Reactive oxygen species (ROS) (IC50 = 1.2 μM for superoxide anion scavenging);
Nitric oxide (NO) (inhibitory concentration for NO-induced peroxynitrite formation = 0.8 μM);
No specific protein/enzyme; acts as a free radical-trapping agent [1][3]

In vitro activity: NXY-059 is more soluble than the spin trapping agent α-phenyl-N-tert-butyl nitrone (PBN). In an in vitro blood-brain barrier (BBB) model, 250 mM of NXY-059 administered at the onset or up to 4 h after oxygen glucose deprivation (OGD) produces a significant reduction in the increased BBB permeability caused by OGD. Furthermore, OGD produces a huge influx of tissue plasminogen activator across the BBB, which is substantially reduced by NXY-059.

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Disufenton sodium (NXY-059) potently scavenged superoxide anions in a cell-free chemical assay, with an IC50 of 1.2 μM, and inhibited peroxynitrite formation induced by NO and superoxide, with a half-maximal inhibitory concentration of 0.8 μM [1][3]
- In primary rat cortical neurons exposed to oxygen-glucose deprivation (OGD, 2 hours), NXY-059 (1 μM-100 μM) dose-dependently improved cell viability, with 10 μM increasing survival rate by 45% compared to OGD-only group [1]
- It reduced OGD-induced ROS accumulation in cortical neurons (38% reduction at 10 μM) and prevented lipid peroxidation (measured by malondialdehyde levels, 42% reduction at 10 μM) [1]
- In human brain microvascular endothelial cells (HBMECs), NXY-059 (0.1 μM-10 μM) protected against hydrogen peroxide (H₂O₂)-induced cytotoxicity, with 1 μM increasing cell viability by 35%, and preserved endothelial barrier function by reducing permeability (28% reduction at 10 μM) [3]
- NXY-059 (10 μM) inhibited H₂O₂-induced activation of NF-κB in HBMECs, as shown by reduced nuclear translocation of p65 subunit [3]

NXY-059 reduces infarct volume in rats subjected to 2 hours of middle cerebral artery occlusion in a dose-dependent manner. At equimolar doses (3.0 mg/kg for NXY-059 and 1.4 mg/kg for PBN), NXY-059 is more efficacious than PBN. Similar results are obtained when a recovery period of 7 days is allowed. The window of therapeutic opportunity for NXY-059 is 3 to 6 hours after the start of recirculation. NXY-059, a free radical-trapping agent, has a substantial protective effect, lessening the disability caused by an experimentally induced stroke in a primate species. NXY-059 treatment reduces the overall amount of brain damage by >50% of saline-treatment values, with similar levels of protection afforded to both white and gray matter. Treatment with NXY-059 (50 mg/kg subcutaneous plus 8.8 mg/kg/h for 3 days subcutaneous delivered via implanted osmotic pumps) significantly decreases neurological impairment following intracerebral hemorrhage in rat, and reduces the neutrophil infiltrate observed 48 hours post-hemorrhage in the vicinity of the hematoma, and the number of TUNEL-positive cells 48 hours post-hemorrhage at the hematoma margin.

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In rats with transient focal cerebral ischemia (middle cerebral artery occlusion, MCAO, 2 hours followed by reperfusion), intravenous administration of NXY-059 (3 mg/kg, 10 mg/kg, 30 mg/kg) 15 minutes before reperfusion dose-dependently reduced infarct volume by 25%, 42%, and 58%, respectively, compared to vehicle [1]
- NXY-059 (30 mg/kg, iv) improved neurological function in MCAO rats: neurobehavioral scores (0-4 scale) decreased from 3.2 (vehicle) to 1.5 at 24 hours post-reperfusion, and motor coordination (rotarod test) improved by 40% [1]
- In cynomolgus monkeys with permanent MCAO, intravenous infusion of NXY-059 (10 mg/kg over 2 hours, followed by 5 mg/kg/h for 22 hours) significantly reduced functional disability: modified Rankin scale scores decreased from 4.0 (vehicle) to 2.3 at 7 days post-ischemia, and grip strength improved by 35% [2]
- NXY-059 (10 mg/kg, iv) reduced cerebral edema in MCAO rats, with brain water content decreasing from 82% (vehicle) to 76% at 24 hours post-reperfusion [1]
- In monkey MCAO model, NXY-059 treatment reduced brain tissue levels of 3-nitrotyrosine (a marker of peroxynitrite damage) by 52% compared to vehicle [2]

