KR20100015014A - Pharmaceutical composition for treating spinal cord injury comprising pregabalin - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing pregabalin for preventing and treating secondary nerve injury after spinal cord injury is provided to reduce apoptosis of neural cells and glial cells and suppress microglial cell activity. CONSTITUTION: A composition for preventing and treating secondary nerve injury caused by traumatic spinal cord injury contains pregabalin. The pregabalin is 3-aminomethyl-5-methyl-hexanoic acid and its derivative. Individual for administering the composition is human and animnal including horse, lamb, pig, camel, and dog.

Description

Pharmaceutical Composition for Treating Spinal Cord Injury Comprising Pregabalin

The present invention relates to a pharmaceutical composition for preventing and treating secondary nerve injury after spinal cord injury, including pregabalin, and to a method of inhibiting secondary nerve injury after spinal cord injury of an individual using the same.

Despite extensive research, clinical advances and improved rehabilitation programs, spinal cord injury (SCI) is the leading cause of disability and death. After the initial traumatic disorder, the injured spinal cord undergoes various secondary pathological changes. These secondary damage mechanisms include ischemia reperfusion, inflammation, lethal free radicals, cytoskeleton degradation, excitotoxicity, and endogenous and exogenous apoptotic pathways (Bracken MB et al., 1997, National Acute Spinal Cord Injury). JAMA 277: 1597-1604; Dumont RJ et al., 2001, Clin Neuropharmacol 24 (5): 265-276; Fehlings MG et al., 2005, Injury 36 (S): B113-122; Shuman SL et al , 1997, J Neurosci Res 50: 798-808; Tator CH, 2006, Neurosurgery 59 (5): 957-987; Tator CH et al., 1991, J Neurosurg 75 (1): 15-26). These mechanisms are complex and interconnected.

Excitotoxicity is one of these secondary mechanisms, and pathological conditions are caused by excessive or prolonged exposure to excitatory neurotransmitters such as glutamate that induce neuronal cell death (Park E et al., J Neurotrauma 21 (6). ): 754-774). Excitotoxicity also applies to neurons and is associated with glial damage (Karadottir R et al., 2007, Neuroscience 146: 1426-1438; Li S et al., 1999, J Neurosci 19 (14): RC 16; Li S et al., 2000, J Neurosci 20 (3): 1190-1198). In particular, in case of traumatic spinal cord injury, excitatory toxicity is emphasized as an important secondary mechanism. The effect of excitatory toxicity on white matter, particularly oligodendrocytes and axons, has been demonstrated (Karadottir R et al., 2005, Nature 438: 1162-1166; Li S et al., 2000, J Neurosci 20 (3): 1190 -1198; Liu XZ et al., 1997, J Neurosci 17: 5395-5406; Park E et al., 2004, J Neurotrauma 21 (6): 754-774; Xu GY et al., 2004, Exp Neurol 187 ( 2): 329-336). Receptors of the cell body and myelin membranes are related to each other and their abnormal activation alters the ion channel and induces apoptosis and dysfunction of myelin and axons. Excessive accumulation of glutamate alters the function of ion channels, particularly Na ⁺ channels, of neurons and glial cells and induces apoptosis (Li S et al., 1999, J Neurosci 19 (14): RC 16; Li S et al., 2000, J Neurosci 20 (3): 1190-1198). Non-N-methyl D-aspartate (NMDA) receptors are known to be involved in this mechanism. As these mechanisms associated with excitatory toxicity have been identified, excitatory toxicity, including the application of NBQX (2,3-dihydroxy-6-nitro-7sulfamoylbenzo (f) quinoxaline), non-NMDA receptor antagonists and anti-exciting toxic agents, etc. Experimental attempts have been made on formulations and the like (Wrathall JR et al., Exp Neurol 137 (1): 119-126; Wrathall JR et al., Exp Neurol 145: 565-573).

