KU-0063794
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
产品质控
批次：
纯度：
99.91%
99.91
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 |
|
| 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 |
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 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
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
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。
技术支持
在订购、运输、储存和使用我们的产品的任何阶段，您遇到的任何问题，均可以通过拨打我们的热线电话400-668-6834，或者技术支持邮箱tech@selleck.cn，直接联系到我们。我们会在24小时内尽快联系您。
如果有其他问题，请给我们留言。
* 必填项
Tags: buy KU-0063794 | KU-0063794 supplier | purchase KU-0063794 | KU-0063794 cost | KU-0063794 manufacturer | order KU-0063794 | KU-0063794 distributor
