| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
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| Other Sizes |
Purity: ≥98%
| Targets |
G9a (IC50=4.7 μM), SUV39H1(H320R) (IC50=1.16 μM), PRMT1 , GLP, ESET, SET7/9 (inhibited at 15 μM). [1]
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|---|---|
| ln Vitro |
In enzyme assays, BIX-01338 inhibited G9a with an IC50 of 4.7 μM and SUV39H1(H320R) with an IC50 of 1.16 μM. It started to affect methylation activity of all tested HMTases at around 5 μM and neutralized almost all enzymes at 15 μM, as determined by mass spectrometry. The compound displayed competitive inhibition with SAM in Eadie-Hofstee analysis. [1]
In cellular assays, treatment of mouse embryonic stem (ES) cells with 3.3 μM BIX-01338 for 2 days resulted in no detectable differences in methylation states at H3K9 and H3K27 compared to mock-treated cells, as measured by quantitative mass spectrometry. No alterations were observed for H3K36 and H3K79 methylation either. Although the compound is known to enter mammalian cells, it did not reduce H3K9me2 levels in vivo. [1] |
| Enzyme Assay |
The high-throughput screen used a DELFIA (dissociation enhanced lanthanide fluoroimmunoassay) with recombinant GST-G9a (amino acids 685-1000) fusion protein, biotinylated histone H3 peptide, and 20 μM SAM. The H3K9me2 reaction product was bound to neutravidin-coated 384-well microtiter plates and detected with an H3K9me2 antibody followed by europium-labeled secondary antibody. Hits were scored as reduction of europium signal below 65% of control. [1]
For dose-response assays, BIX-01338 was tested on G9a, SUV39H1(H320R), and PRMT1 using DELFIA. IC50 values were calculated. [1] To determine mode of action, reaction kinetics were measured at varying SAM concentrations. Eadie-Hofstee plot analysis showed that BIX-01338 acts as a competitive inhibitor with respect to SAM. [1] For mass spectrometry-based validation, HMTase reactions were performed with nonbiotinylated H3(1-20)-cys peptide (for G9a, SUV39H1, GLP, SET7/9) or H4(1-20)-cys (for PRMT1). Reactions were stopped by dilution with 0.1% trifluoroacetic acid. Peptides were separated by C18 reverse-phase HPLC coupled to mass spectrometers, and peak areas of 3+ and 4+ peptide ions were quantified against synthetic peptide standards. [1] |
| Cell Assay |
Mouse ES cells (wild-type and G9a-deficient), mouse embryonic fibroblasts (MEFs), and human HeLa cells were treated with BIX-01338 at 3.3 μM for 2 days. Nuclear extracts were prepared, and histone H3 was isolated. After propionylation of lysines and carbamidomethylation of cysteines, tryptic digestion was performed. Peptides were separated by reverse-phase HPLC coupled to mass spectrometry. Quantification was performed by comparison of peak areas to external synthetic peptide standards containing the same posttranslational modifications. No changes in H3K9me2, H3K9me3, H3K9me1, H3K27me, H3K36me, or H3K79me were detected in BIX-01338-treated cells compared to mock-treated controls. [1]
|
| References | |
| Additional Infomation |
BIX-01338 belongs to a class of inhibitors containing a 2-(N-acyl)-aminobenzimidazole backbone, which is also found in a number of biologically active molecules. It is related to BIX-01294 but acts broadly. The compound was identified from the Boehringer Ingelheim chemical library. Unlike BIX-01294, which specifically inhibits G9a, BIX-01338 is non-selective and competes with SAM. [1]
|
| Molecular Formula |
C32H26F3N3O7
|
|---|---|
| Molecular Weight |
621.559958934784
|
| Exact Mass |
621.172
|
| CAS # |
1228184-65-3
|
| PubChem CID |
71311908
|
| Appearance |
Light blue to blue solid powder
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
10
|
| Heavy Atom Count |
45
|
| Complexity |
990
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
FC(C1C=CC(=CC=1)C(NC1=NC2C=C(C(=O)O)C=CC=2N1CCC1C=CC(=CC=1)OC(C1C=CC(=CC=1)OC)=O)=O)(F)F.O
|
| InChi Key |
LLTPCGBKXOESTJ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C32H24F3N3O6.H2O/c1-43-24-13-6-21(7-14-24)30(42)44-25-11-2-19(3-12-25)16-17-38-27-15-8-22(29(40)41)18-26(27)36-31(38)37-28(39)20-4-9-23(10-5-20)32(33,34)35;/h2-15,18H,16-17H2,1H3,(H,40,41)(H,36,37,39);1H2
|
| Chemical Name |
1-[2-[4-(4-methoxybenzoyl)oxyphenyl]ethyl]-2-[[4-(trifluoromethyl)benzoyl]amino]benzimidazole-5-carboxylic acid;hydrate
|
| Synonyms |
BIX-01338; BIX01338; BIX 01338
|
| 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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
DMSO : ~200 mg/mL (~321.77 mM)
|
|---|---|
| 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 | 1.6089 mL | 8.0443 mL | 16.0886 mL | |
| 5 mM | 0.3218 mL | 1.6089 mL | 3.2177 mL | |
| 10 mM | 0.1609 mL | 0.8044 mL | 1.6089 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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