PROTAC BRD4 Degrader-48
PROTAC BRD4 Degrader-48 is a BRD4 degrader with a Kd of 0.69 μM for human BRD4 and a DC50 of 0.21 μM. PROTAC BRD4 Degrader-48 binds to BRD4 and GID4 to form a ternary complex, and induces ubiquitination and degradation of BRD4 via the ubiquitin-proteasome pathway. PROTAC BRD4 Degrader-48 inhibits proliferation and migration of tumor cells, blocks tumor growth, and maintains degrading activity even in VHL- and CRBN-deficient models. PROTAC BRD4 Degrader-48 can be used in studies related to renal cell carcinoma.
(Pink: BRD4 ligand (HY-78695); Blue: no E3 ligase ligand; Black: linker (HY-W014883)).
For research use only. We do not sell to patients.
- Formula: C49H64ClN9O3S
- Molecular Weight:894.61
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
All PROTACs Isoforms
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Biological Activity
Description
IC50 & Target
[1]|
BRD4 0.69 μM (Kd) |
BRD4 0.21 μM (DC50) |
In Vitro
PROTAC BRD4 Degrader-48 (compound a11) binds to purified GID4 protein with moderate affinity, with a KD value of 4.76 μM; it forms a stable cooperative ternary complex with BRD4-BD1 and GID4, with a ternary complex KD of 0.69 μM and a cooperativity factor of 6.90[1].
PROTAC BRD4 Degrader-48 (0.5-10 μM, 500 nM; 10 h) induces concentration-dependent degradation of BRD4-BD1 in HEK293T reporter cells[1].
PROTAC BRD4 Degrader-48 (14-3333 nM, 3 μM; 24 h, 2-24 h) induces efficient, concentration- and time-dependent degradation of BRD4 in VHL-deficient 786-O renal cell carcinoma cells, with a DC50 of 0.21 μM[1].
PROTAC BRD4 Degrader-48 (3 μM; 24 h, with 3 h pretreatment) mediates the degradation of BRD4 in VHL-deficient 786-O renal cell carcinoma cells via the ubiquitin-proteasome system[1].
PROTAC BRD4 Degrader-48 (3 μM; 24 h, 2 h pretreatment) induces BRD4 degradation in VHL-deficient 786-O renal cell carcinoma cells, a process that requires simultaneous binding to both BRD4 and GID4[1].
PROTAC BRD4 Degrader-48 (3 μM; 16 h) promotes the formation of the BRD4-a11-GID4 ternary complex in 786-O renal cell carcinoma cells and HEK293T cells expressing Flag-GID4[1].
PROTAC BRD4 Degrader-48 (3 μM; 24 h, 48 h siRNA transfection, 24 h transfection) induces strictly GID4-dependent BRD4 degradation in 786-O renal carcinoma cells and HEK293T cells[1].
PROTAC BRD4 Degrader-48 (1.5-3333 nM; 24 h) retains potent BRD4-degrading activity in CRBN-deficient 786-O renal cell carcinoma (RCC) cells, and can overcome resistance to PROTACs targeting CRBN and VHL[1].
PROTAC BRD4 Degrader-48 (0.12-10 μM; 24 h) induces concentration-dependent degradation of BRD4 in MDA-MB-231 breast cancer cells and A549 lung cancer cells, and its potency correlates with the intracellular expression level of GID4[1].
PROTAC BRD4 Degrader-48 exhibits potent antiproliferative activity in both parental and CRBN-deficient 786-O renal cell carcinoma cells, with IC50 values of 4.40 μM and 5.00 μM, respectively[1].
PROTAC BRD4 Degrader-48 (3 μM; 12 h, 24 h) potently inhibits the lateral migration of VHL-deficient 786-O renal cell carcinoma cells at a concentration of 3 μM[1].
