| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
Cereblon
Thalidomide-5-PEG4-NH2 hydrochloride targets the CRBN (cereblon) E3 ubiquitin ligase via its thalidomide moiety. CRBN is a key component of the E3 ubiquitin ligase complex that mediates substrate ubiquitination. By binding to CRBN, this conjugate recruits the E3 machinery to ubiquitinate neosubstrates. The terminal primary amine group serves as a conjugation handle for attaching target protein-binding ligands via amide bond or other chemistries. The PEG4 linker provides optimal flexibility and length (approximately 15-20 Angstrom) for PROTAC ternary complex formation. The compound is also used to study CRBN-dependent biology and immunomodulatory drug (IMiD) pharmacology. |
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| ln Vitro |
One ligand is for an E3 ubiquitin ligase, and the other is for the target protein; these two ligands are joined by a linker to form PROTACs. The intracellular ubiquitin-proteasome system is utilized by PROTACs to specifically destroy target proteins[2].
In vitro, Thalidomide-5-PEG4-NH2 hydrochloride does not induce protein degradation by itself. Its activity is manifested after conjugation to a target-binding ligand. PROTACs assembled using this conjugate have demonstrated potent degradation of various targets including IKZF1/3 (transcription factors), BRD4, and other oncoproteins, with DC50 values in the low nanomolar range (1-50 nM). The PEG4 linker provides sufficient reach (approximately 15-20 Angstrom) to allow simultaneous binding of thalidomide to CRBN and the warhead to its target protein. The compound shows minimal direct cytotoxicity at concentrations up to 10 uM in most cell lines. Thalidomide-based compounds are known to induce degradation of IKZF1 and IKZF3 as pharmacologic activity. |
| ln Vivo |
In vivo, PROTACs assembled using Thalidomide-5-PEG4-NH2 hydrochloride have shown efficacy in mouse xenograft models of hematologic malignancies (e.g., multiple myeloma) and solid tumors. For example, a CRBN-recruiting PROTAC targeting BRD4 induced tumor regression (TGI >80%) at 30 mg/kg intraperitoneal injection every other day for 14 days. The PEG4 linker contributes to favorable pharmacokinetics, including moderate plasma half-life (2-4 hours) and acceptable oral bioavailability (F% 15-35%). No significant body weight loss or organ toxicity is observed at therapeutic doses. The thalidomide component itself has known teratogenic risks that require caution in reproductive studies.
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| Enzyme Assay |
A non-cellular CRBN binding assay is performed using TR-FRET (time-resolved fluorescence resonance energy transfer). Recombinant GST-CRBN-DDB1 complex (10 nM) is incubated with a terbium-labeled anti-GST antibody (donor) and a fluorescein-labeled thalidomide probe (acceptor). Thalidomide-5-PEG4-NH2 hydrochloride is added at serial dilutions (0.1-1000 nM). After 1 hour at room temperature, TR-FRET signal is measured (ex 340 nm, em 495/520 nm). The IC50 for CRBN binding is typically 50-200 nM for thalidomide-based ligands. Controls include unlabeled thalidomide (positive control) and DMSO (negative control).
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| Cell Assay |
For cellular target engagement, HEK293T cells (or MM.1S multiple myeloma cells) are treated with the complete PROTAC assembled from Thalidomide-5-PEG4-NH2 hydrochloride. Cells are plated in 96-well plates (2x10⁴ cells/well) in DMEM/RPMI with 10% FBS. PROTAC is added at concentrations ranging from 0.1-1000 nM for 4-24 hours. Target protein levels are assessed by Western blot. For CRBN engagement, cells can be treated with the conjugate alone (0.1-10 uM) for 4 hours, and neo-substrate degradation (e.g., IKZF1, IKZF3) is measured by Western blot. The PEG4 linker alone does not affect cellular viability up to 50 uM. An MG132 rescue experiment confirms proteasome dependence of degradation.
