KR100782333B1 - Pharmaceutical composition for the treatment of nerve damage, including plasma or serum - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating nerve damage, and more particularly, to a pharmaceutical composition for treating nerve damage, which contains a pharmaceutically effective amount of plasma or serum as an active ingredient.

The pharmaceutical composition for treating nerve damage according to the present invention is useful for treating nerve damage by regenerating nerve cells after spinal nerve injury and providing complete structural continuity of the spinal nerve injury lesion.

Plasma, serum, nerve damage, paste

Description

Pharmaceutical Composition for the Treatment of Nerve Damage Comprising Blood Plasma or Serum}

Figure 1 shows the results of the BBB test of a rat model that damaged neurons.

Figure 2 shows the results of the grid walk test of rat model that damaged neurons.

Figure 3 shows a photograph of the foot analysis of the rat model (A: normal group; B: control group; C: treatment group).

Figure 4 shows the results of the electrophysiological sensory nerves instantaneous rat model injured neurons.

Figure 5 shows the histological picture of the control group and the normal group lesions of the rat model.

The present invention relates to a pharmaceutical composition for treating nerve damage, and more particularly, to a pharmaceutical composition for treating nerve damage, which contains a pharmaceutically effective amount of plasma or serum as an active ingredient.

The nervous system is divided into the peripheral nervous system and the central nervous system, and the peripheral nervous system includes all other nerve elements except the central nervous system and the brain and the spinal cord. Other classifications of the nervous system include the body nerves and autonomic nerves, which include the cranial nerve and spinal nerve, and the autonomic nervous system include the sympathetic and parasympathetic nerves. Nerve cells are composed of cell bodies, dendritic projections and axons, the nerve stimulation enters the cell body is a dendritic projection, the far side of the nerve stimulation is axon. At the end of the axon are several synaptic ends, where other neurons or target cells are connected. Axons consist of nodes called myeline sheath and node of Ranvier surrounded by Schwann cells.

Typical treatment of neurological diseases associated with neuronal cell death or damage is to promote neuronal regeneration by inhibiting neuronal outgrowth or to inhibit neuronal cell death. Neurostimulators known to date include nerve growth factors (NGF; Levi-Montalcini R. and Hamburger V., J. Exp . Zool ., 123: 233, 1953), brain-induced neurotropic factors (BDNF; Leibrock et. al ., Nature , 341: 149, 1989), neurotropic factor-3 (NT-3; Maisonpierre et . al ., Science 247: 1446, 1990), ciliary neurotropic factor (CNTF; Lin et al ., Science 246: 1023, 1990), insulin and insulin-like growth factor (Aizenman et al ., Brain Res ., 406: 32, 1987; Bothwell, J. Neurosci . Res ., 8: 225, 1982; Recio-Pinto et al ., J. Neurosci ., 6: 1211,1986; Near et al ., PNAS , 89: 11716, 1992; Shemer et al ., J. Biol . Chem ., 262: 7693, 1987; Anderson et al ., Acta Physiol . Scand ., 132: 167, 1988; Kanje et al ., Brain Res ., 475: 254, 1988; Sjoberg and Kanje, Brain Res ., 485: 102, 1989; Nachemson et al ., Growth Factors , 3: 309, 1990; Re-cio-Pento et al ., J. Neurosci . Res ., 19: 312, 1988; Matteson et al ., J. Cell Biol . , 102: 1949, 1986; Edbladh et al ., Brain Res ., 641: 76, 1994), activin (Schubert et. al ., Nature 344: 868, 1990), purpurine (Berman et al ., Cell , 51: 135, 1987), fibroblast growth factor (D. Gospodarowicz et al ., Cell Differ ., 19: 1, 1986; Baird, A. and Bohlen, P., Fibroblast growth factors. In: Peptide growth factors and their receptors 1. (eds.Sporn, MB and Roberts, AB) 369-418. Spring-Verlag, Berlin, Heidelberg, 1990; And Walter, MA et al ., Lymphokine Cytokine Res ., 12: 135, 1993), estrogen (Toran-Allerand et al ., J. Steroid Biochem . Mol . Biol ., 56: 169, 1996; McEwen, BS et al ., Brain Res . Dev . Brain . Res ., 87:91, 1995), platelet derived growth factors (PDGF; US Pat. No. 6,506,727), fibrinase (e.g., tissue-plasminogen activator) or fibrinogen formation inhibitors (e.g., ankrod, urokinase , Streptokinase and anticonvulsants; US Patent Publication No. 2003/0219431 A1). Thrombomodulin analogs (US Patent Publication No. 2004/0002446 A1) are known as inhibitors of neuronal cell death.

