KU-0063794

KU-0063794是一种有效的,高度特异性的,作用于mTORC1mTORC2的双重mTOR抑制剂,在无细胞试验中IC50约为~10 nM;对PI3Ks没有作用。

KU-0063794 Chemical Structure

KU-0063794 Chemical Structure

CAS: 938440-64-3

规格 价格 库存 购买数量
10mM (1mL in DMSO) 1464.74 现货
5mg 727.74 现货
10mg 1970 现货
50mg 3857.09 现货
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KU-0063794相关产品

相关信号通路图

细胞实验数据示例

细胞系 实验类型 给药浓度 孵育时间 活性描述 文献信息
H1650 Function Assay 7.61 nM 72 h inhibits phosphorylation of p70S6K 23874880
PC9GR Function Assay 6.21 nM 72 h inhibits phosphorylation of p70S6K 23874880
PC9 Function Assay 10.15 nM 72 h inhibits phosphorylation of p70S6K 23874880
H1975 Function Assay 11.15 nM 72 h inhibits mTOR phosphorylation status  23874880
H1650 Function Assay 7.61 nM 72 h inhibits mTOR phosphorylation status  23874880
PC9GR Function Assay 6.21 nM 72 h inhibits mTOR phosphorylation status  23874880
PC9 Function Assay 10.15 nM 72 h inhibits mTOR phosphorylation status  23874880
HepG3  Function Assay 0.1–50 μM 48 h induces cell autophagy in a dose dependent manner 26278819
HepG2  Function Assay 0.1–50 μM 24 h induces p62 downregulation, Beclin-1 expression and LC3B-I to LC3B-II conversion in a dose dependent manner 26278819
HepG2  Function Assay 5/10 μM 24 h downregulates the levels HIF1α and cyclin D1  26278819
HepG2  Function Assay 5/10 μM 24 h dramatically inhibits phosphorylation of AKT at Ser-473 26278819
HepG2  Apoptosis Assay 0.1–50 μM 48 h induces apoptosis in a dose dependent manner 26278819
HepG2  Colony Formation Assay 1–50 μM 10 d decreases the number of viable HepG2 colonies significantly 26278819
HepG2  Cell Viability Assay 0.1–50 μM 72 h decreases cell viability in a dose dependent manner 26278819
H1975 Function Assay 11.15 nM 72 h inhibits phosphorylation of p70S6K 23874880
LNCaP Cell Viability Assay 0-10 μM 24 h  decreases cell viability in a dose dependent manner 23840605
PC-3 Cell Viability Assay 0-10 μM 24 h  decreases cell viability in a dose dependent manner 23840605
MDA-MB-468  Cell Viability Assay 0-10 μM 24 h  decreases cell viability in a dose dependent manner 23840605
LNCaP Function Assay 200–800 nM 24 h  decreases the phosphorylation level of p70S6K in a dose dependent manner 23840605
PC-3 Function Assay 200–800 nM 24 h  decreases the phosphorylation level of p70S6K in a dose dependent manner 23840605
MDA-MB-468  Function Assay 200–800 nM 24 h  decreases the phosphorylation level of p70S6K in a dose dependent manner 23840605
Caki-1  Function Assay 100-2000 nM 10-180 min inhibits both mTORC1 and mTORC2 as indicated by the decrease in phosphorylation of downstream effectors 23349989
786-O Function Assay 100-2000 nM 10-180 min inhibits both mTORC1 and mTORC2 as indicated by the decrease in phosphorylation of downstream effectors 23349989
Caki-1  Cell Viability Assay 300-4000 nM 24-96 h suppresses the cell viability in both time and dose dependent manner 23349989
786-O Cell Viability Assay 300-4000 nM 24-96 h suppresses the cell viability in both time and dose dependent manner 23349989
Caki-1  Function Assay 2 µM 72 h induces G1 cell cycle arrest and autophagy 23349989
786-O Function Assay 2 µM 72 h induces G1 cell cycle arrest and autophagy 23349989
HBCx-10 Function assay 5 mg/kg Potentiation of irinotecan-induced tumor regression against human HBCx-10 cells xenografted in immunocompromised mouse at 5 mg/kg, po qd administered on days 1 to 3 of weekly cycle 19762236
PC3 Function assay 100 mg/kg Inhibition of Akt phosphorylation at Ser473 in PTEN-deficient human PC3 cells xenograft mouse model at 100 mg/kg, po single dose measured up to 8 hrs 29211480
PC3 Antitumor assay 30 mg/kg Antitumor activity against human PC3 cells xenograft mouse model assessed as inhibition of tumor growth at 30 mg/kg, po bid in presence of 1-aminobenzotriazole 29211480
PC3 Antitumor assay 60 mg/kg Antitumor activity against human PC3 cells xenograft mouse model assessed as inhibition of tumor growth at 60 mg/kg, po bid in presence of 1-aminobenzotriazole 29211480
SW620 Function assay 20 mg/kg Potentiation of irinotecan-induced tumor regression against human SW620 cells xenografted in immunocompromised mouse at 20 mg/kg, po qd administered on days 2 to 4 of weekly cycle 29211480
H1975 Growth Inhibition Assay 72 h IC50=11.15±0.93 nM 23874880
H1650 Growth Inhibition Assay 72 h IC50=7.61±0.62 nM 23874880
PC9GR Growth Inhibition Assay 72 h IC50=6.21±1.30 nM 23874880
PC9 Growth Inhibition Assay 72 h IC50=10.15±0.62 nM 23874880
HEK293 Function assay 2 hrs Inhibition of recombinant FLAG-tagged mTOR expressed in HEK293 cells using biotinylated p70S6K substrate after 2 hrs by alphascreen competition assay, IC50=0.003μM 19762236
U87MG Function assay 2 hrs Inhibition of mTORC1 in human U87MG cells assessed as phosphorylated S6 ribosomal protein (Ser235/236) level after 2 hrs by Western blotting, IC50=0.1μM 19762236
U87MG Function assay 2 hrs Inhibition of mTORC2 in human U87MG cells assessed as phosphorylated AKT (Ser473) level after 2 hrs by Western blotting, IC50=0.15μM 19762236
T47D Antiproliferative assay 120 hrs Antiproliferative activity against human T47D cells after 120 hrs by SRB assay, GI50=0.35μM 19762236
MDA-MB-468 Function assay 2 hrs Inhibition of mTORC2 in human MDA-MB-468 cells assessed as reduction of AKT phosphorylation at Ser473 after 2 hrs, IC50=0.24μM 23375793
MDA-MB-468 Function assay 2 hrs Inhibition of mTORC1 in human MDA-MB-468 cells assessed as reduction of pS6 phosphorylation at Ser235/236 after 2 hrs, IC50=0.66μM 23375793
HEK293 Function assay 30 mins Inhibition of mTORC1 in HEK293 cells using GST-tagged S6K1 or Akt1 as substrate after 30 mins by immunoblotting assay, IC50=0.01μM 29211480
HEK293 Function assay 30 mins Inhibition of mTORC2 in HEK293 cells using GST-tagged S6K1 or Akt1 as substrate after 30 mins by immunoblotting assay, IC50=0.01μM 29211480
NUGC4 Growth Inhibition Assay IC50=2.93 ± 0.31 μM 24597478
MKN45 Growth Inhibition Assay IC50=0.82 ± 0.01 μM 24597478
HGC27 Growth Inhibition Assay IC50=15.0 ± 4.82 μM 24597478
AGS Growth Inhibition Assay IC50=15.0 ± 2.91 μM 24597478
HEK293 Function assay Inhibition of recombinant FLAG-tagged mTOR (1362 to 2549) (unknown origin) expressed in HEK293 cells, IC50=0.0025μM 23375793
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells 29435139
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 29435139
BT-37 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells 29435139
NB-EBc1 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells 29435139
OHS-50 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells 29435139
MG 63 (6-TG R) qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells 29435139
Rh41 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells 29435139
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for A673 cells) 29435139
BT-12 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-12 cells 29435139
DAOY qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for DAOY cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SJ-GBM2 cells 29435139
U-2 OS qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for U-2 OS cells 29435139
Rh41 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh41 cells 29435139
RD qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for RD cells 29435139
Rh18 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh18 cells 29435139
Rh30 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh30 cells 29435139
SK-N-SH qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-SH cells 29435139
点击查看更多细胞系数据