Representative excitatory neurotransmitters, glutatemate receptors, are present in glial cells in the white matter of the spinal cord and apoptosis, particularly apoptosis of oligodendrocytes, is well known. Glutamate receptors consist of three ionotrophic receptors, N-methy-D-aspartate (NMDA), a-amino-3-hydroxy-5-methyl-4- isoxazolepropionate (AMPA), and kainite Park E et al., 2004, J Neurotrauma 21 (6): 754-774. AMPA-Cinate receptors are the most important for excitatory toxicity and the regulation of ion permeability, particularly cell membrane Ca ²⁺ permeability, increases intracellular Ca ²⁺ concentrations, mitochondrial dysfunction and caspase cascade Apoptosis is induced through activation of specific enzymes that stimulate (Agrawal SK et al., 1996, J Neurosci 16 (2): 545-552).

Expression of Glutamate AMPA-Cinate Receptor in Apoptotic Cells (Agrawal SK et al., 1996, J Neurosci 16 (2): 545-552), Protective Effect and Excitatory Toxicity of AMPA-Catenate Antagonists in Experimental Spinal Cord Injury Susceptibility of oligodendrocytes to glutathione cells indicates that glutamate excitatory toxicity is associated with apoptosis of neurons and oligodendrocytes in spinal cord injury. Recently, oligodendrocytes have received much attention as potential neuroprotective targets because of their association with axon function.

Pregabalin has been invented to control nerve pain in clinical situations such as diabetic neuropathy, neuralgia and complex pain syndrome. The chemical structure is similar to GABA, but does not act as GABA, and in particular cannot bind to GABA receptors. Furthermore, pregabalin binds to the α2-δ subunit of the voltage-controlled calcium channel. Subunit binding reduces the influx of calcium ions at presynaptic nerve endings and reduces the secretion of several neurons, including glutamate and noradrenaline (Coderre TJ et al., 2005, J Neurochem 94). 1131-1139; Fehrenbacher JC et al., 2003, Pain 105: 133-141; Joshi I et al., 2006, Eur J Pharmcol 553: 82-88; Shneker BF et al., 2005, Ann Pharmacother 39 (12 ): 2029-2037).

In clinical practice, spinal cord injury is extremely limited in functional recovery after injury, and as a result, early treatment is important because it can continuously cause many problems both personally and socially. However, the treatment that can lead to recovery of function through its early treatment in the actual clinical practice is very limited, and to date, the use of high dose steroids is the only initial drug treatment.

However, although studies have been conducted on the effects of steroids on neuronal and glial apoptosis, the results are controversial. It has been reported that initial treatment of methylprednisolone after spinal cord injury reduces inducible nitric oxide synthase expression and methylprednisolone acts as a neuroprotective factor by inhibiting neuronal apoptosis (Yu Y et al., 2004 Neuroreport 15 (13): 2103-2107). In contrast, Li et al. (Li X et al., 2000, Restor Neurol Neurosci 17 (4): 203-209) induce expression of caspase-3, a key enzyme of apoptosis, when methylprednisolone treatment after spinal cord injury. In conclusion, Diem et al. Reported that steroid treatment had a negative effect on neuronal apoptosis of optic nerve autoimmune diseases (Diem R et al., 2003, J Neurosci 23 (18): 6993). -7000). As mentioned above, doubts about the utility of the use of high doses of steroids and the occurrence of complications following their use, the demand for the development of new therapeutics is increasing. Under this theoretical background, the present invention investigated the possibility of pregabalin, questioning whether pregabalin would act as a neuroprotective factor after spinal cord injury.

The present invention aims to develop a drug that can maximize the functional recovery after spinal cord injury by minimizing the mechanism of secondary injury after spinal cord injury.

In the present invention, it was confirmed whether pregabalin, which inhibits hypersecretion of excitatory neuroconductors, has a protective function against secondary damage after spinal cord injury.

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention relates to a pharmaceutical composition for preventing and treating secondary nerve damage of spinal cord injury, including pregabalin.

