| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
β-Glucosidase[1]
beta-Glucosidase (irreversible covalent inhibitor) |
|---|---|
| ln Vitro |
Cyclophellitol aziridine is a potent inhibitor of beta-glucosidase. It acts as a mechanism-based inactivator, with the aziridine ring opening to form a stable covalent bond with the catalytic nucleophile of retaining beta-glucosidases. This irreversible inhibition allows for activity-based profiling of beta-glucosidase enzymes in complex biological samples. The compound is a cyclophellitol analog where the epoxide of cyclophellitol has been replaced with an aziridine, enhancing its reactivity toward the active site nucleophile.
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| ln Vivo |
No specific in vivo data reported for Cyclophellitol aziridine as a therapeutic agent. As an activity-based probe, it has potential for in vivo imaging of beta-glucosidase activity in animal models of lysosomal storage disorders. Covalent binding to beta-glucosidase allows for labeling and detection of active enzyme in tissues. In vivo studies would involve intravenous or intraperitoneal administration of the probe followed by tissue collection and fluorescence imaging or Western blot analysis of labeled enzyme.
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| Enzyme Assay |
Recombinant human beta-glucosidase (e.g., GBA1, GBA2) or lysosomal beta-glucosidase from tissue homogenates is incubated with Cyclophellitol aziridine (0.1-1000 nM) in citrate-phosphate buffer (pH 5.5-7.0) at 37degC for 10-60 minutes. The irreversible inhibition is time-dependent and concentration-dependent. Remaining enzyme activity is measured using a fluorogenic substrate such as 4-methylumbelliferyl-beta-D-glucopyranoside (4-MU-Glc). The reaction is terminated by adding glycine-NaOH buffer (pH 10.5), and fluorescence is measured at excitation 365 nm, emission 445 nm. Inactivation rate constants (kinact) and Ki values are determined from progress curves. The covalent binding can be confirmed by SDS-PAGE and fluorescence scanning if the probe is fluorescently labeled.
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| Cell Assay |
For activity-based protein profiling (ABPP), cells (e.g., human fibroblasts, HeLa, or Gaucher disease patient cells) are cultured in DMEM with 10% FBS. Cells are treated with Cyclophellitol aziridine conjugated to a fluorophore (e.g., Cy5 or BODIPY) at 0.1-10 uM for 1-4 hours. Cells are washed, lysed, and proteins are separated by SDS-PAGE. Labeled beta-glucosidase is visualized by in-gel fluorescence scanning. Alternatively, cell lysates are pre-incubated with the probe before SDS-PAGE. For competition assays, cells are pre-treated with unlabeled Cyclophellitol aziridine (0.1-100 uM) for 1 hour, followed by probe labeling. Reduction in fluorescence indicates specific binding to beta-glucosidase. Cell viability is assessed by MTT or trypan blue exclusion.
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| Animal Protocol |
No published in vivo animal study for Cyclophellitol aziridine. For activity-based probe studies, the compound (labeled with a fluorophore or biotin) is formulated in saline or PBS and administered intravenously (tail vein injection) or intraperitoneally to mice at doses of 0.1-10 mg/kg. After circulation for 1-24 hours, mice are euthanized, and tissues (brain, liver, spleen, kidney, lung) are collected. Tissues are homogenized, and labeled proteins are detected by in-gel fluorescence scanning or by Western blot using streptavidin-HRP (if biotinylated). For competitive studies, unlabeled Cyclophellitol aziridine can be administered prior to the labeled probe to assess specificity. Fluorescence imaging of whole organs or sections can also be performed.
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| ADME/Pharmacokinetics |
No specific pharmacokinetic data for Cyclophellitol aziridine. As a small, hydrophilic molecule (MW 175.18, LogP -2.3), it is expected to have low oral bioavailability and short plasma half-life due to rapid renal excretion. Distribution to tissues, including the brain, may occur due to its small size. The aziridine ring is reactive and may be subject to nucleophilic attack by biological thiols (glutathione, cysteine), leading to rapid metabolism and clearance. For in vivo studies, intravenous or intraperitoneal administration is preferred over oral dosing.
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| Toxicity/Toxicokinetics |
No specific toxicity data for Cyclophellitol aziridine. The aziridine ring is electrophilic and has the potential to react non-specifically with cellular nucleophiles (proteins, DNA), which could lead to cytotoxicity or genotoxicity. However, as a mechanism-based inhibitor designed for covalent modification of beta-glucosidase active sites, its toxicity profile has likely been characterized in vitro and in vivo. In cell-based studies, cytotoxicity has not been reported at probe concentrations (0.1-10 uM). For research use, handle with standard precautions (gloves, lab coat, fume hood) due to the reactive electrophilic warhead.
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| References | |
| Additional Infomation |
Cyclophellitol aziridine (CAS: 136861-97-7) is a cyclophellitol analog and potent irreversible inhibitor of beta-glucosidase. It functions as an activity-based probe (ABP) for retaining beta-glucosidases, including lysosomal glucocerebrosidase (GBA1), non-lysosomal glucocerebrosidase (GBA2), and other family 1 and family 3 glycosidases. The compound is a valuable research tool for studying glycosidase biology, enzyme mechanism, and for the development of diagnostic assays for Gaucher disease. Molecular formula: C7H13NO4, molecular weight: 175.18. Appearance: light brown to brown solid. Solubility: soluble in DMSO, water. For research use only. Reference: Li KY, et al. Exploring functional cyclophellitol analogues as human retaining beta-glucosidase inhibitors. Org Biomol Chem. 2014;12(39):7786-91. Not approved for clinical use.
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| Molecular Formula |
C7H13NO4
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|---|---|
| Molecular Weight |
175.182422399521
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| Exact Mass |
175.084
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| CAS # |
136861-97-7
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| PubChem CID |
89599365
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| Appearance |
Light brown to brown solid-liquid Mixture
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| LogP |
-2.3
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
12
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| Complexity |
188
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| Defined Atom Stereocenter Count |
6
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| SMILES |
O[C@@H]1[C@H]([C@@H]([C@@H](CO)[C@@H]2[C@H]1N2)O)O
|
| InChi Key |
GPIFFOGPRPKRHS-DRYVTRLFSA-N
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| InChi Code |
InChI=1S/C7H13NO4/c9-1-2-3-4(8-3)6(11)7(12)5(2)10/h2-12H,1H2/t2-,3+,4+,5+,6-,7-/m0/s1
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| Chemical Name |
(1R,2S,3S,4R,5R,6R)-5-(hydroxymethyl)-7-azabicyclo[4.1.0]heptane-2,3,4-triol
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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 | 5.7084 mL | 28.5421 mL | 57.0841 mL | |
| 5 mM | 1.1417 mL | 5.7084 mL | 11.4168 mL | |
| 10 mM | 0.5708 mL | 2.8542 mL | 5.7084 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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