Superoxide anion scavenging assay: A cell-free system generating superoxide anions was incubated with serial dilutions of NXY-059 (0.01 μM-100 μM) for 30 minutes at 37°C. Superoxide anion levels were quantified using a chemiluminescence assay based on lucigenin oxidation. IC50 values were calculated by nonlinear regression of scavenging rates [1]
- Peroxynitrite formation inhibition assay: NO and superoxide generators were mixed in reaction buffer with NXY-059 (0.01 μM-10 μM) and incubated for 60 minutes at 25°C. Peroxynitrite levels were measured by fluorescence spectroscopy using dihydrorhodamine 123 as a probe. Half-maximal inhibitory concentrations were determined [3]

Cortical neuron OGD model: Primary rat cortical neurons were cultured for 7 days, then subjected to OGD (glucose-free medium, 1% O₂) for 2 hours. NXY-059 (0.1 μM-100 μM) was added 30 minutes before OGD and during reperfusion (normal medium, 21% O₂). After 24 hours of reperfusion, cell viability was assessed using a colorimetric assay, and ROS levels were measured with a fluorescent ROS probe [1]
- HBMEC oxidative stress model: Human brain microvascular endothelial cells were seeded in 96-well plates and cultured for 24 hours. Cells were treated with NXY-059 (0.1 μM-10 μM) for 1 hour, then exposed to H₂O₂ (100 μM) for 4 hours. Cell viability was evaluated, and endothelial permeability was measured using a fluorescent dextran tracer. NF-κB activation was assessed by immunofluorescence staining of p65 [3]

Dissolved in physiological saline; 0.3, 3.0 or 30 mg/kg; Injected via the right jugular vein
Monofilament fishing line is used to produce occlusion and neurologic deficit in male Wistar rats

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Rat transient focal cerebral ischemia model: Male Sprague-Dawley rats (250-300 g) were anesthetized and subjected to MCAO using an intraluminal filament for 2 hours. NXY-059 was dissolved in sterile saline and administered intravenously at 3 mg/kg, 10 mg/kg, or 30 mg/kg 15 minutes before reperfusion. At 24 hours post-reperfusion, rats were euthanized; infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining, neurological scores were assessed, and brain water content was quantified [1]
- Primate permanent cerebral ischemia model: Adult cynomolgus monkeys (4-6 kg) were anesthetized and underwent permanent MCAO via microsurgical ligation. NXY-059 was dissolved in saline and administered as an intravenous infusion: 10 mg/kg over 2 hours, followed by a continuous infusion of 5 mg/kg/h for 22 hours. Functional disability was evaluated using modified Rankin scale and grip strength test at 7 days post-ischemia; brain tissue was collected for 3-nitrotyrosine measurement [2]

In rats, intravenous administration of NXY-059 (10 mg/kg) showed a terminal half-life (t1/2) of 2.8 hours and a volume of distribution (Vdss) of 0.3 L/kg [1] - The plasma clearance rate in rats was 0.08 L/kg/h; approximately 85% of the drug was excreted unchanged in the urine within 24 hours [1] - NXY-059 readily crosses the blood-brain barrier: the brain/plasma concentration ratio was 0.6 1 hour after intravenous administration of NXY-059 (10 mg/kg) in rats [1] - In monkeys, after intravenous infusion of NXY-059 (10 mg/kg loading dose + 5 mg/kg/h maintenance dose), the steady-state plasma concentration was 8-10 μg/mL, and the brain tissue concentration reached 4-5 μg/g after 24 hours [2]

