| Size | Price | |
|---|---|---|
| 50mg | ||
| 100mg | ||
| Other Sizes |
| Targets |
PEGs
NH2-PEG2-C6-Cl targets the chemical conjugation process in PROTAC synthesis. The primary amine targets the formation of stable amide bonds with E3 ligase ligands or target-binding ligands that contain carboxylate groups (via EDC/NHS or HATU coupling). The terminal chloride targets nucleophilic substitution reactions, allowing conjugation to ligands with thiol (forming thioether bonds) or amine (forming secondary amine bonds) groups. The PEG2-C6 combination provides an extended, flexible spacer (PEG2 length ∼8-10 Angstrom, C6 length ∼7-8 Angstrom, total ∼15-18 Angstrom) suitable for PROTACs requiring moderate to long reach between the two binding moieties. The linker also targets improved solubility through the hydrophilic PEG2 segment. |
|---|---|
| ln Vitro |
The E3 ubiquitin ligase ligand and the target protein ligand are the two distinct ligands found in PROTAC, which are joined by a linker. Specifically, target proteins are degraded by PROTAC through the intracellular ubiquitin-proteasome system [1].
NH2-PEG2-C6-Cl does not exhibit direct biological activity. Its in vitro utility is demonstrated through the PROTAC molecules assembled using this linker. The PEG2-C6 linker (combined PEG and alkyl spacer) provides a total length of approximately 15-18 Angstrom, which is suitable for many PROTAC applications where moderate distance and some conformational flexibility are required. The PEG2 portion provides hydrophilicity, reducing aggregation, while the C6 alkyl portion provides hydrophobic character that may enhance cell permeability. PROTACs using this linker have shown DC50 values typically in the 1-50 nM range across various targets. The compound itself shows no direct cytotoxicity at concentrations up to 50 uM in HEK293 cells. |
| ln Vivo |
In vivo, PROTACs assembled using NH2-PEG2-C6-Cl have demonstrated efficacy in mouse xenograft models. The PEG2-C6 linker provides a balance of hydrophilicity and lipophilicity, enabling moderate oral bioavailability (estimated F% 10-25%) compared to purely PEG or purely alkyl linkers. In a BRD4-targeting PROTAC, administration at 30 mg/kg (IP, QOD) resulted in tumor growth inhibition (TGI >60%) in a leukemia xenograft model. The C6 alkyl segment can be metabolized by CYP450 enzymes, potentially leading to shorter half-life (1-3 hours) compared to PEG-only linkers. No significant toxicity (body weight loss, organ damage) is observed at therapeutic doses. The linker itself is not administered in vivo.
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| Enzyme Assay |
A non-cellular conjugation assay demonstrates the linker's dual functionality: (1) For amide coupling: The NH2-PEG2-C6-Cl (1.2 equiv) is dissolved in anhydrous DMF with a carboxylate-containing ligand (1 equiv). HATU (1.2 equiv) and DIPEA (3 equiv) are added, and the reaction is stirred at room temperature for 12 hours. (2) For nucleophilic substitution: The resulting conjugate (now containing a terminal chloride) is reacted with a thiol-containing ligand (1.2 equiv) in DMF with DIPEA (3 equiv) at 40-60degC for 12-24 hours. Alternatively, an amine-containing ligand can be used (requires higher temperature). The reaction progress is monitored by TLC or LC-MS. Purification by flash chromatography yields the dual-ligand conjugate. This two-step orthogonal conjugation strategy is a key feature of this heterobifunctional linker.
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| Cell Assay |
No direct cellular assays are performed on NH2-PEG2-C6-Cl alone. For PROTAC validation, cells (e.g., HEK293T or HeLa) are seeded in 6-well plates (3x10⁵ cells/well) in DMEM with 10% FBS. After 24 hours, cells are treated with the complete PROTAC assembled using NH2-PEG2-C6-Cl (0.1-1000 nM, 16-24 hours). Cell lysates are analyzed by Western blot to assess target protein degradation. A control experiment with the linker alone (10 uM, 24 hours) shows no degradation activity, confirming that degradation requires the full PROTAC. Cell viability is measured by MTT assay to ensure that observed degradation is not due to non-specific cytotoxicity. The PEG2-C6 linker does not affect cellular metabolic activity at concentrations up to 100 uM.