生物活性

产品描述 KU-0063794是一种有效的,高度特异性的,作用于mTORC1mTORC2的双重mTOR抑制剂,在无细胞试验中IC50约为~10 nM;对PI3Ks没有作用。
靶点
mTORC1 [1]
(Cell-free assay)
mTORC2 [1]
(Cell-free assay)
~10 nM ~10 nM
体外研究(In Vitro)
体外研究活性

与mTOR抑制剂PP242相比, KU-0063794高特异性作用于mTOR,而对PI3Ks或其他76种激酶则无作用活性。30 nM KU-0063794作用于HEK-293细胞,通过抑制疏水基团(Thr389)磷酸化,及随后的T-环残基(Thr229) 磷酸化,而快速切除S6K1活性。300 nM KU-0063794作用于IGF1刺激的血清饥饿处理的 HEK-293细胞,抑制~90%S6K1活性。KU-0063794 为 100-300 nM时,也完全抑制氨基酸诱导的S6K1和 S6蛋白磷酸化。与S6K1类似, KU-0063794抑制mTORC1在 Ser2448位点磷酸化,也抑制mTORC2 在 Ser22481 位点磷酸化,这种作用存在剂量和时间依赖性。在血清存在时,或者IGF1刺激的情况下, KU-0063794抑制Akt 活性,或者Akt在 Ser473和 Thr308(意外的)位点磷酸化,也抑制Akt底物 PRAS40 在 Thr246位点,GSK3α/GSK3β在 Ser21/Ser9位点及Foxo-1/3a 在 Thr24/Thr32位点磷酸化,以上作用存在剂量依赖性。KU-0063794而不是 Rapamycin 抑制SGK1活性和 在Ser422位点磷酸化,也抑制其生理底物NDGR1,与抑制S6K1 和Akt磷酸化程度相似,这种作用存在剂量依赖性,然而KU-0063794 不抑制佛波酯诱导的ERK 或RSK磷酸化和RSK激活。与Rapamycin相比, KU-0063794更有效诱导4E-BP1在 Thr37, Thr46和 Ser65位点完全去磷酸化。KU-0063794抑制野生型和mLST8-缺陷型MEFs细胞生长,也诱导细胞周期停在G1期,比Rapamycin效果更显著。[1]

