Nitric oxide synthase (NOS) catalyzes the formation of nitric oxide and L-citrulline from L-arginine. 1400W 2HCl is a slow, tight binding, selective inhibitor of human inducible nitric-oxide synthase (iNOS). The apparent binding constant of 1400W 2HCl was 2.3 μM at an L-arginine concentration of 0.5 μM. The substrate L-arginine blocked the binding of 1400W 2HCl to iNOS with a Ks value of 3.0 μM. 1400W 2HCl is also a reversible and relatively inefficient inhibitor of human endothelial NOS (eNOS) and neuronal NOS (nNOS). Inhibition of eNOS and nNOS by 1400W 2HCl was competitive with L-arginine with Ki values of 50 μM and 2.0 μM, respectively. Moreover, 1400W 2HCl showed potent inhibitory activity in rat aortic rings with an EC50 value of 0.8 μM[3]. In RAW264.7 cells treated with LPS/IFNγ for 16h, the increased p53 and GAPDH acetylation was blocked by 1400W 2HCl at the concentration of 100 μM. The augmented p300 acetylation, GAPDH–p300 binding and GAPDH-p53 binding in RAW264.7 cells were also abolished by 100 μM 1400W 2HCl[4]. In a rat model of endotoxin-induced microvascular injury, subcutaneous administration of 1400W 2HCl 3h after endotoxin completely suppressed the leakage into ileum with an ED50 value of 0.16 mg/kg[3].
作用机制
1400W 2HCl is a tight binding, selective inhibitor of NOS. It is competitive with the substrate L-arginine.
1400W 2HCl 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Aortic rings
744 µM (IC 50)
15 minutes
Evaluate the inhibitory effect of 1400W on ACh-evoked cGMP synthesis
Br J Pharmacol. 2013 Mar;168(5):1255-65.
Hippocampal slices
150 µM (IC 50)
15 minutes
Evaluate the inhibitory effect of 1400W on NMDA-evoked cGMP accumulation
Br J Pharmacol. 2013 Mar;168(5):1255-65.
Macrophages
10 nM
15–18 hours
Inhibition of NOS2 activity, reducing Lm spread between macrophages
Immunity. 2012 May 25;36(5):807-20.
HK-2 cells
100 μg/mL
24 hours
To evaluate the effect of 1400W on ROS production in HK-2 cells, results showed that 1400W reduced ROS levels.
Br J Pharmacol. 2020 Jan;177(1):110-127.
Microglia cell line
10, 50, 100, 150 µM
24 hours
To confirm the bioactivity of 1400W by determining the suppression of LPS-induced nitrite levels in the media.
Neurobiol Dis. 2020 Jan;133:104443.
N/TERT keratinocytes
1 µM
8 days
Inhibition of NOS2 significantly improved the skin barrier formation in the HI 3D model, restoring the lipid barrier.
J Clin Invest. 2020 Sep 1;130(9):4798-4810.
COH-BR1 cells
10 µM
8-20 hours
To evaluate the effect of iNOS inhibition on PDT-induced apoptosis, results showed that 1400W significantly enhanced apoptosis.
Free Radic Biol Med. 2013 Apr;57:39-48.
MDA-MB-231 cells
10 µM
To evaluate the effect of 1400W on PDT-induced apoptosis, results showed that 1400W significantly enhanced apoptosis.
Free Radic Biol Med. 2013 Apr;57:39-48.
1400W 2HCl 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Sprague Dawley rats
Aged rat model
Intraperitoneal injection
1 mg/kg
Single dose, sacrificed 30 minutes after treatment
1400W markedly improved ACh-induced vasorelaxation (maximal relaxation: 87%±8%, P<0.05), while having no effect on acidified NaNO2-induced vasorelaxation.
Acta Pharmacol Sin. 2010 Oct;31(10):1324-8
Sprague-Dawley rats
Insulin resistance model
Subcutaneous injection
1 mg/kg
Twice daily for 7 days
1400W significantly reversed or attenuated hyperglycaemia, hyperinsulinaemia, insulin resistance (HOMA index), body weight gain, peroxynitrite formation (nitrotyrosine expression) and the up-regulation of biomarkers of inflammation (B1 receptor, carboxypeptidase M, iNOS and IL-1β) in renal cortex and aorta and to some extent in the liver.