PROTAC BRD4 Degrader-48 (1.5-3 μM; 24 h) inhibits Transwell migration of VHL-deficient 786-O renal cell carcinoma cells in a dose-dependent manner, and exhibits stronger efficacy than MZ1 (HY-107425) at concentrations of 1.5 μM and 3 μM[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:VHL-deficient 786-O renal cell carcinoma (RCC) cells
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Concentration:14, 41, 123, 370, 1111, 3333 nM
3 μM -
Incubation Time:24 h
2, 4, 6, 8, 12, 24 h -
Result:Induced concentration-dependent BRD4 degradation with a DC50 of 0.21 μM after 24 h treatment.
Caused ~50% BRD4 degradation within 8 h and nearly complete degradation after 24 h at 3 μM.
Showed no hook effect at concentrations up to 30 μM.
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Cell Line:VHL-deficient 786-O renal cell carcinoma (RCC) cells
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Concentration:3 μM
100 nM MLN7243 (HY-100487), 1 μM MG132 (HY-13259) (pretreatment) -
Incubation Time:24 h
3 h (pretreatment) -
Result:Completely blocked BRD4 degradation when co-treated with MLN7243 or MG132.
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Cell Line:VHL-deficient 786-O renal cell carcinoma (RCC) cells
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Concentration:3 μM
3 μM JQ1 (HY-13030), 3 μM PFI-7 (HY-D2259), 3 μM compound 2 (pretreatment) -
Incubation Time:24 h
2 h (pretreatment) -
Result:Rescued BRD4 levels from degradation when pretreated with JQ1, PFI-7, or compound 2, with PFI-7 showing stronger blocking efficacy than compound 2.
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Cell Line:VHL-deficient 786-O renal cell carcinoma (RCC) cells, HEK293T cells
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Concentration:3 μM
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Incubation Time:24 h; 48 h (siRNA transfection, 786-O), 24 h (transfection, HEK293T)
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Result:Markedly attenuated BRD4 degradation when GID4 was silenced in 786-O cells.
Fully restored BRD4 degradation after GID4 knockdown when Flag-GID4 was ectopically re-expressed in HEK293T cells.
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Cell Line:786-O cells, CRBN-knockout (KO) 786-O renal cell carcinoma (RCC) cells
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Concentration:0.16, 0.31, 0.63, 1.3, 2.5, 5.0, 10 nM (786-O cells)
1.5, 4.5, 14, 41, 123, 370, 1111, 3333 nM (CRBN-KO 786-O cells) -
Incubation Time:24 h
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Result:Retained robust concentration-dependent BRD4 degradation activity, while MZ1 and dBET1 completely lost degradation activity.
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Cell Line:MDA-MB-231 breast cancer cells, A549 lung cancer cells
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Concentration:0.12, 0.37, 1.1, 3.3, 10 μM
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Incubation Time:24 h
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Result:Induced concentration-dependent BRD4 degradation in both cell lines, with a DC50 of 1.33 μM in A549 cells and 8.80 μM in MDA-MB-231 cells, efficacy correlating with basal GID4 expression levels.
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Cell Line:VHL-deficient 786-O renal cell carcinoma (RCC) cells
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Concentration:3 μM
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Incubation Time:12 h, 24 h
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Result:Significantly delayed scratch closure at 12 and 24 h, showing strong inhibition of lateral cell migration, while MZ1 had little effect.
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Cell Line:VHL-deficient 786-O renal cell carcinoma (RCC) cells
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Concentration:1.5, 3 μM
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Incubation Time:24 h
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Result:Markedly reduced the number of migrating cells in a dose-dependent fashion, with a more potent inhibitory effect than MZ1.
Parmacokinetics
| Species | Dose | Route | T1/2 | Cmax | AUCinf | F |
|---|---|---|---|---|---|---|
| Mice[1] | 15 mg/kg | i.p. | 3.52 h | 4273.43 ng/mL | 15422.47 ng·h/mL | 3.48 % |
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:BALB/c nude (female, 5 weeks old)[1]
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Dosage:15 mg/kg; 30 mg/kg
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Administration:i.p.; every other day; 14 days
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Result:Achieved a tumor growth inhibition (TGI) rate of 54% at 15 mg/kg.