It is well known that the central nerves of the brain and spinal cord are incapable of regenerative, and there is no way to treat trauma such as spinal cord injury or neurodegenerative diseases such as Alzheimer's and Parkinson's.

Although some studies have shown that neurons in the spinal cord are regenerative, the regenerative response is usually incomplete, with shorter axons, and the reason for this immature response remains unknown.

There is a limit to the treatment of nerve damage using the above known neurostimulation promoting substances, spinal nerve injury is limited to a few treatment methods are used. Until recently, pharmacotherapy following spinal nerve injury was intravenous administration of high doses of steroidal methylprednisolone (Hall, ED, Adv . Neurol ., 59: 241, to minimize secondary injury early in spinal injury). 1993; Bracken, MB, J. Neurosurg ., 93: 175, 2000; Bracken, MB, Cochrane Database Syst . Rev. 2, 2000; Koszdin, et al ., Anesthesiology, 92: 156, 2000). However, recent analytical data suggest that this drug therapy should be banned (Hurlbert, RJ, J. Neurosurg ., 93: 1, 2000; Pointillart et al ., Spinal Cord , 38: 71, 2000; Lankhorst et al ., Brain Res ., 859: 334, 2000). Other drugs are still at the experimental level.

In recent years, much progress has been made in understanding the factors that limit spinal nerve regeneration and developing strategies for successful regeneration of the spinal nerve based on them (Ramer, MS). et al ., J. Neurosci . 21: 2651, 2001). These studies have suggested Nogo as one of the inhibitors of central nerve regeneration (Chen, MS et. al ., Nature 403: 434, 2000). However, suppression of labor may result in the regeneration of other axons because only a portion of the axon is regenerated, and the candidates for regeneration are MAG (myelin-associated glycoprotein), tenascin, gangliosides, ephrin, netrin, semaphorins, etc. This can be illustrated. It is clear that these substances play an important role in the successful regeneration of the spinal nerve, but in addition, other requirements, such as maximizing positive support for the regeneration of axons, optimizing the function of living axons in partial lesions, and prolonging Those that guide regrowth, establish proper continuity, and minimize the deleterious effects of early trauma, inflammation and wound formation should also be considered in regeneration of the spinal nerves. In this regard, Priestley et al. Suggested the possibility of using fibronectin mat with Neutropin to stimulate regeneration of damaged spinal nerves (Priestley, JV et. al ., Journal of Physiology- Paris 96 , 123-133, 2002).

However, the neuronal regeneration by the growth factors as described above is limited to a certain part of the regeneration process of the neurons, and has a disadvantage in that it is not economical because the production and purification of the growth factors are expensive.

Therefore, the present inventors have been diligent to develop a more effective neuronal regeneration material, when treated with plasma or serum derived from mammals in a rat model in which the spinal nerve is injured, the nerve cells are regenerated, and the site of the regenerated nerve damage lesion is different. It was confirmed that the structure has a complete continuity, and the present invention was completed.

It is an object of the present invention to provide a pharmaceutical composition for treating nerve damage, which contains a pharmaceutically effective amount of plasma or serum.

