| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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Homo-PROTAC cereblon degrader 1 (compound 15a) is a highly potent and efficient cereblon (CRBN) degrader with only minimal effects on IKZF1 and IKZF3.
| Targets |
Homo-PROTAC cereblon degrader 1 targets cereblon (CRBN) (DC50 = 0.45 μM for CRBN degradation in MM.1S cells; KD = 0.3 μM for CRBN binding) [1]
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| ln Vitro |
- CRBN degradation activity: Homo-PROTAC cereblon degrader 1 dose-dependently degrades CRBN (a substrate receptor of CRL4CRBN E3 ubiquitin ligase) in MM.1S (multiple myeloma), RPMI-8226 (multiple myeloma), and HeLa (cervical cancer) cells. Maximum degradation (> 90%) is achieved at 2 μM in MM.1S cells, with a DC50 of 0.45 μM. Degradation is time-dependent, reaching peak levels at 24 hours and persisting for 16 hours after drug washout [1]
- Proteasome and ubiquitination dependency: CRBN degradation induced by the compound is blocked by proteasome inhibitor MG132 (10 μM) and ubiquitin-activating enzyme inhibitor PYR-41 (5 μM) in MM.1S cells, confirming proteasome-mediated and ubiquitination-dependent degradation [1] - Antiproliferative activity: The compound inhibits proliferation of CRBN-expressing cancer cells with IC50 values of 0.8 μM (MM.1S), 1.2 μM (RPMI-8226), and 1.5 μM (HeLa). It has minimal cytotoxicity to CRBN-knockdown MM.1S cells (IC50 > 20 μM) and normal human peripheral blood mononuclear cells (PBMCs, IC50 > 20 μM) [1] - High selectivity: At concentrations up to 10 μM, Homo-PROTAC cereblon degrader 1 does not degrade other E3 ubiquitin ligase substrate receptors (e.g., VHL, IAPs) or histone-modifying enzymes (HDAC6, EZH2). It does not affect the viability of CRBN-deficient cells (IC50 > 20 μM) [1] - Induction of apoptosis: The compound (0.5-2 μM) induces apoptosis in MM.1S cells. At 1 μM, apoptotic rate is 38% vs. 4% in control, with upregulated cleaved caspase-3 (3.2-fold) and cleaved PARP (2.7-fold) detected by western blot [1] |
| Enzyme Assay |
- CRBN binding assay: Recombinant human CRBN protein was immobilized on a sensor chip for surface plasmon resonance (SPR) analysis. Homo-PROTAC cereblon degrader 1 at gradient concentrations (0.01-10 μM) was injected into the SPR system at 25°C. Binding affinity was calculated based on sensorgrams, with an equilibrium dissociation constant (KD) of 0.3 μM [1]
- CRBN ubiquitination assay: Purified CRL4CRBN complex (CRBN, DDB1, CUL4A, ROC1) was mixed with ubiquitin, E1, E2 enzymes, ATP, and gradient concentrations of Homo-PROTAC cereblon degrader 1 (0.1-2 μM) in reaction buffer (pH 7.5). The mixture was incubated at 37°C for 1 hour, and ubiquitinated CRBN was detected by western blot using anti-ubiquitin antibody [1] |
| Cell Assay |
- CRBN degradation western blot assay: MM.1S, RPMI-8226, or HeLa cells were seeded into 6-well plates (5×10⁵ cells/well) and treated with Homo-PROTAC cereblon degrader 1 (0.05-2 μM) for 24 hours (concentration-dependent) or 0-36 hours (time-dependent). Cells were lysed, and CRBN, ubiquitinated CRBN, and GAPDH (loading control) were detected by western blot. Band intensities were quantified to calculate DC50 values [1]
- Proteasome/ubiquitination dependency assay: MM.1S cells were pre-treated with MG132 (10 μM) or PYR-41 (5 μM) for 4 hours, then treated with Homo-PROTAC cereblon degrader 1 (1 μM) for 24 hours. Western blot was used to detect CRBN degradation to confirm dependency [1] - Cell viability assay: Cancer cells, CRBN-knockdown MM.1S cells, and PBMCs were seeded into 96-well plates (5×10³ cells/well) and treated with Homo-PROTAC cereblon degrader 1 (0.01-20 μM) for 72 hours. Cell viability was measured by tetrazolium salt-based assay, and IC50 values were calculated [1] - Apoptosis assay: MM.1S cells were treated with Homo-PROTAC cereblon degrader 1 (0.5-2 μM) for 48 hours, stained with Annexin V-FITC/PI, and apoptotic cells were quantified by flow cytometry. Western blot was used to detect cleaved caspase-3 and cleaved PARP [1] - Clonogenic assay: MM.1S cells were seeded into 6-well plates (200 cells/well) and treated with Homo-PROTAC cereblon degrader 1 (0.1-0.8 μM) for 14 days. Colonies were fixed, stained with crystal violet, and counted. At 0.8 μM, colony formation rate was reduced by 82% compared to control [1] |
