KR101643737B1 - Composition for Improving, Preventing or Treating Neurological Diseases comprising Brassica oleracea var. botrytis aut italiana L. Extracts - Google Patents

Abstract

The present invention relates to a composition for improving, preventing or treating a neurological disease, which comprises an extract of Nogrel ( Brassica oleracea var. Botrytis aut italiana L. ) as an active ingredient. According to the present invention, the composition is excellent in nerve cell protection and is effective for neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and ischemic brain diseases such as cerebral infarction. In addition, the composition has an effect of improving cognitive function and improving mental disorder.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for improving, preventing or treating a neurological disease including neurotrophic factor, botrytis aut italiana L. Extracts}

The present invention relates to a composition for improving, preventing or treating a neurological disease comprising a neungrin extract.

Brassica oleracea Brassica oleracea is a balsamic plant native to the coasts of Southern Europe and Western Europe. Wild Brassica oleracea is a perennial plant that has been cultivated for thousands of years. Brassica oleracea Acephala Group, Brassica oleracea Alboglabra Group, Cauliflower, Broccoli Flower, Brassica oleracea Botrytis Group, Cabbage (Brassica oleracea Capitata Group), Brassica oleracea sprout, Brassica oleracea Gemmifera Group), cola (Brassica oleracea Gongylodes Group) and broccoli (Brassica oleracea Italica Group).

Domestic patents relating to brassica plants include, but are not limited to, Published Patent No. 10-2011-0122735 " Santomonas Campestris PV. &Quot; Brassica oleracea plant resistant to nasal disease ", Registered patent No. 10-0531679, " Chemotherapeutic effect of green leafy cabbage ", Patent No. 10-2005-0026012, &Quot; Herbicide-Resistant Brassica Plants and Methods of Use ", and US Pat. No. 6,040,784 entitled " Method for selective " , " Brassica plants with high levels of antivarcinogenic glucosinolates "and WO2012033422" Medical cosmetic herbal composition on the basis of herbal mixture and antioxidant activity of Brassica sp. ", US5811536 "Cauliflower floral meristem identity and methods of using same ", US20070033675A1 the procedure for its obtention ".

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have sought to develop a composition capable of improving, preventing, or treating a neurological disorder or mental disorder, which comprises an effective ingredient derived from a natural product, which has less side effects on the human body than a chemical synthetic medicine and can be easily prepared and consumed. As a result, it was confirmed that the extract of Brassica oleracea var. Botrytis aut italiana L. is excellent in nerve cell protecting action, and is effective for neurodegenerative diseases or cognitive function enhancement.

Accordingly, an object of the present invention is to provide a composition for improving, preventing or treating a brain disease.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a composition for improving, preventing or treating a neurological disease comprising an extract of Nogrel ( Brassica oleracea var. Botrytis aut italiana L. ) as an active ingredient.

The present inventors have sought to develop a composition capable of improving, preventing, or treating a neurological disorder or mental disorder, which comprises an effective ingredient derived from a natural product, which has less side effects on the human body than a chemical synthetic medicine and can be easily prepared and consumed. As a result, it was confirmed that the extract of Nogrel ( Brassica oleracea var. Botrytis aut italiana L. ) excelled in nerve cell protection, and was effective in improving neurodegenerative diseases or cognitive function.

When the extract of the present invention is used in the present invention, various extracting solvents may be used. Preferably, a polar solvent or a non-polar solvent can be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o -xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF.

More preferably, the extraction solvent used in the present invention is (a) water, (b) anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.) (E) ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butylene glycol, (i) hexane and (j) diethyl ether. . Most preferably, the extract of the present invention is obtained by treating water, ethanol or a combination thereof with nigrin.

According to one embodiment of the present invention, the nogrin extract of the present invention is obtained by treating a polar solvent with nogrin. According to another embodiment of the present invention, the nogran extract of the present invention is obtained by treating an alcohol (preferably, methanol, ethanol , Butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol).

According to a specific embodiment of the present invention, the above-mentioned nogrin extract is obtained by treating alcohol with nogrin and sonication.

As used herein, the term " extract " means that it is used in the art as a crude extract as described above, but broadly includes fractions obtained by further fractionating the extract. That is, the nogrin extract includes not only those obtained by using the above-described extraction solvent but also those obtained by further applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) The fractions obtained by the purification method are also included in the nogran extract of the present invention.

The neungrin extract used in the present invention can be prepared into a powder state by an additional process such as vacuum distillation and freeze drying or spray drying.

