| 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) |
DMSO: ~100 mg/mL (~1040.7 mM; with ultrasonication)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (26.02 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 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 DMSO stock solution (25.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and 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 (26.02 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 DMSO stock solution (25.0 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well. 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. (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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