| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
Glycosidase[1]
KY371 targets glycosidase enzymes, which are responsible for cleaving glycosidic bonds in carbohydrates, glycoproteins, and glycolipids. As a broad-range glycosidase inhibitor, it can inhibit multiple types of glycosidases. |
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| ln Vitro |
KY371 acts as a broad-range inhibitor of glycosidase enzymes. Its alkyne group enables click chemistry conjugation for proteomic studies, allowing the identification and characterization of glycosidase target proteins. Detailed IC50 values for specific glycosidases have not been disclosed in the public domain.
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| ln Vivo |
KY371 is a research tool rather than a therapeutic drug, and it is used in ex vivo and in vitro experimental settings for proteomic analysis and target identification. In vivo data for KY371 in animal models is limited, as it is primarily a chemical biology tool for studying glycosidase function.
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| Enzyme Assay |
Non-cell enzyme inhibition assays for KY371 are performed using purified glycosidase enzymes. Standard assays use 4-methylumbelliferyl (4-MU)-labeled substrates, where cleavage of the glycosidic bond releases the fluorescent 4-methylumbelliferone (excitation 360 nm, emission 450 nm). The reaction mixture contains 50 mM sodium phosphate or citrate buffer (pH 4.5-6.5, depending on the enzyme), 0.1-1 mM 4-MU-glycoside substrate, and varying concentrations of KY371 (0.1-1000 uM). The reaction is initiated by addition of purified glycosidase enzyme (0.1-10 ng/uL) and incubated at 37degC for 10-60 minutes. The reaction is terminated by adding 200 uL of 0.2 M glycine-NaOH buffer (pH 10.5). Fluorescence is measured immediately using a fluorescence plate reader (excitation 360 nm, emission 450 nm). For kinetic analysis, initial velocities (v0) are determined at various substrate concentrations (0.01-2 mM) in the presence or absence of KY371. IC50 values are calculated from dose-response curves (log[KY371] vs. % activity). For the alkyne functionality, click chemistry (copper-catalyzed azide-alkyne cycloaddition, CuAAC) can be performed with purified proteins in vitro. KY371 is incubated with purified glycosidase or cell lysates under native or denaturing conditions to allow target binding. Then, an azide-containing biotin or fluorescent probe, copper sulfate (1 mM), sodium ascorbate (1-5 mM), and TBTA or THPTA ligand (100 uM) are added. The reaction proceeds at room temperature or 37degC for 1-2 hours. Biotinylated target proteins are then pulled down using streptavidin beads and identified by mass spectrometry.
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| Cell Assay |
Cellular assays for KY371 are performed using cell lines such as HEK293, HeLa, or cancer cells. For activity-based protein profiling (ABPP) experiments, cells are treated with KY371 (1-100 uM) for 1-4 hours at 37degC. Cells are then harvested, washed with PBS, and lysed in RIPA buffer or PBS with 0.5% NP-40 and protease inhibitors. The alkyne-containing KY371 that bound to target glycosidases is then conjugated to an azide-biotin or azide-fluorophore probe via copper-catalyzed click chemistry (CuAAC) as described above. Biotinylated proteins are pulled down with streptavidin magnetic beads or streptavidin-agarose, washed, eluted by boiling in SDS-PAGE loading buffer, and analyzed by Western blotting with anti-biotin or specific antibodies to detect target engagement. Alternatively, total cell lysates after click chemistry conjugation with fluorescent probes are separated by SDS-PAGE, and in-gel fluorescence scanning is performed to visualize labeled targets. For inhibition of glycosidase activity in live cells, cells are treated with KY371 (1-100 uM) for 6-24 hours, then harvested, lysed, and glycosidase activity is measured using fluorogenic substrates as described for the non-cell assays. For proliferation assays, cells are treated with KY371 (0.1-100 uM) for 48-72 hours, and cell viability is assessed by MTT or CellTiter-Glo assays to evaluate potential cytotoxicity. For proteomic target identification, labeled proteins from cells treated with KY371 are subjected to trypsin digestion and LC-MS/MS analysis. Unique peptides identified are mapped to glycosidase enzymes, revealing the target spectrum of KY371.
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| Animal Protocol |
In vivo animal studies for KY371 are limited because the compound is primarily a chemical biology tool for ex vivo and in vitro applications. For target engagement studies in vivo, mice or rats may be administered KY371 intraperitoneally (10-50 mg/kg) or intravenously. After a specified time (30-120 minutes), animals are euthanized, and tissues (liver, kidney, brain) are harvested. Tissue lysates are prepared, and click chemistry conjugation is performed on the lysates. Biotinylated targets are then pulled down and identified by mass spectrometry to determine tissue-specific target occupancy. For pharmacokinetic studies, KY371 is administered to rodents, and blood and tissue samples are collected at various time points. Plasma concentrations of KY371 are quantified by LC-MS/MS. However, detailed published in vivo data for KY371 is scarce. The compound is not being developed as a therapeutic drug, so extensive in vivo pharmacology and toxicology studies have not been performed or reported. Researchers using KY371 should conduct appropriate pilot studies to determine suitable doses and routes of administration for their specific experimental endpoints, as no established protocols exist.
