| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
PEG4 linker functions as a structural component in ADC design rather than targeting a biological receptor or enzyme directly. The linker covalently attaches the cytotoxic payload MMAF to the targeting antibody, ensuring stable conjugation until the ADC reaches its target cells. The MMAF payload is a potent antitubulin agent that inhibits microtubule polymerization, leading to cell cycle arrest and apoptosis in cancer cells. The non-cleavable PEG4 linker provides stability in circulation, preventing premature release of the cytotoxic payload. This linker-payload construct is designed for conjugation to antibodies targeting tumor-associated antigens, such as anti-HER2 antibodies for breast cancer treatment.
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| ln Vitro |
PEG4 linker demonstrates potent in vitro activity as part of the complete ADC construct in which it is incorporated. The MMAF payload linked via PEG4 exhibits potent antitumor activity by inhibiting tubulin polymerization and disrupting microtubule dynamics. In cell-based assays, ADCs containing the PEG4-aminooxy-MMAF construct show potent cytotoxicity against antigen-expressing cancer cell lines. The non-cleavable nature of the PEG4 linker ensures that the MMAF payload remains attached even after internalization, leading to sustained cytotoxic activity within the target cell. The linker's hydrophilic properties contribute to favorable ADC characteristics including reduced aggregation and improved pharmacokinetics.
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| ln Vivo |
In vivo activity of PEG4 linker is evaluated through studies of ADC constructs incorporating this linker in animal models of human cancer. The PEG4 linker has been used in the synthesis of ADCT-502, an anti-HER2 ADC for breast cancer treatment. In tumor-bearing mouse models, ADCs containing the PEG4-aminooxy-MMAF construct demonstrate significant antitumor efficacy with reduced off-target toxicity compared to cleavable linkers. The non-cleavable PEG4 linker contributes to improved stability in circulation, resulting in a longer half-life and reduced premature payload release. The linker's hydrophilic properties also contribute to favorable biodistribution and reduced aggregation, enhancing overall ADC performance in vivo.
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| Enzyme Assay |
In vitro enzyme/receptor binding assays are not directly applicable to PEG4 linker as it is a structural component rather than a traditional enzyme inhibitor or receptor ligand. However, the linker's properties can be characterized using biochemical methods. High-performance liquid chromatography (HPLC) and mass spectrometry are used to verify the molecular weight (923.19 g/mol) and chemical composition (C47H82N6O12). The purity (≥98%) is confirmed by HPLC and NMR analysis. Stability studies assess the linker's integrity in various buffer systems and plasma by incubating the compound in relevant media and monitoring degradation over time using analytical techniques. Conjugation efficiency to antibodies and payloads can be evaluated using UV-Vis spectroscopy or size-exclusion chromatography.
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| Cell Assay |
In vitro cellular assays for PEG4 linker are performed using the complete ADC construct rather than the linker alone. Cancer cell lines expressing the target antigen are cultured in appropriate media and treated with varying concentrations of the ADC. Following incubation, cell viability is measured using standard assays such as MTT, CellTiter-Glo, or resazurin reduction. IC50 values are calculated from dose-response curves to determine the potency of the ADC. Antigen binding and internalization are assessed using flow cytometry or immunofluorescence microscopy. The mechanism of cell death (apoptosis vs. necrosis) can be investigated using annexin V/propidium iodide staining or caspase activity assays. Cytotoxicity is compared between target antigen-positive and antigen-negative cell lines to confirm target specificity.
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| Animal Protocol |
In vivo animal studies for PEG4 linker are conducted using immunodeficient mice bearing human tumor xenografts expressing the target antigen. Typically, 6-8 week old female athymic nude mice or NSG mice are implanted subcutaneously with cancer cells. Once tumors reach a predetermined size (e.g., 100-200 mm3), animals are randomized into treatment groups and administered the ADC construct via intravenous injection at various doses and schedules. Tumor size is measured twice weekly using calipers, and body weight is monitored as a safety indicator. At study termination, tumors are excised, weighed, and processed for histopathological analysis or biomarker assessment. Pharmacokinetic samples are collected at multiple time points to determine ADC stability and exposure in circulation. Efficacy is expressed as tumor growth inhibition (TGI) relative to vehicle control.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of PEG4 linker are characterized as part of the ADC construct. The linker's non-cleavable nature contributes to overall ADC stability in circulation, resulting in a longer half-life and reduced premature payload release compared to cleavable linkers. The PEG4 spacer provides optimal hydrophilicity, reducing ADC aggregation and improving pharmacokinetic properties. The ADC typically exhibits biphasic elimination, with an initial distribution phase followed by a slower elimination phase. The volume of distribution is generally limited to the vascular compartment due to the large molecular weight of the ADC construct. Clearance occurs primarily through proteolytic degradation and elimination of small peptide fragments. The linker's hydrophilicity can influence pharmacokinetic behavior, which is typically assessed by size-exclusion chromatography during ADC development.
