UDP-GlcNAc:polypeptidyltransferase (OGTase). It serves as an unnatural sugar donor substrate for this enzyme. [1]
O-GlcNAcase (OGA / Hexosaminidase C). It is part of substrates for this enzyme. [1]
Enzymes of the GlcNAc salvage pathway: GlcNAc kinase (GNK), Phospho-N-acetylglucosamine mutase (AGM1), and UDP-GlcNAc pyrophosphorylase (AGX1). [1]
UDP-GlcNAz disodium primarily targets O-GlcNAc transferase (OGT) , where it serves as an unnatural sugar donor substrate for the O-GlcNAcylation of nuclear and cytoplasmic proteins . It is also recognized by O-GlcNAcase (OGA) , the enzyme that cleaves O-GlcNAc modifications . Additionally, the compound is a substrate for the three key enzymes of the GlcNAc salvage pathway: GlcNAc kinase (GNK) , phospho-N-acetylglucosamine mutase (AGM1) , and UDP-GlcNAc pyrophosphorylase (AGX1) , which convert GlcNAz to UDP-GlcNAz intracellularly .

UDP-GlcNAc:polypeptidyltransferase (OGTase). It serves as an unnatural sugar donor substrate for this enzyme. [1]
O-GlcNAcase (OGA / Hexosaminidase C). It is part of substrates for this enzyme. [1]
Enzymes of the GlcNAc salvage pathway: GlcNAc kinase (GNK), Phospho-N-acetylglucosamine mutase (AGM1), and UDP-GlcNAc pyrophosphorylase (AGX1). [1]

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In vitro, UDP-GlcNAz disodium effectively serves as a sugar donor substrate for OGTase, enabling the O-GlcNAc modification of target proteins such as p62 . The resulting glycosidic bond can be specifically cleaved by O-GlcNAcase, confirming the specificity of the labeling . When the peracetylated form (Ac₄GlcNAz) is used in cell culture, it efficiently penetrates cell membranes, undergoes deacetylation by intracellular esterases, and enters the GlcNAc salvage pathway to be metabolically converted to UDP-GlcNAz . Studies in Jurkat cells have shown incorporation into nuclear proteins after 3 days of treatment . The azide group is well-tolerated by the enzymatic machinery, and the incorporated GlcNAz can be detected via click chemistry with alkyne-fluorophores or biotin .

Specific in vivo efficacy data for UDP-GlcNAz disodium (e.g., therapeutic endpoints, survival rates) is not available in the search results. As a metabolic labeling probe, its in vivo utility is established through the use of the cell-permeable peracetylated precursor Ac₄GlcNAz, which has been successfully employed in small animal models (such as mice and zebrafish) to metabolically label glycans and study O-GlcNAcylation in living organisms. This approach enables the visualization and tracing of glycoproteins in tissues via bioorthogonal chemistry following systemic administration. Systematic in vivo pharmacokinetic and pharmacodynamic profiling of the UDP-GlcNAz compound itself is not typically performed due to its role as an intracellular metabolite rather than a therapeutic agent.

