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| 1g |
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| Other Sizes |
| Targets |
Cereblon
The primary molecular target of N-Me-Thalidomide 4-fluoride is the E3 ubiquitin ligase complex, specifically the Cereblon (CRBN) protein. By binding to CRBN, this compound induces a conformational change that allows the recruitment of neo-substrates (target proteins). In the context of a PROTAC, one end of the molecule (the thalidomide portion) binds to CRBN, while the other end is designed to bind to a specific protein of interest (POI). This brings the POI into close proximity with the E3 ligase, leading to its ubiquitination and subsequent degradation by the proteasome. The fluorine substitution is often introduced to modulate potency, selectivity, and metabolic stability. The molecular formula is C14H11FN2O4, and it has a molecular weight of 290.25. |
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| ln Vitro |
The in vitro activity of N-Me-Thalidomide 4-fluoride is measured by its ability to induce degradation of a target protein when incorporated into a PROTAC. The molecule itself does not have an intrinsic IC50 against a specific disease target; rather, it is a ligand. In an in vitro ubiquitination assay, a purified system containing CRBN, E1 and E2 ubiquitin enzymes, and a target protein (neo-substrate) is used. The addition of N-Me-Thalidomide 4-fluoride (or a PROTAC containing it) enhances the ubiquitination of the target, which is detected by Western blotting using an anti-ubiquitin antibody. The potency is often described by the concentration that leads to 50% degradation (DC50). This compound is specifically used to synthesize anti-inflammatory agent 70.
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| ln Vivo |
N-Me-Thalidomide 4-fluoride itself has no direct therapeutic in vivo activity. It is a synthetic intermediate and a ligand for cereblon. Its activity is only realized once it is conjugated to a target-binding warhead to form a PROTAC molecule. In vivo, PROTACs containing this ligand have been used in mouse xenograft models to degrade disease-causing proteins. For example, an anti-inflammatory PROTAC using this moiety would be administered intraperitoneally or orally. The endpoint is the reduction in the levels of the target inflammatory protein (e.g., BRD4 or IRAK4) in tissue homogenates, as measured by Western blot, and the subsequent reduction in inflammatory cytokines. The compound is also used to synthesize an anti-inflammatory agent 70.
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| Enzyme Assay |
A standard non-cellular binding assay for N-Me-Thalidomide 4-fluoride is a TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) binding assay to measure its affinity for CRBN. The assay uses recombinant human CRBN protein (often fused to a GST tag) and a terbium-labeled anti-GST antibody (donor). An acceptor dye (e.g., HiLyte Fluor 647) is conjugated to a known high-affinity thalidomide derivative (probe). The test compound (N-Me-Thalidomide 4-fluoride) is serially diluted in a 384-well plate. The CRBN protein and the fluorescent probe are added. The TR-FRET signal is measured after 1-2 hours of equilibration. A decrease in signal indicates displacement of the probe by the test compound. The IC50 is calculated, and the binding affinity (Ki) is derived. This confirms the compound's function as a CRBN binder before it is used to create a PROTAC.
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| Cell Assay |
Cellular assays to validate the activity of N-Me-Thalidomide 4-fluoride are performed indirectly by testing the final PROTAC molecule. A typical cell-based degradation assay uses target-expressing cell lines (e.g., MDA-MB-231 breast cancer cells for an IRAK4-targeting PROTAC). Cells are seeded in 6-well plates and treated with increasing concentrations (0.1 nM to 1 uM) of the PROTAC (which incorporates this ligand) for 6-24 hours. The cells are lysed, and the lysates are subjected to Western blotting. The blot is probed with an antibody against the target protein (POI). A loading control (e.g., GAPDH) is used for normalization. The DC50 (concentration required to degrade 50% of the target) is calculated by densitometric analysis. A lack of degradation when using a PROTAC with an inactive CRBN ligand (negative control) confirms the specific activity of N-Me-Thalidomide 4-fluoride.
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| Animal Protocol |
In vivo experiments to evaluate the efficacy of a PROTAC built with N-Me-Thalidomide 4-fluoride are standard for drug discovery. A protocol uses female BALB/c nude mice bearing subcutaneous xenograft tumors of a target-positive cancer cell line (e.g., MM.1S multiple myeloma). When tumors reach 150-200 mm3, mice are randomized (n=8-10) into treatment groups. The PROTAC is administered intravenously or orally at doses of 1-30 mg/kg, daily or every other day, for 2-4 weeks. Tumor size is measured with calipers twice weekly. At endpoint, tumors are collected, and the target protein levels are measured by Western blot or immunohistochemistry (IHC). A reduction in tumor growth and target protein levels compared to vehicle control indicates the in vivo efficacy of the PROTAC, validating the utility of the thalidomide-based ligand. The compound is also used to synthesize anti-inflammatory agent 70.
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| ADME/Pharmacokinetics |
N-Me-Thalidomide 4-fluoride has a molecular weight of 290.25 and a molecular formula of C14H11FN2O4. It is a solid powder at room temperature. It should be stored as a powder at -20degC (stable for 3 years) and at 4degC for 2 years. Once dissolved in DMSO, it should be stored at -80degC for up to 6 months or -20degC for 1 month. The compound is supplied for research purposes, and its solubility in water is poor, but it is soluble in organic solvents like DMSO, and also in methanol and ethanol. A typical stock solution is prepared at 10-50 mM in DMSO. The compound features a piperidinedione ring, which is characteristic of immunomodulatory imide drugs (IMiDs). The CAS number is 2244520-92-9.
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| Toxicity/Toxicokinetics |
The toxicology of N-Me-Thalidomide 4-fluoride is not fully characterized, but it is expected to be similar to other thalidomide derivatives. Thalidomide is known for its teratogenicity (causing birth defects). Therefore, this compound must be handled with extreme caution in the laboratory. Researchers should wear double gloves, a lab coat, and eye protection. It is classified as a research chemical and is not for human consumption. Inhalation and skin contact should be avoided. While the methyl and fluorine substitutions may alter the toxicological profile, the compound's mechanism of action (cereblon binding) still carries the potential risk for immunomodulatory and developmental effects. Safety data sheets for this class of compounds typically include hazard statements for reproductive toxicity. It is essential to prevent exposure for pregnant women working in the lab.
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| References | |
| Additional Infomation |
N-Me-Thalidomide 4-fluoride is a key component in the rapidly advancing field of targeted protein degradation (TPD). Unlike traditional inhibitors that block the active site of a protein, degraders (PROTACs) remove the protein entirely. This offers the potential to target "undruggable" proteins and overcome drug resistance. The thalidomide core is currently the most widely used CRBN ligand in PROTAC design due to its small size and high binding affinity. The specific modifications (N-methyl and 4-fluoro) are examples of medicinal chemistry optimization to improve the drug-like properties of the ligand, such as increasing its metabolic stability or fine-tuning its binding to CRBN. This compound is an intermediate in the synthesis of anti-inflammatory agent 70. It is not a drug itself but a sophisticated tool for creating next-generation therapies for inflammation and cancer. It is for research use only and is not approved for clinical use.
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| Molecular Formula |
C14H11FN2O4
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| Molecular Weight |
290.25
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| CAS # |
2244520-92-9
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| Appearance |
Solid powder
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| SMILES |
FC1=CC=CC2=C1C(N(C2=O)C1C(N(C)C(CC1)=O)=O)=O
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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) |
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 | 3.4453 mL | 17.2265 mL | 34.4531 mL | |
| 5 mM | 0.6891 mL | 3.4453 mL | 6.8906 mL | |
| 10 mM | 0.3445 mL | 1.7227 mL | 3.4453 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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