The active ingredient of the present invention for preventing and treating secondary nerve injury after spinal cord injury of the composition of the present invention is pregabalin. Pregabalin means a compound known as 3-aminomethyl-5-methyl-hexanoic acid and derivatives thereof, and is bipolar with epilepsy, pain, inflammation, gastrointestinal damage, alcoholism, insomnia and mania It is known to have a therapeutic effect against psychosis such as disorder. The present invention relates to a novel activity of pregabalin as a neuroprotective factor that inhibits the progression of secondary nerve injury after spinal cord injury, which has not been revealed to date.

In the present invention, the term “secondary nerve injury” refers to the widespread loss of nerves that are caused by primary injury directly after primary traumatic spinal nerve injury, and this secondary injury process is responsible for the initial traumatic injury. It gets worse. After injury to the spinal cord, primary injury often becomes complex with progressive secondary damage of adjacent nerves that are not apparently damaged by the initial injury or that are only peripherally damaged. The primary damage causes extracellular ion concentrations, an increase in the amount of free radicals, release of neurotransmitters, depletion of growth factors, and local inflammations, and these changes were originally released from primary damage. It triggers a series of destructive reactions in adjacent nerves. These secondary damage mechanisms include ischemia reperfusion, inflammation, lethal free radicals, cytoskeletal degradation, excitotoxicity and endogenous and exogenous apoptosis. Thus, pharmacologically controlling secondary nerve injury may be a new therapeutic strategy for spinal cord injury. As part of this strategy, the study confirmed that pregabalin protects, prevents, or inhibits nerve damage from the effects of secondary nerve damage that persists even after primary risk factors are eliminated or weakened.

Pregabalin administration promotes neurological recovery, reduces spinal cord cavitation, reduces apoptosis of neurons and glial cells, and decreases the activation of microglial cells. Could.

Pregabalin of the present invention may be in chemically modified form. Such chemical variants may have different pharmacokinetic profiles than the parent compound, with changes in absorbency, solubility and stability (eg, increased plasma half-life).

In addition, the pregabalin of the present invention may be in free form or in pharmaceutically acceptable complexes, salts, solvates, hydrates and polymorphic forms. The term "pharmaceutically acceptable" means within the scope of medical judgment, a substance having a reasonable benefit to risk ratio and effective for the desired use, without causing excessive toxicity, irritation, allergic reactions.

Specifically. Pregabalin or variants thereof of the present invention may include pharmaceutically acceptable salts of organic and inorganic acids or bases. Pharmaceutically acceptable acid addition salts of basic compounds useful in the methods of the invention include non-toxic salts derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, and the like, Non-toxic salts derived from organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids and the like. The salts may also be sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromide, iodides, Acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suverate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methyl Benzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfonate and the like. Examples of the amino acid salts include alginate, gluconate, galacturonate, and the like.

The composition of the present invention may be formulated in various forms of preparations containing a pharmaceutically acceptable carrier and suitable for oral, parenteral, topical administration, etc. for human or veterinary use. At this time, short-term, rapid-offset, controlled release, sustained release, delayed release, and pulsatile relase). Solid agents for oral administration of powders, granules, tablets, capsules and the like can be used binders, suspending agents, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavorings and the like. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents, water and liquid paraffin. In the case of injections, buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers and the like can be used in combination.

As another aspect, the present invention relates to a method for inhibiting secondary injury after spinal cord injury in a subject using a pharmaceutical composition that prevents and treats secondary injury after spinal cord injury comprising pregabalin.

Individuals who can administer the compositions of the present invention include humans with spinal cord injury, as well as animals such as horses, sheep, pigs, goats, camels, antelopes, dogs, and the like. By administering to a subject a composition comprising the pregabalin of the present invention, it is possible to effectively prevent and treat secondary damage due to spinal cord injury. The composition of the present invention can be administered in parallel with existing therapeutic agents for spinal cord injury.

As used herein, the term "administration" means introducing a predetermined substance into a patient by any suitable method, and the route of administration of the composition of the present invention is oral or parenteral via any general route as long as the target tissue can be reached. May be administered. In addition, the composition may be administered by any device in which the active agent may migrate to the target cell.