激酶实验 mTOR 复合物激酶检测实验
HEK-293细胞新鲜溶解在Hepes裂解液中。溶解产物(1-4 mg) 通过与5-20 μL G蛋白的琼脂糖凝胶结合免疫前抗体IgG温育而预先清除。裂解抽提物与5-20 μL G蛋白的琼脂糖凝胶结合的 5-20 μg抗Rictor或抗Raptor抗体的 免疫前抗体IgG温育。所有抗体共价结合到蛋白G蛋白琼脂糖凝胶上。在振动转载台上4oC下进行 免疫沉淀1小时。使用Hepes 裂解液冲洗免疫沉淀物四次,随后使用Hepes激酶buffer冲洗两次。使用 Raptor免疫沉淀物磷酸化 S6K1, 两次冲洗的初始步骤中,buffer 含0.5 M NaCl,确保最佳激酶活性。血清饥饿的 HEK-293细胞中分离的GST-Akt1与PI-103(1 μM 进行 1 小时)温育。从血清饥饿的HEK-293 细胞中纯化的GST-S6K1与Rapamycin (0.1 μM进行1小时)温育。加入0.1 mM ATP和 10 mM MgCl2 ,在不同浓度KU-0063794和GST-Akt1 (0.5 μg)或GST-S6K1 (0.5 μg)存在时,开始mTOR反应。反应在 30oC下在振动转载台上进行30分钟,然后加入SDS样本缓冲液终止反应。反应混合物在0.22-μm-孔径大小 Spin-X过滤器上过滤,样本进行电泳处理,然后使用指定抗体进行免疫印迹分析。
细胞实验 细胞系 野生型和mLST8缺陷型MEFs
浓度 溶于DMSO,终浓度为~3 μM
孵育时间 24, 48, 和72 小时
方法

使用KU-0063794 处理细胞 24, 48, 和72 小时,每24小时更换一次培养基,加入新鲜溶解的KU-0063794。为了测量细胞生长,使用PBS冲洗细胞一次,然后与4% (v/v) 多聚甲醛在 PBS混合15分钟。用水冲洗一次后,使用溶于 10% 乙醇的0.1% 结晶紫对细胞染色20分钟,然后用水冲洗三次。 使用0.5 mL 10% (v/v) 乙酸从细胞中抽提结晶紫20分钟。然后洗脱液在水中按1:10稀释,然后在 590 nm 处测量吸光值。为了测量细胞周期分布,通过胰蛋白酶化作用收集细胞,然后在PBS中冲洗一次,再悬浮于冰冻70% (v/v)乙醇中。细胞在 PBS和 1% (w/v) BSA中冲洗两次,再在 PBS 和含 50 g/mL 碘化丙啶和 50 g/mL RNase A的0.1% (v/v) Triton X-100中染色 20分钟。使用FACSCalibur流式细胞仪和CellQuest软件检测细胞中DNA含量。在线性标度上测量红色荧光(585 nm),进行脉冲宽度分析用于排除双峰值。使用FlowJo软件测定细胞周期分布。

实验图片 检测方法 检测指标 实验图片 PMID
Western blot p-S6K / S6K / p-4E-BP1 / E7 / E6 / p53 p-mTOR 28115701
体内研究(In Vivo)
体内研究活性

Ku0063794在肾脏上皮肾细胞癌的临床前模型中,抑制肿瘤生长和mTOR信号[2]

动物实验 Animal Models Nu/Nu裸鼠
Dosages 8 mg/kg
Administration i.p.

化学信息&溶解度

分子量 465.54 分子式

C25H31N5O4

CAS号 938440-64-3 SDF Download KU-0063794 SDF
Smiles CC1CN(CC(O1)C)C2=NC3=C(C=CC(=N3)C4=CC(=C(C=C4)OC)CO)C(=N2)N5CCOCC5
储存条件(自收到货起)

体外溶解度
批次:

DMSO : 16 mg/mL ( (34.36 mM) ;DMSO吸湿会降低化合物溶解度,请使用新开封DMSO)

Ethanol : 5 mg/mL (10.74 mM)

Water : Insoluble

摩尔浓度计算器

体内溶解度
批次:

现配现用,请按从左到右的顺序依次添加,澄清后再加入下一溶剂

动物体内配方计算器

实验计算

摩尔浓度计算器

质量 浓度 体积 分子量

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。

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