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| 1mg |
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| Targets |
The compound targets cereblon (CRBN), a component of the CUL4-RBX1-DDB1-CRBN E3 ubiquitin ligase complex. The thalidomide-derived moiety binds to CRBN, recruiting the E3 ubiquitin ligase machinery. The 4-position attachment site orients the linker in a different direction relative to the CRBN binding pocket compared to 5-position analogs. When conjugated to a target protein ligand, the resulting PROTAC molecule brings the target protein into proximity with the E3 ligase, facilitating target ubiquitination and subsequent proteasomal degradation. The C7 alkyl linker is short and hydrophobic.
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| ln Vitro |
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 induces target protein ubiquitination and proteasome-dependent degradation. The ligand-linker conjugate alone exhibits minimal intrinsic biological activity. In cell-based assays, PROTACs constructed with 4-O-C7 linkers have been evaluated and may show different degradation profiles compared to 5-O-C7 analogs due to altered geometry of the ternary complex. Typical validation involves treating cells with the PROTAC at concentrations ranging from 0.001-10 uM for 4-24 hours, followed by Western blot analysis to quantify target protein levels.
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| ln Vivo |
In vivo efficacy studies with PROTACs incorporating the 4-O-C7 linker are typically conducted in murine xenograft models. Complete PROTAC molecules are administered to tumor-bearing immunodeficient mice via intraperitoneal (IP) or intravenous (IV) routes at doses ranging from 1-30 mg/kg. Dosing frequency is typically daily or every other day for 2-4 weeks. The hydrophobic C7 alkyl linker requires formulation with co-solvents such as DMSO, PEG300, Tween 80, or cyclodextrins. Efficacy is assessed by tumor volume measurements, Western blot analysis of target protein degradation in harvested tumor tissues, and evaluation of pharmacodynamic biomarkers.
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| Enzyme Assay |
CRBN-binding affinity can be measured using fluorescence polarization (FP) competitive binding assays or surface plasmon resonance (SPR). The 4-position attachment site may result in slightly different binding affinity compared to 5-position analogs, as the substituent orientation relative to the CRBN binding pocket differs. For FP assays, purified CRBN-DDB1 complex is incubated with a fluorescently labeled thalidomide probe and varying concentrations of the test compound. Typical assay conditions: 50 mM HEPES (pH 7.4), 150 mM NaCl, 1 mM DTT, 0.01% Tween-20, 0.1 mg/mL BSA. Thalidomide-based CRBN ligands typically exhibit KD values in the low micromolar range, with positional isomers showing comparable affinity.
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| Cell Assay |
To construct a PROTAC molecule, the terminal primary amine is conjugated to a target protein ligand containing a carboxylic acid via standard amide coupling chemistry. General protocol: The target ligand carboxylic acid (1.0 equiv) is dissolved in anhydrous DMF or DMSO, then HATU or EDCI/HOBt (1.2 equiv) and DIPEA (2-3 equiv) are added and the mixture is stirred for 10-30 minutes. The amine linker (1.0-1.2 equiv) is then added, and the reaction is stirred for 2-12 hours. The resulting PROTAC is purified by preparative HPLC and characterized by LC-MS. For cellular activity validation, cells are treated with the PROTAC (0.001-10 uM, 4-24 h), then lysed and analyzed by Western blotting to assess target protein degradation.
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| Animal Protocol |
In vivo animal studies with PROTACs containing the 4-O-C7 linker follow standard xenograft protocols. Immunodeficient mice (nude or NSG, 6-8 weeks old, n=5-10 per group) bearing subcutaneous human tumor xenografts (approximately 100-200 mm3) are randomized into treatment groups. PROTACs are formulated in a vehicle suitable for hydrophobic compounds (e.g., 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline; or 10% DMSO, 90% corn oil) and administered IP or IV at 1-30 mg/kg. Tumor volumes and body weights are recorded every 2-3 days. At study termination (2-4 weeks), tumors are harvested for Western blot or IHC analysis of target protein degradation, and plasma is collected for PK analysis.
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| ADME/Pharmacokinetics |
For the standalone ligand-linker conjugate, the C7 alkyl linker is hydrophobic, resulting in low aqueous solubility. The 4-O attachment site may influence the overall shape and lipophilicity of the conjugate. When incorporated into PROTACs, the C7 linker affects pharmacokinetic properties including limited aqueous solubility (requiring formulation), high plasma protein binding, potentially moderate half-life. The hydrochloride salt form improves handling solubility. No dedicated PK studies are available for this specific positional isomer. PROTACs containing 4-O linkers may show different distribution and clearance profiles compared to 5-O isomers due to altered overall molecular shape and protein binding characteristics.
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| Toxicity/Toxicokinetics |
Toxicology data specific to Thalidomide-4-O-C7-NH2 hydrochloride is not publicly available. Thalidomide derivatives are known teratogens causing severe birth defects via CRBN binding. The 4-O-C7 modification does not eliminate this risk. Researchers must handle this compound with extreme caution: work in a chemical fume hood, wear appropriate PPE (gloves, lab coat, safety glasses), avoid skin contact, inhalation, and ingestion. Pregnant or potentially pregnant individuals should avoid handling. This product is for research use only; not for human or therapeutic use.
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| Additional Infomation |
Thalidomide-4-O-C7-NH2 hydrochloride is a research tool for PROTAC-based targeted protein degradation where attachment at the 4-position may enable degradation of target proteins that are inaccessible using 5-position linked PROTACs due to different ternary complex geometries. The 4-O attachment site offers distinct vector orientation for the linker, expanding the chemical space for PROTAC optimization. The compound is part of a series including 4-O-C7, 4-O-C11, 4-O-C12, and 4-O-C14 variants. This product is strictly for laboratory research applications. It has not been evaluated in clinical trials and is not an approved drug. Storage: powder at -20degC, protected from light and moisture, under inert atmosphere.
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| Molecular Formula |
C20H26CLN3O5
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| Molecular Weight |
423.89
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| Related CAS # |
Thalidomide-4-O-C7-NH2;2093536-11-7
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| Appearance |
Off-white to light 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 2.3591 mL | 11.7955 mL | 23.5910 mL | |
| 5 mM | 0.4718 mL | 2.3591 mL | 4.7182 mL | |
| 10 mM | 0.2359 mL | 1.1796 mL | 2.3591 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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