| Size | Price | Stock | Qty |
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| 5mg |
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| 25mg |
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| Targets |
5-A-RU HCl activates mucosal-associated invariant T (MAIT) cells via non-enzymatic reactions with metabolites such as methylglyoxal, generating potent MAIT-activating antigens that target the MR1 protein . MR1 (MHC class I-related protein 1) is essential for MAIT cell activation. The compound can be used to generate human and mouse MR1 tetramers for MAIT cell identification .
5-A-RU HCl targets mucosal-associated invariant T (MAIT) cells, a subset of innate-like T cells that recognize microbial metabolites. The compound is a precursor to riboflavin, which is metabolized by bacteria to produce MAIT cell antigens. 5-A-RU HCl reacts non-enzymatically with dihydroxy acetone to form potent MAIT cell antigens that are presented by the MHC class I-related protein MR1. Activation of MAIT cells leads to the production of cytokines and cytotoxic molecules, contributing to the immune response against microbial infections and cancer. |
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| ln Vitro |
In vitro cellular assays demonstrate that 5-A-RU HCl (2 μM) can be used to stimulate human MAIT cells and to produce human and mouse MR1 tetramers for MAIT cell identification . The compound activates primary mouse MAIT cells after reacting with bacterial metabolites.
In vitro, 5-A-RU HCl is used to stimulate human MAIT cells and produce MR1 tetramers for the identification of human and mouse MAIT cells. At a concentration of 2 µM, 5-A-RU HCl can efficiently activate MAIT cells. The compound is also used in studies of MAIT cell biology, including their development, function, and role in disease. By activating MAIT cells, 5-A-RU HCl provides a tool for studying the immune response to microbial infections and cancer. |
| ln Vivo |
In in vivo mouse studies, 5-A-RU HCl (100 nM/animal) pre-mixed with methylglyoxal (MeG) is administered via intraperitoneal injection. After 18 hours, effective activation of MAIT cells is observed in iVα19 Cα−/−-Tg transgenic mice .
In vivo, 5-A-RU HCl has been used to study the role of MAIT cells in various disease models. Activation of MAIT cells by 5-A-RU HCl has been shown to have protective effects against bacterial infections. The compound has also been investigated for its potential in cancer immunotherapy, as MAIT cells can exert cytotoxic activity against tumor cells. However, the in vivo use of 5-A-RU HCl is limited by its rapid metabolism and the complexity of the immune response. |
| Enzyme Assay |
The activity of 5-A-RU HCl depends on its non-enzymatic reaction with 1,2-dicarbonyl compounds such as methylglyoxal and glyoxal. In a typical assay, 5-A-RU HCl is incubated with methylglyoxal in phosphate-buffered saline (pH 7.4) to generate potent MAIT cell antigens. The reaction products bind to immobilized recombinant MR1 protein, and MAIT cell-specific recognition is detected by flow cytometry using MR1 tetramers .
In vitro enzyme/receptor binding assays for 5-A-RU HCl involve studying its role in the formation of MAIT cell antigens. The compound is incubated with dihydroxy acetone in a buffer at physiological pH and temperature. The formation of the antigen is monitored by HPLC or mass spectrometry. The binding of the antigen to MR1 and its presentation to MAIT cells can be assessed using T cell activation assays. These assays are essential for understanding the mechanism of MAIT cell activation. |
| Cell Assay |
MAIT Cell Stimulation Assay: Human MAIT cells are isolated from peripheral blood mononuclear cells and cultured in medium containing 2 μM 5-A-RU HCl for 5-7 days. Cytokines such as IL-2 are added to maintain cell growth. MAIT cell activation status (e.g., CD69 upregulation) is detected by flow cytometry using anti-TCR Vα7.2 and anti-CD161 antibodies .
MR1 Tetramer Production: 5-A-RU HCl is used to treat antigen-presenting cells to produce antigen-loaded MR1 molecules, which are then used to construct human and mouse MR1 tetramers for MAIT cell identification .
In vitro cellular experiments for 5-A-RU HCl are performed using MAIT cell lines or primary MAIT cells isolated from human blood. Cells are treated with varying concentrations of 5-A-RU HCl, and MAIT cell activation is assessed by measuring the expression of activation markers (e.g., CD69, CD25) by flow cytometry. Cytokine production (e.g., IFN-γ, TNF-α) is measured by ELISA or intracellular staining. The cytotoxicity of activated MAIT cells against target cells is assessed using a chromium release or lactate dehydrogenase (LDH) release assay. |
| Animal Protocol |
In the iVα19 Cα−/−-Tg transgenic mouse model, 5-A-RU HCl (100 nM/animal) is pre-mixed with methylglyoxal and administered via intraperitoneal injection. After 18 hours, mice are euthanized, and splenic and hepatic lymphocytes are isolated. The proportion of MAIT cells and the expression of activation markers among TCR-positive cells are assessed by flow cytometry to evaluate the in vivo MAIT cell activation efficacy of the compound .
