| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Ki: 2 µM (nNOS), 50 µM (eNOS)[1]
|
|---|---|
| ln Vitro |
1400W (60 μM, 1 h) reduces the production of NO, 3-NT and MDA in primary adult microglia and prevents apoptosis of cerebral cortical neurons [2].
|
| ln Vivo |
1400W (0.1-10 mg/kg, subcutaneous injection, once) inhibits rat ileal leakage with an EC50 of 0.16 mg/kg[1].
In rats exposed to LPS-induced iNOS, 1400W potently (ED50=0.3 mg/kg) decreases the delayed vascular injury, but when administered in conjunction with LPS, it does not worsen acute vascular leakage[1]. Every experimental group's NOx levels are reduced by the administration of 1400W. Furthermore, the late post-hypoxia period (48 hours and 5 days) is marked by lipid peroxidation, the proportion of apoptotic cells, and nitrated protein expression[3]. Nitric oxide (NO(*)) from inducible NO(*) synthase (iNOS) has been reported to either protect against, or contribute to, hypoxia/re-oxygenation lung injury. The present work aimed to clarify this double role in the hypoxic lung. With this objective, a follow-up study was made in Wistar rats submitted to hypoxia/re-oxygenation (hypoxia for 30 min; re-oxygenation of 0 h, 48 h, and 5 days), with or without prior treatment with the selective iNOS inhibitor 1400W (10 mg/kg). NO(*) levels (NOx), lipid peroxidation, apoptosis, and protein nitration were analysed. This is the first time-course study which investigates the effects of 1400W during hypoxia/re-oxygenation in the rat lung. The results showed that the administration of 1400W lowered NOx levels in all the experimental groups. In addition, lipid peroxidation, the percentage of apoptotic cells, and nitrated protein expression fell in the late post-hypoxia period (48 h and 5 days). Our results reveal that the inhibition of iNOS in the hypoxic lung reduced the damage observed before the treatment with 1400W, suggesting that iNOS-derived NO(*) may exert a negative effect on this organ during hypoxia/re-oxygenation. These findings are notable, since they indicate that any therapeutic strategy aimed at controlling excess generation of NO(*) from iNOS may be useful in alleviating NO(*)-mediated adverse effects in hypoxic lungs[3]. |
| Enzyme Assay |
Reverse Phase Chromatography of [14C]1400 W Incubated with iNOS[1]
[14C]1400 W (15 μM) was incubated with iNOS (at a concentration that would convert 2 μM/min of 10 μML-arginine), and the reaction was analyzed by HPLC at 10, 20, and 40 min. Reactions were as described above for NOS except L-arginine was not included. Control reactions were without enzyme or without NADPH. 50-μl aliquots were filtered through Ultrafree MC filters and applied to a Waters Symmetry C18 HPLC column. The column was developed isocratically with 5 mM 1-octanesulfonic acid in 22% acetonitrile at a flow rate of 1 ml/min. 1400W was eluted from the column at 15 min. NO production assay[2] The nitrate/nitrite concentration was considered an indicator of NO production and was measured as previously described using a commercially available Nitric Oxide Fluorometric Assay Kit according to the manufacturer's instructions. Fluorescence was measured at 360 nm excitation/450 nm emission using the Thermo Scientific Varioskan Flash fluorescence reader. The fluorescence was an indicator of the concentration of sodium nitrite in the solution, and sodium nitrite concentrations were used to draw a standard curve, from which the concentration of nitrite was calculated. Microglia culture medium and cerebral cortex tissues homogenate were used to assess NO production. The values of NO production were expressed in nmol/mg protein. |
| Cell Assay |
Cytotoxicity assay[2]
Cell viability was evaluated using an MTT assay as previously described. Cells were seeded into 96 well plates and maintained at 37 °C for 24 h. The cells were exposed to various concentrations of 1400 W (20, 40, 60, 80, and 100 μM). After 24 h exposure, 0.5 mg/ml MTT in DPBS was added to each well and incubated for further 4 h. Then 150 μl of DMSO was added to the wells to dissolve the formazan crystals, and absorbance was measured at 490 nm using the Thermo Scientific Varioskan Flash microplate reader. The cellular viability was determined from the absorbance value and compared with that of the untreated control group. Detection of apoptosis using flow cytometry[2] Cells were seeded into 96-well plates at a density of 4 × 104 cells/cm2 and maintained at 37 °C for 24 h. Cells were then cultured in complete DMEM/F12 medium supplemented with 500 μM arginine, and placed in a