As used herein, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve efficacy or activity of the following neurogenic extracts. Since the present invention is a composition extracted from a natural plant material, nogrin, there is no adverse effect on the human body even when administered in an excessive amount, so that the quantitative upper limit of the amount of the nogrin extract contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.

The composition of the present invention is used for ameliorating, preventing or treating cranial nerve diseases.

The term " cranial nerve disease " as used herein includes neurodegenerative diseases, diseases caused by ischemia or reperfusion, and mental disorders.

According to one embodiment of the present invention, the neurodegenerative diseases are Parkinson's disease, Huntington's disease, Alzheimer's disease, mild cognitive impairment, senile dementia, amyotrophic lateral sclerosis, Spinocerebellar Atrophy, Tourette's Syndrome, Friedrich's Ataxia, Machado-Joseph`s disease, Lewy Body Dementia, Dystonia, Progressive Nucleus Progressive Supranuclear Palsy, and Frontotemporal Dementia.

According to one embodiment of the present invention, the disease caused by ischemia or reperfusion is a disease selected from the group consisting of ischemic stroke, cerebral hemorrhage, cerebral infarction, head injury and cerebral circulatory metabolic disorder.

According to one embodiment of the invention, the mental disorder is selected from the group consisting of anxiety, depression, mood disorder, insomnia, delusional disorder, obsessive compulsive disorder, migraine, stress, memory disorder, cognitive disorder, senile dementia, Alzheimer & Parkinson ' s disease related disorders, attention disorders, insomnia disorders and ischemic or brain trauma related disorders.

The neungrin extract of the present invention protects nerve cells.

According to one embodiment of the present invention, the nogrin extract inhibits the death of nerve cells and protects nerve cells.

According to another embodiment of the present invention, the present invention inhibits neuronal cell death by beta amyloid, which is one of the major causes of mild cognitive impairment such as Alzheimer's disease and senile dementia.

The protective action of the nerve cells of the present invention is usually a protective action of the brain nerve cells. Cerebral nerve cell protection refers to the action of alleviating or ameliorating damage to nerve cells or nerve tissue caused by metabolic causes, causes of toxicity, causes of neurotoxicity, and physical or chemical causes, or to protect or restore neurons affected by such damage do.

As used herein, the term " cranial nerve cell " refers to neurons, neural support cells, glia, and Schumann cells constituting the brain, brainstem and spinal cord. The protection of cranial nerve cells by the composition of the present invention can be accomplished through various mechanisms, for example, by inhibiting apoptosis of neuronal cells.

The neungrin extract of the present invention improves cognitive function and memory.

In this specification, the term 'cognition' refers to all the processes of accepting and storing certain information in the brain and finding and using stored information, and involves all the processes of thinking, speaking, remembering, judging and executing. Cognitive function refers to all the processes by which the brain receives information, stores it, and finds and uses the stored information. These cognitive functions can be divided into attention, language, time and space, memory, and execution (or management) functions.

The cognitive functions of the present invention include learning, memory and concentration.

According to one embodiment of the invention, the neurogenic extract alleviates cognitive function 1.5-4.0 times, 1.8-3.8 times, or 2.0-3.5 times in the memory decay model.

According to another embodiment of the present invention, the said extract of neugrin alleviates cognitive function 1.0-2.0 times, 1.2-1.8 times, or 1.3-1.7 times in the normal experimental group.

The neungrin extract of the present invention alleviates anxiety or stress.

According to one embodiment of the present invention, the nogrin extract alleviates anxiety or stress 0.2-2.5 times, 0.3-2.3 times, or 0.4-2.1 times.

The composition of the present invention can be provided as a food composition. When a composition for preventing, ameliorating or treating a cerebral disorder, comprising the active ingredient of the present invention as an active ingredient, is prepared from a food composition, it contains not only the nogrin extract as an active ingredient, but also ingredients normally added in food production, For example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, have.

The composition for preventing, ameliorating or treating a cerebral disorder, which comprises the extract of Newark of the present invention as an active ingredient, can be prepared from a functional food composition. When the composition of the present invention is prepared with a functional food composition, it includes components that are conventionally added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when it is made of a drink, a flavoring agent or a natural carbohydrate may be added as an additional ingredient in addition to the nogrin extract as an active ingredient. For example, natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.); Disaccharides (e.g., maltose, sucrose, etc.); oligosaccharide; Polysaccharides (e.g., dextrin, cyclodextrin and the like); And sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). Natural flavoring agents (e.g., tau marin, stevia extract, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) may be used as flavorings.

