UMP synthetase (UMPS) inhibitor. It is a prodrug that is metabolized to 6-azauridine, an inhibitor of orotidine monophosphate decarboxylase (OMPD), a key enzyme in the de novo pyrimidine synthesis pathway. [1]

To MDCK cells (MTT: CC50=19.66 μM), asazibine (2',3',5'-Tri-O-acetyl-6-azauridine; 0-2 μM; 48 hours) is cytotoxic[1]. The proliferation of MDCK cells (EC50=0.29 μM) and A549 cells (EC50=0.55 μM) is significantly inhibited by it [1]. After being added to MDCK cells, azaribenem had inhibitory effects on H1N1 and H3N2 IAV and IBV, with EC50 values of 0.60 μM, 0.77 μM, and 0.80 μM, in that order [1]. After 16HBE cells are treated, azaribidin exhibits an anti-NC H1N1 action with an EC50 of 1.58 μM [1].

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Azaribine demonstrated potent inhibitory activity against influenza A virus (IAV) and influenza B virus (IBV) infection in vitro. [1]
In posttreatment assays using Madin-Darby canine kidney (MDCK) cells infected with a bireporter influenza A/Puerto Rico/8/34 H1N1 virus (BIRFLU), azaribine exhibited a median effective concentration (EC50) of 0.29 μM. The selective index (SI), calculated as the ratio of CC50 to EC50, was 67.79 based on MTT assay (CC50 = 19.66 μM) and >172.41 based on XTT assay (CC50 > 50.00 μM). [1]
In pretreatment experiments (cells treated 24h before infection), azaribine also inhibited BIRFLU infection with an EC50 of 0.14 μM. The SI-MTT was >357.14, and SI-XTT was >357.14. [1]
In cotreatment experiments (compound mixed with virus during adsorption), azaribine showed an EC50 of 0.45 μM against BIRFLU. The SI-MTT was 43.69, and SI-XTT was >111.11. [1]
Azaribine inhibited the production of infectious IAV progeny in a dose- and time-dependent manner in multicycle virus growth kinetics assays in MDCK cells infected with BIRFLU at an MOI of 0.01 and treated with 0.1x, 1x, and 10x EC50 concentrations. [1]
In a minigenome (MG) assay in human 293T cells, which measures viral polymerase replication and transcription activity, azaribine reduced levels of MG-directed GFP and Gaussia luciferase (Gluc) expression with an EC50 of 0.21 μM, suggesting its antiviral effect is mediated through inhibition of the viral ribonucleoprotein complex (vRNP) activity. [1]
Azaribine did not inhibit host cellular RNA polymerase II activity, as shown by its lack of effect on Gluc expression from a polymerase II-dependent promoter (pCAGGS-Gluc) in MDCK cells (EC50 > 50 μM), indicating its antiviral effect is not due to general toxicity to the host transcriptional machinery. [1]
Azaribine demonstrated broad-spectrum antiviral activity against seasonal influenza viruses. In MDCK cells, it inhibited Venus-expressing pandemic A/California/04/09 H1N1 (pH1N1) with an EC50 of 0.60 μM (SI-MTT 32.77, SI-XTT >83.33), A/Wyoming/3/03 H3N2 with an EC50 of 0.77 μM (SI-MTT 25.53, SI-XTT >64.94), and B/Brisbane/60/08 IBV (Victoria lineage) with an EC50 of 0.80 μM (SI-MTT 24.57, SI-XTT >62.50). [1]
In human alveolar A549 cells infected with BIRFLU, azaribine showed an EC50 of 0.84 μM, with SI-MTT >59.52 and SI-XTT >59.52. [1]
In primary human bronchial epithelial immortalized (16HBE) cells infected with A/New Caledonia/20/1999 H1N1 (NC H1N1), azaribine exhibited an EC50 of 1.58 μM, with SI-MTT >31.65 and SI-XTT >31.65. [1]

Cytotoxicity assay [1]
Cell Types: MDCK cells
Tested Concentrations: 0-2 μM
Incubation Duration: 48 h
Experimental Results: Cytotoxic to MDCK cells (CC50=19.66 μM).

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The cytotoxicity of azaribine was evaluated in various cell lines using MTT and XTT assays to assess cell viability and proliferation. [1]
In MDCK cells, the 50% cytotoxic concentration (CC50) for azaribine was 19.66 μM in the MTT assay and greater than 50.00 μM in the XTT assay. [1]
In human A549 cells, the CC50 for azaribine was greater than 50.00 μM in both MTT and XTT assays. [1]
In human 16HBE cells, the CC50 for azaribine was greater than 50.00 μM in both MTT and XTT assays. [1]

[1]. Identification and Characterization of Novel Compounds with Broad-Spectrum Antiviral Activity against Influenza A and B Viruses. J Virol. 2020 Mar 17;94(7):e02149-19.

[2]. Identification of Inhibitors of ZIKV Replication. Viruses. 2020 Sep 18;12(9):1041.

Azaribine is an N-glycosyl-1,2,4-triazine compound with acetylated 2', 3', and 5' positions on its sugar ring. It is a prodrug of 6-azouridine used to treat psoriasis. It is both an anti-psoriatic drug and a prodrug. Azaribine is an acetate compound belonging to the N-glycosyl-1,2,4-triazine family. Functionally, it is related to 6-azouridine. Azaribine is the triacetate salt of azouridine, a synthetic triazine nucleoside derivative with antitumor and anti-psoriatic activities. After 6-azouridine is metabolized to 6-azouridine-5' monophosphate, it inhibits de novo pyrimidine synthesis, and its 5' triphosphate metabolite can be incorporated into RNA, thereby preventing RNA synthesis.

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Azaribine is a prodrug that is metabolized to 6-azauridine. [1]
It functions as an inhibitor of UMP synthetase (UMPS), an enzyme in the de novo pyrimidine biosynthesis pathway. By inhibiting this pathway, it depletes nucleotide pools required for viral RNA replication, thereby exerting its antiviral effects. [1]
This study identified azaribine as one of eight compounds with potent anti-influenza virus activity, initially discovered from screening the ReFRAME drug repurposing library for inhibitors of mammarenaviruses. [1]

C14H17N3O9

371.3

371.096

2169-64-4

16574

White to off-white solid powder

1.6g/cm3

99-101ºC(lit.)

1.621

-0.2

1

10

8

26

662

4

CC(=O)OC[C@@H]1[C@H]([C@H]([C@@H](O1)N2C(=O)NC(=O)C=N2)OC(=O)C)OC(=O)C

QQOBRRFOVWGIMD-OJAKKHQRSA-N

InChI=1S/C14H17N3O9/c1-6(18)23-5-9-11(24-7(2)19)12(25-8(3)20)13(26-9)17-14(22)16-10(21)4-15-17/h4,9,11-13H,5H2,1-3H3,(H,16,21,22)/t9-,11-,12-,13-/m1/s1

[(2R,3R,4R,5R)-3,4-diacetyloxy-5-(3,5-dioxo-1,2,4-triazin-2-yl)oxolan-2-yl]methyl acetate

NSC 67239 NSC 67239 AzaribineNSC 67239

2934.99.9001

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.

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

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

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.)