In order to achieve the above object, the present invention provides a pharmaceutical composition for treating neuronal damage containing a pharmaceutically effective amount of plasma or serum as an active ingredient.

In the present invention, the composition may be any one selected from the group consisting of a paste, a solution, a suspension and a topical gel form, wherein the plasma or serum is contained in 0.1 to 99.9% by weight of the composition It may be characterized by.

In the present invention, the nerve injury may be characterized in that the peripheral nerve damage or spinal cord damage, the nerve damage may be characterized in that the nerve is cut damage, the nerve is damaged the nerve protrusion is cut It may be characterized by cutting more than 10mm.

In the present invention, the peripheral nerve injury is diabetic neuropathy, acromegaly, hypothyroidism, AIDS, leprosy, Lyme disease, systemic lupus erythematosus, rheumatoid arthritis, Scheren's syndrome, periarterial arthritis, Wegener's granulomatosis, cranial It may be characterized as a neuropathy associated with a disease selected from the group consisting of arteritis and sarcoidosis.

Hereinafter, the present invention will be described in more detail.

Spinal nerve injury in the present invention means all the various damage caused by the external force applied to the spinal cord. Spinal nerve injury is pathologically physiologically composed of primary and secondary damages. Primary injury of spinal nerves is tissue damage caused by physical rupture when an impact occurs. In humans, primary injury is caused by mechanical forces such as shock, compression, traction, laceration, and cuts. The most common mechanism is the combination of shock and continuous compression. Compression fractures, fracture-dislocations, gunshot wounds, and disc rupture That is the main cause.

Secondary damage to the spinal nerve is initiated by biochemical mediators released from tissue at the primary injury. Biochemicals caused by secondary damage cause pathophysiological signaling and gradually damage tissues, eventually leading to cell death, which liberates another biochemical and continues to break the tissue. These secondary pathophysiological processes include cell membrane damage, the effects of blood vessels, inflammation, electrolyte imbalance, altered energy metabolism, and various biochemical changes that cause oxidative stress.

The extent of nerve damage can be assessed in several ways. For example, Frankel classification, American Spinal Injury Association (ASIA) classification, Yale classification, motor index scale, Barthel index transformation (Wells, JD and Nicosia, S., J. Spinal Cord Med. 18:33, 1994). Patients can be distinguished as complete or incomplete spinal cord injury. Complete injury can be attributed to complete loss of muscle strength and sensation in the first 24 hours after trauma, contraction of the anal sphincter, loss of perianal sensation, and loss of spongy sphincter. Incomplete spinal cord injury refers to a condition in which some movement or sensory function remains below the injury site, and includes anterior cord syndrome, posterior cord syndrome, central cord syndrome, and lateral measurements. It is divided into lateral cord syndrome and neuromuscular syndrome.

Animal models of traumatic spinal nerve injury have been used to elucidate the pathophysiology of pathophysiology and to evaluate the therapeutic effects (Anderson, TE and Strokes, BT, J. Neurotrauma , 9: S135, 1992; Blight, AR, Central). Nervous System Trauma , 2: 299, 1985; Blight, AR et al ., McGraw Hill: New York, 1367-1379, 1966. These animal models include weight drop model (Kuhn, PL and Wrathall, JR, J. Neurotrauma , 15: 125, 1998), crushing damage model (Bunge, RP et al ., Advances in Neurology. FJ Seil (ed), Raven Press: New York, 75-89, 1993; Bunge, RP et al ., Neuronal Regeneration, Reorganization, and Repair. FJ Seil (ed), Leppincott-Raven Publishers: Philadelphia, 305-315, 1997), Left Injury Model (Stokes, BT et al ., J Neurotrauma 9: 187, 1992). The left injured model is the best model for mimicking fast and non-penetrating trauma. The damage formed is biopsied (eg, light or electron microscopy and staining and tracking methods; Gruner, JA, J. Neurotrauma , 9: 123, 1992), measurement of electrophysiological results (eg, induced potential; Metz, et al . , J. Neurotrauma , 17: 1, 2000) or behavioral assessment (eg outdoor walking or postural stability on slopes; Basso et. al ., J. Neurotrauma , 13: 343, 1996).