The composition of the present invention is administered in a pharmaceutically effective amount.

As used herein, the term “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level refers to an individual's sexually transmitted disease, age, severity, and drug activity. , Drug sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs, and other factors well known in the medical arts. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. It may be single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, and can be easily determined by those skilled in the art. Dosage levels for an individual can vary depending on gender, age, health condition, diet, time of administration, method of administration, drug mixture and severity of disease.

Pregabalin inhibits and protects secondary neuronal damage caused after spinal cord injury, restoring motor function after spinal cord injury, effectively reducing spinal cord cavitation and apoptosis of neurons and glial cells, and effectively reducing microglial activation. .

Hereinafter, the present invention will be described in detail by examples. However, the following examples are merely provided to more easily understand the present invention, and the present invention is not limited by the following examples.

Example  1: materials and methods

1-1: Experiment Animal

A total of 32 male white rats (adult male Sprague-Dawley rats: 300-350 g body weight) were used. Animals were maintained at averaged conditions (4 rats / cage, 20-24 ° C., 45-65% relative humidity, 12 hour light / dark cycle) and were given food and water randomly. Rats were randomized to control group (n = 8, no drug injection after spinal cord injury), methylprednisolone (MP) treatment group (n = 8, peritoneal injection of MP after spinal cord injury), MC (minocycline) treated group (n = 8, MC intraperitoneal injection after spinal cord injury), GP (pregabalin) treated group (n = 8, GP intraperitoneal injection after spinal cord injury) Classified.

1-2: working technology

Mice were anesthetized using ketamine (ketamine, 50 mg / kg) and rumpun (rompun, 2 mg / kg). The back was shaved and sterilized with antiseptic betadine and excision was performed at T9 after exposing T8-10 paraspinal muscles. Spinal cord injury was induced using a NYU impactor (New York University impactor, 10 gm rod dropped from a height of 2.5 cm). NYU impactors using electronically released flangers induced reproducible damage. A 25g-cm lesion was selected to investigate the neuroprotective effects of severe central nerve injury that resulted in insufficient motor nerve regeneration for several weeks. After surgery, 5 mg of gentamycin was administered intramuscularly. Post-operative care processes include providing water to drink, soft foods, and regular pellets. The bladder was manually emptied twice a day during the experiment. All processes of surgery, preoperative and postoperative animal care were based on the Experimental Animal Welfare Act and guidelines and policies for rodent surviving animal testing provided by the Animal Research Council of the Catholic University of Korea.

1-3: Pharmaceutical Administration

MP group was intraperitoneally administered 30 mg / kg methylprednisoline at 30 minutes, 12 hours and 2 hours after SCI. The MC group received peritoneal doses of 30 mg / kg of minocycline (Sigma, St Louis, MO) 30 minutes, 12 hours, and 24 hours after SCI, and the GP group received 30 mg / kg pregabalin (Pfizer Pharm, New). York, NY) was intraperitoneally administered for 30 minutes and every 2 hours every 12 hours for 2 days. Since solubilization of minocycline was known to be difficult, 9.6 ml of minocycline (400 mg) was dissolved in sterile distilled water and sonicated.

1-4: Behavioral Assessment

(1) motor function scores

Motor function scores (Gale K et al., 1985, Exp Neurol 88 (1): 123-134) were used to assess motor function (Table 1). The hip, knee and ankle joints were recorded by allowing the animals to move freely and noticing the observer and observing at least 1 minute every day.

(2) Inclined plane test

All rats underwent a slope test to assess the animal's ability to maintain its position on the board increased by 5 °. This test was used as a measure of hind leg strength. The maximum angle that can hold the position for up to 5 seconds was defined as the slope score. Mice underwent this test for 7 days after surgery until sacrifice.

