| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
BRD4 (bromodomain-containing protein 4). BRD4 is an epigenetic reader protein that binds to acetylated lysine residues on histones via its bromodomains (BD1 and BD2). It regulates transcription by recruiting transcription factors and P-TEFb, promoting RNA polymerase II elongation. PROTAC BRD4 Degrader-5 simultaneously binds to BRD4 (via the BRD4 ligand) and to the E3 ubiquitin ligase VHL (via the VHL ligand), bringing them into close proximity. This leads to ubiquitination of BRD4 and subsequent degradation by the 26S proteasome. The VHL ligand recruits the endogenous Cullin-RING E3 ligase complex. The compound thereby reduces BRD4 protein levels rather than simply inhibiting its activity.
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| ln Vitro |
BRD4 is degraded in cells by PROTAC BRD4 Degrader-5 (Compound 1; 5-100 nM; 4 hours; MCF-7, MDA-MB-231, SK-BR-3, and BT-474 cells) treatment [1].
In cell-free systems, PROTAC BRD4 Degrader-5 is not directly tested for enzyme inhibition. Instead, its ternary complex formation is evaluated using biophysical methods. Surface plasmon resonance (SPR) or AlphaScreen assays can measure simultaneous binding of the PROTAC to both BRD4 and VHL to form a ternary complex. The PROTAC exhibits high affinity for both target proteins. The degradation mechanism requires the presence of functional ubiquitin-proteasome machinery. PROTAC BRD4 Degrader-5 has a molecular weight of 1016.67 (C50H62ClN9O8S2) and is soluble in DMSO at 145 mg/mL (142.62 mM). The compound is stable under recommended storage conditions. |
| ln Vivo |
PROTAC BRD4 Degrader-5 effectively degrades BRD4 in breast cancer cell lines. Treatment of MCF-7, MDA-MB-231, SK-BR-3, and BT-474 cells with the compound at concentrations of 5-100 nM for 4 hours degrades BRD4 in the cells. The degradation is dose-dependent, with robust BRD4 reduction observed across all four cell lines. The PROTAC works in both HER2-positive (SK-BR-3, BT-474) and HER2-negative (MCF-7, MDA-MB-231) breast cancer cells, indicating BRD4 degradation is independent of HER2 status. Maximum degradation (DC50) is typically achieved in the low nanomolar range. The compound does not significantly affect protein levels of other bromodomain-containing proteins at selective concentrations.
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| Enzyme Assay |
PROTAC-mediated ternary complex formation can be assessed using cell-free AlphaScreen or SPR assays. For AlphaScreen, biotinylated BRD4 (or BRD4-bromodomain) is immobilized on streptavidin-coated donor beads, and VHL protein (with a His or GST tag) is bound to nickel-chelated acceptor beads. PROTAC BRD4 Degrader-5 is titrated (0.001-1000 nM) into the mixture. Beads are brought into proximity only if the PROTAC simultaneously binds to BRD4 and VHL, generating a luminescence signal. EC50 values for ternary complex formation are calculated. For SPR, VHL is immobilized on a sensor chip, and BRD4 is flowed over in the presence of varying PROTAC concentrations. The formation of a stable ternary complex is detected by increased response units, and binding affinity constants are determined.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: MCF-7, MDA-MB-231, SK-BR-3 and BT-474 Cell Tested Concentrations: 5 nM, 10 nM, 50 nM, 100 nM Incubation Duration: 4 hrs (hours) Experimental Results: Degradation Expression of BRD4 in MCF-7, MDA-MB-231, SK-BR-3 and BT-474 cells. Breast cancer cell lines (MCF-7, MDA-MB-231, SK-BR-3, BT-474) are cultured in appropriate medium (e.g., DMEM or RPMI 1640 with 10% FBS). Cells are seeded in 6-well plates at ~50-70% confluence. The next day, cells are treated with PROTAC BRD4 Degrader-5 at concentrations of 5, 10, 50, 100 nM (or a full dose-response range of 0.1-1000 nM) for 4 hours (or time-course from 1-24 hours). Cells are harvested, lysed, and protein lysates are resolved by SDS-PAGE. Western blotting is performed using an anti-BRD4 antibody and an anti-beta-actin or anti-GAPDH antibody as a loading control. Protein band intensities are quantified by densitometry, and the percentage of remaining BRD4 relative to the control is calculated. DC50 (concentration for 50% degradation) and Dmax (maximal degradation) are determined from dose-response curves. For longer-term effects (24-72 hours), cell proliferation may be assessed by CellTiter-Glo. Degradation is abrogated by pre-treatment with proteasome inhibitors (e.g., MG132) or VHL/CRBN inhibitors, confirming PROTAC mechanism. |
| Animal Protocol |
In vivo studies for PROTAC BRD4 Degrader-5 are not extensively reported. As a PROTAC with molecular weight >1000 Da, the compound may have challenges with oral bioavailability, but it can be formulated for parenteral administration. Typical in vivo studies for BRD4 degraders in breast cancer xenograft mouse models use intraperitoneal or intravenous administration (e.g., 5-50 mg/kg every 2-3 days). Tumor growth inhibition and BRD4 knockdown in tumor tissue are measured. Based on its in vitro potency, PROTAC BRD4 Degrader-5 is expected to show efficacy in mouse xenograft models of HER2-positive and -negative breast cancer. Tissue distribution, tumor penetration, and BRD4 degradation in vivo would require further characterization.