The compositions of the present invention may be prepared with pharmaceutical compositions.

According to a preferred embodiment of the present invention, the composition of the present invention comprises (a) a pharmaceutically effective amount of the above-described nigrin extract of the present invention; And (b) a pharmaceutically acceptable carrier. As used herein, the term " pharmaceutically effective amount " means an amount sufficient to achieve efficacy or activity of the above-described nogrin extract.

When the composition of the present invention is manufactured from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably parenterally administered.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . Typical dosages of the pharmaceutical compositions of this invention are in the range of 0.001-100 mg / kg on an adult basis.

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a composition for improving, preventing or treating a neurological disease comprising an extract of Nogrel ( Brassica oleracea var. botrytis aut italiana L. ) as an active ingredient.

(b) Since the composition of the present invention has an excellent neuronal protective action, it can be effectively applied to neurodegenerative diseases.

(c) The composition of the present invention has excellent efficacy in enhancing cognitive function.

FIG. 1 shows the results of confirming the neuronal cell death inhibition effect of the neuroblastoma extract by treating beta amyloid protein (A?) With neuroblastoma.
FIG. 2 shows the result of confirming the neuronal cell death inhibition effect on the nissl and hippocampal region by administering the nogrin extract to the animal model of Alzheimer's.
FIG. 3 shows the result of unilateral rotatory experiment by administering a Newrin extract to a Parkinson's animal model.
FIG. 4 shows the results of a memory retention experiment using a Morris water maze in a mouse induced decline in memory by scopolamine.
FIG. 5 shows the results of passive avoidance ability test in mice induced to decrease memory by scopolamine.
FIG. 6 shows the result of the spatial memory ability test by the Morris water maze experiment in a normal mouse.
Figure 7 shows the results of the manual avoidance ability test in normal mice.
FIG. 8 shows changes in staying time as a result of confirming the anti-anxiety improving effect through the cross-lab experiment.
FIG. 9 shows the change in the number of accesses to the open space as a result of confirming the anti-anxiety improvement effect through the cross-lab experiment.

The present invention will be described in more detail. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example 1: Preparation of nogrin extract

Newgrin ( Brassica oleracea var. Botrytis aut italiana L. ) was pulverized into powders and dried in an electric dryer. Newgrin is pulverized selectively, and it is extracted 5 times with ultrasonic wave for 10 hours with 8 times volume (w / v) aqueous 50% ethyl alcohol solution of pulverized material. The extract is immediately filtered and concentrated in vacuo, and then mixed with dextrin by a blending ratio and lyophilized to obtain an extract. At this time, the temperature during the concentration was kept at 45 캜 or lower so as to prevent decomposition and hydrolysis of the constituents. All the substituents such as the sugar (polysaccharide in the monosaccharide) and the methyl group (-CH ₃ ) bonded to the aglycon can be extracted by the above-mentioned extraction method.

Example 2: Alzheimer's disease prevention effect

Cell culture

SK-N-SH cells, a neuroblastoma cell, were cultured in DMEM supplemented with 10% FBS (fetal bovine serum). (DMEM with 1% FBS) for 2 hours before the experiment.

Beta amyloid protein (A? _1-42 )

The beta amyloid protein (Aβ _1-42 ) was purchased from Biosource (California, USA) and dissolved in phosphate buffered saline (PBS, pH 7.4). Aβ _1-42 was incubated at 37 ° C. for 2 hours before use and subjected to an aging step. Aβ _1-42 was used as 10, and this concentration showed cell viability of about 55% at 36 hours.

Cell viability assay (Alamarblue assay)

SK-N-SH cells were seeded in 96-well (Nunc, Slangerup, Denmark) at a density of 15,000 cells / well and cultured for 24 hours. The cell culture medium was changed to DMEM containing 1% FBS and further cultured for 2 hours. To the cultured cells, 10% concentration of neuron extract was added and the same amount of phosphate buffered saline (pH 7.4) was added as a negative control for 2 hours. After that, A? _1-42 was added to the culture solution of each group to 20, and the culture was further cultured for 2 hours. As a negative control for the Aβ _{42 1-} treatment it was cultured by the addition of phosphate buffered saline (pH 7.4) in the same amount.