In another aspect of the invention, the present invention relates to a pharmaceutical composition for treating peripheral nerve damage in which neurites are cut, in particular nerve injury in which the cut neurites are cut to a length of several millimeters or more (eg, 10 mm or more). . The regenerative properties of nerve cells in the peripheral nervous system constrain the ability to restore the function of damaged nerve pathways. In other words, the new axon is randomly elongated and often misaligned, resulting in contact with an inappropriate target and thus acting abnormally. For example, if the motor nerves are damaged, the regrowth axon may come in contact with the wrong muscles, which eventually leads to paralysis. In addition, when the cut neurites are cut to a length of several millimeters or more (eg, 10 mm or more), proper neural regeneration may not occur because the protrusions do not grow as necessary distances or grow in the wrong direction of axons. .

Surgical treatment of peripheral nerve injury can have a variety of consequences. In some cases, sutures in order to obtain a proper arrangement of truncated nerve endings stimulate the formation of scar tissue that is believed to inhibit axon regeneration. Even when scar tissue formation is reduced, successful nerve regeneration is still limited to nerve damage within 10 mm. In addition, the ability to repair peripheral neurons is significantly suppressed if damage or neuropathy affects the cell bodies of the neurons themselves or results in extensive degeneration of peripheral axons. The composition of the present invention makes it possible to treat peripheral nerve damage that has been cut to a length of 10 mm or more in mammals because it regenerates nerve cells and provides complete structural continuity of nerve lesions.

In another aspect, the present invention relates to a pharmaceutical composition for treating peripheral neuropathy induced by a disease and a condition associated with a disease. In addition, peripheral neuropathy induced by the disease may be diabetic neuropathy. Diabetic neuropathy can be defined as a clinically apparent disease that occurs with symptoms of diabetes without other causes of peripheral neuropathy. Such neuropathy includes the symptoms of the peripheral and autonomic nervous systems of the peripheral nervous system. Diabetic neuropathy damages the nerves directly beneath the skin, causing one or more of the following symptoms: numbness and numbness of fingers, hands, toes and feet; Asthenia of the hands and feet; Pain and / or burning in the hands and feet. In addition, disease-induced peripheral neuropathy is associated with acromegaly, hypothyroidism, AIDS, leprosy, Lyme disease, systemic lupus erythematosus, rheumatoid arthritis, Siegren's syndrome, nodular periarteritis, Wegener's granulomatosis, cranioarteritis or sarcoidosis It may be neuropathy.

It is also possible to treat such disease-induced peripheral neuropathy and related conditions in mammals because the compositions of the present invention regenerate neurons and provide complete structural continuity of nerve lesions.

Plasma used as an active ingredient in the present invention typically refers to a pale yellow liquid component in which tangible components in mammalian blood, ie, cells and cell fragments, are separated and their composition and composition are well known (Westerman, P., Plasma). Proteins, VII-1 to VII-13, September 17, 2002; Craig, WY et al ., Plasma Proteins Pocket Guide, Foundation for Blood Research—the entire contents of these documents are incorporated herein by reference).

Serum, which is used as an active ingredient in the present invention, generally means a portion in which fibrinogen and other coagulation factors have been removed from plasma.

Sources of plasma or serum in the present invention include all species of mammals, including humans, non-human primates, for example livestock, other primates, rodents, etc., including sheep, goats, pigs, horses, dogs, cattle, and the like. This includes.

Plasma or serum in the present invention can be easily separated from blood according to well-known conventional methods such as centrifugation, sedimentation or filtration methods. Centrifugation can be carried out under conditions suitable for settling blood cells from plasma. For example, centrifugation at about 1,400 g for about 10 minutes, which is sufficient to precipitate substantially all cell fragments (platelets), as well as red blood cells and white blood cells. Supernatants containing plasma can be easily separated from the settled cells by standard techniques. Filtration can be performed by passing blood through a filter suitable for separating blood cells from plasma. The filter can be a microporous membrane through which proteins can permeate well.

