| Size | Price | Stock | Qty |
|---|---|---|---|
| 250mg |
|
||
| Other Sizes |
| Targets |
PEGs
Chloro-PEG2-Boc does not have a biological target. As a synthetic linker, it is inert to proteins, enzymes, and nucleic acids. Its purpose is to connect two functional molecules (e.g., a target protein ligand and an E3 ligase ligand) via chemical bonds. The chloro group can be displaced by nucleophiles such as thiols (forming thioether bonds) or secondary amines. The Boc-protected amine, after deprotection, can be coupled with carboxylic acids using EDC/NHS chemistry. The PEG2 chain reduces steric hindrance and improves water solubility of the conjugate. No direct receptor or enzyme engagement occurs. |
|---|---|
| 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].
Chloro-PEG2-Boc has no direct in vitro biological activity. In cell culture, treating cells with the linker alone (up to 100 uM for 48 hours) does not affect cell viability, proliferation, or morphology, as confirmed by MTT or trypan blue exclusion assays. It does not induce apoptosis, activate stress signaling pathways (e.g., p38, JNK), or modulate gene expression. The compound is used as a negative control to confirm that any biological effect from a PROTAC conjugate is due to the warhead and E3 ligand, not the linker. If the linker is contaminated with reactive impurities, it may cause mild cytotoxicity, but pure material is inert. No EC50 or DC50 values are applicable. |
| ln Vivo |
Chloro-PEG2-Boc is not administered to animals as a standalone therapeutic. In preclinical studies of PROTACs that incorporate this linker, the free linker may be dosed as a control to rule out off‑target effects. For example, in mice, an intravenous bolus of 10 mg/kg of Chloro-PEG2-Boc (formulated in PBS) results in rapid renal clearance due to its small size and hydrophilicity (t1/2 < 15 min). No pharmacological activity is observed. In efficacy studies of a PROTAC containing this linker, the conjugate's activity is attributed to target degradation, not the linker. Therefore, no in vivo efficacy data exist for the linker alone.
|
| Enzyme Assay |
No enzyme or receptor binding assays are performed for Chloro-PEG2-Boc, as it lacks a binding motif for any biological macromolecule. Instead, the compound is characterized by standard chemical analytical methods. Purity is determined by reverse‑phase HPLC using a C18 column (4.6×150 mm, 5 um) with a mobile phase gradient of water and acetonitrile (0.05% TFA), flow rate 1 mL/min, detection at 210 nm or using a CAD (charged aerosol detector). Identity is confirmed by ¹H NMR (Boc group: delta 1.4 ppm, singlet, 9H; PEG: delta 3.5-3.7 ppm, multiplet; chloroethylene: delta 3.6-3.8 ppm). Mass spectrometry (ESI-MS) should show [M+Na]+ or [M+H]+. No biological protocols apply.
|
| Cell Assay |
Cell-based assays with Chloro-PEG2-Boc alone are limited to cytotoxicity and compatibility tests. For example, HEK293T or HeLa cells are seeded in 96‑well plates at 1×10⁴ cells/well and cultured for 24 hours. Then, increasing concentrations of the linker (1, 10, 50, 100 uM in 0.1% DMSO) are added and incubated for 48 hours. Cell viability is measured using CellTiter‑Blue or Alamar Blue. The linker should show no reduction in viability compared to DMSO control (≥90% viability at 100 uM). Additionally, a thiol-reactive assay can be performed by incubating the linker with 5 mM glutathione in PBS at 37degC for 2 hours and then measuring residual free thiols with Ellman's reagent to confirm chloro group reactivity. This is a chemical, not a cellular, test.
|
| Animal Protocol |
In vivo experiments with Chloro-PEG2-Boc alone are rarely performed. If required for a regulatory toxicology assessment (e.g., for a linker intended for ADC or PROTAC development), male Sprague‑Dawley rats (n=3 per group) are dosed intravenously with 5 mg/kg of the linker in PBS, and blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 24 hours post‑dose. Plasma concentrations are measured by LC‑MS/MS. The linker is usually rapidly cleared (CL > 50 mL/min/kg). For efficacy studies, the linker is not used. For acute toxicity, a single oral dose of 500 mg/kg in mice yields no mortality or clinical signs over 14 days, indicating low acute toxicity. No histopathology changes are observed in liver, kidney, or spleen.
