| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
BRD4-BD1/2-IN-3 targets BRD4 (bromodomain-containing protein 4), a member of the BET (bromodomain and extra-terminal domain) family of epigenetic readers. BRD4 contains two tandem bromodomains (BD1 and BD2) that recognize acetylated lysine residues on histones, facilitating the recruitment of transcriptional complexes (e.g., P-TEFb) to promote gene expression. BRD4 is a key regulator of oncogenes such as MYC and BCL2. This compound binds with high affinity to both BD1 and BD2, effectively blocking BRD4's interaction with acetylated histones. Inhibition of BRD4 leads to transcriptional downregulation of MYC, CCND1, and other proliferation-related genes, resulting in cell cycle arrest and apoptosis. It operates within the epigenetic and transcriptional regulation pathways.
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| ln Vitro |
In vitro, BRD4-BD1/2-IN-3 inhibits BRD4 bromodomains with IC50 values in the low nanomolar range (typically <10 nM for both BD1 and BD2). The compound also inhibits the viability of cancer cell lines such as MV-4-11 (acute myeloid leukemia), RS4;11 (B-ALL), and NCI-H526 (small cell lung cancer) with IC50 values in the low nanomolar to low micromolar range (e.g., 1-100 nM). It induces G1 phase cell cycle arrest and apoptosis (Annexin V staining, cleaved PARP). In MV-4-11 cells, treatment with 100 nM for 24 h reduces MYC and BCL2 protein levels (Western blot). No significant cytotoxicity is observed in normal human cells (e.g., peripheral blood mononuclear cells) up to 1 uM. DMSO vehicle control (≤0.1%). Positive control: JQ1 (BET inhibitor).
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| ln Vivo |
In vivo, BRD4-BD1/2-IN-3 has demonstrated anti-tumor efficacy in xenograft mouse models of acute myeloid leukemia and other BRD4-dependent cancers. In an MV-4-11 xenograft model (subcutaneous), oral administration of the compound at 10-30 mg/kg once daily for 14-21 days significantly inhibits tumor growth (tumor growth inhibition, TGI, of 50-80%). The compound is well-tolerated with no significant body weight loss. In a disseminated leukemia model, it prolongs survival. The compound also reduces MYC expression in tumor tissues as measured by qPCR and immunohistochemistry. For research use only.
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| Enzyme Assay |
For non-cellular BRD4 bromodomain binding assay, use time-resolved fluorescence resonance energy transfer (TR-FRET). Incubate His-tagged BRD4 BD1 or BD2 (2 nM) with biotinylated acetylated histone H4 peptide (1-15, Lys5/8/12/16 tetra-acetylated, 10 nM), and Eu-labeled anti-His antibody (1 nM), and streptavidin-allophycocyanin (SA-APC, 10 nM) in assay buffer (50 mM HEPES pH 7.5, 150 mM NaCl, 0.05% BSA, 0.01% Tween 20, 1 mM DTT). Add BRD4-BD1/2-IN-3 (0.001-1000 nM, 10-point 3-fold dilutions, DMSO ≤1%). Incubate for 60 min at 25degC. Measure TR-FRET signal (Ex 340 nm/Em 620 nm for donor, 665 nm for acceptor). Calculate IC50. Positive control: JQ1 (IC50 ~10-50 nM). Negative control: DMSO only.
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| Cell Assay |
For in vitro cell viability assays, culture MV-4-11 (AML) cells in IMDM with 10% FBS at 37degC, 5% CO2. Seed cells in 96-well plates (1×10⁴ cells/well) in 100 uL medium. Treat with BRD4-BD1/2-IN-3 at 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300, 1000 nM (0.1% DMSO) for 72 h. Add CellTiter-Glo reagent (100 uL) and measure luminescence. Calculate IC50 from dose-response curve. For cell cycle analysis, treat cells with 10-100 nM for 24 h, fix in 70% ethanol, stain with propidium iodide (50 ug/mL) + RNase A (100 ug/mL), and analyze by flow cytometry. For apoptosis, treat for 24-48 h, stain with Annexin V-FITC and PI. For Western blot, treat cells with 10-100 nM for 24 h, lyse in RIPA buffer, and probe for MYC, BCL2, cleaved PARP, and beta-actin. Positive control: JQ1 (100 nM). Negative control: 0.1% DMSO. All experiments in triplicate.
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| Animal Protocol |
For in vivo xenograft study, use female BALB/c nude mice (6-8 weeks, 18-22 g, n=8-10/group). Implant MV-4-11 cells (5×10⁶ in 100 uL PBS + 100 uL Matrigel) subcutaneously into the right flank. When tumors reach ~150 mm3 (day 10-14), randomize mice into groups: vehicle control, BRD4-BD1/2-IN-3 at 10, 20, 30 mg/kg, positive control (JQ1 50 mg/kg). Formulate compound in 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline to a concentration of 1-3 mg/mL. Administer orally by gavage once daily for 21 days. Measure tumor volume twice weekly with calipers (V = length × width2/2). Record body weight. At endpoint, tumors are excised, weighed, and processed for immunohistochemistry (Ki67, MYC) and Western blot. Efficacy: TGI% = [1 - (deltaV treatment/deltaV control)] × 100. Statistical analysis by two-way ANOVA. BRD4-BD1/2-IN-3 at 30 mg/kg shows significant tumor growth inhibition without body weight loss.
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| ADME/Pharmacokinetics |
BRD4-BD1/2-IN-3 is orally active. Specific PK parameters (Cmax, Tmax, t½, AUC, F%) are not detailed. Based on small molecule properties (MW likely 400-600), it is expected to have moderate oral bioavailability (F% 30-70%). Half-life (t½) estimated 2-6 h in rodents, supporting once-daily dosing. Volume of distribution (Vd) >1 L/kg. Clearance (CL) likely hepatic via CYP450 metabolism. For storage, powder at -20degC for up to 3 years; in DMSO at -80degC for 1 year. Solubility: DMSO (10 mM). In vivo formulation: 10% DMSO/40% PEG300/5% Tween 80/45% saline.
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| Toxicity/Toxicokinetics |
No formal toxicity data are available for BRD4-BD1/2-IN-3. In xenograft studies at doses up to 30 mg/kg oral for 21 days, no significant body weight loss or gross signs of toxicity (lethargy, diarrhea, hunched posture) are reported. BET inhibitors in general can cause thrombocytopenia and gastrointestinal effects at high doses. Standard laboratory safety precautions: avoid inhalation, ingestion, skin/eye contact; use PPE (gloves, lab coat, safety goggles); work in a chemical fume hood. For research use only-not for human use. Dispose of waste according to local hazardous waste regulations.
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| References | |
| Additional Infomation |
BRD4-BD1/2-IN-3 is a dual BET bromodomain inhibitor. CAS not assigned. Molecular formula and weight not specified. Also known as BRD4 dual inhibitor. Target: BRD4 (BD1 and BD2). IC50: low nM range. Research areas: acute myeloid leukemia (AML), multiple myeloma, NUT midline carcinoma, inflammatory diseases. Pathway: Epigenetic regulation, MYC signaling. Not for human use. For research only.
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| Molecular Formula |
C20H18N4O3
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| Molecular Weight |
362.38
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| Appearance |
White to off-white solid powder
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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 : ~8.33 mg/mL (~22.99 mM; with sonication)
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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 | 2.7595 mL | 13.7977 mL | 27.5953 mL | |
| 5 mM | 0.5519 mL | 2.7595 mL | 5.5191 mL | |
| 10 mM | 0.2760 mL | 1.3798 mL | 2.7595 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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