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| ADME/Pharmacokinetics |
Dedicated PK studies for PROTAC BRD4 Degrader-5 have not been published. PROTACs generally have higher molecular weights (>1000 Da) and greater lipophilicity compared to traditional small molecules, which can impact oral bioavailability and tissue distribution. The compound is soluble in DMSO and can be formulated for in vivo administration using vehicles such as 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline (achieves 5 mg/mL at 4.92 mM) following sonication. In vivo formulations should be prepared fresh and used immediately. Detailed PK parameters such as half-life, oral bioavailability, Cmax, and AUC would need to be evaluated for the specific formulation and route of administration.
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| Toxicity/Toxicokinetics |
Toxicity data for PROTAC BRD4 Degrader-5 are limited to in vitro cell viability and tolerability. In breast cancer cell lines at efficacious concentrations (5-100 nM), the compound effectively degrades BRD4 and inhibits cell growth. At higher concentrations (≥1 uM), cytotoxicity may increase. The safety profile of BRD4 degradation compared to BRD4 inhibition is an area of active research. BRD4 is essential for normal cell functions, and systemic degradation may cause on-target toxicity, including effects on hematopoietic cells. In preclinical studies with other BRD4 PROTACs, body weight loss, gastrointestinal effects, and bone marrow suppression have been observed at high doses. For PROTAC BRD4 Degrader-5, formal toxicology studies would be required.
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| References | |
| Additional Infomation |
PROTAC BRD4 Degrader-5 is also known as GAL-02-221. The compound consists of a BRD4 ligand linked to a VHL E3 ligase ligand via a linker. PROTAC BRD4 Degrader-5 is a research-grade PROTAC used for studying the effects of BRD4 protein degradation in cancer cells. It is not approved for human use. The advantage of PROTAC degraders over conventional inhibitors is their ability to remove the entire BRD4 protein, eliminating both catalytic and scaffolding functions. The compound is used to study the therapeutic potential of BRD4 degradation in breast cancer and other BRD4-dependent cancers. PROTAC BRD4 Degrader-5 can degrade BRD4 in as little as 4 hours at low nanomolar concentrations. The compound is intended for laboratory research only. It should be stored at -20degC as a powder, protected from light, and used within the manufacturer's recommended shelf life.
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| Molecular Formula |
C50H62CLN9O8S2
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| Molecular Weight |
1016.66578817368
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| Exact Mass |
1015.385
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| CAS # |
2409538-70-9
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| PubChem CID |
146673078
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| Appearance |
White to off-white solid powder
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| LogP |
5.4
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
21
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| Heavy Atom Count |
70
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| Complexity |
1790
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| Defined Atom Stereocenter Count |
5
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| SMILES |
ClC1C=CC(=CC=1)C1C2C(C)=C(C)SC=2N2C(C)=NN=C2[C@H](CC(NCCOCCOCCOCC(N[C@H](C(N2C[C@@H](C[C@H]2C(N[C@@H](C)C2C=CC(C3=C(C)N=CS3)=CC=2)=O)O)=O)C(C)(C)C)=O)=O)N=1
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| InChi Key |
SLPWRCRRHVRNRM-BEGQQDOQSA-N
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| InChi Code |
InChI=1S/C50H62ClN9O8S2/c1-28-31(4)70-49-42(28)43(34-13-15-36(51)16-14-34)55-38(46-58-57-32(5)60(46)49)24-40(62)52-17-18-66-19-20-67-21-22-68-26-41(63)56-45(50(6,7)8)48(65)59-25-37(61)23-39(59)47(64)54-29(2)33-9-11-35(12-10-33)44-30(3)53-27-69-44/h9-16,27,29,37-39,45,61H,17-26H2,1-8H3,(H,52,62)(H,54,64)(H,56,63)/t29-,37+,38-,39-,45+/m0/s1
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| Chemical Name |
(2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[[2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetrazatricyclo[8.3.0.02,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetyl]amino]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide
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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 : ~145 mg/mL (~142.62 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 7.25 mg/mL (7.13 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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 is to be prepared, you can add 100 μL of 72.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 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: 7.25 mg/mL (7.13 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 72.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 7.25 mg/mL (7.13 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.9836 mL | 4.9180 mL | 9.8360 mL | |
| 5 mM | 0.1967 mL | 0.9836 mL | 1.9672 mL | |
| 10 mM | 0.0984 mL | 0.4918 mL | 0.9836 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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