1-5: Preparing the Organization

On day 7 post-op, mice were sacrificed under deep sleep by intraperitoneal administration of ketamine and lumps. Pericardial perfusion was performed in 100 ml of buffered saline, treated with 500 ml of 4% paraformaldehyde, and a 1.5-cm spinal cord region containing an aqueous phase was removed. Samples were fixed overnight in cold 4% paraformaldehyde, incubated in 30% sucrose solution for 4 hours at room temperature and stored at -70 ° C.

1-6: Immunofluorescence  dyeing

The tissue was cut at right angles to the long axis of the spinal cord at a thickness of 10 μm. Samples were enclosed in gelatin-coated slides and washed three times for 10 minutes with PBS, followed by blotting with 0.15% Triton X-100 and 5% NHS (normal horse serum) in 0.1 mM PBS for 30 minutes. Tissue samples were treated overnight with CC-1 (Calbiochem, Darmstadt, Germany), a mouse monoclonal antibody specific for oligodendrocytes diluted 1:20 with 0.1 M PBS + 0.15% Triton X-100 + 1% NHS. The following day, the slides were incubated for 2 hours with a Texas red-conjugated anti mouse IgG (Jackson ImmunoResearch, West Grove, PA), a secondary antibody diluted 1: 100. Slides were washed five times with 0.1 mM PBS, covered and stored in the freezer. After thawing, slides were taken using an Olympus Microscope Confocal / Image Analysis system, Tokyo, Japan. Cells showing red fluorescence in the 75 × 250 μm area were counted at the center of the spinal cord injury. In addition, in order to detect apoptotic cell apoptosis, apoptosis detection kit (Roche, Mannheim, Germany) using in situ Double staining was done using nick-end labeling (TUNEL) staining technique. Deparaffinized tissue sections were incubated with proteinase K (20 μg / mL). To inhibit endogenous peroxide activity, tissue sections were treated with 3% H ₂ O ₂ and incubated with 1 × buffer for 30 minutes at room temperature. Next, it was treated with digosigenin-labeled deoxynucleotide transferase at 37 ° C. for 1 hour. The slide was washed in stop / wash buffer for 10 minutes at room temperature. Tissue sections were incubated for 30 minutes at room temperature using anti-digosigenin-peroxidase antibody and stained with anti-fluorescein isothiocyanate (Jackson ImmunoResearch). Staining overnight with dilution monoclonal antibody OX-42 (Serotec, Oxford, UK) at 1: 500 in 0.1 M PBS + 0.15% Triton + 1% NHS. The following day, slides were stained for 2 hours with secondary antibody, FITC conjugated anti-mouse IgG (Jackson ImmunoResearch) diluted 1: 100.

1-7: Immunofluorescence  Quantification of Cells

Morphological analysis of fluorescence positive cells was performed in a blind fashion under high power magnification. In each slide, six fields were randomly selected and positive cells were counted around the cystic lesions to eliminate possible bias by hemorrhage and necrosis. To quantify the degree of apoptosis, the number of TUNEL positive cells in each group was recorded. Finally, the total mean number for each set of samples was calculated and the mean values were compared. Cells double stained with TUNEL and CC-1 antibodies appear yellow and were counted in injured spinal cord stool. The average number recorded in all samples and groups was compared.

1-8: Statistical Analysis

For statistical analysis, all data were subjected to the Kruskal-Wallis test (a nonparametric test) and pair-wise multiple comparisons using the Mann-Whitney U test. P values lower than 0.05 were considered to indicate statistical significance.

Example  2: SCI  after Pregabalin  Function recovery Improve

2-1: motor skills score

The effect of pregabalin on recovery after SCI was compared with the MP and MC used to treat acute SCI. The hind limbs of all rats were incapacitated immediately after injury, and the hind limb movements recovered over time in four groups. The hourly route of functional recovery was recorded using the Gale rating score (FIG. 1). Motor neuron scores are presented as mean values and SEM (Table 2). According to the Kruskal Wallis test, the mean motor function scores of the four groups were significantly different ( P = 0.01). According to the Man-Wittney test, the mean motor neuron scores were significantly different between the control group and the GP group ( P = 0.01). Recovery of hind limb motor function scores in the GP treated group was significantly higher than in the MP or MC treated group ( P <0.05).

