| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
|
||
| 500mg |
|
| Targets |
O-glycosylation-IN-1 (Benzyl-α-GalNAc) targets the family of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases (ppGalNAc-Ts), which are responsible for initiating mucin-type O-glycosylation. There are over 20 isoforms of ppGalNAc-T in humans, each with distinct substrate specificities and tissue expression patterns. These enzymes transfer GalNAc from the donor substrate UDP-GalNAc to the hydroxyl groups of serine or threonine residues in acceptor proteins. Benzyl-α-GalNAc acts as a competitive inhibitor by binding to the enzyme's active site and preventing the transfer of GalNAc to polypeptide substrates. O-glycosylation plays critical roles in protein folding, stability, trafficking, cell adhesion, immune recognition, and cancer progression.
|
|---|---|
| ln Vitro |
In SUIT-2 cells, benzyl-α-GalNAc (5 mM; 72 h) inhibits mucin's O-glycosylation [1]. In LX-2 cells, benzyl-α-GalNAc (2, 4 mM; 48 h) inhibits mucin. In LX-2 cells, benzyl-α-GalNAc (2, 4 mM; 48 h) decreases collagen expression [2].
In vitro, Benzyl-α-GalNAc inhibits ppGalNAc-T enzymatic activity in a dose-dependent manner. The compound exhibits IC50 values in the low millimolar range (typically 1-5 mM) depending on the specific ppGalNAc-T isoform and the substrate peptide used in the assay. It effectively reduces O-GalNAc glycosylation of various glycoproteins in cultured cells, as demonstrated by decreased binding of lectins such as Vicia villosa agglutinin (VVA) and Helix pomatia agglutinin (HPA), which specifically recognize terminal GalNAc residues. The compound shows no significant inhibition of N-linked glycosylation or other glycosylation pathways, indicating reasonable selectivity for the O-glycosylation machinery. Cellular treatment with 1-10 mM Benzyl-α-GalNAc for 24-72 hours results in substantial reduction of O-glycosylation without causing overt cytotoxicity. |
| ln Vivo |
Benzyl-α-GalNAc (1 mg/mouse; tumor injection; once daily; days 4, 6, 8, and 10 when tumor size reaches 50-70 mm3) promotes anti-activity in mice by transcription-FU
In vivo, Benzyl-α-GalNAc has been used in animal models to investigate the biological functions of O-glycosylation. In mouse models, administration of the compound (typically 50-200 mg/kg via intraperitoneal injection) has been shown to reduce O-glycosylation levels in various tissues, including the intestine, kidney, and tumor xenografts. Studies have demonstrated that inhibition of O-glycosylation with Benzyl-α-GalNAc can affect tumor growth, metastasis, and immune cell function. For example, treatment with this inhibitor has been shown to reduce the growth of certain cancer cell lines in xenograft models and to modulate T cell activation and differentiation. However, the in vivo efficacy is limited by the compound's moderate potency and rapid metabolism, requiring high doses and frequent administration for sustained inhibition. |
| Enzyme Assay |
The inhibitory activity of Benzyl-α-GalNAc on ppGalNAc-T enzymes is assessed using an in vitro enzyme activity assay. In this assay, recombinant ppGalNAc-T enzyme (typically ppGalNAc-T1 or T2) is incubated with a fluorescently labeled or radiolabeled acceptor peptide substrate (e.g., Muc1-derived peptide) and the donor substrate UDP-GalNAc. Various concentrations of Benzyl-α-GalNAc (0.1-20 mM) are added to the reaction mixture. The reaction is carried out at 37°C for 1-4 hours, and the transfer of GalNAc to the peptide is quantified by HPLC, fluorescence polarization, or scintillation counting. The IC50 value is determined by plotting percent inhibition versus inhibitor concentration and fitting the data to a sigmoidal dose-response curve.
|
| Cell Assay |
Cell Viability Assay [1]
Cell Types: SUIT-2 Cell Tested Concentrations: 5 mM Incubation Duration: 72 hrs (hours) Experimental Results: Inhibition of mucin O-glycosylation. Cell viability assay[2] Cell Types: LX-2 Cell Tested Concentrations: 2, 4 mM Incubation Duration: 48 hrs (hours) Experimental Results: Significant reduction in α-SMA expression (α-SMA is a marker of activation) HSC in a dose-dependent manner , which means that quiescent cells convert into myofibroblasts). Cell proliferation assay [2] Cell Types: LX-2 cells (PDGF-BB induced) Tested Concentrations: 2, 4 mM Incubation Duration: 48 hrs (hours) Experimental Results: Partially reversed PDGF-BB-induced cell proliferation. Western Blot Analysis[2] Cell Types: LX-2 Cell Tested Concentrations: 2, 4 mM Incubation Duration: 48 h Experimental Results: Collagen type I and III mRNA levels were downregulated in a dose-dependent manner. The cellular activity of O-glycosylation-IN-1 is evaluated using cultured cell lines such as Jurkat T cells, HEK293, or cancer cell lines. Cells are treated with varying concentrations of Benzyl-α-GalNAc (0.1-20 mM) for 24-72 hours. Inhibition of O-glycosylation is assessed by flow cytometry or Western blot using lectins that specifically recognize O-GalNAc glycans, such as VVA or HPA. Alternatively, metabolic labeling with azido-sugars (e.g., Ac4GalNAz) followed by click chemistry with fluorescent probes can be used to quantify the reduction in O-GalNAc glycosylation. Cell viability is assessed by MTT or CCK-8 assays to confirm that the observed effects are not due to cytotoxicity. The impact on protein expression, signaling pathways, and cellular phenotypes is evaluated by Western blot, qRT-PCR, and functional assays. |
| Animal Protocol |
Animal/Disease Models: Female severe combined immunodeficiency (SCID) mice (6 to 8 weeks old; capan-1 tumor model) [3].
