| Size | Price | Stock | Qty |
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| 100mg |
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| 500mg |
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| 1g | |||
| Other Sizes |
| Targets |
EN450 targets NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells), a family of transcription factors that play a central role in immune responses, inflammation, and cell survival. Specifically, EN450 is a cysteine-reactive covalent molecular glue that binds to the allosteric cysteine residue C111 in the E2 ubiquitin-conjugating enzyme UBE2D. This binding induces a ternary complex between UBE2D and NFKB1 (the p105/p50 subunit of NF-κB), leading to ubiquitination and proteasomal degradation of NF-κB1.
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| ln Vitro |
In vitro, EN450 (50 µM; 24 h) significantly inhibits the proliferation of HAP1 leukemia cancer cells and HEK293T cells. It significantly reduces the levels of the p105 and p50 subunits of the oncogenic transcription factor NF-κB1. EN450 (50 µM; 1 h) enhances the ubiquitination of NF-κB1 mediated by UBE2D1 in the presence of the full CUL4A/RBX1/NEDD8 complex. The anti-proliferative effects of EN450 are exerted through a Cullin E3 ligase and proteasome-dependent mechanism.
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| ln Vivo |
In vivo activity of EN450 has been demonstrated in preclinical models, with its mechanism of action involving targeted degradation of NF-κB through covalent molecular glue interaction. The compound disrupts NF-κB signaling, which plays a crucial role in cancer cell survival, inflammation, and immune regulation. While detailed in vivo efficacy data are limited in publicly available sources, EN450 represents a promising tool for studying NF-κB-dependent pathologies, particularly in leukemia and other cancers.
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| Enzyme Assay |
The in vitro enzyme/receptor binding assay for EN450 typically involves binding studies to assess its interaction with UBE2D and NFKB1. Surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) can be used to measure the binding affinity between EN450 and the UBE2D E2 ubiquitin ligase. Covalent binding to cysteine residue C111 can be confirmed by mass spectrometry or using cysteine-reactive probe displacement assays. Ternary complex formation between UBE2D and NFKB1 in the presence of EN450 can be assessed using co-immunoprecipitation or pull-down assays.
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| Cell Assay |
The in vitro cell-based assay for EN450 is performed using HAP1 leukemia cells or HEK293T cells. Cells are seeded in multi-well plates and treated with EN450 at 50 µM for 24 hours. Cell proliferation is assessed using standard viability assays such as CellTiter-Glo or MTT. For mechanistic studies, cells are treated with EN450 (50 µM; 1 h) and NF-κB1 ubiquitination is analyzed by immunoprecipitation followed by Western blotting. Protein levels of NF-κB1 p105 and p50 subunits are measured by Western blot after 24 hours of treatment.
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| Animal Protocol |
In vivo animal studies for EN450 would typically involve mouse xenograft models of leukemia or other NF-κB-dependent cancers. Tumor-bearing mice are administered EN450 via oral gavage or intraperitoneal injection at doses determined from pharmacokinetic studies. Tumor volume is measured over time, and tumors are collected at study termination for analysis of NF-κB degradation by Western blot and immunohistochemistry. Detailed protocols for EN450 in vivo studies are not extensively published, but standard approaches for molecular glue degraders apply.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of EN450 are characteristic of small-molecule molecular glue degraders. It is soluble in DMSO at 100 mg/mL (346.31 mM) and has been formulated for in vivo administration using vehicles such as 10% DMSO/40% PEG300/5% Tween-80/45% saline or 10% DMSO/90% corn oil. As a covalent binder, its pharmacokinetics may be influenced by target engagement and off-target reactivity. Detailed PK parameters including bioavailability, half-life, and tissue distribution require further characterization.
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| Toxicity/Toxicokinetics |
The toxicity profile of EN450 is primarily derived from in vitro cell-based assays. At 50 µM, EN450 significantly inhibits proliferation of HAP1 and HEK293T cells, indicating potential cytotoxicity at high concentrations. As a covalent molecular glue degrader, off-target reactivity with other cysteine-containing proteins is a theoretical concern. No comprehensive toxicology data are publicly available, and the compound is for research use only. Standard safety assessments would include cytotoxicity screening, hERG channel inhibition, and in vivo tolerability studies.
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| References | |
| Additional Infomation |
EN450 is a research compound and has not been approved for clinical use. It is a cysteine-reactive covalent molecular glue degrader targeting NF-κB via interaction with UBE2D E2 ubiquitin ligase. The compound displays anti-proliferative effects in HAP1 leukemia cells and HEK293T cells and acts through a Cullin E3 ligase and proteasome-dependent mechanism. EN450 is a valuable tool for studying targeted protein degradation of transcription factors and has potential applications in cancer research.
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| Molecular Formula |
C11H13CLN2O3S
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|---|---|
| Molecular Weight |
288.750520467758
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| Exact Mass |
288.03
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| Elemental Analysis |
C, 45.76; H, 4.54; Cl, 12.28; N, 9.70; O, 16.62; S, 11.10
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| CAS # |
793719-01-4
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| PubChem CID |
4877041
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| Appearance |
White to off-white solid powder
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| LogP |
1.3
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
18
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| Complexity |
417
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC1C=CC(=CC=1NC(C=C)=O)S(N(C)C)(=O)=O
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| InChi Key |
OXFXDOLAQBMOPH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H13ClN2O3S/c1-4-11(15)13-10-7-8(5-6-9(10)12)18(16,17)14(2)3/h4-7H,1H2,2-3H3,(H,13,15)
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| Chemical Name |
N-(2-chloro-5-(N,N-dimethylsulfamoyl)phenyl)acrylamide
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| Synonyms |
EN450; EN-450; EN 450; starbld0003560; GTPL12636
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.4632 mL | 17.3160 mL | 34.6320 mL | |
| 5 mM | 0.6926 mL | 3.4632 mL | 6.9264 mL | |
| 10 mM | 0.3463 mL | 1.7316 mL | 3.4632 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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