In vivo animal studies for 5-A-RU HCl are conducted using mouse models. The compound is administered via intraperitoneal or intravenous injection. MAIT cell activation in tissues such as the spleen, liver, and lungs is assessed by flow cytometry. The effects of MAIT cell activation on bacterial clearance or tumor growth are evaluated. The compound's safety and toxicity are also assessed by monitoring body weight, clinical signs, and histopathological changes in major organs. |
| ADME/Pharmacokinetics |
The pharmacokinetic properties of 5-A-RU HCl are characterized by its rapid absorption and metabolism. The compound is a precursor to riboflavin, and its metabolism is linked to the riboflavin biosynthesis pathway. The compound's half-life in circulation is short, and it is rapidly cleared from the body. The compound's pharmacokinetic profile supports its use in acute in vivo studies.
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| Toxicity/Toxicokinetics |
The toxicity of 5-A-RU HCl has not been extensively characterized in the literature. As a nucleoside analog and a precursor to riboflavin, it is expected to have a low toxicity profile. However, high doses may lead to adverse effects related to MAIT cell activation, such as cytokine release syndrome. Standard toxicology studies would be required to fully characterize its safety profile.
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| References |
[1]. https://pubchem.ncbi.nlm.nih.gov/compound/146026065
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| Additional Infomation |
5-A-RU HCl (CAS: 134452-11-2) is different from 5-A-RU (CAS: 17014-74-3, the free base form). Its hydrochloride salt form offers better water solubility and stability, making it more convenient for experimental manipulation. This compound is currently a common tool molecule for activating MAIT cells in immunology research, particularly holding significant application value in the field of studying the interaction between bacterial metabolites and host immune cells. Its potential application in breast cancer and prostate cancer research has also garnered attention.
5-A-RU HCl is a precursor to bacterial riboflavin and an activator of MAIT cells. It reacts non-enzymatically with dihydroxy acetone to form potent MAIT cell antigens. This compound is used in research to study MAIT cell activation, immunology, and cancer. 5-A-RU HCl has been investigated for its potential in cancer immunotherapy, as MAIT cells can exert cytotoxic activity against tumor cells. It is also used to study the role of MAIT cells in microbial infections. |
| Molecular Formula |
C₉H₁₇CLN₄O₆
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|---|---|
| Molecular Weight |
312.71
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| Exact Mass |
312.083
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| CAS # |
134452-11-2
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| Related CAS # |
17014-74-3
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| PubChem CID |
146026065
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
9
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
20
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| Complexity |
398
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| Defined Atom Stereocenter Count |
3
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| SMILES |
Cl.O[C@H]([C@@H](CO)O)[C@H](CNC1=C(C(NC(N1)=O)=O)N)O
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| InChi Key |
VGAFXPBLNINPEY-ZUOBHZEMSA-N
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| InChi Code |
InChI=1S/C9H16N4O6.ClH/c10-5-7(12-9(19)13-8(5)18)11-1-3(15)6(17)4(16)2-14;/h3-4,6,14-17H,1-2,10H2,(H3,11,12,13,18,19);1H/t3-,4+,6-;/m0./s1
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| Chemical Name |
5-amino-6-[[(2S,3S,4R)-2,3,4,5-tetrahydroxypentyl]amino]-1H-pyrimidine-2,4-dione;hydrochloride
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| Synonyms |
5ARU HCl; 134452-11-2; 5-amino-6-(((2S,3S,4R)-2,3,4,5-tetrahydroxypentyl)amino)-1,2,3,4-tetrahydropyrimidine-2,4-dione hydrochloride; 5-amino-6-{[(2S,3S,4R)-2,3,4,5-tetrahydroxypentyl]amino}-1,2,3,4-tetrahydropyrimidine-2,4-dione hydrochloride; RefChem:532344; 989-079-9; 5 A RU HCl
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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 |
| 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 | 3.1979 mL | 15.9893 mL | 31.9785 mL | |
| 5 mM | 0.6396 mL | 3.1979 mL | 6.3957 mL | |
| 10 mM | 0.3198 mL | 1.5989 mL | 3.1979 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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