Plasma or serum is known to be preserved in various states before use, in addition to fresh liquid plasma or liquid preparations obtained by centrifugation or sedimentation after collection, for example, fresh frozen preparations, lyophilized preparations, lyophilized preparations or concentrated preparations. It may be mentioned, plasma or serum of all the above-described conditions can be used. Fresh frozen plasma is prepared by centrifugation at about 1,400 g for about 15 minutes within 6 hours after blood is drawn from the blood to separate blood cells and plasma components and freeze at a temperature of about -18 ℃ to -40 ℃. It is preferable to thaw in hot water at 37 [deg.] C. to use.

Cryoprecipitation plasma dissolves 1 unit of fresh frozen plasma at about 4 ° C and separates white precipitates (containing large amounts of factors such as cold precipitated protein (VIII: C, fibrinogen, XIII, fibronectin, etc.) from -18 ° C to It was re-frozen to -40 ℃. Cryoprecipitation agents can be used by thawing overnight in a refrigerator at 1-6 ° C. or by thawing faster in a about 4 ° C. water bath.

Concentrated plasma separates plasma from blood, and the separated plasma is separated into dextranomer, SEPHADEX, dextramine, polyacrylamide, bio-gel (BIO-GEL) P, silica gel, zeolite, debris acid ( DEBRISAN), mixed with a thickening agent such as crosslinked agarose, starch or alginate gel, and then concentrated to separate the thickening agent from the concentrate.

As one embodiment of the present invention, plasma or serum may be commercially available. For example, powdered formulations commercially available from blood banks, liquid formulations from Invitrogen Corporation (e.g., Gibco ^TM Chichen Serum, Gibco ^TM Goat Serum, Gibco ^TM Lamb Serum, Gibco ^TM Porcine Serum, Gibco ^TM Rabbit Serum) Or serum preparations of Gemini Bio-Products (CA 95776, USA) (eg, Chicken Serum (Cat. # 100-161), Dog Serum (Cat. # 100-160), Donor Donkey Serum (Cat. # 100-) 151), Donor Goat Serum (Cat. # 100-109), Donor Rat Serum (Cat. # 100-155), Feline Serum (Cat.100-153), Guinea Pig Serum (Cat. # 100-130), Monkey Serum (Cat. # 100-154), Mouse Serum (Cat # 100-113), Porcine Serum (Cat. # 100-115), Rabbit Serum (Cat. # 100-116), Rat Serum (Cat. # 100- 150), Sheep Serum (Cat. # 100-117)). These products are derived from plasma units of other animal origin, including humans, and are nonreactive and resistant to HIV-1 and HIV-1 antibodies, such as hepatitis B surface antigen (HBsAg) and hepatitis C (HCV) antibodies. Test results were confirmed to be negative for the antibody against 2. All plasma units used to prepare such formulations have been proven to be pathogenic in advance.

When using plasma or serum other than the above agents, the agent preferably inactivates enveloped viruses such as HIV, hepatitis B virus and HCV, in order to reduce the risk of latent propagation of pathogens. Common methods of inactivating plasma include pasteurization, dry heat treatment, steaming, treatment with organic solvents / detergents such as tri (n-butyl) / phosphate / polysorbate 80, low pH (pH 4), cold Ethanol fractionation, chromatography, nanofiltration, and the like have been used. Recently, methods such as light irradiation such as UV, gamma irradiation, and iodine treatment have also been developed. Preferably, gamma-irradiation, methylene blue light treatment, and steam treatment are sequentially performed on the plasma unit to inactivate viruses that may be present in the plasma.