2-2: motor skills score

(1) slope test

The results of the slope test were shown by the mean value and SEM. According to the Kruskal Wallis test, the slope scores of all groups differed significantly ( P = 0.01). In addition, for the Mann-Wittney test, there was a statistical significance ( P = 0.01) comparing the slope of the slope between the control group and the GP group. The results for the GP group were higher than for the MP and MC groups. (P <0.05).

(2) Pregabalin reduces cystic lesions

Morphological features of the hematoclinin-eosin stained site showed that cystic lesions were mainly located in the dorsal region of the spinal cord, and the lesion size was larger in the control group and the other three groups (Fig. 2).

Example  3: Pregabalin  Nerves and In glia cells Apoptosis  Increase

Traumatic injury induces widespread apoptosis of neurons and glial cells in the spinal cord. After SCI, it was observed by TUNEL staining whether pregabalin inhibited neuronal and glia cell death. TUNEL positive cells (TPC) were distributed mainly around the damaged area and extended to the abdomen of cystic lesions. The average number of TPCs in each group was 63.5 ± 7.4, 53.6 ± 4.0, 44.2 ± 3.9, and 36.5 ± 3.6, respectively. TPC in the control group was significantly higher than in the other three groups ( P <0.05), and TPC numbers in the GP group were significantly lower than the MP and MC groups ( P <0.05) (FIG. 3). Apoptotic indexes (AI) were calculated as follows in double staining (TUNEL and anti-CC1) for apoptotic liposomal cells. AI = (number of double stained cells / number of cells positive for anti-CC1) × 100. The mean AI was 88.6% in the control group, 46.7% in the MP group, 82.1% in the MC group and 70.3% in the GP group. AI values were significantly lower in MP and GP than in the other groups (p <0.05) (FIG. 4).

Example  4: Pregabalin Microglia  Activation Restrain

Anti-OX-42 positive cells were detected mainly around the site of injury (FIG. 5). The average number of anti-OX-42 positive cells was 29.8 ± 3.9 in the control group, 22.7 ± 4.1 in the MP group, 21.0 ± 3.9 in the MC group and 17.8 ± 4.3 in the GP group. The average number of high anti-OX-42 positive cells in the control group was statistically significant ( P = 0.01). The number of anti-OX-42 oriented cells in the GP group was significantly lower than the MP and MC groups (p <0.05). This means that pregabalin significantly reduces the activity of microglial cells and the effect is comparable to minocycline treatment.

Since the pharmaceutical composition comprising the pregabalin of the present invention can effectively inhibit, protect, and prevent secondary damage after spinal cord injury, wide application in veterinary and medical fields can be expected.

Figure 1 shows the evaluation of the recovery of exercise capacity over time after spinal cord injury, the pregabalin administration group shows that the recovery of exercise ability was significantly promoted compared to the control and steroid administration group.

Figure 2 is a histological comparison of the cavitation of the injury site after spinal cord injury, the plazavaline administration group reduced the spinal cord cavitation area compared to the control and other experimental groups (MP; methylprednisolone, MC: minocycline, GP: pregabalin).

FIG. 3 shows that TUNEL staining for apoptosis of neurons and glial cells was statistically analyzed and compared, indicating that TUNEL positive cells were smaller than those of the control group.

4 is a result of TUNEL and anti-CC1 staining to confirm the apoptosis of lipophilic cells, the apoptosis of the lipophilic cells shows a significantly lower apoptosis rate in the steroid administration group and the fluzavaline administration group.

5 shows the activation of microglial cells, the microglial cell activation is significantly lower than the control group.

Claims (4)

A composition for preventing and treating secondary nerve damage caused by traumatic spinal cord injury, comprising pregabalin. The composition of claim 1, wherein the composition inhibits apoptosis of neurons and glial cells. The composition of claim 1, wherein the composition inhibits microglial activity. A method of inhibiting secondary nerve damage caused by traumatic spinal cord injury using the composition of claim 1.

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