hypoxic humidified incubator (1% O2). After 12 h hypoxia, cells were cultured in normoxic conditions for reoxygenation for 0, 6, or 24 h 1400 W (60 μM) dissolved in PBS was added to cell cultures 1 h before H/R, and control cultures received only vehicle (PBS). After H/R, cells were harvested and washed three times with ice-cold PBS. Cells were resuspended at a concentration of 4 × 105 cells per 500 μl binding buffer, and incubated with Annexin V-FITC and propidium iodide (PI) in the dark for 15 min at room temperature. The samples were analyzed using BD FACSCanto II flow cytometer. Apoptosis ratio was defined as the ratio between Annexin V positive/PI negative cells (right lower quadrant) and total cells. |
| Animal Protocol |
Endotoxin-induced Vascular Leakage in Rats[1]
The effects of 1400 W on plasma leakage were assessed in rats by determining the leakage of [125I]human serum albumin from plasma into organs essentially as described. 1400 W (0.1-10 mg/kg, subcutaneous) was dissolved in isotonic saline and administered either concurrently with endotoxin or 3 h following LPS administration (E. coli LPS, 3 mg/kg intravenously). Plasma leakage was then assessed 1 or 5 h after delivery of 1400W. The intravascular volumes were subtracted, and the results were expressed as Δμl g−1 tissue. Animals were randomly assigned to one of four experimental groups: vehicle-treated normoxia group, 1400 W-treated normoxia group, vehicle-treated hypoxia group, and 1400 W-treated hypoxia group. The 1400 W-treated groups were pretreated with ip injections of 1400 W (20 mg/kg, optimum dose) at 12 h intervals as previously described. 1400 W was dissolved in sterile distilled water at a concentration of 20 mg/ml. Vehicle-treated groups were pretreated with ip injections of an equal volume of sterile distilled water. Two hours after administration of vehicle or 1400 W, normoxia groups were maintained in a normoxic environment while hypoxia groups were exposed to simulated hypobaric hypoxia (HH) and reoxygenation as previously described. In brief, rats were exposed to simulated HH for 12 h at 8000 m (267 Torr) in an animal decompression chamber with the temperature and humidity maintained at 22 ± 2 °C and 30 ± 5%, and animals were provided with food and water ad libitum. After 12 h of HH, the hypoxia groups were brought down to sea level. Subjects from each experimental group were assessed at 0, 1 or 3 days post-HH with behavioral experiments or by resection of the cerebral cortex for embedding in paraffin and preparing tissue homogenate. Treatment of all 1400 W treated animals was stopped prior to spatial memory retention trial or resection of the cerebral cortex. |
| References |
|
| Additional Infomation |
N-[3-(aminomethyl)benzyl]acetamidine is an aralkylamine that is Nbenzylacetamidine substituted at position 3 on the benzene ring by an aminomethyl group. An inhibitor of nitric oxide synthase. It has a role as an EC 1.14.13.39 (nitric oxide synthase) inhibitor, a geroprotector and an angiogenesis inhibitor. It is a carboxamidine, an aralkylamine and a primary amino compound.
N-(3-(Aminomethyl)benzyl)acetamidine has been reported in Crotalaria pallida with data available. |
| Molecular Formula |
C10H15N3
|
|---|---|
| Molecular Weight |
177.25
|
| Exact Mass |
177.127
|
| Elemental Analysis |
C, 67.76; H, 8.53; N, 23.71
|
| CAS # |
180001-34-7
|
| Related CAS # |
180001-34-7; 214358-33-5 (HCl)
|
| PubChem CID |
1433
|
| Appearance |
Typically exists as solid at room temperature
|
| Density |
1.09 g/cm3
|
| Boiling Point |
329ºC at 760 mmHg
|
| Melting Point |
208-220ºC
|
| Flash Point |
152.7ºC
|
| Vapour Pressure |
0.000183mmHg at 25°C
|
| LogP |
2.423
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
13
|
| Complexity |
177
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
RODUKNYOEVZQPR-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C10H15N3/c1-8(12)13-7-10-4-2-3-9(5-10)6-11/h2-5H,6-7,11H2,1H3,(H2,12,13)
|
| Chemical Name |
N'-[[3-(aminomethyl)phenyl]methyl]ethanimidamide
|
| Synonyms |
180001-34-7; 1400W; N-(3-(AMINOMETHYL)BENZYL)ACETAMIDINE; n-[3-(aminomethyl)benzyl]acetamidine; N-{[3-(aminomethyl)phenyl]methyl}ethanimidamide; W 1400; CHEBI:90721; UNII-M1VB8VP8OH;
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
Typically soluble in DMSO (e.g. 10 mM)
|
|---|---|
| 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 | 5.6417 mL | 28.2087 mL | 56.4175 mL | |
| 5 mM | 1.1283 mL | 5.6417 mL | 11.2835 mL | |
| 10 mM | 0.5642 mL | 2.8209 mL | 5.6417 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