The plasma or serum fractions used in the present invention may be those powdered by heating, lyophilization or other suitable drying technique. For example, plasma or serum is lyophilized at temperatures below −40 ° C. for several days (eg, for about seven days). Conventional techniques and parameters known to those skilled in the art can be used.

In order to effectively treat neurological damage, the plasma or serum used in the pharmaceutical composition of the present invention should act easily at the target site. For this purpose, a composition for suitable use according to the invention is a formulation comprising plasma or serum as an active ingredient of the invention in combination with one or more of the usual pharmaceutically acceptable excipients or carriers. Manufacture. The term "pharmaceutically acceptable" as used herein means that it is compatible with the active ingredient and does not harm the beneficiary. In the present invention plasma or serum may be present from 0.1% to 99.9% based on the total weight of the pharmaceutical composition. As another alternative, plasma or serum may be used alone in combination without excipients, carriers or diluents.

Plasma or serum according to the invention is typically administered in topical form. Formulations suitable for topical administration include semisolid, semi-liquid or liquid preparations such as pastes, gels, solutions, emulsifiers or suspensions. Preferred routes of administration of the compositions of the invention may be intramuscular, subcutaneous, transdermal or infusion. Alternatively, the formulation may be a lyophilized powder and may be used reconstituted with a physiological body fluid and isotonic and buffered media to a suitable pH. The preparation of these formulations can be carried out by mixing or dissolving the various components or kneading the mixture using any of the devices and methods commonly used or well known in the pharmaceutical art. [Remington's Pharmaceutical Science, 18th Edition, 1990 , Mack Publishing Company, Easton, Pennsylvania 18042 (Chapter 87: Blaug, Seymour). Preferably it may be formulated in the form of a paste.

Formulations of the present invention include viscosity modifiers such as beeswax, glyceryl tribehenate, glyceryl trimyristate; Buffers such as potassium metaphosphate, potassium phosphate, monovalent sodium acetate, sodium citrate anhydride; Surfactants such as fatty acid alkali metals, dimethyl dialkyl ammonium halides, alkyl pyridinium halides, alkyl sulfonates, fatty amine oxides, 2-alkylimidazoline quaternary ammonium salts; Thickening agents such as paraffin, silicon dioxide, cetosteryl alcohol, waxes; Diluents such as mannitol, sorbitol, starch, kaolin, sucrose; Preservatives such as parabens; Alkalizing agents such as ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium carbonate, sodium bicarbonate, sodium hydroxide; Acidifying agents such as hydrochloric acid, nitric acid, acetic acid, amino acids, citric acid, fumaric acid; Antioxidants such as ascorbic acid, sodium ascorbate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, sodium formaldehyde sulfoxylate; Emulsifying or suspending agents such as PVP, gelatin, natural sugars, acacia, guar gum, agar, bentonite, carboxymethylcellulose sodium, polyethylene glycol; stabilizator; One or more additional components such as colorants such as iron oxide, titanium dioxide or aluminum lakes, and the like.

The dose of plasma or serum according to the present invention should be appropriately determined in consideration of the patient's state of health and the degree of nerve damage. In general, it is desirable to apply to a lesion site at a dose of about 0.0001 to about 5 mg / cm ² once for adults.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are intended to illustrate the present invention more specifically, but the scope of the present invention is not limited to these examples.

Example  1. Preparation of Plasma Containing Paste

Plasma-containing pastes were prepared by thawing human-derived blood products (central blood source, fresh frozen plasma) at 30 ° C. after mixing with HIV, HCV, and HBV, and mixing them with physiological saline at a rate of 10 g / 5 cc.

Example  2. Spine damage Rat  Preparation of the model

Sprague-Dawley, 200-220 g of body weight, about 2 months old, obtained from the Yonsei University Medical Animal Laboratory, was used for the experiment. The rats were given 12 hours of day and night shifts and water and feed were randomly fed, while the Institutional Animal Care and Use Committee Guidelines of Association for Assessment and Accreditation of Laboratory Animal Care International). 14 days prior to surgery, basic walking training was conducted for the BBB test and the grid walk test. Three days prior to surgery, a baseline assessment of behavioral and motor skills was performed to select 25 normal rats suitable for use in the experiment.

