| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| 50mg |
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| 100mg | |||
| Other Sizes |
| Targets |
1. BRD4(BD1) (targets Met149 residue in the bromodomain binding pocket, IC50 for BRD4(BD1) = 12 nM) [1]
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|---|---|
| ln Vitro |
1. Covalent binding activity: ZEN-3219 selectively forms a covalent bond with Met149 in BRD4(BD1) via its epoxide warhead; thermal denaturation assays confirmed that the compound increased the melting temperature of BRD4(BD1) by 12 °C, while no significant thermal shift was observed for other bromodomains (e.g., BRD2(BD1), BRD3(BD1)), indicating high target selectivity [1]
2. Bromodomain binding inhibition: ZEN-3219 inhibited the interaction between BRD4(BD1) and acetylated lysine (AcLys) peptides with an IC50 of 12 nM; MALDI-TOF mass spectrometry verified the formation of a covalent adduct between the compound and BRD4(BD1), with no detectable adducts formed with other BET family bromodomains [1] 3. Antiproliferative activity: In MV4-11 acute myeloid leukemia cells (BET-dependent cell line), ZEN-3219 exhibited durable antiproliferative effects; after a 24 h treatment with 50 nM of the compound followed by washout, cell proliferation remained inhibited by 78% at 72 h post-washout, whereas non-covalent BET inhibitors showed only 22% inhibition under the same conditions [1] 4. Transcriptional inhibition: ZEN-3219 suppressed the transcription of BRD4-dependent genes (e.g., MYC, BCL2) in MV4-11 cells; qRT-PCR results showed that 50 nM of the compound reduced MYC mRNA levels by 65% and BCL2 mRNA levels by 58% at 24 h, with the inhibitory effect persisting for 48 h post-compound washout [1] |
| Enzyme Assay |
1. Thermal denaturation assay: Purified BRD4(BD1) protein was incubated with serial concentrations of ZEN-3219 (0.1-1000 nM) in a buffer system for 1 h at room temperature. The sample was subjected to a temperature gradient (25-95 °C) in a thermal shift instrument, and the melting temperature (Tm) of the protein was calculated based on fluorescence signals from a dye that binds to unfolded protein. The change in Tm was used to evaluate the compound’s binding affinity and covalent interaction with BRD4(BD1) [1]
2. MALDI-TOF mass spectrometry assay for covalent adduct detection: Purified BRD4(BD1) was incubated with ZEN-3219 (100 nM) for 2 h at 37 °C, then desalted to remove unbound compound. The protein sample was analyzed via MALDI-TOF mass spectrometry, and the mass shift of BRD4(BD1) was calculated to confirm the formation of a covalent adduct (mass increase consistent with the molecular weight of ZEN-3219 minus the epoxide ring-opening moiety) [1] 3. X-ray crystallography assay: ZEN-3219 was co-crystallized with BRD4(BD1) protein, and the crystal structure was resolved at 1.8 Å resolution. The structure confirmed that the epoxide warhead of ZEN-3219 forms a covalent bond with the sulfur atom of Met149 in BRD4(BD1), while the core scaffold of the compound occupies the AcLys binding pocket, blocking the interaction with acetylated histones [1] |
| Cell Assay |
1. Cell proliferation assay: MV4-11 cells were seeded in 96-well plates at a density of 5×10³ cells per well and incubated for 12 h to attach. The cells were treated with serial dilutions of ZEN-3219 (1-1000 nM) for 24 h, followed by washing to remove unbound compound and further incubation for 48 h. A cell viability detection reagent was added to each well and incubated for 3 h at 37 °C, and absorbance was measured to calculate cell proliferation inhibition rates and evaluate the durability of the compound’s antiproliferative effect [1]
2. qRT-PCR for gene transcription detection: MV4-11 cells were treated with ZEN-3219 (50 nM) for 24 h, with a non-covalent BET inhibitor as a control (same concentration). Total RNA was extracted from the cells, reverse-transcribed into cDNA, and amplified using specific primers for MYC, BCL2, and reference genes. The relative mRNA expression levels were calculated via the 2^(-ΔΔCt) method to assess the compound’s effect on BRD4-dependent gene transcription [1] |
| References | |
| Additional Infomation |
1. ZEN-3219 is a novel covalent BET inhibitor designed by linking an epoxy group to a non-covalent BRD4(BD1) inhibitor backbone, which targets the Met149 residue in the BRD4(BD1) binding pocket[1]. 2. The covalent binding mechanism of ZEN-3219 is the reaction of the epoxy group with the sulfur-containing side chain of Met149 to form a stable covalent adduct, which irreversibly blocks the AcLys binding pocket of BRD4(BD1), thereby persistently inhibiting BRD4-mediated transcriptional regulation[1]. 3. ZEN-3219 represents a new BET inhibitor development strategy, namely, by covalently modifying methionine residues, this strategy has higher target selectivity and more durable efficacy compared with non-covalent BET inhibitors[1].
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| Molecular Formula |
C19H18N2O3
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|---|---|
| Molecular Weight |
322.36
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| Exact Mass |
322.131
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| CAS # |
1952264-34-4
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| PubChem CID |
121414852
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| Appearance |
White to off-white solid powder
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| LogP |
1.8
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
24
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| Complexity |
551
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1(=CC=C(CN2C=C(C3C(=NOC=3C)C)C=CC2=O)C=C1)C1OC1
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| InChi Key |
IELJHNIVCXDKQD-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H18N2O3/c1-12-19(13(2)24-20-12)16-7-8-18(22)21(10-16)9-14-3-5-15(6-4-14)17-11-23-17/h3-8,10,17H,9,11H2,1-2H3
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| Chemical Name |
5-(3,5-dimethyl-1,2-oxazol-4-yl)-1-[[4-(oxiran-2-yl)phenyl]methyl]pyridin-2-one
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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 : ~25 mg/mL (~77.55 mM)
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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 | 3.1021 mL | 15.5106 mL | 31.0212 mL | |
| 5 mM | 0.6204 mL | 3.1021 mL | 6.2042 mL | |
| 10 mM | 0.3102 mL | 1.5511 mL | 3.1021 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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