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| Toxicity/Toxicokinetics |
PEG4 linker is intended for laboratory research use only and has not undergone comprehensive toxicology testing. As a non-cleavable ADC linker, the compound is not expected to be directly toxic, but it influences the toxicity profile of the ADC through its impact on stability, pharmacokinetics, and payload release. Standard in vitro cytotoxicity assays in cell lines are typically performed alongside efficacy studies to rule out nonspecific toxicity. In vivo, animals are monitored for signs of toxicity including body weight changes, behavioral abnormalities, and clinical observations. The non-cleavable nature of the linker may reduce off-target toxicity by minimizing premature release of the MMAF payload in circulation. However, on-target toxicity can still occur due to antigen expression on normal tissues. The compound is not approved for human use and is strictly intended for research purposes.
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| References | |
| Additional Infomation |
PEG4 linker (PEG4-aminooxy-MMAF, Amberstatin, AS269) is a drug-linker conjugate designed for antibody-drug conjugate (ADC) synthesis. It consists of the potent antitubulin agent MMAF linked via a non-cleavable PEG4 spacer. The compound has a molecular formula of C47H82N6O12 and a molecular weight of 923.19 g/mol. PEG4 linker has been used in the synthesis of ADCT-502, an anti-HER2 ADC for breast cancer treatment. The linker's non-cleavable nature and hydrophilic PEG4 spacer contribute to ADC stability and favorable pharmacokinetics. The compound has not entered clinical trials and has not received regulatory approval for any indication. It is available from research chemical suppliers for non-clinical research purposes only.
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| Molecular Formula |
C47H82N6O12
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|---|---|
| Molecular Weight |
923.186994075775
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| Exact Mass |
922.599
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| CAS # |
1415246-35-3
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| Related CAS # |
2364345-05-9 (bis);1415246-35-3;
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| PubChem CID |
89283237
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| Appearance |
White to off-white solid powder
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| LogP |
1.2
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
33
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| Heavy Atom Count |
65
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| Complexity |
1400
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| Defined Atom Stereocenter Count |
9
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| SMILES |
O(C)[C@H]([C@H](C(N[C@H](C(=O)O)CC1C=CC=CC=1)=O)C)[C@@H]1CCCN1C(C[C@H]([C@H]([C@@H](C)CC)N(C)C([C@H](C(C)C)NC([C@H](C(C)C)N(C)CCOCCOCCOCCON)=O)=O)OC)=O
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| InChi Key |
OJYZGXVPCDZLCK-KVEFUIBJSA-N
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| InChi Code |
InChI=1S/C47H82N6O12/c1-12-33(6)42(52(9)46(57)40(31(2)3)50-45(56)41(32(4)5)51(8)21-22-62-23-24-63-25-26-64-27-28-65-48)38(60-10)30-39(54)53-20-16-19-37(53)43(61-11)34(7)44(55)49-36(47(58)59)29-35-17-14-13-15-18-35/h13-15,17-18,31-34,36-38,40-43H,12,16,19-30,48H2,1-11H3,(H,49,55)(H,50,56)(H,58,59)/t33-,34+,36-,37-,38+,40-,41-,42-,43+/m0/s1
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| Chemical Name |
(2S)-2-[[(2R,3R)-3-[(2S)-1-[(3R,4S,5S)-4-[[(2S)-2-[[(2S)-2-[2-[2-[2-(2-aminooxyethoxy)ethoxy]ethoxy]ethyl-methylamino]-3-methylbutanoyl]amino]-3-methylbutanoyl]-methylamino]-3-methoxy-5-methylheptanoyl]pyrrolidin-2-yl]-3-methoxy-2-methylpropanoyl]amino]-3-phenylpropanoic acid
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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: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). 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) |
H2O : ≥ 100 mg/mL (~108.32 mM)
DMSO : ~100 mg/mL (~108.32 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.71 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 (2.71 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 (2.71 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 | 1.0832 mL | 5.4160 mL | 10.8320 mL | |
| 5 mM | 0.2166 mL | 1.0832 mL | 2.1664 mL | |
| 10 mM | 0.1083 mL | 0.5416 mL | 1.0832 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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