| Size | Price | Stock | Qty |
|---|---|---|---|
| 500mg |
|
||
| 1g |
|
||
| 5g | |||
| Other Sizes |
| Targets |
Boc-Gly-Gly-Phe-Gly-OH does not have a specific biological target as a drug itself; rather, it serves as a cleavable linker molecule in ADC constructs. The peptide sequence GGFG is recognized and cleaved by specific proteases, including cathepsin B and other lysosomal proteases that are upregulated in tumor microenvironments. When incorporated into ADC molecules, the linker connects the antibody (which targets tumor-associated antigens) to the cytotoxic payload (drug). Upon ADC internalization into target cells via receptor-mediated endocytosis, the linker is cleaved by proteases in the lysosomal compartment, releasing the active drug payload specifically within tumor cells. This protease-cleavable mechanism ensures targeted drug delivery and reduces systemic toxicity.
|
|---|---|
| ln Vitro |
In vitro studies of Boc-Gly-Gly-Phe-Gly-OH focus on its properties as a linker rather than as a pharmacologically active compound. The peptide's ability to be cleaved by proteases is typically assessed using purified enzymes in cell-free assays. Boc-Gly-Gly-Phe-Gly-OH shows good stability in plasma, preventing premature drug release in circulation, while being efficiently cleaved by lysosomal proteases such as cathepsin B in the acidic tumor microenvironment. The peptide's self-assembly properties have also been studied. As an ADC linker, Boc-Gly-Gly-Phe-Gly-OH contributes to the overall stability and efficacy of the ADC construct. Its GGFG sequence is one of the most commonly used cleavable linkers in ADC development, providing a balance of stability in circulation and efficient release of payload in target cells. In cellular studies, Boc-Gly-Gly-Phe-Gly-OH is evaluated as part of complete ADC molecules rather than as a standalone compound. When incorporated into ADCs, the linker enables targeted delivery and release of cytotoxic payloads to cancer cells. In vitro efficacy studies involve treating cancer cells expressing the target antigen with the ADC construct, followed by assessment of cell viability, proliferation, and apoptosis. The specificity of the linker is demonstrated by comparing the activity of ADCs containing the cleavable linker versus non-cleavable linkers or free drug. In antigen-negative cells, the ADC shows minimal activity, confirming target specificity. Intracellular trafficking studies using fluorescently labeled ADCs demonstrate internalization via receptor-mediated endocytosis, followed by lysosomal trafficking and proteolytic cleavage of the GGFG linker, resulting in release of the active payload.
|
| ln Vivo |
In vivo studies of Boc-Gly-Gly-Phe-Gly-OH are conducted in the context of ADC therapeutics. The linker is incorporated into ADC constructs and evaluated in murine xenograft models of human cancer. Immunodeficient mice are implanted subcutaneously with tumor cells expressing the target antigen. When tumors reach a certain size, animals are treated intravenously with the ADC at various dose levels (e.g., 1-30 mg/kg). Tumor volumes are measured twice weekly, and body weights are monitored. Pharmacodynamic endpoints include tumor growth inhibition, tumor regression, and survival prolongation. Toxicity is assessed by monitoring body weight loss, clinical signs, and histopathological examination of major organs. The GGFG linker has been validated in multiple ADC programs, demonstrating favorable pharmacokinetic and safety profiles.
|
| Enzyme Assay |
For protease cleavage assays, Boc-Gly-Gly-Phe-Gly-OH or its ADC conjugate is incubated with purified cathepsin B or other relevant proteases in assay buffer (e.g., 50 mM sodium acetate, pH 5.0, containing 2 mM DTT and 1 mM EDTA) at 37°C for various time points (0, 0.5, 1, 2, 4, 8, 24 hours). The reaction is terminated by addition of protease inhibitors or by heat inactivation. Cleavage products are analyzed by HPLC or LC-MS to quantify the release of the drug payload or to monitor the disappearance of the intact linker. For stability studies in plasma, the compound is incubated in human or mouse plasma at 37°C for up to 24-48 hours, and samples are analyzed at various time points to assess linker stability and resistance to non-specific cleavage. For enzymatic kinetics, Michaelis-Menten parameters (Km and kcat) may be determined using varying substrate concentrations.
|
| Cell Assay |
For cellular uptake and trafficking studies, cancer cells expressing the target antigen are cultured in appropriate medium and treated with fluorescently labeled ADC constructs containing the Boc-Gly-Gly-Phe-Gly-OH linker. Cells are incubated with the ADC (0.01-10 µg/ml) at 37°C for 0.5-24 hours. Cells are then fixed and processed for confocal microscopy to visualize ADC internalization and intracellular trafficking. Co-localization with lysosomal markers (LAMP-1, LAMP-2) is assessed to confirm lysosomal delivery. For efficacy studies, cells are seeded in 96-well plates (5,000-10,000 cells/well) and treated with serial dilutions of ADC (0.001-100 µg/ml) for 72-96 hours. Cell viability is measured by MTT, CCK-8, or CellTiter-Glo assays. IC50 values are calculated from dose-response curves. For apoptosis assays, cells are stained with Annexin V/PI and analyzed by flow cytometry.
