| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
|
||
| 500mg |
|
| Targets |
NH-bis(PEG4-Boc) functions as a chemical linker rather than a pharmacologically active drug. Its "targets" are the functional groups on molecules to which it conjugates. In PROTAC applications, the linker connects an E3 ubiquitin ligase ligand to a target protein ligand, enabling selective protein degradation via the ubiquitin-proteasome system. The amino groups serve as handles for conjugation to carboxylic acids (via amide bond formation), activated NHS esters, or carbonyls (via reductive amination). The two Boc-protected amines provide protected nucleophiles that can be deprotected under acidic conditions (e.g., TFA) to yield free amines for further conjugation to carboxylic acids or other electrophiles. The PEG4 spacer improves aqueous solubility and reduces aggregation of the final conjugates. This linker does not bind to biological receptors or enzymes but serves as a structural bridge.
|
|---|---|
| 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[1].
As a synthetic linker molecule, NH-bis(PEG4-Boc) does not exhibit pharmacological activity in cell-based assays. Its in vitro utility is demonstrated through the successful synthesis and characterization of PROTAC constructs incorporating this linker. Researchers evaluate the linker's performance by assessing conjugation efficiency (via HPLC, LC-MS, or NMR), the stability of resulting conjugates in cell culture media, and the biological activity of final PROTAC constructs in target cell lines. For PROTACs, target protein degradation is quantified by Western blot or ELISA following treatment of cells with the construct. The branched PEG architecture provides enhanced solubility and may improve the pharmacokinetic properties of the final construct. The compound is soluble in DMSO and other organic solvents, facilitating its use in standard bioconjugation workflows. |
| ln Vivo |
No direct in vivo pharmacological activity is attributed to NH-bis(PEG4-Boc) itself. Its in vivo relevance is demonstrated through the performance of PROTAC constructs synthesized using this linker in animal models. For in vivo administration, PROTAC constructs are typically formulated in vehicles such as DMSO/PEG300/Tween-80/saline or other standard formulations. The linker's stability in biological matrices, its contribution to the construct's pharmacokinetic profile, and its ability to maintain construct integrity are key parameters evaluated in preclinical studies. The Boc-protected amines are stable at physiological pH and are only deprotected under acidic conditions, ensuring that the construct remains intact during circulation. The PEG4 spacer enhances aqueous solubility and may prolong circulation time.
|
| Enzyme Assay |
In vitro enzyme/receptor binding assays are not applicable to NH-bis(PEG4-Boc) as it is a chemical linker with no intrinsic affinity for biological macromolecules. Quality control and characterization are performed using standard analytical methods. Purity is assessed by HPLC (typically ≥97%) and structural integrity is confirmed by ¹H NMR, ¹³C NMR, and mass spectrometry. The amino group content and the presence of Boc-protected amines are verified through spectroscopic analysis. For researchers using this linker, conjugation reactions (e.g., amide bond formation with carboxylic acids) are monitored by TLC or HPLC. Deprotection of Boc groups is confirmed by NMR or LC-MS following acid treatment (e.g., TFA in DCM), indicated by the disappearance of tert-butyl signals. Solubility testing in various solvents is performed to guide formulation development.
|
| Cell Assay |
Cell-based assays are not performed directly on NH-bis(PEG4-Boc) because the compound is a synthetic linker lacking biological activity. However, the biological activity of PROTAC constructs containing this linker is evaluated in relevant cell lines. Typical protocols involve treating cells with the PROTAC construct for 4-48 hours, then assessing target protein degradation by Western blot or ELISA. Cell viability, proliferation, and apoptosis are monitored using standard assays such as MTT, CCK-8, or flow cytometry. The linker's contribution to the construct's cellular uptake, stability, and efficacy is inferred from structure-activity relationship studies comparing different linker architectures. DMSO stock solutions are prepared and diluted in cell culture media to achieve desired final concentrations. Control groups include vehicle-treated cells and cells treated with the individual ligand components.