Achieved a tumor growth inhibition (TGI) rate of 67% at 30 mg/kg.
Induced extensive necrosis, marked disruption of tumor structural integrity, pronounced reduction in Ki67-positive proliferating cells, and significant BRD4 degradation in tumor tissues.
Caused no significant effect on mouse body weight.
Showed normal tissue architecture without pathological alterations in major organs (heart, liver, spleen, lung, kidney) via histopathological analysis.
Chemical Information
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Molecular Weight 894.61
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Formula C49H64ClN9O3S
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SMILES
O=C(C[C@H]1C2=NN=C(N2C3=C(C(C4=CC=C(C=C4)Cl)=N1)C(C)=C(S3)C)C)NCCCCCCCCCCCCNC([C@@H]5CC[C@@H](CC5)NC(CNCC6=CC7=C(C=CC=C7)N6)=O)=O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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Cell migration
Cell migration is a method that plays an important role in wound healing, cell differentiation, embryonic development, etc.
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Patient-Derived Xenograft (PDX)
Patient-derived xenograft (PDX) models are generated by engrafting primary human tumor tissue directly into immunodeficient mice, allowing in vivo propagation of patient tumor biology without initial in vitro adaptation. These models are used to preserve key histopathological and molecular characteristics of the original tumor and enable assessment of tumor growth dynamics and therapeutic response in a living organism. The biological readout is tumor engraftment and subsequent growth in the murine host, which reflects the ability of human tumor cells to survive, vascularize, and expand in an immunocompromised microenvironment.
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Patient-Derived Orthotopic Xenograft (PDOX)
Patient-derived orthotopic xenograft (PDOX) modeling implants fresh patient tumor tissue or patient-derived tumor cells into the anatomically corresponding organ or tissue site of immunodeficient mice, usually by surgical orthotopic implantation, to preserve patient tumor histology, local microenvironmental context, invasion, metastatic behavior, and treatment-response features better than subcutaneous implantation. PDOX readouts include tumor engraftment, orthotopic tumor growth, local invasion, metastasis, recurrence after resection, histologic similarity to the donor tumor, biomarker retention, molecular concordance, survival, and response or resistance to therapy. PDOX models are used for preclinical drug testing and individualized therapy evaluation, but engraftment success varies by tumor type and specimen quality.
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Subcutaneous Cell-Line-Derived Xenograft
Subcutaneous cell-line-derived xenograft (CDX) models are established by implanting cultured human cancer cell lines into immunodeficient mice, where the injected cells form localized tumors that can be monitored in vivo as a measure of tumorigenic potential, growth kinetics, and treatment response. These models are widely used in oncology research because they allow reproducible tumor formation and enable comparative assessment of tumor growth between different cell lines or genetic manipulations in a controlled in vivo microenvironment. Subcutaneous implantation of cancer cells in immunodeficient mice is a standard approach for evaluating tumor growth behavior and therapeutic response across multiple cancer types, including prostate, esophageal, pancreatic, and colon cancer models.
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Orthotopic Cell-Line Xenograft
Orthotopic cell-line xenograft models involve implantation of human cancer cell lines into the anatomically corresponding organ of immunodeficient mice to reproduce tumor growth within a native microenvironment, enabling more clinically relevant tumor behavior compared with subcutaneous models. These models are widely used because orthotopic placement better recapitulates tumor progression, including invasion and metastatic spread, which are often underrepresented in heterotopic implantation systems. Compared with conventional xenografts, orthotopic implantation is described as more technically complex but provides improved simulation of tumor-microenvironment interactions and metastatic behavior, making it particularly valuable for translational oncology research. Surgical orthotopic implantation approaches have been emphasized as enabling faithful reproduction of clinical cancer features, including metastasis and disease progression patterns that align with the tumor’s organ of origi
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Cell Viability Determination by MTT Colorimetric Assay
The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.
Purity & Documentation
References
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)