|
| Animal Protocol |
For in vivo ADC efficacy studies, 6-8 week old female immunodeficient mice (e.g., nude or SCID) are used. Mice are subcutaneously implanted with 5 × 10⁶ cancer cells expressing the target antigen in the flank. When tumors reach 100-200 mm³, animals are randomized into treatment groups (n = 6-10 per group). ADC containing the Boc-Gly-Gly-Phe-Gly-OH linker is administered intravenously via tail vein injection at doses of 1, 3, 10, or 30 mg/kg, typically on a q4d × 4 or q7d × 3 schedule. Control groups receive vehicle, non-targeting ADC, or free drug. Tumor volumes are measured twice weekly using calipers (volume = length × width² / 2). Body weights are monitored for toxicity. At study termination (day 21-28 or when tumors reach 2000 mm³), tumors are excised, weighed, and processed for histopathology or biomarker analysis. Blood samples may be collected for pharmacokinetic analysis of the ADC and released payload.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties of Boc-Gly-Gly-Phe-Gly-OH are studied as part of ADC constructs. The linker's stability in circulation contributes to the overall ADC pharmacokinetic profile. ADCs containing the GGFG linker typically show a biphasic pharmacokinetic profile with an initial distribution phase followed by a slower elimination phase. The linker is designed to be stable in plasma (half-life > 24 hours) to prevent premature drug release, while being rapidly cleaved by lysosomal proteases upon ADC internalization. The released drug payload is then subject to its own pharmacokinetic properties, including metabolism and excretion. As a peptide linker, Boc-Gly-Gly-Phe-Gly-OH itself is cleared through proteolytic degradation and renal excretion. No dedicated ADME studies for the linker alone are typically performed.
|
| Toxicity/Toxicokinetics |
Toxicological data for Boc-Gly-Gly-Phe-Gly-OH are derived from ADC toxicology studies rather than studies of the linker alone. The linker is considered a non-toxic component of ADC constructs, as it is a small peptide that is metabolized to non-toxic amino acid degradation products. The safety and tolerability of ADCs containing the GGFG linker are evaluated in animal toxicology studies, typically using rats and cynomolgus monkeys. ADCs are administered at various dose levels, and parameters assessed include clinical observations, body weight, food consumption, clinical pathology (hematology, serum chemistry, coagulation), organ weights, and histopathology. The GGFG linker has demonstrated acceptable safety in multiple ADC programs, with toxicity primarily attributed to the payload or the antibody rather than the linker itself.
|
| Additional Infomation |
Boc-Gly-Gly-Phe-Gly-OH is a tetrapeptide linker used in the development of antibody-drug conjugates (ADCs). Its sequence GGFG is one of the most well-validated protease-cleavable linkers in the ADC field. The Boc-protected N-terminus allows for selective conjugation to antibodies, while the free C-terminal carboxyl group enables attachment of drug payloads. Upon ADC internalization into target cells and trafficking to lysosomes, the GGFG linker is cleaved by cathepsin B and other lysosomal proteases, releasing the cytotoxic payload specifically within tumor cells. This approach improves the therapeutic index of cytotoxic drugs by reducing systemic toxicity and enhancing tumor-specific delivery. Boc-Gly-Gly-Phe-Gly-OH is strictly for research use and has not been approved as a standalone therapeutic. It is a fundamental building block for next-generation ADC development.
|
| Molecular Formula |
C20H28N4O7
|
|---|---|
| Molecular Weight |
436.45892
|
| Exact Mass |
436.196
|
| Elemental Analysis |
C, 55.04; H, 6.47; N, 12.84; O, 25.66
|
| CAS # |
187794-49-6
|
| Related CAS # |
Boc-Gly-Gly-Phe-Gly-OH TFA;2450273-39-7
|
| PubChem CID |
10873788
|
| Appearance |
White to off-white solid powder
|
| LogP |
2.28
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
12
|
| Heavy Atom Count |
31
|
| Complexity |
654
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
CC(C)(OC(NCC(NCC(N[C@H](C(NCC(O)=O)=O)CC1=CC=CC=C1)=O)=O)=O)C
|
| InChi Key |
PTUJJIPXBJJLLV-AWEZNQCLSA-N
|
| InChi Code |
InChI=1S/C20H28N4O7/c1-20(2,3)31-19(30)23-10-15(25)21-11-16(26)24-14(18(29)22-12-17(27)28)9-13-7-5-4-6-8-13/h4-8,14H,9-12H2,1-3H3,(H,21,25)(H,22,29)(H,23,30)(H,24,26)(H,27,28)/t14-/m0/s1
|
| Chemical Name |
2-[[(2S)-2-[[2-[[2-[(2-methylpropan-2-yl)oxycarbonylamino]acetyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]acetic acid
|
| Synonyms |
Boc-Gly-Gly-Phe-Gly-OH;
|
| 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: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
DMSO : ≥ 100 mg/mL (~229.12 mM)
H2O : ~25 mg/mL (~57.28 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.73 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 (5.73 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. View More
Solubility in Formulation 3: 33.33 mg/mL (76.36 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2912 mL | 11.4558 mL | 22.9116 mL | |
| 5 mM | 0.4582 mL | 2.2912 mL | 4.5823 mL | |
| 10 mM | 0.2291 mL | 1.1456 mL | 2.2912 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