GlcNAc Salvage Pathway Enzyme Kinetics: The specificity of each enzyme in the GlcNAc salvage pathway towards azido analogues was determined in vitro. [1]
GlcNAc Kinase (GNK) Assay: The enzyme was incubated with varying concentrations of GlcNAc or GlcNAz. The kinetic parameters were determined: for GlcNAc, Km was 50 ± 10 μM and Vmax/[E]t was 1.5 ± 0.1 μmol•min⁻¹•mg⁻¹; for GlcNAz, Km was 210 ± 40 μM and Vmax/[E]t was 2.2 ± 0.1 μmol•min⁻¹•mg⁻¹. [1]
AGM1 Assay: The enzyme was assayed with GlcNAc-6-P or GlcNAz-6-P. For GlcNAc-6-P, Km was 30 ± 10 μM and Vmax/[E]t was 0.12 ± 0.01 μmol•min⁻¹•mg⁻¹; for GlcNAz-6-P, Km was 120 ± 20 μM and Vmax/[E]t was 0.13 ± 0.01 μmol•min⁻¹•mg⁻¹. [1]
UDP-GlcNAc Pyrophosphorylase (AGX1) Assay: The enzyme was tested with GlcNAc-1-P or GlcNAz-1-P. For GlcNAc-1-P, Km was 200 ± 50 μM and Vmax/[E]t was 1.00 ± 0.05 μmol•min⁻¹•mg⁻¹; for GlcNAz-1-P, Km was 700 ± 90 μM and Vmax/[E]t was 0.90 ± 0.03 μmol•min⁻¹•mg⁻¹. [1]
O-GlcNAcase Assay: To determine the tolerance of O-GlcNAcase for the azide, a chromogenic assay was performed using para-nitrophenyl glycosides. The enzyme was incubated with pNP-GlcNAc or pNP-GlcNAz, and the release of p-nitrophenol was monitored spectrophotometrically. For pNP-GlcNAc, Km was 1,500 ± 100 μM and Vmax/[E]t was 0.77 ± 0.01 μmol•min⁻¹•mg⁻¹. For pNP-GlcNAz, Km was 1,100 ± 100 μM and Vmax/[E]t was 0.36 ± 0.01 μmol•min⁻¹•mg⁻¹. [1]
In Vitro OGTAse Activity Assay with UDP-GlcNAz: Recombinant OGTAse was incubated with recombinant p62 in a buffer containing Tris (pH 7.5), 12.5 mM MgCl₂, and 1 mM 2-mercaptoethanol, along with either UDP-GlcNAz or UDP-GlcNAc. The reaction proceeded overnight at room temperature. After dilution, the mixture was reacted with a biotin-phosphine probe (1 mM final concentration) in 20% dimethylformamide at 37°C for 1.5-2 hours. Samples were then analyzed by SDS-PAGE and Western blot using streptavidin-HRP for detection. [1]
Ni-NTA Plate OGT Assay: A high-throughput assay was developed using UDP-GlcNAz. 6×His-tagged substrate proteins (e.g., Nup62 or CKII-α) were immobilized on Ni-NTA-coated 96-well plates. After blocking, OGT enzyme (from bacterial lysate or partially purified) and UDP-GlcNAz in assay buffer were added and incubated (e.g., 25 min at 37°C for inhibition studies). The transferred azido groups were then reacted via copper-catalyzed click chemistry with TAMRA-Alkyne or via Staudinger ligation with Biotin-Phosphine. After washes, the incorporated probe was detected with an appropriate antibody (e.g., anti-TAMRA) conjugated to a fluorophore (e.g., IRDye800CW) and quantified using an infrared imaging system. [2]

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For non-cellular OGT activity assays, the reaction mixture typically contains purified OGT enzyme, a peptide substrate (e.g., a casein kinase II peptide or a derived peptide from a known O-GlcNAc protein), and UDP-GlcNAz as the sugar donor in a buffer containing 50 mM HEPES (pH 7.4), 10 mM MnCl₂, and 1 mM DTT . After incubation at 37°C for 1-2 hours, the transfer of GlcNAz to the peptide is detected via click chemistry. An alkyne-biotin or alkyne-fluorophore probe, along with CuSO₄, THPTA ligand, and sodium ascorbate as a reducing agent, is added to the reaction and incubated for 1 hour at room temperature. The labeled peptide can then be analyzed by SDS-PAGE with in-gel fluorescence scanning, Western blotting with streptavidin-HRP, or mass spectrometry . This assay avoids the use of radioactivity and is suitable for high-throughput screening of OGT inhibitors .

