| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
VHL
EED (Embryonic Ectoderm Development), a core subunit of the polycomb repressive complex 2 (PRC2). It binds to EED with a pKD of 9.02 +/- 0.09 and functions as a PRC2 inhibitor with a pIC50 of 8.17 +/- 0.24, targeting the EED subunit. |
|---|---|
| ln Vitro |
PROTAC EED degrader-1 (compound PROTAC 2) binds to EED with a pKD of 9.02±0.09 and has a pIC50 of 8.17±0.24 to inhibit PRC2 function [1]. The EZH2 mutant DLBCL cell line Karpas422 can be successfully inhibited from proliferating by PROTAC EED degrader-1 (0.01-100 μM; 14 days)[1]. In Karpas422 cells, PROTAC EED degrader-1 (0.1-3 μM; 48 hours) lowers the protein levels of EED, EZH2, and H3K27me3 [1]. In Karpas422 cells, PROTAC EED degrader-1 (1 μM; 1–24 hours) stimulates the breakdown of EED, EZH2, and SUZ12. Reduced proliferation was the outcome of the cell proliferation assay [1] using Karpas422 cells as the cell line. Concentration: 0.01, 0.1, 1, 10, 100μM. Incubation time: 4, 8, 12, 14 days. GI50 on Day 14 is 0.045 uM. Results of Western Blot Analysis [1]: Protein levels of EED, EZH2, and H3K27me3 were decreased. Cell Line: Karpas422 cells; Concentration: 0.1, 1, and 3 μM; Incubation Time: 48 hours.
In Karpas422 (EZH2 mutant DLBCL) cells, PROTAC EED degrader-1 (0.01-100 microM; 14 days) successfully inhibits proliferation with a GI50 of 0.045 microM on Day 14. At 0.1-3 microM for 48 hours, it lowers the protein levels of EED, EZH2, and H3K27me3. At 1 microM for 1-24 hours, it stimulates the breakdown of EED, EZH2, and SUZ12. |
| ln Vivo |
No specific in vivo data is available. As a VHL-based PROTAC, when administered to mice, it is expected to be formulated in 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline for IP injection at doses of 10-50 mg/kg, qd or qod, to evaluate degradation of PRC2 proteins in tumor xenografts (e.g., Karpas422) by Western blot and IHC.
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| Enzyme Assay |
The binding affinity to EED is evaluated using surface plasmon resonance (SPR) or fluorescence polarization (FP) assays. For SPR: immobilize EED protein on a sensor chip, then inject PROTAC EED degrader-1 at various concentrations (0.1 nM - 10 uM) to determine the KD. The pKD (negative log of KD) is calculated as 9.02, indicating sub-nanomolar affinity.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: Karpas422 cells Tested Concentrations: 1 μM Incubation Duration: 1, 2, 4, 6, 8, 24 hrs (hours) Experimental Results: EED protein levels diminished within 1-2 h of treatment. (1) Seed Karpas422 cells (5,000-10,000 cells/well) in 96-well plates overnight. (2) Treat with PROTAC EED degrader-1 at 0.01-100 uM for 14 days, changing medium every 2-3 days. (3) Add CellTiter-Glo reagent, measure luminescence to calculate GI50 (0.045 uM). (4) For Western blot: treat cells with 0.1, 1, 3 uM for 48 h, lyse, run SDS-PAGE, blot for EED, EZH2, H3K27me3. (5) For degradation time-course: treat with 1 uM for 1, 2, 4, 6, 8, 24 h and blot for EED. |
| Animal Protocol |
No in vivo animal protocol has been published. A standard protocol for PROTACs would be: (1) 6-8 week old female SCID-beige mice bearing Karpas422 xenografts (150-200 mm3). (2) Administer PROTAC EED degrader-1 IP at 10-50 mg/kg daily or every other day. (3) Formulation: 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. (4) Monitor tumor volume and body weight for 2-4 weeks. (5) Collect tumors for Western blot (EED, EZH2, H3K27me3) and IHC (Ki-67, cleaved caspase-3).
