| Size | Price | Stock | Qty |
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| 10mg |
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| 100mg |
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| Targets |
Glycosidase Cleavable Linker Cleavable Linker
Me-triacetyl-beta-D-glucopyranuronate-Ph-CH2OH-Fmoc itself is not a biologically active drug but a linker. Its intended target, when used in an ADC, is the lysosomal enzyme beta-glucuronidase. The glucuronide moiety of the linker is a specific substrate for this enzyme, which is overexpressed in the necrotic regions of many solid tumors. Upon cleavage of the glucuronide by beta-glucuronidase, the linker undergoes a spontaneous 1,6-elimination (self-immolation), releasing the conjugated payload. The ultimate pharmacological target of the released drug varies but is typically a component of the cancer cell's division machinery or survival pathways. |
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| ln Vitro |
ADCs are comprised of an antibody to which is attached an ADC cytotoxin through an ADC linker[1].
No direct in vitro activity is reported for Me-triacetyl-beta-D-glucopyranuronate-Ph-CH2OH-Fmoc alone. Its function is evaluated as part of an ADC. In vitro, the cleavability of this linker is confirmed by incubating the ADC with purified human beta-glucuronidase and monitoring payload release via mass spectrometry. The efficacy of the ADC is measured using cell viability assays (e.g., CellTiter-Glo) on cancer cell lines with varying beta-glucuronidase expression levels. Co-incubation with a beta-glucuronidase inhibitor serves as a control to confirm mechanism-based activity. |
| ln Vivo |
No in vivo activity is reported for the linker alone. For an ADC that incorporates this linker, in vivo efficacy is evaluated in mouse xenograft models. The linker enables targeted delivery and release of the cytotoxic payload specifically within the tumor, where high levels of beta-glucuronidase are found. This has been shown to result in potent anti-tumor activity, often superior to non-cleavable linkers or other cleavable technologies. The mechanism is validated by measuring the concentration of the released payload in the tumor versus healthy tissues, confirming tumor-selective release.
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| Enzyme Assay |
A typical non-cellular enzyme cleavage assay for this linker involves incubating the linker-payload conjugate (0.5 mg/mL) with 10 units of human beta-glucuronidase in 100 uL of acetate buffer (pH 5.0) at 37degC. After 2, 4, 8, and 24 hours, the reaction is quenched with acetonitrile. The reaction mixture is then analyzed by UPLC-MS to quantify the amount of released payload versus intact conjugate. The half-life of cleavage is calculated from the kinetic data. A control incubation without the enzyme is performed simultaneously to confirm chemical stability in the buffer.
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| Cell Assay |
A standard in vitro cell-based assay for an ADC containing this linker uses flow cytometry and confocal microscopy to track its fate. Cells are incubated with the ADC at 37degC for 1-24 hours. After fixation and permeabilization, an anti-payload antibody is used to detect the intracellular localization of the released payload. Colocalization studies with a LAMP1 antibody (a lysosomal marker) are performed to confirm that the ADC reaches the lysosomes, the primary site of beta-glucuronidase activity. The effect on cell cycle is assessed by propidium iodide staining and flow cytometry.
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| Animal Protocol |
Animal studies for this linker are conducted as part of the ADC evaluation. For a beta-glucuronidase-responsive ADC, a common model is mice bearing human HT-1080 fibrosarcoma or U87 glioblastoma xenografts, which are known to have high beta-glucuronidase expression. 5-6-week-old female BALB/c nude mice are injected subcutaneously with 5 × 10⁶ tumor cells. When tumors reach an appropriate volume, mice are randomized and treated intravenously with the ADC at 5-10 mg/kg, every 4 days for 3 doses. Tumors are collected at 24h, 48h, and 96h post-dose for immunohistochemistry (IHC) to detect the released payload, as well as analysis by LC-MS/MS to quantify payload concentrations.
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| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties are not determined for the linker alone. For an ADC that employs a glucuronide linker, the PK profile of the ADC is typically characterized by a long half-life (days), low clearance, and a limited volume of distribution. The released payload, upon enzymatic cleavage, often has a very short plasma half-life (e.g., 1-2 hours) and low systemic exposure, which is beneficial for reducing off-target toxicity. The favorable PK properties of the ADC enable it to circulate for extended periods, allowing for efficient tumor accumulation via the enhanced permeability and retention (EPR) effect.
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| Toxicity/Toxicokinetics |
No toxicity data is available for Me-triacetyl-beta-D-glucopyranuronate-Ph-CH2OH-Fmoc alone. The toxicity profile of an ADC using this linker is expected to be related to the conjugated payload. Preclinical toxicology studies are performed in rats or cynomolgus monkeys. The aim of these studies is to identify the MTD and dose-limiting toxicities. Common findings associated with maytansinoid or auristatin payloads include peripheral neuropathy, neutropenia, and hepatotoxicity. The glucuronide linker is designed to minimize these systemic toxicities by restricting payload release to the tumor site, thereby improving the therapeutic window of the ADC.
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| References | |
| Additional Infomation |
Me-triacetyl-beta-D-glucopyranuronate-Ph-CH2OH-Fmoc is not a drug and has no clinical status. It is an advanced, research-grade ADC linker designed for the development of cleavable ADCs that are activated by beta-glucuronidase in the tumor microenvironment. The linker's mechanism relies on the presence of the enzyme for payload release and includes a self-immolative spacer to ensure efficient liberation of the active drug. No clinical trials are registered for this linker. For research use only; not for human therapeutic or diagnostic use.
|
| Molecular Formula |
C38H40N2O14
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|---|---|
| Molecular Weight |
748.729211807251
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| Exact Mass |
748.247
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| CAS # |
894096-02-7
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| PubChem CID |
59816548
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| Appearance |
White to light yellow solid powder
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| LogP |
2.6
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
18
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| Heavy Atom Count |
54
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| Complexity |
1320
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| Defined Atom Stereocenter Count |
5
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| SMILES |
C(C1C2=CC=CC=C2C2C=CC=CC1=2)OC(=O)NCCC(=O)NC1=CC(CO)=CC=C1O[C@@H]1O[C@H](C(=O)OC)[C@@H](OC(=O)C)[C@H](OC(=O)C)[C@H]1OC(=O)C
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| InChi Key |
COFWDBMZXNIYIR-DVEMFWCFSA-N
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| InChi Code |
InChI=1S/C38H40N2O14/c1-20(42)50-32-33(51-21(2)43)35(52-22(3)44)37(54-34(32)36(46)48-4)53-30-14-13-23(18-41)17-29(30)40-31(45)15-16-39-38(47)49-19-28-26-11-7-5-9-24(26)25-10-6-8-12-27(25)28/h5-14,17,28,32-35,37,41H,15-16,18-19H2,1-4H3,(H,39,47)(H,40,45)/t32-,33-,34-,35+,37+/m0/s1
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| Chemical Name |
methyl (2S,3S,4S,5R,6S)-3,4,5-triacetyloxy-6-[2-[3-(9H-fluoren-9-ylmethoxycarbonylamino)propanoylamino]-4-(hydroxymethyl)phenoxy]oxane-2-carboxylate
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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 |
| 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: 125 mg/mL (166.95 mM)
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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 | 1.3356 mL | 6.6780 mL | 13.3559 mL | |
| 5 mM | 0.2671 mL | 1.3356 mL | 2.6712 mL | |
| 10 mM | 0.1336 mL | 0.6678 mL | 1.3356 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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