Metabolic Labeling of Jurkat Cells with Ac₄GlcNAz: Jurkat cells were cultured in RPMI medium with 5-10% FBS. Ac₄GlcNAz (the peracetylated form of GlcNAz) was added to the culture flasks at a final concentration of 40 μM. Cells were seeded at 2.5 x 10⁵ cells/mL and incubated for 2-3 days at 37°C until they reached a density of ~1.0 x 10⁶ cells/mL. Control cultures were grown without Ac₄GlcNAz. [1]
Isolation of Nuclei and Detection of Labeled Proteins: Nuclei were isolated from the Jurkat cells by hypotonic swelling and Dounce homogenization, followed by differential centrifugation through a sucrose gradient. Nuclear proteins were extracted by sonication in denaturing conditions. The protein extracts were reacted with a FLAG-phosphine probe (0.25 mM) for 6 hours at 37°C. Samples were then subjected to SDS-PAGE and Western blot. Detection with an anti-FLAG-HRP conjugate showed a large number of modified proteins only in the sample from Ac₄GlcNAz-treated cells, indicating metabolic incorporation of GlcNAz. [1]
Immunoprecipitation and Analysis of p62: Jurkat cells cultured with or without Ac₄GlcNAz were harvested and lysed in a buffer containing 1% SDS by heating and sonication. After adding a nonionic detergent solution, the lysate was cleared and incubated with mAb 414 (anti-p62) bound to protein G-Sepharose. The immunoprecipitated p62 was either treated with O-GlcNAcase overnight at 37°C or left untreated. All samples were then reacted with FLAG-phosphine and analyzed by Western blot. Probing with anti-FLAG-HRP showed a signal only in p62 from cells treated with Ac₄GlcNAz, and this signal was markedly reduced by O-GlcNAcase digestion. Reprobing the same blot with anti-p62 antibody confirmed equal loading. [1]

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For cellular metabolic labeling, cells (e.g., Jurkat, HeLa, or CHO cells) are cultured in complete medium supplemented with the peracetylated precursor Ac₄GlcNAz at concentrations ranging from 20-200 µM for 24-72 hours to allow metabolic incorporation . The peracetylated form is essential for efficient cell permeability. After incubation, cells are washed extensively, lysed (for protein analysis), or fixed with 4% paraformaldehyde (for imaging). Click chemistry is then performed on cell lysates or fixed cells using either copper-catalyzed conditions (CuAAc: 50-100 µM CuSO₄, 200-500 µM THPTA, 1-5 mM sodium ascorbate) with an alkyne-fluorophore or alkyne-biotin probe for 30-60 minutes, or copper-free conditions using a DBCO-conjugated probe for faster and gentler labeling . Labeled proteins can be visualized by in-gel fluorescence, Western blot, or immunofluorescence microscopy .

Based on the methodology for metabolic glycan labeling in animal models, the typical approach involves intraperitoneal (IP) or intravenous (IV) injection of the peracetylated precursor Ac₄GlcNAz (e.g., 50-200 mg/kg) dissolved in PBS or saline, often with 5-10% DMSO to aid solubility, into mice or other model organisms . For zebrafish embryos, microinjection is used. Tissues (liver, brain, heart, etc.) are harvested 24-72 hours post-injection. Visualizing labeled glycoproteins is achieved by performing click chemistry on tissue sections or extracted proteins using fluorescent or biotinylated alkyne probes. Control groups should receive vehicle (PBS/DMSO) alone or an equivalent dose of non-reactive sugar analog to assess background signal.

Metabolism (Cellular Uptake and Conversion): The peracetylated form of GlcNAz (Ac₄GlcNAz) was used in cell feeding experiments because it penetrates the cell membrane more efficiently. Once inside the cell, it is readily deacetylated by intracellular esterases to yield GlcNAz. The liberated GlcNAz then enters the GlcNAc salvage pathway, where it is converted by a series of enzymes (GNK, AGM1, AGX1) to UDP-GlcNAz. This UDP-GlcNAz can then be used by OGTAse to modify nuclear and cytoplasmic proteins with O-GlcNAz. [1]

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As a charged nucleotide sugar analog, UDP-GlcNAz itself has limited cell membrane permeability. Therefore, it is typically not administered directly to animals. Instead, the neutral, cell-permeable peracetylated form (Ac₄GlcNAz) is used to achieve in vivo delivery . Once inside cells, Ac₄GlcNAz is rapidly deacetylated by intracellular esterases to yield GlcNAz, which then enters the GlcNAc salvage pathway and is converted to UDP-GlcNAz by the sequential actions of Gnk, AGM1, and AGX1 . UDP-GlcNAz is a normal intracellular metabolite and is expected to be rapidly turned over and cleared.

