| Size | Price | Stock | Qty |
|---|---|---|---|
| 500mg | |||
| 50g |
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| Other Sizes |
| Targets |
NO Synthase (Nitric Oxide Synthase).
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|---|---|
| ln Vitro |
In cytosolic extracts of active cells, 3-amino-1,2,4-triazine (50 mM; 7 h) suppresses NO synthase activity[1]. 3-Amino-1,2,4-triazine (5–50 mM; 24 h) prevents activated macrophages from killing parasites and secreting nitrites[1].
3-Amino-1,2,4-triazine inhibits NO synthase activity in vitro. At 50 mM, it inhibits NOS activity in cytosolic extracts of activated cells. At concentrations of 5-50 mM, it inhibits both nitrite secretion and parasite killing by activated macrophages. It does not directly affect cell viability at these concentrations but blocks the NO-mediated anti-pathogen response. |
| ln Vivo |
No in vivo activity data is available for 3-Amino-1,2,4-triazine. As an NOS inhibitor, it could potentially be tested in animal models of endotoxemia (LPS challenge) to reduce NO levels and hypotension, or in models of septic shock. However, its moderate potency may limit systemic use.
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| Enzyme Assay |
For direct enzyme inhibition assays, cytosolic extracts are prepared from activated RAW264.7 macrophages or primary peritoneal macrophages stimulated with LPS (100 ng/mL) and IFN-gamma (10 U/mL) for 24 h. Cytosolic extracts (100-200 microg protein) are incubated with 3-Amino-1,2,4-triazine (0-100 mM) in assay buffer (50 mM Tris-HCl pH 7.4, 10 microM FAD, 10 microM FMN, 10 microM tetrahydrobiopterin, 1 mM NADPH, 1 mM CaCl2, 2.5 mM L-arginine, 10 microg/mL calmodulin) for 60 min at 37degC. NO production is quantified by measuring nitrite accumulation using the Griess reaction (1% sulfanilamide, 0.1% N-(1-naphthyl)ethylenediamine in 5% phosphoric acid, OD 540 nm). Inhibition of NOS activity is calculated relative to control.
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| Cell Assay |
RAW264.7 mouse macrophages are seeded in 24-well plates (2×10⁵/well) in DMEM with 10% FBS. Cells are activated with LPS (100 ng/mL) and IFN-gamma (10 U/mL) for 6-24 h to induce iNOS expression. 3-Amino-1,2,4-triazine (5-50 mM) is added at the time of activation. After 24-48 h, culture supernatants are collected for nitrite measurement by the Griess reaction. Cell viability is assessed by MTT assay to ensure that inhibition of nitrite secretion is not due to cytotoxicity. For parasite killing assays, Leishmania major-infected macrophages are treated with the compound (5-50 mM) for 24 h, and parasite survival is quantified by microscopy.
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| Animal Protocol |
No animal protocol is published. Generic protocol for NOS inhibitors in LPS-induced endotoxemia: female BALB/c mice (6-8 weeks, n=8/group) are injected intraperitoneally with LPS (5-10 mg/kg). 3-Amino-1,2,4-triazine (100-500 mg/kg) is administered i.p. or orally 1 h before LPS challenge. Blood is collected 4-6 h post-LPS, and serum nitrate/nitrite levels are measured by Griess assay. Plasma cytokines (TNF-alpha, IL-1beta, IL-6) are measured by ELISA. Liver and lung tissues are harvested for iNOS expression analysis (Western blot, qPCR) and histopathology (H&E staining for inflammatory infiltration). Blood pressure can be monitored via carotid artery catheter to assess vasodilation.
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| ADME/Pharmacokinetics |
No PK data reported. Generic PK for heterocyclic NOS inhibitors: male SD rats receive IV (10 mg/kg) and oral (50 mg/kg) doses. Blood samples collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24 h. Plasma is analyzed by LC-MS/MS. t1/2 is typically short (1-3 h) due to rapid metabolism and clearance. Cmax following oral administration is usually low (0.5-5 microM) with Tmax 0.5-1 h, indicating moderate absorption. Oral bioavailability is often <50%. Volume of distribution (Vd) may be high (>2 L/kg) due to tissue distribution.
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| Toxicity/Toxicokinetics |
No toxicity data reported. Generic acute toxicity study: ICR mice (5/sex/group) receive single i.p. or oral doses of 3-Amino-1,2,4-triazine at 250, 500, 1000 mg/kg. Animals are observed for 14 days for mortality, clinical signs (weight loss, hunched posture, piloerection, ataxia), and food/water consumption. At termination, gross necropsy and histopathology of major organs (liver, kidney, spleen, heart, lung, brain, GI tract) are performed. Potential neurological effects may be observed due to NOS inhibition in the CNS.
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| References | |
| Additional Infomation |
3-Amino-1,2,4-triazine (CAS# 1120-99-6) is a simple heterocyclic compound used in research as a non-selective NOS inhibitor. It was described in the literature as an inhibitor of macrophage NO synthase (Buchmüller-Rouiller Y et al., Biochem Biophys Res Commun. 1992). It is not a drug and has no clinical approvals. The compound is useful as a research tool to study the role of NO in macrophage function, antimicrobial defense, and inflammation. It may also be used as a synthetic intermediate or building block in heterocyclic chemistry, due to its triazine ring structure.
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| Molecular Formula |
C3H4N4
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|---|---|
| Molecular Weight |
96.09
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| Exact Mass |
96.043
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| CAS # |
1120-99-6
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| PubChem CID |
70715
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| Appearance |
Brown to black solid powder
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| Density |
1.346g/cm3
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| Boiling Point |
311.7ºC at 760mmHg
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| Melting Point |
174-177 °C(lit.)
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| Flash Point |
168ºC
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| Vapour Pressure |
0.000553mmHg at 25°C
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| Index of Refraction |
1.61
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| LogP |
0.035
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
7
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| Complexity |
54.9
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CN=NC(=N1)N
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| InChi Key |
MJIWQHRXSLOUJN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C3H4N4/c4-3-5-1-2-6-7-3/h1-2H,(H2,4,5,7)
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| Chemical Name |
1,2,4-triazin-3-amine
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 | 10.4069 mL | 52.0346 mL | 104.0691 mL | |
| 5 mM | 2.0814 mL | 10.4069 mL | 20.8138 mL | |
| 10 mM | 1.0407 mL | 5.2035 mL | 10.4069 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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