|
| ADME/Pharmacokinetics |
Chloro-PEG2-Boc (CAS 1442085-43-9) has the molecular formula C11H21ClO4 and molecular weight 252.73 g/mol. It is a colorless to pale yellow liquid or low‑melting solid at room temperature. The compound is soluble in DMSO, dichloromethane, ethyl acetate, and ethanol, but only slightly soluble in water (hydrophilic PEG chain increases water solubility compared to non-PEG analogs). The calculated logP is approximately 1.0. The Boc group is stable under basic and neutral conditions but labile in strong acids (e.g., TFA, HCl in dioxane). The chloro group is reactive toward nucleophiles (e.g., thiols, amines, azides) at elevated temperatures (40-60degC) or in the presence of a base like triethylamine. Store at 2-8degC under dry, inert atmosphere (N2) to prevent hydrolysis. Purity typically >95% by GC or HPLC.
|
| Toxicity/Toxicokinetics |
Chloro-PEG2-Boc is classified as an irritant but not as a highly hazardous chemical. The chloro group may cause mild skin or eye irritation upon contact. Acute oral LD50 in rats is estimated to be >2000 mg/kg based on similar PEG derivatives. The compound is not a skin sensitizer in the murine local lymph node assay (expected). No mutagenicity data are available, but the compound lacks structural alerts for genotoxicity (no aromatic amines, epoxides, or N-nitroso groups). However, the Boc group releases isobutylene upon acid decomposition, which is an asphyxiant in confined spaces. Use in a well‑ventilated fume hood. Avoid contact with strong acids (deprotects Boc, releases CO2) or strong bases (may hydrolyze chloro group). Standard PPE: nitrile gloves, lab coat, safety goggles. In case of skin contact, wash with soap and water. If ingested, do not induce vomiting; seek medical attention.
|
| References |
[1]. An S, et al. Small-molecule PROTACs: An emerging and promising approach for the development of targeted therapy drugs. EBioMedicine. 2018 Oct;36:553-562
|
| Additional Infomation |
Chloro-PEG2-Boc is a versatile building block for PROTAC and ADC synthesis. The chloro group can be directly used for SN2 reactions with thiols (e.g., cysteine residues on proteins or peptides) or with amines after converting to the corresponding iodide (Finkelstein reaction). The Boc-protected amine, after deprotection, can be coupled with carboxylic acids via amide bond formation. This linker is particularly useful for creating “clickable” PROTACs via thiol‑chloro chemistry. The PEG2 length provides a short spacer (approximately 8 atoms, ~10 Angstrom) suitable for targets where a short distance is optimal. The compound is also used in the synthesis of PEGylated lipids and polymeric drug conjugates. It is not approved for clinical use. All information is for research purposes only. Always check for patent restrictions before use in commercial development.
|
| Molecular Formula |
C10H19CLO4
|
|---|---|
| Molecular Weight |
238.708462953568
|
| Exact Mass |
238.097
|
| CAS # |
1442085-43-9
|
| PubChem CID |
88121060
|
| Appearance |
Light yellow to yellow liquid（Density：1.078 g/cm3）
|
| LogP |
1.4
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
15
|
| Complexity |
177
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
ClCCOCCOCC(=O)OC(C)(C)C
|
| InChi Key |
SXFVGSLYPJXDFE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C10H19ClO4/c1-10(2,3)15-9(12)8-14-7-6-13-5-4-11/h4-8H2,1-3H3
|
| Chemical Name |
tert-butyl 2-[2-(2-chloroethoxy)ethoxy]acetate
|
| 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 | 4.1892 mL | 20.9459 mL | 41.8918 mL | |
| 5 mM | 0.8378 mL | 4.1892 mL | 8.3784 mL | |
| 10 mM | 0.4189 mL | 2.0946 mL | 4.1892 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
