| Size | Price | Stock | Qty |
|---|---|---|---|
| 50mg |
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| Other Sizes |
| Targets |
Cereblon
The compound targets cereblon (CRBN), a substrate receptor component of the CUL4-RBX1-DDB1-CRBN E3 ubiquitin ligase complex. The thalidomide-derived moiety binds to CRBN, leading to recruitment of the E3 ligase machinery. This binding alters the substrate specificity of the ligase complex, enabling the ubiquitination and subsequent proteasomal degradation of target proteins when the ligand-linker conjugate is incorporated into a PROTAC molecule. The piperazine ring contributes to structural rigidity and favorable binding geometry for CRBN interaction. |
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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 realized through its use as a PROTAC building block. When conjugated to a target protein-binding ligand, the resulting PROTAC molecule induces ubiquitination and proteasome-dependent degradation of the target protein. The ligand-linker conjugate alone exhibits minimal intrinsic biological activity. In vitro validation involves treating cells with PROTACs constructed using this linker at concentrations typically ranging from 0.001-10 uM, followed by assessment of target protein levels via Western blotting. Degradation potency (DC50) is determined from dose-response curves. The short PEG1 linker provides minimal flexibility and may be optimal for certain target protein-binding geometries where proximity between the E3 ligase and target is critical. |
| ln Vivo |
In vivo studies with PROTACs incorporating this linker are typically conducted in rodent tumor xenograft models. Complete PROTAC molecules (including a target protein ligand conjugated to this compound) are administered via intraperitoneal or intravenous routes at doses of 1-50 mg/kg, often daily or every other day for 2-4 weeks. Efficacy is assessed by tumor volume measurements, Western blot analysis of target protein degradation in harvested tumor tissues, and evaluation of pharmacodynamic biomarkers. The short PEG1 linker offers advantages in creating highly compact PROTAC molecules, which may exhibit different pharmacokinetic and cell penetration properties compared to longer PEG variants.
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| Enzyme Assay |
CRBN-binding affinity of the thalidomide-piperazine component can be measured using fluorescence polarization (FP) competitive binding assays or surface plasmon resonance (SPR). Purified CRBN-DDB1 protein complex is incubated with a fluorescently labeled thalidomide probe and varying concentrations of the test compound. Binding (IC50 or KD) is determined by measuring displacement of the probe. Alternatively, isothermal titration calorimetry (ITC) can directly measure thermodynamic binding parameters. Typical assay buffer: 50 mM HEPES (pH 7.5), 150 mM NaCl, 1 mM DTT, 0.1% BSA. Thalidomide-derived CRBN ligands typically exhibit KD values in the low micromolar to sub-micromolar range.
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| Cell Assay |
To construct a PROTAC, the terminal amine of this compound is conjugated to a target protein ligand bearing a carboxylic acid group via standard amide bond formation using coupling reagents (HATU, EDCI/HOBt) in DMF or DMSO with DIPEA as base. The resulting PROTAC is purified by preparative HPLC and characterized by LC-MS. For cellular activity assays, cells are treated with the PROTAC at concentrations ranging from 0.1 nM to 10 uM for 4-24 hours, then lysed for Western blotting to assess target protein degradation. Control conditions include treatment with the unconjugated target ligand alone and the CRBN ligand alone to confirm target engagement is required for degradation.
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| Animal Protocol |
In vivo animal studies employ complete PROTAC molecules synthesized using this linker. Male or female immunodeficient mice (e.g., nude or NSG) bearing subcutaneous xenografts of human cancer cell lines are randomized into treatment groups (n=5-10 per group). PROTACs are typically formulated in vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline) and administered IP or IV once daily or every other day at doses of 1-30 mg/kg. Tumor volumes and body weights are measured every 2-3 days. At study endpoint (typically 2-4 weeks), tumors are collected for Western blot analysis, IHC, and histopathology. Pharmacokinetic studies involve serial blood sampling post-dose for LC-MS/MS analysis.
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| ADME/Pharmacokinetics |
For the standalone ligand-linker conjugate (molecular weight approximately 657.5 g/mol), the PEG1 linker (one ethylene oxide unit) is the shortest flexible spacer in this series, resulting in minimal hydrophilic character. However, as a PROTAC building block, the compound is not typically dosed alone. When incorporated into PROTACs, the overall molecular weight typically ranges from 800-1200 Da. PROTAC pharmacokinetics are characterized by generally poor oral bioavailability (due to high molecular weight and polar surface area), short plasma half-lives (1-4 hours in rodents), and extensive hepatic metabolism. Intravenous administration often achieves higher exposure and is preferred in efficacy studies. The diTFA salt aids solubility during synthesis and formulation.
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| Toxicity/Toxicokinetics |
This compound is strictly for research use with no clinical applications. Trifluoroacetate (TFA) salt requires standard chemical safety precautions-use fume hoods, gloves, and eye protection, as TFA is corrosive. The thalidomide core structure is associated with known teratogenic effects (birth defects) due to its cereblon-binding activity, which alters developmental gene regulation. Although this compound is a research building block and not a therapeutic agent, researchers should be aware of the potential developmental toxicity risks associated with thalidomide-derived compounds and handle them with appropriate caution, particularly female researchers of childbearing potential. All work should be conducted following institutional safety guidelines for controlled substances.
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| References | |
| Additional Infomation |
Thalidomide-Piperazine-PEG1-NH2 diTFA is a research-grade chemical building block exclusively for PROTAC development and targeted protein degradation studies. Compared to the PEG2 variant, the shorter PEG1 linker reduces molecular flexibility and overall molecular weight, which may affect PROTAC cell permeability and degradation efficiency depending on the target protein. PROTACs exploit the intracellular ubiquitin-proteasome system to selectively degrade target proteins, representing an emerging therapeutic modality in precision medicine. This compound has not been evaluated in clinical trials and is not approved for human use. It is intended for laboratory research applications only, including cancer biology studies, validation of protein targets, and investigation of E3 ligase mechanisms. Store at -20degC, protected from light and moisture.
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| Molecular Formula |
C25H29F6N5O9
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|---|---|
| Molecular Weight |
657.52
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| Appearance |
White to 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) |
DMSO :~100 mg/mL (~152.09 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (3.80 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (3.80 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5209 mL | 7.6043 mL | 15.2087 mL | |
| 5 mM | 0.3042 mL | 1.5209 mL | 3.0417 mL | |
| 10 mM | 0.1521 mL | 0.7604 mL | 1.5209 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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