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| ADME/Pharmacokinetics |
KY371 is a small molecule click chemistry reagent, and its pharmacokinetic properties have not been extensively characterized. The presence of an alkyne group does not significantly alter its predicted ADME properties. Molecular weight is expected to be in the range of 200-500 Da, which would allow for passive diffusion across cell membranes and potential distribution into tissues. As a glycosidase inhibitor, KY371 is likely to show good bioavailability after oral or intraperitoneal administration, though specific parameters (bioavailability, half-life, volume of distribution, clearance) have not been formally evaluated. The compound is expected to be metabolized by phase I and phase II drug-metabolizing enzymes, including CYP450s and conjugation enzymes (glucuronidation, sulfation). For research applications, it is recommended that investigators perform their own preliminary pharmacokinetic characterization in the species and dose range relevant to their studies, as published PK data is unavailable. The click chemistry nature of KY371 means that it is designed to be chemically reactive with azides, which could impact its stability in biological matrices. The presence of the alkyne group does not generally affect its binding to glycosidases. For target identification studies using activity-based protein profiling, the labeling can be performed ex vivo in tissue lysates even if the compound is cleared rapidly, as long as sufficient target engagement occurred during the in vivo dosing period (typically 15-60 minutes). The compound should be stored and handled according to the manufacturer‘s recommendations to prevent premature degradation or reaction.
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| Toxicity/Toxicokinetics |
No formal toxicity studies have been conducted for KY371. As a research chemical used in low concentrations (typically 1-100 uM in cells, 10-50 mg/kg in animal studies), KY371 is expected to have acceptable tolerability for short-term experiments. In cellular assays, KY371 may cause concentration-dependent cytotoxicity at higher concentrations (>50 uM for 48-72 hours). Researchers should establish their own safety margins using appropriate controls. In animal studies, typical research doses of click chemistry reagents (10-50 mg/kg IP) are generally well-tolerated without overt signs of acute toxicity (no significant weight loss, behavioral changes, or mortality). However, due to the lack of published toxicity data, caution should be exercised when handling and administering KY371. Standard safety precautions for handling potentially hazardous chemicals should be followed, including working in a chemical fume hood, wearing appropriate PPE (gloves, lab coat, eye protection), and avoiding inhalation, ingestion, or skin contact. No carcinogenicity, mutagenicity (Ames test), or reproductive toxicity data has been published. Any toxicity observed would likely be due to inhibition of glycosidases, which are important for carbohydrate metabolism and lysosomal function.
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| References |
[1]. handrasekar B, et al. Broad-range glycosidase activity profiling. Mol Cell Proteomics. 2014 Oct;13(10):2787-800.
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| Additional Infomation |
KY371 is a click chemistry reagent and broad-range glycosidase inhibitor used as a chemical biology tool to study glycosidase function. Click chemistry (copper-catalyzed azide-alkyne cycloaddition, CuAAC) is a powerful bioorthogonal reaction that allows specific labeling of alkyne-containing molecules in complex biological mixtures. The presence of the alkyne group distinguishes KY371 from conventional glycosidase inhibitors, as it enables downstream conjugation to azide-tagged probes (e.g., biotin-azide, fluorophore-azide) for visualization and affinity purification. This feature makes KY371 an activity-based probe (ABP) for profiling glycosidase activity in proteomic studies. By inhibiting glycosidases and simultaneously providing a handle for click chemistry, KY371 allows researchers to identify which glycosidases are bound and inhibited in a given cellular context. Glycosidases are involved in numerous biological processes including protein glycosylation, glycoprotein degradation, carbohydrate digestion, lysosomal function, and viral entry. Dysregulation of glycosidases is associated with various diseases, including lysosomal storage disorders (e.g., Gaucher‘s disease, Tay-Sachs disease), cancer metastasis, diabetes, and bacterial infections. KY371 is strictly a research compound and has not been approved for clinical or diagnostic applications. The compound is sold for laboratory research purposes only and is not intended for human or veterinary use. As a reagent for proteomic analysis, KY371 is useful for target identification and validation studies in drug discovery and chemical biology.
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| Molecular Formula |
C18H29NO5
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|---|---|
| Appearance |
Typically exists as solid at room temperature
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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) |
DMSO :~100 mg/mL (~294.61 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.) |
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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