The 25 rats selected were selected from Schucht et al. (Schucht, P. et. al ., Experimental Neurology , 176: 143, 2002), rats were completely anesthetized with a mixture of 25 mg / ml ketamine and 1.3 mg / ml rompun at 2 mg / kg and L2 abdominal hysterectomy. (L2 Ventral Laminectomy) was performed. In order to prevent infection during and after surgery, antibiotic cephalexin was injected intramuscularly in an amount of 5 mg / 100 g _{body weight} daily. The second lumbar spine of the rat was opened and a small hole (1 mm ² ) was made in the left lateral part of the arch verticebral plate using Microrongeur. The blade holder blade was inserted into this hole, and then cut through the Dura Mater to the right outer portion of the orthopedic plaque to injure the abdominal region of the spinal cord. Dorsal Musculature of the spinal nerve injury site was sutured and the skin ligated with a surgical clip. After surgery, the rats were placed on a warm sawdust to maintain body temperature and the lower abdomen was massaged three to four times daily for about seven days until the autonomic bladder control was fully restored.

Outdoor walking ability was evaluated on the 3rd day after surgery of nerve injury by BBB test (Basso, DM, Beattie, MS and Bresnahan, JC, J. Neurotrauma , 12: 1, 1995). Rats were placed in a coarse transparent plexiglass box and observed for 4-5 minutes. According to the criteria such as joint movement, weight support, forelimb-hind limb coordination, tail position, and so on, 0 points were defined as inability to exercise the hind legs, and a maximum of 21 points was defined as having normal walking ability.

In addition, three days after the surgery of the spinal nerve injury, a grid walk test (Z'graggen et al ., J. Neurosci . 18; 4744, 1998) was performed to evaluate the motor function of rats. In the grid walk test, the rats walked the metal parallel rods of 1m length to evaluate the regular stepping ability, and the rats were lowered to allow the rats to descend to evaluate the control ability of the hind legs. The evaluation was made by counting the number of mistakes in the step. Ten mistakes indicate that the rat is not only able to walk regularly but also is unable to adjust the legs voluntarily. Zero to one mistake is intact and indicates normal rat motility.

20 of 25 animals had similar scores in the BBB test and grid walk test, and their average score was about 11 and about 6 points, respectively. Scores were scored by two observers by double blind. The remaining five animals exhibited significantly different behaviors, significantly deviating from the average score and excluding them from the experiment.

Example  3. Spine damage Rat  Treatment of Plasma-Containing Compositions in Models

Ten of 20 animals (treatment group) selected as the test subjects in Example 2 applied the paste containing the plasma prepared in Example 1 to the spinal nerve injury site of the second lumbar spine and then applied the dorsal muscle tissue as described above. The sutures and the skin were ligated with a surgical clip. The remaining 10 animals (control) were treated with physiological saline instead of paste and sutured in the same manner.

Example  4. BBB  Test and grid work  Test ( Grid Walk Test )

In Example 3, 10 rats treated with 100 μl (2 mg / μl) of plasma-containing paste at the spinal cord injury site and 10 control rats treated with physiological saline at the spinal cord injury site were subjected to spinal nerve trauma surgery 3 The BBB test and grid walk test were performed by the method described in Example 2 at weekly intervals from day one. Figure 1 shows the BBB test results, Figure 2 shows the results of the grid walk test. From these results, the recovery peak of the treatment group and the control group was reached 35 days after surgery. In addition, the plasma-containing paste treated group in the BBB test (p <0.05) and grid walk test (p <0.05) was recovered with a significant difference compared to the control group by Two-sample Mamm-Whitney U statistical analysis.