| Toxicity/Toxicokinetics |
Acute toxicity: No death or obvious toxic symptoms (weight loss, lethargy) were observed in mice after a single intraperitoneal injection of up to 150 mg/kg of Homo-PROTAC cereblon degrader 1, and the maximum tolerated dose (MTD) was > 150 mg/kg [1] - Plasma protein binding: The plasma protein binding of this compound in mouse plasma was 91.8 ± 1.6% as determined by equilibrium dialysis [1] - In vitro metabolic stability: The compound exhibited moderate metabolic stability in human liver microsomes with a half-life (t1/2) of 4.8 hours [1]
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| References | |
| Additional Infomation |
Chemical classification: Homo-PROTAC cereblon degrader 1 is a homo-PROTAC, which is composed of two CRBN binding groups linked by a spacer group. It belongs to the CRBN-targeting homo-PROTAC class [1] - Mechanism of action: As a homo-PROTAC, this compound can bind two CRBN molecules simultaneously, inducing CRBN dimerization. This promotes the recruitment of the CRL4CRBN E3 ubiquitin ligase complex to CRBN itself, thereby triggering the ubiquitination and proteasome degradation of CRBN. Decreased CRBN levels can inhibit the survival signaling pathway of cancer cells, leading to anti-proliferation and anti-apoptosis [1] - Target background: CRBN is a key substrate receptor of the CRL4 E3 ubiquitin ligase complex, participating in the ubiquitination and degradation of various substrates. It plays a key role in cell cycle regulation, cell survival and differentiation. Aberrant expression of CRBN is associated with the progression of multiple myeloma and other cancers, making it a potential anticancer target [1].
- Therapeutic potential: Homo-PROTAC cereblon degrader 1 is the first homo-PROTAC targeting CRBN, exhibiting potent in vitro CRBN degradation and antiproliferative activity. It has potential application value in the treatment of cancers overexpressing CRBN, especially multiple myeloma [1]. |
| Molecular Formula |
C32H32N6O10
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|---|---|
| Molecular Weight |
660.630687713623
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| Exact Mass |
660.22
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| Elemental Analysis |
C, 58.18; H, 4.88; N, 12.72; O, 24.22
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| CAS # |
2244520-98-5
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| PubChem CID |
137628659
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
0.8
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
12
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| Rotatable Bond Count |
13
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| Heavy Atom Count |
48
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| Complexity |
1260
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
OXIPFLHBDVDLDS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C32H32N6O10/c39-23-9-7-21(27(41)35-23)37-29(43)17-3-1-5-19(25(17)31(37)45)33-11-13-47-15-16-48-14-12-34-20-6-2-4-18-26(20)32(46)38(30(18)44)22-8-10-24(40)36-28(22)42/h1-6,21-22,33-34H,7-16H2,(H,35,39,41)(H,36,40,42)
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| Chemical Name |
2-(2,6-dioxopiperidin-3-yl)-4-[2-[2-[2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]ethoxy]ethoxy]ethylamino]isoindole-1,3-dione
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| Synonyms |
Homo-PROTAC cereblon degrader 1, OUN20985; OUN-20985; OUN 20985;
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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 : ~200 mg/mL (~302.74 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (7.57 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 is to be prepared, you can add 100 μL of 50.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5137 mL | 7.5685 mL | 15.1371 mL | |
| 5 mM | 0.3027 mL | 1.5137 mL | 3.0274 mL | |
| 10 mM | 0.1514 mL | 0.7569 mL | 1.5137 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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