| ln Vitro |
EN1033 (6.25-100 μM) does not exhibit cytotoxicity in THP1 macrophage leukemia cancer cells, nor does it induce stress granules formation[1]. EN1033 (1, 10, 100 µM, 24 h) induces on-pathway inhibition of IRF5 transcriptional activity[1]. EN1033 (100 μM, 18 h) degrades IRF8 in EN1033 (12.5-100 μM, 2-24 h) degrades both IRF8 and IRF5, in a dose-responsive and time-dependent manner[1]. EN1033 (100 μM, 6 h) engages IRF8 more robustly than IRF5, while EN1033 and its analog TH3-189 exert only moderate effects across the proteome[1]. EN1033 (50 μM, 18, 24 h) causes the degradation of IRF5 and IRF8 by targeting C28 and C223, respectively[1]. EN1033 (12.5-100 μM, 24 h) degrades IRF8 in IRF5 Knockout cells, suggesting IRF5 does not regulate IRF8. In contrast, IRF8 knockdown markedly reduces IRF5 levels, indicating IRF8 regulates IRF5 in THP1 cells[1].
|
|---|---|
| Cell Assay |
Western Blot Analysis[1]
Cell Types: THP1 cells Tested Concentrations: 12.5, 25, 50, 75, 100 μM Incubation Duration: 24 h Experimental Results: Decreased the levels of IRF8 and IRF5 gradually as the concentration increased. Triggered the most prominent degradation of IRF8 and IRF5 at a concentration of 100 μM. Western Blot Analysis[1] Cell Types: THP1 cells Tested Concentrations: 100 μM Incubation Duration: 0, 2, 4, 8, 20, 24 h Experimental Results: Induced the degradation of IRF8 and IRF5 starting at 8 h, and this effect was gradually strengthened over time. Western Blot Analysis[1] Cell Types: HEK293T cells Tested Concentrations: 50 μM Incubation Duration: 18, 24 h Experimental Results: Induced IRF5 degradation, which was fully rescued in C28S and C28S/C121S variants, but not in the C121S mutant. Triggered IRF8 loss, which was fully rescued in C223S and C223S/C385S IRF8 mutants, but not in the C385S mutant-expressing cells. |
| References |
| Molecular Formula |
C15H18N2O
|
|---|---|
| Molecular Weight |
242.32
|
| CAS # |
2193367-28-9
|
| Appearance |
Colorless to light yellow liquid（Density: 1.178±0.10 g/cm3）
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 4.1268 mL | 20.6339 mL | 41.2677 mL | |
| 5 mM | 0.8254 mL | 4.1268 mL | 8.2535 mL | |
| 10 mM | 0.4127 mL | 2.0634 mL | 4.1268 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