Br J Pharmacol. 2016 Jun;173(12):1988-2000
Mice
Murine model of oxygen-induced retinopathy of prematurity
Subcutaneous injection
1 mg/ml
Every 8 hours from P12 to P17
To evaluate the potential utility of iNOS inhibitors for therapeutic use in ischemic retinopathy, the results showed that 1400W significantly reduced the size of the central capillary-free area and inhibited intravitreal neovascularization.
J Clin Invest. 2001 Mar;107(6):717-25
Mice
GSNOR-deficient mice
Intraperitoneal injection
1 μg/g
Twice daily for 4 days
Inhibition of iNOS activity, prevention of AGT depletion, reduction of O6-etdG lesions, and decrease in hepatocarcinogenesis
Cancer Res. 2013 May 1;73(9):2897-904
C57BL/6 mice
Systemic Lm infection model
Intraperitoneal injection
10 mg/kg
Once at 0 and 24 hours post infection
Inhibition of NOS2 activity, reducing Lm spread and burden in the liver
Immunity. 2012 May 25;36(5):807-20.
Mice
High-fat diet-induced obesity model
Oral
10 mg/kg
Once daily for 28 days
To evaluate the effect of 1400W on obesity-induced chronic kidney disease, results showed that 1400W did not significantly improve kidney function.
Br J Pharmacol. 2020 Jan;177(1):110-127.
Wistar rats
Hypoxia/re-oxygenation model
Intraperitoneal injection
10 mg/kg
Single administration, observed for 48 hours and 5 days
To investigate the effects of 1400W on lung injury under hypoxia/re-oxygenation conditions in rats. Results showed that 1400W reduced NOx levels, decreased lipid peroxidation, apoptotic cell percentage, and nitrated protein expression, indicating a negative role of iNOS-derived NO in lung hypoxia/re-oxygenation.
Redox Rep. 2010;15(4):169-78
C57 BL6/J mice
Bleomycin-induced lung injury model
Subcutaneous osmotic pump
10 mg/kg/h
Continuous administration starting six days prior to ITB
Continuous administration of 1400 W via osmotic pump significantly attenuated bleomycin-induced lung inflammation, decreased chemotactic activity of the bronchoalveolar lavage (BAL), and reduced BAL cell count and nitrogen oxide production. Additionally, 1400 W inhibited the formation of S-nitrosylated SP-D (SNO-SP-D) and the structural disruption of SP-D.
Free Radic Biol Med. 2016 Feb;91:293-301
Sprague Dawley rats
DFP-induced long-term neurotoxicity model
Intramuscular
20 mg/kg
Twice daily for the first three days
1400W significantly reduced DFP-induced iNOS and 3-NT upregulation in the hippocampus and piriform cortex, and the serum nitrite levels at 24h post-exposure. 1400W also prevented DFP-induced mortality in <24h. The brain immunohistochemistry (IHC) at 7d post-exposure revealed a significant reduction in gliosis and neurodegeneration (NeuN+ FJB positive cells) in the 1400W-treated group. 1400W, in contrast to the vehicle, caused a significant reduction in the epileptiform spiking and spontaneous recurrent seizures (SRS) during 12 weeks of continuous video-EEG study.
Neurobiol Dis. 2020 Jan;133:104443.
Sprague Dawley rats
Rat soman (GD) model of epilepsy
Intramuscular injection
20 mg/kg
Once daily for 2 weeks
1400W significantly reduced GD-induced motor and cognitive dysfunction; reduced nitrooxidative stress (nitrite, ROS; increased GSH:GSSG ratio); reduced proinflammatory cytokines in serum and cerebrospinal fluid (CSF); reduced epileptiform spikes and spontaneously recurring seizures (SRS) in males; reduced neuroinflammatory markers (GFAP + C3 and IBA1 + CD68-positive glia) and neurodegeneration (especially parvalbumin-positive neurons) in some brain regions.
J Neuroinflammation. 2023 Jul 12;20(1):163
Mice
Obesity model induced by high-fat high-sucrose diet
Implanted osmotic pump
30 mg/kg/day
Continued for 8 weeks
Reversed established CMD and oxidative stress, and preserved systolic/diastolic function in mice
JACC Basic Transl Sci. 2023 Feb 1;8(5):501-514
MRL/mpj mice
Autoimmune disease model
Intraperitoneal injection
5 mg/kg
Once daily, starting the day before SEB instillation and continuing throughout the experiment
1400W significantly inhibited lipid radical production in the lung, reduced the number of macrophages and neutrophils in BAL fluid, and alleviated inflammatory cell infiltration and collagen fiber synthesis in lung tissue.
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