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Tr-PEG4

Cat No.:V16884 Purity: ≥98%
Tr-PEG3-OH is a non-cleavable (non-degradable) ADC linker containing 3 Polyethylene glycol (PEG) units, which may be utilized to prepare active Antibody-drug conjugates (ADC).
Tr-PEG4
Tr-PEG4 Chemical Structure CAS No.: 133699-09-9
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
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Product Description
Tr-PEG3-OH is a non-cleavable (non-degradable) ADC linker containing 3 Polyethylene glycol (PEG) units, which may be utilized to prepare active Antibody-drug conjugates (ADC).
Tr-PEG4 is a polyethylene glycol (PEG) derivative containing a trityl protecting group and a terminal hydroxyl group. It is a non-cleavable linker used in the synthesis of antibody-drug conjugates (ADCs). The PEG4 chain is key to its hydrophilicity, providing high water solubility and good biocompatibility, which allows Tr-PEG4 to disperse stably in aqueous solutions and reduce molecular interactions and aggregation. Its primary targets are the antibodies to which it is attached.
Biological Activity I Assay Protocols (From Reference)
Targets
As a chemical linker, Tr-PEG4 does not have a specific biological target. Its primary role is as a linker in ADCs. The antibody guides the ADC to cancer cells, and once bound to the antigen, the ADC is internalized, and the cytotoxic drug is released to kill the cancer cell. The exact biochemical pathways affected depend on the specific antibody and cytotoxic drug used in the ADC.
ln Vitro
An ADC cytotoxin is connected to an antibody by use of an ADC linker to form an ADC.
Tr-PEG4 itself does not exhibit direct biological activity. Its activity is dependent on the final conjugated ADC molecule. The linker's properties, such as length and hydrophilicity, can influence the pharmacokinetics and efficacy of the final construct. The PEG linker can enhance the solubility and stability of the ADC.
ln Vivo
Tr-PEG4 is not a therapeutic agent and does not have direct in vivo activity. Its function is as a chemical intermediate in the synthesis of active pharmaceutical ingredients. The in vivo properties of the final ADC are determined by the entire molecule, including the antibody, payload, and linker. ADCs are designed to have a long circulation time to maximize the chance of reaching target cancer cells.
Enzyme Assay
Non-cell based assays are not applicable for Tr-PEG4 as it is not a direct enzyme inhibitor or receptor ligand. Its identity and purity are confirmed using analytical techniques such as Nuclear Magnetic Resonance (NMR) spectroscopy and Mass Spectrometry. Its utility as a linker is assessed in chemical synthesis, where the successful formation of the desired conjugate is verified.
Cell Assay
Cell-based assays are not applicable for Tr-PEG4, as it is not a biologically active compound. Its use is strictly limited to chemical synthesis. It is not tested in cell-based assays for direct biological activity. The biological activity is evaluated on the final ADC molecule that incorporates this linker.
Animal Protocol
In vivo animal experiments are not applicable for Tr-PEG4 as it is not a therapeutic agent. It is not administered to animals for pharmacological or toxicological studies. Its use is limited to the synthesis of other compounds. The pharmacokinetics of the final ADC can be studied in animal models.
ADME/Pharmacokinetics
Pharmacokinetic (PK) properties are not applicable for Tr-PEG4 as a standalone drug substance. However, as part of an ADC, a PEG linker can enhance the solubility and stability of the conjugate, potentially improving its PK properties. The compound has a molecular weight of 392.5 g/mol and a molecular formula of C25H28O4. It should be stored at -20°C in a dry, dark place.
Toxicity/Toxicokinetics
Toxicology data are not relevant for Tr-PEG4 as a standalone drug substance, as it is not administered to patients. As a chemical reagent, it should be handled with appropriate safety precautions. It may be an irritant if ingested, inhaled, or absorbed through the skin.
References

[1]. Glycan-Directed Grafting-from Polymerization of Immunoglobulin G: Site-Selectively Modified IgG-Polymer Conjugates with Preserved Biological Activity. Biomacromolecules. 2018 Jul 9;19(7):3086-3095.

Additional Infomation
Other information: Tr-PEG4 is a PEG linker used in the synthesis of antibody-drug conjugates (ADCs). It is also known as 2-[2-(2-trityloxyethoxy)ethoxy]ethanol. The compound is available from chemical suppliers for research purposes. Its CAS number is 133699-09-9.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C25H28O4
Molecular Weight
392.48700
Exact Mass
392.199
CAS #
133699-09-9
PubChem CID
11825346
Appearance
Colorless to light yellow viscous liquid
Density
1.126±0.06 g/cm3(Predicted)
Boiling Point
521.8±45.0 °C(Predicted)
LogP
4.02
Hydrogen Bond Donor Count
1
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
12
Heavy Atom Count
29
Complexity
364
Defined Atom Stereocenter Count
0
SMILES
C1=CC=C(C=C1)C(C2=CC=CC=C2)(C3=CC=CC=C3)OCCOCCOCCO
InChi Key
CKYIMQGYUMGSEO-UHFFFAOYSA-N
InChi Code
InChI=1S/C25H28O4/c26-16-17-27-18-19-28-20-21-29-25(22-10-4-1-5-11-22,23-12-6-2-7-13-23)24-14-8-3-9-15-24/h1-15,26H,16-21H2
Chemical Name
2-[2-(2-trityloxyethoxy)ethoxy]ethanol
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 Data
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
(e.g. IP/IV/IM/SC)
Injection Formulation 1: DMSO : Tween 80: Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution 50 μL Tween 80 850 μL Saline)
*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 : PEG300Tween 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).
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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO 100 μLPEG300 200 μL castor oil 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10: 10 : 80 (i.e. 100 μL Ethanol 100 μL Cremophor 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH 400 μLPEG300 50 μL Tween 80 450 μL 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).
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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders


Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.5478 mL 12.7392 mL 25.4784 mL
5 mM 0.5096 mL 2.5478 mL 5.0957 mL
10 mM 0.2548 mL 1.2739 mL 2.5478 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.

Calculator

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

  • Calculate the Mass of a compound required to prepare a solution of known volume and concentration
  • Calculate the Volume of solution required to dissolve a compound of known mass to a desired concentration
  • Calculate the Concentration of a solution resulting from a known mass of compound in a specific volume
An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
  • Enter 350.26 in the Molecular Weight (MW) box
  • Enter 10 in the Concentration box and choose the correct unit (mM)
  • Enter 5 in the Volume box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
  • Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
  • Enter 25 into the Concentration (End) box and select the correct unit (mM)
  • Enter 25 into the Volume (End) box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
  • To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.
Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
  • Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.
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Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

  • Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
  • Click the “Calculate” button
  • The answer appears in the Volume (to add to vial) box
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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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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