After incubation for 3 hours, 10 μl of alamarblue (Serotec, Oxford, UK) was treated and absorbance was measured at 570 nm using an ELISA reader (Molecular Divices, Sunnycale, CA, USA). The background absorbance (blank count) was measured at 600 nm.

Beta amyloid protein (A [beta]) was added to neuroblastoma to confirm the neuronal protection effect of the mixed composition of the extract of Brassica oleracea var. Botrytis aut italiana L. Apoptotic cell death was induced by adding beta amyloid protein (Aβ) to neuroblastoma. However, treatment with the extract of Nogrel ( Brassica oleracea var. Botrytis aut italiana L. ) showed a similar degree of cell survival to that of the untreated beta amyloid protein (Aβ). As a result of measuring the cell viability in order to quantitatively confirm the above results, as shown in FIG. 1, there was no difference in the survival rate of the cells when the beta amyloid protein was not treated, but the survival rate of the cells was decreased when the beta amyloid protein was treated Respectively. However, it was confirmed that the survival rate of neurons to apoptosis by beta amyloid protein was recovered to about 83% when the extract was treated with neurogenic extract.

Alzheimer's disease induction model production

Male ICR mice weighing 25 g were anesthetized with ketamine (40 ㎎ / ㎏) and xylazine (20 ㎎ / ㎏), followed by stereotaxic frame , David Kopf, USA), and Aβ _1-42 capable of inducing Alzheimer's disease was injected into the lateral ventricle to produce an Alzheimer's disease-induced model. The nogrin extract prepared in Example 1 was diluted in physiological saline and administered by intraperitoneal injection once daily for 3 days at a dose of 50 mg / kg from 3 days before administration of A? _1-42 .

Through brain tissue staining Newgrin  Identifying the effect of extracts on Alzheimer's disease

Example An animal model produced in the 'Alzheimer's disease induction model production' was anesthetized using ketamine (40 mg / kg) and xylazine (20 mg / kg), followed by perfusion through physiological saline solution After 4% paraformaldehyde dissolved in phosphate buffer (pH 7.4) was perfused and fixed, the brain was extracted.

The cells were fixed in the same fixative at 4 ° C for 24 hours, transferred to 15% sucrose dissolved in 1 M phosphate buffer (pH 7.4), and dehydrated for 24 hours. Samples were further dehydrated in 20%, 25% and 30% sucrose for 24 hours, respectively. The dehydrated samples were cut into 35 ㎛ using a freezing sectioning machine (Leica, Germany). The sectioned tissue was subjected to nissl staining using a cresyl violet staining the nisse body, and ApopTag (CHEMICON, CA, USA), a kit for cell death confirmation, was used to observe the nerve cell death in the hippocampal region TUNEL staining was performed.

As a result, as shown in Fig. 2, it was confirmed that the damage of nisse body caused by A? _1-42 was protected by administration of the nogrin extract. In order to observe the protective effect of the hippocampal neuronal cell death, the result of FIG. 2, which was stained with the kits for cell death, also showed that neuronal apoptosis due to Aβ _1-42 was effectively protected by the administration of the nogrin extract.

Example  3: Parkinson's disease prevention effect

Construction of an Advanced Model of Parkinson's Disease

(Joo WS et al., 2004), which induces gradual degeneration of dopaminergic neurons by injecting 6-hydroxydopamine (6-OHDA), widely used as a Parkinson's disease-inducing drug, into the striatum of the rat brain. , Neuroreport, 9 (18), 4123-4126, 1998).

Male Sprague - Dawley rats weighing 200-250 g were anesthetized by intraperitoneal injection of ketamine (100 mg / kg) and xylazine (50 mg / kg). Anesthetized animals were punctured using a stereotaxic frame (David Kopf, USA), and 0.2 ㎎ / ㎖ ascorbic acid was used in the sham group and dopamine hydroxylated in the lesioned group 20 μg / 5 μl free base in 0.2 mg / ml ascorbic acid) was injected into the right striatum using a Hamilton syringe (10 μl, 26G needle) at a rate of 1 μl per minute (Paxinos et al., J. Neurosci. Methods. 3 (2), 129-249, 1980). After the drug injection, the needle was left for 5 minutes, the needle was pulled out at a rate of 1 mm per minute, and the incision was closed. Three days before the administration of 6-OHDA, the extract of neuron prepared in Example 1 was diluted in physiological saline and administered by intraperitoneal injection once a day for three days at a dose of 50 mg / kg.

