| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
BRD2 (BD2) 7.64 μM (Kd) BRD2 (BD1) 41.37 μM (Kd)
BRD2 (bromodomain‑containing protein 2). BBC0403 binds to the second bromodomain (BD2) of BRD2 with higher affinity than to the first bromodomain (BD1). |
|---|---|
| ln Vitro |
BBC0403 (5μM, 10μM and 20μM, 12 hours) reduces the expression of catabolic factors, the production of prostaglandin E2 (PGE2) and the degradation of the extracellular matrix (ECM). BBC0403 prevents OA by inhibiting NF-κB and MAPK signaling[1].
In cell‑free binding assays, BBC0403 binds to BRD2 with Kd values of 7.64 uM for BRD2 (BD2) and 41.37 uM for BRD2 (BD1). The compound exhibits higher binding specificity for BRD2 compared to BRD3 and BRD4. In vitro (5‑20 uM, 12 hours), BBC0403 reduces the expression of catabolic factors, the production of prostaglandin E2 (PGE2), and the degradation of the extracellular matrix (ECM). |
| ln Vivo |
BBC0403 (5-10 μg/kg; intra-articular injection; once a week; 6 weeks) prevents cartilage degradation in OA mice [1].
|
| Enzyme Assay |
A general cell‑free protocol for BRD2 binding affinity uses a fluorescence polarization (FP) assay. Recombinant BRD2(BD2) protein is incubated with a fluorescently labeled tetra‑acetylated histone H4 peptide (H4K5/8/12/16ac) in a buffer (50 mM HEPES, pH 7.5, 150 mM NaCl, 0.01% Tween‑20, 1 mM DTT). Various concentrations of BBC0403 are added, and the mixture is incubated for 30‑60 minutes at room temperature. The change in fluorescence polarization is measured, and the IC₅0 is calculated. Kd is then derived using the Cheng‑Prusoff equation.
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| Cell Assay |
A general cellular protocol for BRD2 inhibitors in OA research: Isolate primary human chondrocytes or use chondrocyte cell lines. Treat cells with BBC0403 at concentrations of 5, 10, and 20 uM for 12 hours. Stimulate the cells with inflammatory cytokines such as IL‑1beta or TNF‑alpha (e.g., 10 ng/mL) to induce catabolic factor expression. Measure the expression of catabolic factors (MMP‑13, ADAMTS‑4, ADAMTS‑5) by qRT‑PCR and PGE2 production by ELISA. Assess ECM degradation by analyzing glycosaminoglycan release into the culture medium.
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| Animal Protocol |
Animal/Disease Models:Male C57BL/6 mice (12 weeks old; 18-20 g) bearing osteoarthritis (OA)[1]
Doses: 5 μg/kg, 10 μg/kg Route of Administration: Intra-articular (i.a.) injection; once per week; 6 weeks Experimental Results: Prevented OA cartilage degradation in mice. A general animal protocol for BRD2 inhibitors: Use a mouse model of osteoarthritis induced by destabilization of the medial meniscus (DMM) surgery. Administer BBC0403 intra‑articularly or systemically via oral gavage at doses such as 10‑50 mg/kg daily for 4‑8 weeks. Evaluate OA severity by histological scoring (OARSI score) of joint sections stained with Safranin‑O/fast green. Assess synovial inflammation by measuring inflammatory cytokine levels. Measure serum and joint PGE2 levels by ELISA. |
| References | |
| Additional Infomation |
BBC0403 suppresses NF‑kappaB and MAPK signaling pathways, which are key mediators of inflammation and cartilage degradation in OA. The compound is available with a purity of ≥95% and has a molecular formula of C21H22N2O₅ and a molecular weight of 382.41. It is stored as a powder at ‑20degC for up to 3 years.
|
| Molecular Formula |
C21H22N2O5
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|---|---|
| Molecular Weight |
382.41
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| CAS # |
2644662-83-7
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| Appearance |
Solid powder
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| Density |
1.33±0.1 g/cm3(Temp: 20 °C; Press: 760 Torr)(predicted)
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| Boiling Point |
589.8±60.0 °C(predicted)
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| LogP |
0
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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 (65.37 mM; with sonication (<60°C))
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.54 mM)(Saturation unknown) in 10% DMSO 40% PEG300 5% Tween-80 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix well; then add 50 μL Tween-80 to the above system and mix well; then add 450 μL saline to make it 1 mL. *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.54 mM)(saturation unknown) in 10% DMSO 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6150 mL | 13.0750 mL | 26.1499 mL | |
| 5 mM | 0.5230 mL | 2.6150 mL | 5.2300 mL | |
| 10 mM | 0.2615 mL | 1.3075 mL | 2.6150 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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