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| ADME/Pharmacokinetics |
Standard formulation for in vivo: 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline for IP injection (clear solution). Solubility: soluble in DMSO (≥ 50 mg/mL, 47.44 mM). LogP: 6.7 (predicted). PK in rodents (predicted from similar VHL-based PROTACs): IV administration (10 mg/kg) in mice, t1/2 ~ 2-4 h, CL ~ 20-40 mL/min/kg, Vd ~ 2-4 L/kg, oral bioavailability ~ 10-30%.
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| Toxicity/Toxicokinetics |
For in vitro toxicity: CCK-8 assay on HEK293 cells, IC50 > 50 uM for the PROTAC alone. For in vivo toxicity: MTD studies in CD-1 mice, single IP dose up to 200 mg/kg; repeated dosing at 50-100 mg/kg IP qd × 14 days, monitor body weight loss, lethargy, and clinical chemistry (ALT, AST, BUN, creatinine). Histopathology: assess liver and kidney. The compound is for research use only, not for human therapeutic use.
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| References | |
| Additional Infomation |
PROTAC EED degrader-1 is a first-in-class VHL-based PROTAC targeting the EED subunit of PRC2. It efficiently degrades EED, EZH2, and SUZ12 proteins, leading to loss of H3K27me3 and anti-proliferative effects in EZH2-mutant DLBCL cells. This compound is not FDA-approved and is strictly for research use in chemical epigenetics and cancer biology.
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| Molecular Formula |
C55H60FN11O8S
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|---|---|
| Molecular Weight |
1054.20
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| Exact Mass |
1053.433
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| CAS # |
2639882-72-5
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| PubChem CID |
145925663
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| Appearance |
White to off-white solid powder
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| LogP |
6.7
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
16
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| Rotatable Bond Count |
21
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| Heavy Atom Count |
76
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| Complexity |
1910
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| Defined Atom Stereocenter Count |
3
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| SMILES |
CC1=C(C=CC(=N1)CCC(=O)NCCOC2=CC=C(C=C2)OCC(=O)N[C@H](C(=O)N3C[C@@H](C[C@H]3C(=O)NCC4=CC=C(C=C4)C5=C(N=CS5)C)O)C(C)(C)C)C6=CN=C(N7C6=NN=C7)NCC8=C(C=CC9=C8CCO9)F
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| InChi Key |
FSFJUWLIBSALJI-FENRPDQVSA-N
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| InChi Code |
InChI=1S/C55H60FN11O8S/c1-32-40(43-27-60-54(67-30-62-65-51(43)67)59-26-42-41-20-22-74-46(41)18-17-44(42)56)16-10-36(63-32)11-19-47(69)57-21-23-73-38-12-14-39(15-13-38)75-29-48(70)64-50(55(3,4)5)53(72)66-28-37(68)24-45(66)52(71)58-25-34-6-8-35(9-7-34)49-33(2)61-31-76-49/h6-10,12-18,27,30-31,37,45,50,68H,11,19-26,28-29H2,1-5H3,(H,57,69)(H,58,71)(H,59,60)(H,64,70)/t37-,45+,50-/m1/s1
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| Chemical Name |
(2S,4R)-1-[(2S)-2-[[2-[4-[2-[3-[5-[5-[(5-fluoro-2,3-dihydro-1-benzofuran-4-yl)methylamino]-[1,2,4]triazolo[4,3-c]pyrimidin-8-yl]-6-methylpyridin-2-yl]propanoylamino]ethoxy]phenoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]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 :~150 mg/mL (~142.29 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3.75 mg/mL (3.56 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 37.5 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: 3.75 mg/mL (3.56 mM) in 10% DMSO + 90% Corn Oil (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 37.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.9486 mL | 4.7429 mL | 9.4859 mL | |
| 5 mM | 0.1897 mL | 0.9486 mL | 1.8972 mL | |
| 10 mM | 0.0949 mL | 0.4743 mL | 0.9486 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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