Apomorphine  by Unilateral  Rotational reaction experiment

Behavioral changes in Parkinson 's disease model using hydroxyapatite were investigated by subcutaneous injection of apomorphine (0.5 ㎎ / ㎏) on day 14 after lesion preparation and unilateral rotation reaction for 60 minutes Respectively.

In this experiment, dopaminergic neurons were killed and the concentration of dopamine in the striatum was reduced, resulting in the hypersensitivity of the dopamine receptor in the striatum. As a result, apomorphine acted as an agonist on dopamine receptors, which caused excessive stimulation of the hypersensitive striatum. (Ungerstedt, Brain Res. 24, 485-493, 2970). The rotation was performed using an automated rotometer as described in Ungerstedt, supra, and the number of revolutions was [net turns = non-lesion side (contralateral) - lesion side (ipsilateral) ].

As a result, as shown in FIG. 3, unilateral net ratios of the normal control group treated with 0.2 mg / ml ascorbic acid were not significantly changed, but in the lesion control group in which only the dopamine hydroxide was administered, . On the other hand, the number of unilateral net revolutions was decreased in the group treated with the Newrin extract compared to the lesion control group. Thus, it was confirmed that the nogrin extract of the present invention has an inhibitory effect on Parkinson's disease.

Example  4: Prevention of memory decline

Animal model of memory decline and Newgrin  Administration of the extract

When the scopolamine, the antagonist of the muscarinic receptor, is administered to rats, acetylcholine, a neurotransmitter liberated in presynapse, binds to muscarinic receptors, receptors in postsynaps, And thus it is commonly used to test learning and memory-enhancing effects by creating an animal model of memory loss by administering scopolamine.

In the present invention, to examine the memory enhancing effect of the nogrin extract mixture, scopolamine 1 mg / kg was administered to mice to prepare a memory decay animal model. After 2 hours from the administration of scopolamine, the extract of neuron prepared in Example 1 was diluted in physiological saline and administered by intraperitoneal injection at a dose of 50 mg / kg for 4 days.

Morris's Underwater maze  Used memory retention test

In order to evaluate the effect of the nogrin extract obtained in Example 1 on the memory of the experimental rats induced to decline in memory by scopolamine, an experiment of an underwater maze against the mice of the above-mentioned 'memory decay animal model' Respectively.

The Morris Underwater Maze Experiment is a commonly used experiment to view spatial capacity memory in general. Experimental methods were as follows: 28 ± 1 ℃ water was filled in a round black water tank (60 ㎝ diameter and 40 ㎝ depth) to a depth of 30 ㎝, and the platform was placed on one side of the quadrant of the water tank. We measured the time taken to put the animal in each of the four directions of the tank and to swim to find the platform. After reaching the platform, they rested for 15 seconds and then rescued from the tank. If the platform was not found after more than 300 seconds, the experimenter handed over the animal to the platform. After four days of learning, one day of resting and six days of returning to the platform were measured (Manschot et al., Brain Res . 966 (2) 274-282. 2003).

As a result, as shown in FIG. 4, it was confirmed that the decrease of memory by scopolamine was effectively inhibited by the nogrin extract. These results confirmed that the extracts of Newgreen were highly effective in the protection of memory decline.

Passive avoidance ability test

To evaluate the effect of the nogrin extract obtained in Example 1 on the memory of the experimental rat induced to decrease the memory by scopolamine, a manual avoidance ability test was conducted against the mouse of the 'memory decay animal model' Respectively.

The passive avoidance test apparatus is divided into two sections, which are movable through the middle door. The floor is made of wire mesh. After the adaptation period of 90 seconds in the dark place, the light and noise elements I made it possible to move to another dark area through the open door to avoid light and noise. During the training, light and noise were stimulated to move to the dark area, and a current of 1.5 mA was applied to the footshock for 2 seconds. Twenty-four hours after the training, the mice were placed in the first zone and the time taken to move to the dark zone after stimulation with light and noise was measured after 90 seconds of adaptation period. The maximum time of measurement was 300 seconds (Sonkusare et al. Life Sci . 77 (1). 1-14. 2005).

As a result, as shown in FIG. 5, mice administered with scopolamine alone forget the memory of the training immediately and visited the dark room, whereas mice administered with the extract of Newrin remained a longer time, or when the maximum measurement time elapsed I did not go to the dark room until. As a result, it was confirmed that the degradation of memory by scopolamine was effectively inhibited by the nogrin extract. These results confirm that ingestion of Newgrin extract is very effective for the protection of memory decline.