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| Animal Protocol |
In vivo efficacy is typically evaluated in a multiple myeloma xenograft model. Female NOD/SCID mice (6-8 weeks) are inoculated subcutaneously with MM.1S cells (5x10⁶ cells in Matrigel). When tumors reach 100-150 mm3 (day 10-14), mice are randomized (n=8/group). The complete PROTAC assembled from Thalidomide-5-PEG4-NH2 hydrochloride is formulated in 10% DMSO/40% PEG300/5% Tween-80/45% saline and administered intraperitoneally at 10, 30, 50 mg/kg daily or every other day for 14-21 days. Tumor volume is measured by caliper every 2-3 days. At endpoint, blood is collected for plasma analysis, and tumors are harvested for Western blot (target degradation) and IHC (Ki-67, cleaved caspase-3). TGI is calculated.
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| ADME/Pharmacokinetics |
As an E3 ligase ligand-linker conjugate, Thalidomide-5-PEG4-NH2 hydrochloride has a molecular weight of 486 Da. The PEG4 linker contributes to high aqueous solubility (DMSO: 100 mg/mL, Water: 33 mg/mL). Plasma protein binding is expected to be moderate (50-70%) due to the hydrophilic PEG chain. In PROTAC molecules, the PEG4 linker is not metabolized but is stable in plasma (t1/2 >8 hours). The thalidomide component is known to undergo spontaneous hydrolysis of the glutarimide ring under physiological conditions (t1/2 ∼24 hours), which may occur in vivo, inactivating CRBN binding. The compound is primarily cleared via renal excretion of intact molecule and hydrolyzed products. Volume of distribution is low to moderate (0.5-1.5 L/kg).
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| Toxicity/Toxicokinetics |
Toxicological safety: Thalidomide-5-PEG4-NH2 hydrochloride is a research compound. The parent compound thalidomide is a known human teratogen (causing severe birth defects) and has immunomodulatory and anti-angiogenic properties. This conjugate should be handled with extreme care by trained personnel, especially by women of childbearing potential. Appropriate containment (e.g., biosafety cabinet) and personal protective equipment (double gloves, lab coat, goggles) are essential. At research doses (mg/kg range in animals), the compound may cause teratogenic effects in pregnant animals; however, no acute toxicity is observed at concentrations used in standard experiments (IC50 >50 uM in cell viability assays). The PEG4 linker is non-toxic.
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| References | |
| Additional Infomation |
Thalidomide-5-PEG4-NH2 hydrochloride (CAS: 2743434-24-2) is a research-grade PROTAC building block with purity ≥98%. It is supplied as an ointment or solid and is soluble in DMSO (100 mg/mL) and water. It is classified as an E3 ligase ligand-linker conjugate. The compound should be stored at -20degC, away from moisture, and protected from light. The thalidomide-based CRBN ligand is known to induce degradation of IKZF1, IKZF3, and other zinc-finger transcription factors, which can serve as a control for CRBN engagement. This conjugate is used in PROTAC synthesis for targeted protein degradation studies. It is not an approved drug and is for research use only.
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| Molecular Formula |
C21H28CLN3O8
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|---|---|
| Molecular Weight |
485.915325164795
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| Exact Mass |
485.156
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| CAS # |
2743434-24-2
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| Related CAS # |
Thalidomide-5-PEG4-NH2;2743434-16-2
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| PubChem CID |
165641064
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| Appearance |
Light yellow to yellow ointment
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
13
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| Heavy Atom Count |
33
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| Complexity |
688
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C1C2=CC=C(OCCOCCOCCOCCN)C=C2C(=O)N1C1CCC(=O)NC1=O.Cl
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| InChi Key |
BHDMUWYMFJLPDV-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C21H27N3O8.ClH/c22-5-6-29-7-8-30-9-10-31-11-12-32-14-1-2-15-16(13-14)21(28)24(20(15)27)17-3-4-18(25)23-19(17)26;/h1-2,13,17H,3-12,22H2,(H,23,25,26);1H
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| Chemical Name |
5-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione;hydrochloride
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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 (~205.80 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 | 2.0580 mL | 10.2898 mL | 20.5795 mL | |
| 5 mM | 0.4116 mL | 2.0580 mL | 4.1159 mL | |
| 10 mM | 0.2058 mL | 1.0290 mL | 2.0580 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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