|
| Animal Protocol |
In vivo animal studies are conducted with PROTAC constructs incorporating NH-bis(PEG4-Boc), not with the linker alone. Typical protocols utilize rodent models (e.g., mice bearing tumor xenografts or disease-relevant transgenic models). The construct is administered via intravenous, intraperitoneal, or oral gavage at doses determined by preliminary pharmacokinetic and tolerability studies. Efficacy is assessed by measuring disease progression endpoints such as tumor volume, biomarker levels, or survival. Pharmacodynamic endpoints include target protein degradation in tissues (measured by Western blot or IHC) and downstream pathway modulation. The linker's stability in circulation and its ability to maintain the integrity of the PROTAC construct are evaluated through plasma sampling and LC-MS/MS analysis. Formulation vehicles typically include DMSO, PEG300, and saline or other standard excipients.
|
| ADME/Pharmacokinetics |
As a chemical linker, NH-bis(PEG4-Boc) does not have a conventional pharmacokinetic profile. However, the pharmacokinetic properties of PROTAC constructs incorporating this linker are evaluated in preclinical studies. Following administration in rodents, key parameters such as half-life, clearance, volume of distribution, and bioavailability are determined from plasma concentration-time data. The PEG4 spacer contributes to enhanced aqueous solubility, reduced protein binding, and potentially prolonged circulation time of the conjugate. The Boc-protected amines are stable at physiological pH and are only deprotected under acidic conditions, ensuring that the construct remains intact during circulation. Linker stability in plasma is assessed by measuring intact construct concentrations over time using LC-MS/MS. The compound itself is not administered systemically; therefore, its absorption, distribution, metabolism, and excretion are not independently characterized.
|
| Toxicity/Toxicokinetics |
Toxicological data for NH-bis(PEG4-Boc) are limited because it is a research-grade reagent not intended for human use. Standard laboratory safety precautions should be followed when handling this compound: use of personal protective equipment (gloves, safety goggles, lab coat) and handling in a well-ventilated fume hood. The compound should be stored at -20°C or as recommended by the supplier. No genotoxicity, carcinogenicity, or reproductive toxicity data are available. The compound is not classified as a hazardous drug but should be treated with care. Researchers should consult the safety data sheet (SDS) before handling and follow institutional chemical safety guidelines.
|
| References | |
| Additional Infomation |
Additional information for NH-bis(PEG4-Boc): The compound has a CAS number of 2182601-75-6. Its molecular formula is C₃₀H₆₁N₃O₁₂ and molecular weight is 655.82 g/mol. The compound is a PEG-based PROTAC linker. It is a bifunctional molecule containing two amine groups and four ethylene glycol units. The amino groups are reactive with carboxylic acids, activated NHS esters, and carbonyls. The two Boc-protected amines provide protected nucleophiles. The PEG spacer improves solubility and enhances drug delivery. The product is for research use only and is not approved for clinical applications. No FDA approvals or investigational new drug applications exist. The compound is used as a synthetic building block for the preparation of PROTAC protein degraders.
|
| Molecular Formula |
C30H61N3O12
|
|---|---|
| Molecular Weight |
655.818250417709
|
| Exact Mass |
655.425
|
| CAS # |
2182601-75-6
|
| PubChem CID |
129627648
|
| Appearance |
Colorless to light yellow liquid
|
| LogP |
0
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
13
|
| Rotatable Bond Count |
34
|
| Heavy Atom Count |
45
|
| Complexity |
634
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC(C)(C)OC(=O)NCCOCCOCCOCCOCCNCCOCCOCCOCCOCCNC(=O)OC(C)(C)C
|
| InChi Key |
NRRXRYVNQACVQL-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C30H61N3O12/c1-29(2,3)44-27(34)32-9-13-38-17-21-42-25-23-40-19-15-36-11-7-31-8-12-37-16-20-41-24-26-43-22-18-39-14-10-33-28(35)45-30(4,5)6/h31H,7-26H2,1-6H3,(H,32,34)(H,33,35)
|
| Chemical Name |
tert-butyl N-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[(2-methylpropan-2-yl)oxycarbonylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethyl]carbamate
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 1.5248 mL | 7.6240 mL | 15.2481 mL | |
| 5 mM | 0.3050 mL | 1.5248 mL | 3.0496 mL | |
| 10 mM | 0.1525 mL | 0.7624 mL | 1.5248 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
