬

Following reconstitution, aliquot and freeze (-20&degC). Stock solutions are stable for up to 3 months at -20°C.

其他说明

Liu, Q., et al. 2010. J Med Chem.53, 7146.
Thoreen, C., et al. 2009. J Biol Chem.284, 8023.

法律信息

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

基本信息

产品性质

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