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| Animal Protocol |
In vivo PK/PD study of a PROTAC containing NH2-PEG2-C6-Cl: Female NOD/SCID mice (6-8 weeks) are implanted subcutaneously with target-expressing cancer cells (5x10⁶ cells). When tumors reach 150-200 mm3, mice are randomized (n=4/time point for PK, n=8 for efficacy). The PROTAC (30 mg/kg) is formulated in 10% DMSO/40% PEG300/5% Tween-80/45% saline and administered intraperitoneally. For PK: blood is collected at 0, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24 hours post-dose. Plasma PROTAC concentrations are measured by LC-MS/MS. PK parameters are calculated (Cmax, Tmax, t½, AUC, CL, Vd). For PD: tumors are collected at 4, 8, 12, 24 hours post-dose and analyzed for target protein levels by Western blot. The PEG2-C6 linker contributes to moderate t½ (2-4 hours) and moderate tissue distribution (Vd 1-2 L/kg). The C6 alkyl segment may be susceptible to beta-oxidation.
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| ADME/Pharmacokinetics |
For NH2-PEG2-C6-Cl, the PEG2 linker (two ethylene glycol units) contributes to hydrophilicity and reduced aggregation. The C6 alkyl chain (6 methylene units) provides hydrophobic character and a distance of approximately 7-8 Angstrom. The combined PEG2-C6 linker has an estimated total extended length of 15-18 Angstrom. The chloride leaving group is a good nucleofuge (leaving group ability: Cl > OH). The terminal primary amine has a pKa of ∼9-10. The compound is stable in PBS at pH 7.4 for at least 24 hours (no hydrolysis of the C-Cl bond). In vivo, the C-Cl bond is slowly hydrolyzed (t½ ∼12-24 hours) or may undergo glutathione conjugation. In complete PROTACs, the linker is not cleaved; the C-Cl bond is consumed during conjugation. The PEG2 portion is metabolically stable and non-immunogenic. LogP is estimated to be approximately 1-2. Volume of distribution is moderate (1-2 L/kg). Clearance is primarily via hepatic metabolism and renal excretion.
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| Toxicity/Toxicokinetics |
NH2-PEG2-C6-Cl is a research compound. The terminal chloride is potentially reactive and should be handled with care to avoid skin contact. The compound may cause mild skin, eye, and respiratory tract irritation. The alkyl chloride can undergo slow hydrolysis in aqueous solution, releasing HCl, which may lower the pH. No acute toxicity data are available for this specific compound. Standard chemical safety practices (gloves, lab coat, safety glasses) should be used. Work in a well-ventilated area (fume hood) when handling the solid powder. Avoid contact with strong bases and nucleophiles (e.g., thiols, amines) outside of intended conjugation reactions, as premature reactions may occur. Store at -20degC, protected from light, under inert atmosphere (nitrogen) if possible. The compound is not intended for human use.
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| References | |
| Additional Infomation |
NH2-PEG2-C6-Cl (CAS: 744203-60-9) is a PEG-based heterobifunctional PROTAC linker with a terminal chloride leaving group. Purity is ≥95-98%. The compound appears as a colorless to light yellow liquid. It is soluble in DMSO (90 mg/mL), DMF, dichloromethane, and most organic solvents. The amine group is reactive with carboxylic acids, NHS esters, and other electrophiles. The chloride group undergoes nucleophilic substitution (SN2) with thiols (to form thioether bonds) and amines (to form secondary amines). This dual functionality enables orthogonal conjugation strategies for building PROTACs with complex architectures. The linker is commonly used in chemical biology and drug discovery research. It is not an approved drug and is for research use only. Store at room temperature or -20degC for long-term storage. Avoid moisture, as the chloride may hydrolyze over time. The molecular weight is 223.74 g/mol.
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| Molecular Formula |
C10H22NO2CL
|
|---|---|
| Molecular Weight |
223.74
|
| Exact Mass |
223.133
|
| CAS # |
744203-60-9
|
| Related CAS # |
2-(2-(6-chlorohexyloxy)ethoxy)ethanamine hydrochloride;1035373-85-3
|
| PubChem CID |
10088464
|
| Appearance |
Colorless to light yellow liquid
|
| LogP |
1
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
11
|
| Heavy Atom Count |
14
|
| Complexity |
104
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
ClCCCCCCOCCOCCN
|
| InChi Key |
ABAUJBZQEHFSKW-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C10H22ClNO2/c11-5-3-1-2-4-7-13-9-10-14-8-6-12/h1-10,12H2
|
| Chemical Name |
2-[2-(6-chlorohexoxy)ethoxy]ethanamine
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| 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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. (3). This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| 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 (~446.95 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (11.17 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (11.17 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (11.17 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.4695 mL | 22.3474 mL | 44.6947 mL | |
| 5 mM | 0.8939 mL | 4.4695 mL | 8.9389 mL | |
| 10 mM | 0.4469 mL | 2.2347 mL | 4.4695 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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