Cytotoxicity: Prolonged exposure of Jurkat cells to 40 μM Ac₄GlcNAz had no apparent toxic effects. The cells appeared normal in all respects, including morphology and growth rate. [1]

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The compound and its peracetylated precursor are widely reported to have low cytotoxicity at working concentrations used for metabolic labeling (20-200 µM in cell culture, 50-200 mg/kg in animal models) . All suppliers emphasize that the product is for research use only and not intended for human or veterinary use . The azide modification is generally biocompatible and well-tolerated by cells. Long-term toxicity, reproductive toxicity, and comprehensive safety pharmacology studies have not been reported. Standard laboratory protective equipment should be used when handling.

[1]. A chemical approach for identifying O-GlcNAc-modified proteins in cells. Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9116-21.

[2]. Versatile O-GlcNAc transferase assay for high-throughput identification of enzyme variants, substrates, and inhibitors. Bioconjug Chem. 2014 Jun 18;25(6):1025-30.

Role as a Chemical Tool: UDP-GlcNAz is an analog of the natural sugar donor UDP-GlcNAc, where the N-acetyl group is modified to contain an azide. This azide serves as a bioorthogonal chemical handle, allowing the transferred sugar (GlcNAz) to be specifically detected via the Staudinger ligation with phosphine probes or copper-catalyzed click chemistry with alkynes, without interference from other cellular components. [1][2]
Application in Studying O-GlcNAc Modification: The metabolic labeling strategy using Ac₄GlcNAz, which leads to the intracellular generation of UDP-GlcNAz and subsequent incorporation into proteins by OGT, provides a powerful method for identifying and studying O-GlcNAc-modified proteins. This approach overcomes the limitations of traditional methods that rely on antibodies or lectins. [1]
Application in High-Throughput OGT Assays: UDP-GlcNAz is a key reagent in the Ni-NTA Plate OGT Assay. Its use enables a non-radioactive, direct measurement of OGT activity. This assay is versatile, cost-effective, and does not require enzyme purification, making it suitable for high-throughput screening of OGT inhibitors and for characterizing enzyme variants and substrate specificities. [2]
Dynamic Modification: The observation that O-GlcNAcase can cleave GlcNAz from modified proteins suggests that the dynamic nature of the O-GlcNAc modification is preserved when using this analog. This was supported by the marked reduction in signal from immunoprecipitated p62 after digestion with O-GlcNAcase. [1]

C17H24N6NA2O17P2

692.3

692.046

1611490-64-2

UDP-GalNAz disodium;653600-61-4

169543669

White to off-white solid powder

7

19

12

44

1230

9

O[C@@H]1[C@@H]([C@@H](COP(O)(=O)OP(O)(=O)O[C@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@H]2NC(=O)CN=[N+]=[N-])O[C@H]1N1C=CC(=O)NC1=O)O.[NaH]

XGTPLFSPWJKVQB-XZUZBEIXSA-L

InChI=1S/C17H26N6O17P2.2Na/c18-22-19-3-9(26)20-10-13(29)11(27)6(4-24)38-16(10)39-42(34,35)40-41(32,33)36-5-7-12(28)14(30)15(37-7)23-2-1-8(25)21-17(23)31;;/h1-2,6-7,10-16,24,27-30H,3-5H2,(H,20,26)(H,32,33)(H,34,35)(H,21,25,31);;/q;2*+1/p-2/t6-,7-,10-,11-,12-,13-,14-,15-,16-;;/m1../s1

disodium;[(2R,3R,4R,5S,6R)-3-[(2-azidoacetyl)amino]-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl] [[(2R,3S,4R,5R)-5-(2,4-dioxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-oxidophosphoryl] phosphate

1611490-64-2; Uridine-5'-disphopho-N-acetylazidoglucosamine disodium salt, 95%; UDP-GlcNAz.2Na?;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O: 125 mg/mL (180.6 mM)

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)