| Size | Price | Stock | Qty |
|---|---|---|---|
| 250mg |
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| Other Sizes |
| Targets |
Cleavable Linker
Fmoc-Gly-Gly-D-Phe-OH does not target a specific biological receptor. It functions as a cleavable ADC linker scaffold. The Gly-Gly-D-Phe sequence is designed to be recognized and cleaved by specific proteases, such as cathepsin B, in the lysosomal compartment of target cells upon ADC internalization. The D-Phe residue may enhance stability toward proteolytic degradation in circulation. Biological activity resides in the full ADC, not the linker alone. |
|---|---|
| ln Vitro |
ADCs are made up of an antibody and an ADC cytotoxin that are joined together by an ADC linker[1].
No independent in vitro biological activity is attributed to the linker itself. In the context of an ADC, the linker connects the antibody to the cytotoxic payload. When conjugated to a potent payload (e.g., MMAE, DM1, Duocarmycin, or PBD dimer) and an antibody targeting a cancer cell-surface antigen, the resulting ADC shows target-dependent cytotoxicity. In vitro assays demonstrate that ADCs containing this linker kill antigen-positive cancer cells with high potency (IC50 in pM to nM range) while having minimal effect on antigen-negative cells. |
| ln Vivo |
In vivo studies are not performed with the linker alone. ADCs constructed using this linker have demonstrated antitumor efficacy in xenograft mouse models. The D-isomer may provide enhanced in vivo stability compared to the L-isomer, potentially improving the therapeutic window. Efficacy studies in subcutaneous and orthotopic tumor models show dose-dependent tumor growth inhibition, regression, and improved survival with ADCs utilizing this linker, attributed to stable circulation and efficient lysosomal cleavage.
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| Enzyme Assay |
Cell-free enzymatic cleavage assays: Conjugate Fmoc-Gly-Gly-D-Phe-OH to a model fluorescent or cytotoxic payload. Incubate with purified cathepsin B (1-10 U/mL) in acetate buffer (50 mM, pH 5.0, 1 mM EDTA, 5 mM DTT) for 1-24 hours at 37degC. Quench, extract, and analyze by HPLC or LC-MS to monitor payload release. For stability studies, incubate the linker-payload conjugate in human plasma (pH 7.4) at 37degC for up to 48 hours and measure intact conjugate. For Fmoc deprotection assays, treat with 20% piperidine in DMF for 20 minutes at room temperature and confirm Fmoc removal by HPLC-MS.
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| Cell Assay |
Not applicable for the linker alone. For ADC studies, culture target antigen-positive and antigen-negative cancer cell lines in appropriate medium. Treat cells with ADCs synthesized using this linker at concentrations of 0.01-100 nM for 72-96 hours. Assess cell viability using CellTiter-Glo (luminescence) or MTT (absorbance). For internalization studies, treat cells with Alexa Fluor 488-labeled ADC (10 ug/mL) for 1-24 hours at 37degC, fix, and image by confocal microscopy. Co-stain with LysoTracker Red (lysosomes) to track ADC trafficking. For competition assays, pre-incubate cells with 100-fold excess unconjugated antibody for 30 minutes before ADC addition to confirm target-specific killing. Perform apoptosis assays by Annexin V/PI staining after 48-72 hours of ADC treatment.
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| Animal Protocol |
Not applicable for the linker alone. For ADC in vivo studies, use subcutaneous xenograft models in immunodeficient mice (e.g., NSG, nude). Inoculate target antigen-positive cancer cells (5 × 10^6 cells per mouse). When tumors reach ~100-150 mm3, randomize mice into treatment groups (n=8-10). Administer ADC via intravenous injection at doses of 1-10 mg/kg (based on antibody content), once weekly for 2-4 doses. Control groups: vehicle, isotype control ADC, and payload alone (at MTD). Measure tumor volume by caliper bi-weekly; calculate tumor growth inhibition (TGI %). Monitor body weight for toxicity. For PK/PD studies, collect blood at multiple time points post-dose (0.5, 1, 6, 24, 48, 72, 96, 120, 168 hours). Analyze plasma concentrations of total antibody, ADC, and released payload by ELISA or LC-MS/MS. At study endpoint, collect tumors for IHC (Ki-67, cleaved caspase-3, gamma-H2AX). For survival studies, continue dosing until humane endpoints and plot Kaplan-Meier survival curves.
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| ADME/Pharmacokinetics |
The linker is a synthetic peptide derivative not intended for in vivo administration. PK data apply to the full ADC, not the linker alone. The D-isomer (D-Phe) is designed to resist proteolytic degradation compared to L-isomers. The Fmoc-Gly-Gly-D-Phe-OH linker has a molecular weight of 501.53 and formula C28H27N3O6. The compound is soluble in DMSO. Storage: powder at -20degC (3 years) or -80degC (1 year for solution); in solvent at -80degC (1 year). Protect from light and moisture. For in vivo formulations, ADC is typically formulated in PBS or saline.
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| Toxicity/Toxicokinetics |
The linker itself is not administered in vivo; toxicity data apply to the full ADC. The Fmoc-Gly-Gly-D-Phe-OH linker is a research chemical for in vitro use only. The compound is not intended for human consumption. As a peptide derivative, it is expected to have low acute toxicity. Standard laboratory safety precautions (gloves, lab coat, safety glasses) should be used. Avoid inhalation, ingestion, and skin contact. For ADCs containing potent payloads, follow institutional guidelines for handling highly potent compounds. Dispose of waste according to hazardous chemical waste protocols. In case of accidental exposure, wash affected area with soap and water for 15 minutes and seek medical attention if irritation persists.
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| Additional Infomation |
Fmoc-Gly-Gly-D-Phe-OH is a cleavable ADC linker used in the synthesis of antibody-drug conjugates. It is the D-isomer of Fmoc-Gly-Gly-Phe-OH. The Fmoc (9-fluorenylmethoxycarbonyl) group is an acid-labile protecting group used in solid-phase peptide synthesis. The Gly-Gly dipeptide provides a flexible spacer. The D-Phe (D-phenylalanine) is a non-natural amino acid that may enhance resistance to proteases. The C-terminal carboxylic acid (-OH) can be activated for conjugation to a payload or to additional linker/spacer units. The compound is for research use only and not for diagnostic or therapeutic applications.
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| Molecular Formula |
C28H27N3O6
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|---|---|
| Molecular Weight |
501.53
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| Appearance |
White to off-white solid powder
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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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
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
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| 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 | 1.9939 mL | 9.9695 mL | 19.9390 mL | |
| 5 mM | 0.3988 mL | 1.9939 mL | 3.9878 mL | |
| 10 mM | 0.1994 mL | 0.9969 mL | 1.9939 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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