KU-0063794 |
Catalog No.GC15167 |
KU-0063794는 10nM의 IC50으로 mTORC1 및 mTORC2 복합체를 모두 억제하는 강력하고 특이적 mTOR 억제제입니다.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 938440-64-3
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Kinase experiment [1]: | |
mTOR complexes kinase assays |
HEK-293 cells were freshly lysed in Hepes lysis buffer. Lysate (1 ~ 4 mg) was pre-cleared by incubating with 5 ~ 20 μL of Protein G-Sepharose conjugated to pre-immune IgG. The lysate extracts were then incubated with 5 ~ 20 μL of Protein G-Sepharose conjugated to 5 ~ 20 μg of either anti-Rictor or anti-Raptor antibody, or pre-immune IgG. All antibodies were covalently conjugated to Protein G-Sepharose. Immunoprecipitations were carried out for 1 hr at 4 °C on a vibrating platform. The immunoprecipitates were washed 4 times with Hepes lysis buffer, followed by two washes with Hepes kinase buffer. For Raptor immunoprecipitates used for phosphorylating S6K1, for the initial two wash steps the buffer included 0.5 M NaCl to ensure optimal kinase activity. GST-Akt1 was isolated from serum-deprived HEK-293 cells incubated with PI-103 (1 μM for 1 hr). GST-S6K1 was purified from serum-deprived HEK-293 cells incubated with Rapamycin (0.1 μM for 1 hr). mTOR reactions were initiated by adding 0.1 mM ATP and 10 mM MgCl2 in the presence or absence of KU-0063794 and GST-Akt1 (0.5 μg) or GST-S6K1 (0.5 μg). Reaction were carried out for 30 mins at 30 °C on a vibrating platform and stopped by addition of SDS sample buffer. Reaction mixtures were then filtered through a 0.22-μm-poresize Spin-X filter and samples were subjected to electrophoresis and immunoblot analysis. |
Cell experiment [1]: | |
Cell lines |
Wild-type and mLST8-deficient MEFs |
Preparation method |
The solubility of this compound in DMSO is limited. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20 °C for several months. |
Reaction Conditions |
~ 3 μM; 24, 48 and 72 hrs |
Applications |
In wild-type and mLST8-deficient MEFs, KU-0063794 inhibited cell growth. |
References: [1]. Garcia-Martinez J, Moran J, Clarke R, et al. Ku-0063794 is a specific inhibitor of the mammalian target of rapamycin (mTOR)[J]. Biochem. J, 2009, 421: 29-42. |
Ku-0063794 is a specific mTOR inhibitor, which inhibits both mTORC1 and mTORC2 with IC50 values of ~ 10nM.[1]
The mTOR (mammalian target of rapamycin) protein kinase lies at the nexus of signaling pathways that regulate cell growth and proliferation. Various cancer-driving mutations in genes such as receptor tyrosine kinases, Ras, PI3K (phosphoinositide 3-kinase) and PTEN (phosphatase and tensin homologue deleted on chromosome 10) stimulate proliferation, growth and survival by the activation of mTOR kinase. MTOR stimulates cell growth by phosphorylating and promoting activation of AGC (protein kinase A/protein kinase G/protein kinase C) family kinases such as Akt (protein kinase B), S6K (p70 ribosomal S6 kinase) and SGK(serum and glucocorticoid protein kinase). MTORC1 phosphorylates the hydrophobic motif of S6K, while mTORC2 phosphorylates the hydrophobic motif of Akt and SGK. Ku-0063794 inhibits both mTORC1 and mTORC2 in vitro and in vivo and can be used to dissect cellular functions of the mTOR pathway.[1]
In HEK-293 cells, Ku-0063794 inhibited the activity of endogenous immunoprecipitated mTORC1 and mTORC2, employing S6K1 and Akt as substrates, with IC50 values of ~10nM. The ability of Ku-0063794 to suppress S6K1 activity and phosphorylation was rapid,Concentrations
of 100–300 nM Ku-0063794 were required to fully suppress amino-acid-induced phosphorylation of S6K1 and S6 protein. Ku-0063794 inhibits phosphorylation of Akt at Thr 308. Ku-0063794 was also assessed for other targets. However, it does not significantly inhibit 76 other protein kinases tested as well as seven lipid kinases, including class 1a PI3K α and PI3K β.[1,2]
The antileukemic effects of Ku-0063794 of the PI3K signaling pathway were evaluated by methylcellulose colony assays with primary murine CML cells obtained from mice that underwent bone marrow reconstitution with hematopoietic cells transduced with a retrovirus vector-expressing BCR/ABL. It showed that Ku-0063794 fully suppressed colony formation of primary murine CML at 1000nM. Moreover, KU-0063794 strikingly reduces cell proliferation in BA/F3 cells expressing BCR/ABL independently of mutation status, which indicates Ku-0063794 is a desirable therapeutic strategy to affect oncogenes signaling downstream of BCR/ABL.[2]
References:
1.Garcia-Martinez J, Moran J, Clarke R, et al. Ku-0063794 is a specific inhibitor of the mammalian target of rapamycin (mTOR)[J]. Biochem. J, 2009, 421: 29-42.
2.Schuster K, Zheng J, Arbini A A, et al. Selective targeting of the mTORC1/2 protein kinase complexes leads to antileukemic effects in vitro and in vivo[J]. Blood cancer journal, 2011, 1(9): e34.
Cas No. | 938440-64-3 | SDF | |
Chemical Name | [5-[2-[(2R,6S)-2,6-dimethylmorpholin-4-yl]-4-morpholin-4-ylpyrido[2,3-d]pyrimidin-7-yl]-2-methoxyphenyl]methanol | ||
Canonical SMILES | CC1CN(CC(O1)C)C2=NC3=C(C=CC(=N3)C4=CC(=C(C=C4)OC)CO)C(=N2)N5CCOCC5 | ||
Formula | C25H31N5O4 | M.Wt | 465.54 |
Solubility | ≥ 11.65mg/mL in DMSO | Storage | Store at -20°C |
General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. |
Prepare stock solution | |||
1 mg | 5 mg | 10 mg | |
1 mM | 2.148 mL | 10.7402 mL | 21.4804 mL |
5 mM | 0.4296 mL | 2.148 mL | 4.2961 mL |
10 mM | 0.2148 mL | 1.074 mL | 2.148 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
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