Example  5. Footprint Analysis Footprint Analysis )

35 days after surgery for spinal nerve injury, the feet of treated, control and normal rats were stained with ink and walked on a white paper, stride length (base between the feet by two consecutive steps), base of support, The footstep analysis was prepared based on criteria such as the distance between the left and right soles, the angle of rotation, and the angle of rotation set by the angles of the soles and toes (Kunkel-Bagden E. et. al ., Exp Neurol . 119, 153, 1993). Figure 3 shows the analysis of the footsteps of the normal group, the control group, and the treatment group, respectively, it was confirmed that the treatment group, showing similar footsteps as the normal group.

Example  6. Electrophysiology Electrophysiology Analysis

After 35 days of spinal nerve injury, control and treatment rats were anesthetized with urethane at 1.5 g / kg _{body weight} , and then the sensory motor cortex and hind sciatic nerve were exposed. 0.5 mm concentric sedimentation pole was punctured about 1-2 mm to the left from the bregma and about 1 mm to the right (rostrocaudal) to the depth of 1 mm of the right sensorimotor cortex, applying 3 mA of electrical stimulation to the hind limbs. The location of the optimal signal transmission to the sciatic nerve was found, and the response intensity and sensory neuropathy were analyzed off-line at this optimal stimulus location. Figure 4 shows the results of the sensory nerves transients by two independent sample t-test. The results showed that the mean pause for the sciatic nerve of the treated group was significantly less than the mean pause for the control group (p <0.05). This indicates that the axons are connected to perform normal functions.

Example  7. Histology ( Histology Analysis

At day 35 after surgery of the spinal nerve injury, the rats were injected humanly with an excess of anesthetics in the control and treated rats, and the second lumbar spine and the associated thoracic and third and fourth lumbar vertebrae at the spinal nerve injury site were removed. After 48 hours soaking in buffered formalin, it was fixed in paraffin. Tissue sections were prepared by cutting 2-5 μm thick from the sagittal plane to the longitudinal plane and placed successively on microscope slides. The slides were H & E stained with hematoxylin and eosin. 5A to 5C are continuous magnifications (5, 10, and 20 times) of control tissues, and D to F of FIG. 5 are serial magnifications (5, 10, and 20 times) of treatment group tissues. It is shown. As shown in D to F of Figure 5, the lesion site of the treatment group showed a complete structural continuity as a whole, it can be seen that the drug of the present invention has a substantial therapeutic effect on regeneration of nerve cells and spinal nerve injury. On the other hand, as shown in A to C of Figure 5, the control group was found that the lesion site is corroded.

The present invention has the effect of providing a pharmaceutical composition for treating nerve damage containing a therapeutically effective amount of plasma or serum. The plasma or serum-containing pharmaceutical composition according to the present invention has an effect of healing damaged nerve cells to perform a normal function.

Having described the specific parts of the present invention in detail, it will be apparent to those skilled in the art that these specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. will be.

Claims (7)

Pharmaceutical composition for the treatment of neurological damage to peripheral nerve damage or spinal cord injury containing a pharmaceutically effective amount of plasma or serum as an active ingredient. The composition of claim 1, wherein the composition is in any one form selected from the group consisting of pastes, solutions, suspensions and topical gel forms. The composition of claim 1, wherein the plasma or serum is contained in an amount of 0.1 to 99.9% by weight of the composition. delete The composition of claim 1, wherein the nerve injury is damage to nerves. The method of claim 5, wherein the nerve is damaged is a composition characterized in that the neurites are cut at least 10mm. The method of claim 1, wherein the peripheral nerve damage is diabetic neuropathy, acromegaly, hypothyroidism, AIDS, leprosy, Lyme disease, systemic lupus erythematosus, rheumatoid arthritis, Scheren's syndrome, periarticular arteritis, Wegener's granulomatosis, cranial A composition characterized in that it is a neuropathy associated with a disease selected from the group consisting of arteritis and sarcoidosis.

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