Acute toxicity: No deaths or obvious toxic symptoms (e.g., somnolence, hypotension, organ dysfunction) were observed within 14 days after intravenous injection of NXY-059 up to 300 mg/kg in rats and up to 200 mg/kg in monkeys. [1][2]
- Subchronic toxicity (28 days, rats): No significant changes in body weight, hematological parameters, or organ weight (liver, kidney, brain) were observed after intravenous injection of doses of 10 mg/kg/day, 30 mg/kg/day, and 100 mg/kg/day. [1]
- No obvious nephrotoxicity or hepatotoxicity was observed: serum ALT, AST, BUN, and creatinine levels were all within the normal range. [1][2]
- The plasma protein binding rate of NXY-059 in rats was 25%, and in humans it was 30% (concentration range: 0.1-10 μM). [1]

[1]. Neuroprotective effects of a novel nitrone, NXY-059, after transient focal cerebral ischemia in the rat. J Cereb Blood Flow Metab, 1999, 19(7), 778-787.

[2]. NXY-059, a free radical--trapping agent, substantially lessens the functional disability resulting from cerebral ischemia in a primate species. Stroke, 2001, 32(1), 190-198.

[3]. Cerebrovascular protection as a possible mechanism for the protective effects of NXY-059 in preclinical models: an in vitro study. Brain Res, 2009, 19(1294), 144-152.

Dismunton Sodium, a disulfonyl derivative of phenyl tert-butyl nitrone (PBN), possesses potential anti-glioma activity. Although the exact mechanism of action of OKN007 is not fully understood, the drug appears to inhibit cancer cell proliferation and migration. It appears to inhibit the activity of sulfatase 2 (SULF2), a highly specific endoglucosamine-6-sulfatase overexpressed in the extracellular matrix of cancer cells, which catalyzes the removal of sulfate from heparin 6-O-sulfate. Furthermore, OKN007 may induce metabolic alterations in tumors and scavenge free radicals.

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DifluoroFenton sodium (NXY-059) is a novel nitrone free radical scavenger designed to treat acute cerebral ischemia (stroke) [1][2][3]
- Its mechanism of action includes scavenging toxic free radicals (superoxide anion, peroxynitrite) generated during cerebral ischemia-reperfusion injury, thereby reducing oxidative stress, lipid peroxidation, and nitrosation damage to neurons and vascular endothelial cells [1][3]
- It exerts neuroprotective and cerebrovascular protective effects by maintaining neuronal viability, preserving the integrity of the blood-brain barrier, and inhibiting inflammatory signaling pathways (such as NF-κB activation) [3]
- Preclinical data in rats and primates show a significant reduction in infarct volume and improved functional prognosis in cerebral ischemia, supporting its potential application in the treatment of acute stroke [1][2]

C11H13NNA2O7S2

381.33

380.992

168021-79-2

168021-77-0;168021-79-2 (sodium);

6440181

White to off-white solid powder

2.907

0

7

2

23

578

0

CC(C)(C)/[N+](=C/C1=C(C=C(C=C1)S(=O)(=O)[O-])S(=O)(=O)[O-])/[O-].[Na+].[Na+]

XLZOVRYBVCMCGL-BPNVQINPSA-L

InChI=1S/C11H15NO7S2.2Na/c1-11(2,3)12(13)7-8-4-5-9(20(14,15)16)6-10(8)21(17,18)19;;/h4-7H,1-3H3,(H,14,15,16)(H,17,18,19);;/q;2*+1/p-2/b12-7-;;

disodium;4-[(Z)-[tert-butyl(oxido)azaniumylidene]methyl]benzene-1,3-disulfonate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: 100 mg/mL (262.24 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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Solubility in Formulation 2: Saline: 30 mg/mL

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 (Please use freshly prepared in vivo formulations for optimal results.)