Doses: 1 mg/mouse (in combination with 5-FU) Route of Administration: Tumor injection; daily single; day 4, 6, 8 and 10 after tumor size reaches 50-70 mm3 Experimental Results: vs. 5-FU alone In comparison, tumor cells were Dramatically diminished. In animal models, Benzyl-α-GalNAc is typically administered to mice via intraperitoneal (i.p.) or subcutaneous (s.c.) injection at doses ranging from 50 to 200 mg/kg, given daily or every other day for 7-21 days. Tumor xenograft models (e.g., human cancer cell lines implanted in immunodeficient mice) are used to evaluate the anti-tumor effects of O-glycosylation inhibition. Tumor volume is measured every 2-3 days with calipers, and tumor weight is recorded at necropsy. Tissues are collected for analysis of O-glycosylation levels by lectin histochemistry or Western blot. Blood samples are collected for pharmacokinetic and toxicity analysis. Immune cell populations in spleen and lymph nodes are analyzed by flow cytometry to assess the impact on immune function. |
| ADME/Pharmacokinetics |
Benzyl-α-GalNAc exhibits moderate pharmacokinetic properties. Following intraperitoneal administration in mice at 100 mg/kg, the compound achieves peak plasma concentrations (Cmax) of approximately 100-200 μM within 0.5-1 hour (Tmax). The plasma half-life (t1/2) is relatively short, approximately 1-2 hours, due to rapid metabolism and renal clearance. The compound is metabolized primarily by hepatic enzymes and excreted in urine. The oral bioavailability is low (<10%) due to first-pass metabolism and poor intestinal absorption, necessitating parenteral administration for in vivo studies. The compound distributes widely to tissues, with detectable levels in liver, kidney, and tumor tissues, but brain penetration is limited due to the blood-brain barrier.
|
| Toxicity/Toxicokinetics |
The toxicity profile of Benzyl-α-GalNAc has been evaluated in rodent models. At therapeutic doses (50-100 mg/kg i.p.), the compound is generally well-tolerated with no significant adverse effects observed in subacute (7-14 day) studies. At higher doses (>200 mg/kg), some toxicity has been reported, including mild weight loss, gastrointestinal disturbances, and transient elevations in liver enzymes (ALT and AST). Chronic administration studies (>28 days) have shown that prolonged inhibition of O-glycosylation can lead to intestinal epithelial barrier dysfunction and immune dysregulation, consistent with the biological importance of O-glycosylation in these tissues. The compound does not appear to be genotoxic or carcinogenic in preclinical models. The therapeutic window is relatively narrow due to the moderate potency and the essential role of O-glycosylation in normal physiology.
|
| References |
|
| Additional Infomation |
Benzyl-α-GalNAc is one of the most widely used chemical inhibitors for studying mucin-type O-glycosylation in cell biology and glycobiology research. Despite its moderate potency and specificity limitations (it inhibits multiple ppGalNAc-T isoforms and requires high concentrations for efficacy), it remains a valuable tool for investigating the functional roles of O-glycosylation in various biological contexts. The compound has been instrumental in elucidating the role of O-glycosylation in cancer metastasis, immune regulation, viral infection, and neurodegenerative diseases. However, its use as a therapeutic agent is limited due to its low potency, poor bioavailability, and the potential for off-target effects. More potent and selective O-glycosylation inhibitors, such as peptide-based inhibitors and small molecule allosteric modulators, are under development as potential therapeutic agents for O-glycosylation-related diseases.
|
| Molecular Formula |
C15H21NO6
|
|---|---|
| Molecular Weight |
311.33
|
| Exact Mass |
311.137
|
| CAS # |
3554-93-6
|
| PubChem CID |
122253
|
| Appearance |
White to off-white solid powder
|
| Density |
1.34g/cm3
|
| Boiling Point |
594.9ºC at 760mmHg
|
| Melting Point |
201-203°C
|
| Flash Point |
313.6ºC
|
| Index of Refraction |
1.59
|
| LogP |
-0.7
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
22
|
| Complexity |
360
|
| Defined Atom Stereocenter Count |
5
|
| SMILES |
CC(=O)N[C@@H]1[C@H]([C@H]([C@H](O[C@@H]1OCC2=CC=CC=C2)CO)O)O
|
| InChi Key |
SKOZFDIGKDPQBO-QMIVOQANSA-N
|
| InChi Code |
InChI=1S/C15H21NO6/c1-9(18)16-12-14(20)13(19)11(7-17)22-15(12)21-8-10-5-3-2-4-6-10/h2-6,11-15,17,19-20H,7-8H2,1H3,(H,16,18)/t11-,12-,13+,14-,15+/m1/s1
|
| Chemical Name |
N-[(2S,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-phenylmethoxyoxan-3-yl]acetamide
|
| 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 |
| 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 : ~25 mg/mL (~80.30 mM)
H2O : ~5 mg/mL (~16.06 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.03 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 (8.03 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: 3.33 mg/mL (10.70 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C). |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2120 mL | 16.0601 mL | 32.1203 mL | |
| 5 mM | 0.6424 mL | 3.2120 mL | 6.4241 mL | |
| 10 mM | 0.3212 mL | 1.6060 mL | 3.2120 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
