| Size | Price | Stock | Qty |
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| 50mg |
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| Other Sizes |
| Targets |
The compound targets cereblon (CRBN), a component of the E3 ubiquitin ligase complex. The thalidomide-derived moiety binds to CRBN, which is part of the CUL4-RBX1-DDB1-CRBN E3 ubiquitin ligase. Binding of the thalidomide-based ligand to CRBN induces a conformational change that promotes the recruitment of neo-substrates to the ubiquitin-proteasome system for degradation. This targeting mechanism is the foundation of PROTAC technology, where bifunctional molecules containing both a CRBN ligand and a target protein ligand induce selective degradation of the target protein.
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| ln Vitro |
A linker separates the two ligands that make up PROTACs; one ligand is for an E3 ubiquitin ligase, and the other is for the target protein. Target proteins are selectively degraded by PROTACs by taking advantage of the intracellular ubiquitin-proteasome system[2].
In vitro activity of this compound is demonstrated primarily within PROTAC conjugates. When conjugated to a target protein ligand, the resulting PROTAC molecule recruits the CRBN E3 ligase to the target protein, leading to its ubiquitination and subsequent degradation by the proteasome. The ligand-linker conjugate itself shows minimal standalone biological activity. In vitro validation studies use cell-based assays to measure degradation of target proteins (e.g., BRD4, BET family proteins) by Western blotting after treatment with PROTACs constructed using this linker. Concentrations typically range from 0.001-10 uM, with degradation potency (DC50, degradation concentration for 50% reduction) determined by dose-response curves. |
| ln Vivo |
In vivo activity studies are performed with complete PROTAC molecules built using this compound as the linker-cereblon ligand component. Animal studies (typically in murine xenograft models of cancer) involve administration of the PROTAC via IP or IV routes at doses ranging from 1-50 mg/kg. Efficacy endpoints include tumor growth inhibition, assessment of target protein degradation in tumor tissues by immunohistochemistry or Western blot, and evaluation of downstream pharmacodynamic markers. The PEG2 linker provides moderate hydrophilicity that improves aqueous solubility while maintaining sufficient cell permeability for in vivo efficacy.
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| Enzyme Assay |
For in vitro binding assays, the cereblon-binding affinity of the thalidomide-piperazine component can be measured using fluorescence polarization (FP) or surface plasmon resonance (SPR). Purified CRBN-DDB1 complex is immobilized, and varying concentrations of the compound are flowed over the chip to calculate KD values. Alternatively, competitive binding assays using a fluorescently labeled thalidomide probe can determine IC50 values. Typical assay buffer: 50 mM HEPES (pH 7.4), 150 mM NaCl, 1 mM DTT, and 0.01% Tween-20. Reported KD values for thalidomide-derived CRBN ligands are typically in the low micromolar to nanomolar range, with variations depending on the specific linker modifications.
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| Cell Assay |
To construct a PROTAC molecule using this compound, the free amine group is conjugated to a target protein ligand that contains a carboxylic acid group via standard amide coupling chemistry using HATU or EDCI/HOBt in DMF or DMSO with a base such as DIPEA. The resulting PROTAC is purified by preparative HPLC. For cell-based validation, cells are treated with the PROTAC (0.001-10 uM) for 4-24 hours, followed by lysis and Western blotting to assess target protein levels. Control treatments include the PROTAC molecule, the target ligand alone, the CRBN ligand alone, and the combination of both ligands without conjugation (negative control).
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| Animal Protocol |
For in vivo efficacy studies, complete PROTAC molecules incorporating this linker are typically administered to mice bearing subcutaneous tumor xenografts. Standard dosing involves IP injection at 5-30 mg/kg, once daily or every other day, for 2-4 weeks. Tumor volumes are measured every 2-3 days using calipers. At study termination, tumors and other tissues (liver, kidney, spleen) are harvested for analysis of target protein degradation by Western blot and histopathological examination. Pharmacokinetic blood sampling is performed at various time points post-dose to measure PROTAC concentrations by LC-MS/MS. Baseline control groups include vehicle-treated, target-ligand-only, and CRBN-ligand-only treated animals.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of this compound when incorporated into PROTACs depend on the attached target ligand and the overall molecular properties. For the ligand-linker conjugate alone, the molecular weight is approximately 701.6 g/mol, and the PEG2 linker (2 ethylene oxide units) provides modest hydrophilicity that contributes to aqueous solubility. As a building block, this compound is not intended for direct in vivo administration; instead, it is a synthetic intermediate. PROTAC molecules typically exhibit intermediate pharmacokinetic profiles, often with short half-lives (1-6 hours in rodents) due to rapid clearance and metabolism, though optimization of the linker can improve these properties. The diTFA salt form enhances solubility but does not affect in vivo PK once the compound is formulated.
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| Toxicity/Toxicokinetics |
The diTFA salt form is generally safe for laboratory handling following standard chemical safety practices. Trifluoroacetate (TFA) is considered moderately toxic and should be handled with appropriate precautions including fume hood use and personal protective equipment. No specific toxicity data is available for this ligand-linker conjugate as a standalone entity. When incorporated into PROTACs, toxicity profiles vary widely depending on the target protein. Thalidomide-derived CRBN ligands share some structural similarity with thalidomide, which is known for teratogenic effects. Therefore, researchers should handle these compounds with caution, particularly regarding potential developmental toxicity. These compounds are strictly for research use only, not for human use, and are not intended for therapeutic or diagnostic applications.
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| References | |
| Additional Infomation |
Thalidomide-Piperazine-PEG2-NH2 diTFA is part of a library of E3 ligase ligand-linker conjugates used for PROTAC synthesis. The piperazine ring provides structural rigidity that may enhance binding to CRBN compared to flexible alkyl chains, while the PEG2 spacer balances solubility and permeability. PROTACs contain two different ligands connected by a linker; one ligand is for an E3 ubiquitin ligase, and the other is for the target protein. PROTACs exploit the intracellular ubiquitin-proteasome system to selectively degrade target proteins. This compound is not an approved drug and is not in clinical trials; it is exclusively a chemical tool for research applications in targeted protein degradation, cancer biology, and chemical biology. The compound should be stored at -20degC, protected from light, under inert atmosphere.
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| Molecular Formula |
C27H33F6N5O10
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| Molecular Weight |
701.57
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| Related CAS # |
Thalidomide-Piperazine-PEG2-NH2;2357110-14-4
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| Appearance |
Light yellow to green yellow solid powder
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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, 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) |
H2O :~50 mg/mL (~71.27 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.4254 mL | 7.1269 mL | 14.2537 mL | |
| 5 mM | 0.2851 mL | 1.4254 mL | 2.8507 mL | |
| 10 mM | 0.1425 mL | 0.7127 mL | 1.4254 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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