| Size | Price | Stock | Qty |
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| 500mg |
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| Other Sizes |
| Targets |
The primary molecular target of nAS-E is the KIX domain of the transcriptional coactivator CREB-binding protein (CBP). nAS-E directly binds to the KIX domain with a dissociation constant (Kd) of 8.6 µM. This binding event blocks the interaction between the KIX domain of CBP and the KID (kinase-inducible domain) of the transcription factor CREB. By disrupting this critical protein-protein interaction, nAS-E inhibits CREB-mediated gene transcription. The KIX-KID interaction is essential for the recruitment of CBP to CREB, which is necessary for the expression of genes involved in cell growth and survival.
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| ln Vitro |
In several signaling pathways originating from G protein-coupled receptors or receptor tyrosine kinases, CREB (cyclic AMP response element binding protein) functions as a downstream transcription factor. Only phosphorylation at Ser133 may activate CREB, which then attaches to CREB-binding protein (CBP) via the KID (kinase-inducing domain) in CBP and the KID-interacting (KIX) domain in CREB. With an IC50 of 2.29 µM, naphthol AS-E suppresses CREB-mediated gene transcription in a cell-based CREB Renilla luciferase reporter experiment. Using recombinant KIX (Kd ~8.6 µM), naphthol AS-E directly binds to the KIX domain of CBP in a dose-dependent manner to block Renilla luciferase activity in a HEK293T-based complementation test. All of these cancer cells' growth was inhibited by naphthol AS-E with low µM activity, which is consistent with its cellular CREB inhibitory potency. Approximately 2.9μM, 2.81μM, 2.35μM, and 1.46μM are the average GI50 values of A549, MCF-7, MDA-MB-231, and MDA-MB-468, respectively. The anti-apoptotic protein Bcl-2's expression is decreased by naphthol AS-E (2.5 µM–10 µM; 48 hours). There is also a decrease in VEGF expression.
In vitro, nAS-E exhibits potent inhibitory activity against the KIX-KID interaction with an IC₅₀ of 2.26 µM. This activity is a direct result of its binding to the KIX domain, as confirmed by biophysical assays. Furthermore, nAS-E has been shown to inhibit the proliferation of various cancer cell lines, with average GI₅₀ values that are consistent with its cellular CREB inhibition potency. This demonstrates that the compound's anti-proliferative effects are likely mediated through its inhibition of CREB-dependent transcription. The compound's cell permeability allows it to effectively reach its intracellular target. |
| ln Vivo |
The in vivo activity of nAS-E has been explored in the context of cancer research. As a CREB inhibitor, it has the potential to suppress tumor growth by blocking the expression of pro-survival and pro-proliferative genes. However, detailed in vivo data, including specific dosing regimens and efficacy in animal models, are not extensively documented in the provided search results. The compound is primarily used as a research tool to validate the KIX-KID interaction as a drug target. Its in vivo efficacy would be a subject of further investigation to determine its therapeutic potential.
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| Enzyme Assay |
For in vitro enzyme/receptor binding assays, the activity of nAS-E is typically assessed using biophysical methods such as surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) to measure its direct binding affinity (Kd) to the KIX domain of CBP. Additionally, fluorescence polarization (FP) or AlphaScreen assays can be employed to measure its ability to disrupt the KIX-KID interaction. In these competitive binding assays, a fluorescently labeled KID peptide is incubated with the KIX domain in the presence of varying concentrations of nAS-E. The displacement of the labeled peptide results in a change in the fluorescence signal, allowing for the calculation of the IC₅₀ for inhibition.
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| Cell Assay |
Cell-based in vitro assays for nAS-E involve treating cancer cell lines with the compound and assessing its effects on cell proliferation and CREB-mediated transcription. Cells are typically treated with nAS-E at concentrations ranging from 1 to 10 µM for 24 to 72 hours. Cell viability and proliferation are measured using standard assays such as MTT or CellTiter-Glo. To specifically measure CREB activity, a luciferase reporter gene assay can be used, where the expression of luciferase is driven by a CREB-responsive promoter. The inhibition of luciferase activity by nAS-E directly reflects its ability to inhibit CREB-mediated transcription in a cellular context.
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| Animal Protocol |
In vivo animal studies for nAS-E would typically involve xenograft models, where human cancer cells are implanted into immunodeficient mice. After tumors are established, nAS-E would be administered, often via intraperitoneal or oral routes, at various doses. Tumor growth would be monitored over time, and endpoints would include tumor volume, tumor weight, and survival. However, as nAS-E is primarily a research tool and not a clinical candidate, such studies may not be as extensive as for more advanced drug candidates. The compound's role is more focused on target validation in a preclinical setting.
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| ADME/Pharmacokinetics |
Pharmacokinetic (PK) data for nAS-E are not readily available in the provided search results. As a small molecule with a molecular weight of 297.74 g/mol, it may have reasonable oral bioavailability and tissue distribution. However, detailed parameters such as half-life, Cmax, and clearance would need to be determined through specific PK studies. The compound is likely metabolized by hepatic enzymes and eliminated via renal or biliary routes. Its cell permeability, as demonstrated by its activity in cell-based assays, suggests it can cross cellular membranes to reach its intracellular target.
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| Toxicity/Toxicokinetics |
Toxicology data for nAS-E are also limited in the provided sources. As a research compound, its safety profile would be characterized in preclinical studies to assess potential off-target effects and general toxicity. The compound is structurally related to azo dyes, which may have implications for its safety. However, specific toxicological endpoints, such as acute toxicity, genotoxicity, or organ-specific toxicity, are not detailed. Researchers handling nAS-E should follow standard laboratory safety practices.
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| References | |
| Additional Infomation |
In summary, nAS-E (Naphthol AS-E) is a potent and cell-permeable inhibitor of the KIX-KID interaction, which is a critical protein-protein interaction for CREB-mediated gene transcription. It is a valuable tool compound for cancer research, used to study the role of CREB in cell proliferation and survival. The compound binds directly to the KIX domain of CBP with a Kd of 8.6 µM and inhibits the KIX-KID interaction with an IC₅₀ of 2.26 µM. Its anti-proliferative effects have been demonstrated in various cancer cell lines. nAS-E is available for research use only and is not approved for therapeutic applications. No clinical trials or regulatory approvals exist for this compound.
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| Molecular Formula |
C17H12CLNO2
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| Molecular Weight |
297.74
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| Exact Mass |
297.056
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| CAS # |
92-78-4
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| PubChem CID |
66720
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| Appearance |
Light brown to brown solid powder
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| Density |
1.399 g/cm3
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| Boiling Point |
416.5ºC at 760 mmHg
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| Melting Point |
255°C
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| Flash Point |
205.7ºC
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| Index of Refraction |
1.735
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| LogP |
4.524
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
21
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| Complexity |
368
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
OHAXNCGNVGGWSO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H12ClNO2/c18-13-5-7-14(8-6-13)19-17(21)15-9-11-3-1-2-4-12(11)10-16(15)20/h1-10,20H,(H,19,21)
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| Chemical Name |
N-(4-chlorophenyl)-3-hydroxynaphthalene-2-carboxamide
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| Synonyms |
nAS E; nAS-E; Naphthol AS-E
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~41.67 mg/mL (~139.95 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (8.40 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (8.40 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3586 mL | 16.7932 mL | 33.5864 mL | |
| 5 mM | 0.6717 mL | 3.3586 mL | 6.7173 mL | |
| 10 mM | 0.3359 mL | 1.6793 mL | 3.3586 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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