HTH-01-015 |
| Catalog No.GC13185 |
HTH-01-015 es un inhibidor selectivo de NUAK1/ARK5 (IC50 es 100 nM). HTH-01-015 inhibe NUAK1 con una potencia >100 veces mayor que NUAK2 (IC50 de >10 μM).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 1613724-42-7
Sample solution is provided at 25 µL, 10mM.
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HTH-01-015 is a highly specific inhibitor of NUAK1 with IC50 value of 100 nM 1.
HTH-01-015 showed extreme selectivity. It only inhibited the NUAK1 (NUAK family SNF1-like kinase-1) isoform of NUAK kinases and showed no significant inhibition of 139 other tested kinases. HTH-01-015 inhibited the phosphorylation of the NUAK1 substrate, MYPT1. The phosphorylated site was identified as Ser445. In HEK cells overexpressing drug-resistant NUAK1, HTH-01-015 displayed no more inhibition effect on MYPT1. In MEF cells, HTH-01-015 markedly reduced cell migration in the wound-healing assay. Besides that, HTH-01-015 impaired the proliferation at concentration of 10 μM both in U2OS cells and in MEF cells. Moreover, 10 μM HTH-01-015 also significantly inhibited the invasiveness of U2OS cells in a 3D matrigel transwell invasion assay 1.
References:
1. Sourav B, Sara J B, Hai-Tsang H, et al. Characterization of WZ4003 and HTH-01-015 as selective inhibitors of the LKB1-tumour-suppressor-activated NUAK kinases. Biochemical Journal, 2014, 457(1): 215-225.
| Kinase experiment [1]: | |
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Kinase activity assays |
In vitro activities of purified GST–NUAK1 and GST–NUAK1 [A195T] were measured using Cerenkov counting of incorporation of radioactive 32P from [γ -32P] ATP into Sakamototide substrate peptide. Reactions were carried out in a 50 μl reaction volume for 30 min at 30 μC and reactions were terminated by spotting 40 μl of the reaction mix on to P81 paper and immediately immersing in 50 mM orthophosphoric acid. Samples were washed three times in 50 mMorthophosphoric acid followed by a single acetone rinse and air drying. The kinase-mediated incorporation of [γ -32P] ATP into Sakamototide was quantified by Cerenkov counting. One unit of activity was defined as that which catalyzed the incorporation of 1 nmol of [32P]phosphate into the substrate over 1 h. |
| Cell experiment [1]: | |
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Cell lines |
HEK293, MEFs and U2OS |
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Preparation method |
Limited solubility. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
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Reaction Conditions |
37oC |
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Applications |
In HEK-293 cells expressing wild-type NUAK1, 3–10 μM HTH-01-015 significantly inhibits phosphorylation of MYPT1. Treatment of NUAK1+/+ MEFs with 10 μM HTH-01-015 prominently suppresses cell migration in the wound-healing assay. In U2OS cells, HTH-01-015 blocks proliferation and phosphorylation of MYPT1 to the same extent as shRNA-mediated NUAK1 knockdown. |
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References: 1. Sourav B, Sara J B, Hai-Tsang H, et al. Characterization of WZ4003 and HTH-01-015 as selective inhibitors of the LKB1-tumour-suppressor-activated NUAK kinases. Biochemical Journal, 2014, 457(1): 215-225. | |
| Cas No. | 1613724-42-7 | SDF | |
| Chemical Name | 4,5,13-trimethyl-2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)-5H-naphtho[2,3-e]pyrimido[5,4-b][1,4]diazepin-6(13H)-one | ||
| Canonical SMILES | CN1C(C2=CC3=CC=CC=C3C=C2N(C)C4=NC(NC5=CN(C6CCNCC6)N=C5)=NC(C)=C14)=O | ||
| Formula | C26H28N8O | M.Wt | 468.55 |
| Solubility | ≥ 23.45mg/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. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.1342 mL | 10.6712 mL | 21.3424 mL |
| 5 mM | 0.4268 mL | 2.1342 mL | 4.2685 mL |
| 10 mM | 0.2134 mL | 1.0671 mL | 2.1342 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
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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.
Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 38 reference(s) in Google Scholar.)
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