| Size | Price | Stock | Qty |
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| 1mL |
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| Other Sizes |
| Targets |
TLR4 Alkyl/ether
The TLR4 inhibitor component (TLR4-IN-C34) is the active warhead that targets the TLR4 protein on cells. TLR4 is a key pattern recognition receptor (PRR) that plays a central role in the innate immune response by recognizing bacterial lipopolysaccharide (LPS) and other damage-associated molecular patterns (DAMPs). When this inhibitor is incorporated into a PROTAC molecule, the PROTAC binds to TLR4 via the warhead, while the other end of the molecule (attached through this linker) recruits an E3 ubiquitin ligase. This brings the ubiquitin ligase close to TLR4, leading to its ubiquitination and subsequent degradation by the proteasome. The target pathway is the TLR4 signaling pathway, a key driver of inflammation. |
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| ln Vitro |
The in vitro activity of the PROTAC built with this linker is measured by its ability to degrade TLR4 protein. In cell-based assays, cells expressing TLR4 (e.g., enterocytes or macrophages) are treated with the complete PROTAC. Degradation of the TLR4 protein is then measured by Western blot analysis using an anti-TLR4 antibody. This shows a time- and concentration-dependent decrease in the TLR4 protein band, indicating successful degradation. The reduction in TLR4 protein levels can also be confirmed by immunofluorescence staining and flow cytometry.
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| ln Vivo |
The in vivo activity of the PROTAC containing this linker is derived from the activity of the TLR4-IN-C34 warhead, but with a potentially longer-lasting effect due to protein degradation. The parent compound, TLR4-IN-C34, has been shown in vivo to inhibit TLR4 in enterocytes and macrophages, reducing systemic inflammation in mouse models of endotoxemia and necrotizing enterocolitis. It is expected that a PROTAC designed to degrade TLR4 could have enhanced potency and duration of action in these models by not only inhibiting but also reducing the total amount of the receptor protein [37L18-L19].
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| Enzyme Assay |
The binding of the TLR4-IN-C34 warhead (and therefore the complete PROTAC) to the TLR4 protein can be studied using surface plasmon resonance (SPR) or a competitive binding assay. A typical SPR protocol involves immobilizing recombinant TLR4/MD-2 complex on a sensor chip. The PROTAC linker (or the full PROTAC) is then flowed over the chip at increasing concentrations to determine its binding affinity (KD). Alternatively, a cell-based competitive binding assay can be performed using a fluorescently labeled TLR4 ligand (e.g., LPS). Cells expressing TLR4 are incubated with the labeled LPS and increasing concentrations of the PROTAC. The decrease in fluorescence signal as the PROTAC displaces the LPS is measured by flow cytometry.
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| Cell Assay |
For in vitro cellular assays to validate the efficacy of the complete PROTAC, the cells of interest (e.g., THP-1 macrophages or enterocytes) are cultured in appropriate medium. The PROTAC molecule is added to the cells at concentrations ranging from 0.001-10 uM. Cells are incubated for 4-24 hours. After incubation, the cells are lysed, and the lysates are subjected to SDS-PAGE and Western blotting. The blot is probed with an anti-TLR4 antibody and an anti-GAPDH or beta-actin antibody as a loading control. A decrease in the TLR4 band intensity in the treatment group compared to the control group indicates successful PROTAC-mediated degradation.
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| Animal Protocol |
In vivo animal studies for a TLR4-targeting PROTAC would be performed in mouse models of TLR4-driven disease, such as the endotoxemia or necrotizing enterocolitis models. Mice are injected with LPS to induce a systemic inflammatory response. The PROTAC (e.g., 5-20 mg/kg) is administered via intraperitoneal (IP) or intravenous (IV) injection, either prophylactically or therapeutically. At various time points post-treatment, blood samples are collected to measure serum levels of pro-inflammatory cytokines (e.g., IL-6, TNF-alpha) by ELISA. At the end of the study, mice are sacrificed, and tissues (e.g., colon, liver, spleen) are harvested for Western blot analysis to confirm TLR4 degradation and for histopathological examination to assess tissue damage.
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| ADME/Pharmacokinetics |
This compound is a PROTAC linker, not a standalone drug, so its pharmacokinetic (PK) properties are not studied in isolation. The overall PK of a PROTAC containing this linker will be determined by the properties of the whole molecule. The linker itself is designed to be non-cleavable, meaning it remains attached to the antibody or targeting ligand. The C2-amide-C6-OH structure and the alkyl/ether composition are chosen to provide the appropriate length and rigidity to facilitate the formation of a stable ternary complex between the target protein (TLR4) and the E3 ubiquitin ligase. The compound is typically stored at 4degC, protected from light, to maintain its stability [37L6-L7].
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| Toxicity/Toxicokinetics |
Specific toxicity data for TLR4-IN-C34-C2-amide-C6-OH is not publicly available. As a chemical tool for research, its safety profile is not fully established. Toxicity would primarily be a concern if the compound were administered in vivo as a PROTAC. Chronic inhibition or degradation of TLR4 could potentially lead to immunosuppression and increased susceptibility to infections. However, in the acute models of inflammation (e.g., endotoxemia), a short-term reduction in TLR4 signaling is the intended therapeutic goal. As with all research chemicals, standard laboratory safety practices should be followed. It is for research use only and not for human or clinical use.
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| References | |
| Additional Infomation |
TLR4-IN-C34-C2-amide-C6-OH is a PROTAC linker containing a small molecule TLR4 inhibitor (TLR4-IN-C34) and is used in PROTAC technology for targeted protein degradation. It falls under the categories of PROTAC Linkers and Immunology/Inflammation research. This compound is used to synthesize PROTAC molecules that can degrade TLR4, a key protein in the innate immune system and a validated therapeutic target for inflammatory diseases. This product is strictly for research use only. It is not a therapeutic drug, not for clinical use, and has not been approved by any regulatory agency. It should be stored at 4degC, protected from light, to ensure its stability for research applications [37L6-L7].
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| Molecular Formula |
C25H42N2O11
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| Appearance |
Colorless to light yellow liquid
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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.) |
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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