KU-0063794

目录号：S1226 Purity: 99.91%

KU-0063794是一种有效的，高度特异性的，作用于mTORC1和mTORC2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

CAS: 938440-64-3

规格	价格	库存
10mM (1mL in DMSO)	RMB 1464.74	现货
5mg	RMB 727.74	现货
10mg	RMB 1405.09	现货
50mg	RMB 3857.09	现货
更大包装有超大折扣
400-668-6834 info@selleck.cn

客户使用Selleck的KU-0063794发表文献72篇

产品质控

批次：纯度： 99.91%

99.91

KU-0063794相关产品

相关靶点	mTORC1 mTORC2	点击展开
相关化合物库	激酶抑制剂库 PI3K/Akt 抑制剂库凋亡分子化合物库细胞周期化合物库 NF-κB信号通路库	点击展开

mTOR抑制剂选择性比较

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mTORC1和mTORC2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
激酶实验	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4^oC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 30^oC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
实验图片	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。然而，在动物研究中，Ku0063794没有temsirolimus有效，可能的原因是temsirolimus对肿瘤的微环境有重要的作用，能在异种移植瘤中减少血管生成，而Ku0063794无此效果。经Temsirolimus处理的肿瘤组织比Ku0063794处理的肿瘤组织更少地表达VEGF和PDGF^[2]。
动物实验	Animal Models	Nu/Nu裸鼠
	Dosages	8 mg/kg
	Administration	i.p.

参考文献
[1] García-Martínez JM, et al. Biochem J, 2009, 421(1), 29 [2] Zhang H, et al. PLoS One. 2013, 8(1):e54918.

参考文献

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)	3年-20°C粉状	储备液保存和期限建议分装储备液，避免反复冻融！	1年-80°C溶于溶剂
储存条件(自收到货起)	3年-20°C粉状	储备液保存和期限建议分装储备液，避免反复冻融！	1个月-20°C溶于溶剂

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

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
=	×	×

稀释计算器分子量计算器

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）

给药剂量 mg/kg 动物平均体重 g 每只动物给药体积 μL 动物数量只

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，注：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：取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：取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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