Example  5: In normal animals Newgrin  Enhancement of memory and cognitive function of extract

The animal behavioral experiments conducted in Example 4 were performed in the normal mice in the same manner. However, the maximum measurement time of the example 'manual avoidance ability test' was 800 seconds.

As a result of administering the above-mentioned extract of Example 1 to normal mice, the spatial memory ability (FIG. 6) and the memory power (FIG. 7) obtained by the Morris water maze experiment in normal mice (Figs. 6 and 7). This study confirms that the extracts of Newgrin are effective for improving memory and cognitive function in normal rats.

Example  6: Elevated  Through the cross-lab experiment Newgrin  Extract Anxiety  To improve mental health

EPM  For animal behavior experiments Newgrin  Mental health improvement effect of extract

(Elevated Plus-Maze (EPM)) was used as a way to identify behavior associated with anxiety, stress and mental health (GR Dawson, Trends Pharmacol. Sci. 16, pp33-36, 1995) .

In order to examine the anti-anxiety and mental health promoting effect of the extract of the present invention, the extract of the present invention prepared in Example 1 was diluted in physiological saline so that administration of the drug was completed 2 hours before the animal behavior test, Lt; / RTI > Negative controls were orally administered 0.9% NaCl. Each experimental group consisted of 10 animals.

After the administration of the drug, the anti-anxiety effect was measured by using the manufactured EPM. The EPM was installed in a dark room (20 Lux) with indirect lighting, and each arm length of the EPM was crossed at a right angle of 30 cm, The wall was covered with synthetic resin and the wall height was 20 cm and the other side was opened. Each plate was 7 cm wide, 7 cm wide and 7 cm wide, and each arm was adjusted at 50 ㎝ away from the ground. The head of the mouse was allowed to freely navigate through the maze, from the central platform toward the closed arm, high and then the maze. The behavior was monitored for 5 minutes and the time spent in the open and closed arms of the mouse, the number of times each arm went out, and the total distance traveled were measured with the EthoVision program (Noldus Ingomation Technology, Wageningen, Netherlands) Were measured.

All behavioral test results were statistically analyzed using one - way ANOVA and significance was tested at P <0.05 level using SPSS when significance was recognized.

Measurement of anxiety reduction efficacy by measurement of changes in staying time

The time spent in the open space of the experimental animals was 38.85 ± 4.32 seconds for the negative control group, while 79.19 ± 9.39 for the control group, 8.33 seconds (P <0.001), indicating that the time spent staying in the open space significantly increased. This result is confirmed to be that the anxiety is reduced by about 2 times as compared with the negative control group when ingested with the extract of New Guinea (Fig. 8)

On the other hand, the duration of stay in the closed space was 182.16 ± 6.21 seconds in the negative control group, and 108.33 ± 4.19 seconds in the Newgrain extract group, indicating that the time spent staying in the closed space was decreased (P <0.05). This result shows that the time spent in the closed space is reduced by about 40.5% when the nogrin extract is consumed as compared with the control group (FIG. 8)

From the above results, it was confirmed that the time spent in the open space and the time spent in the closed space were significantly decreased in the group to which the nogrin extract was administered, and it was confirmed that there was an anti-anxiety, antidepressant and antidepressant effect .

Measurement of change in access frequency to open space

The number of times the animals went into the open space was an indicator of antianxiety and stress reduction efficacy. In the control group, 18.12 ± 1.26 times, while in the experimental group, 25.72 ± 1.93 times, (P <0.001), respectively. Which was significantly increased to about 41.9% when compared with the negative control group (FIG. 9).

On the other hand, the number of accesses to the closed space was 31.05 ± 1.08 times in the negative control group, and 20.02 ± 0.81 times in the number of the accession of the new extract, indicating a significant decrease (P <0.001). Which was about 25.5% lower than that of the negative control (Fig. 9).

From the above results, the number of access to the open space in the experimental group administered with the nogrin extract was remarkably increased, while the number of access to the closed space was dramatically decreased, thereby confirming the effect of improving mental health such as stress and anxiety I could.

Measurement of motility activity in open space

In this experiment, the locomotor activity of the study mouse was measured through an open fileld test to determine whether the behavioral change of the nogrin extract was due to mobility change rather than mental health improvement.

Rocomoto activity was measured in the prepared open space test box, which was 41.5 cm wide, 41.5 cm high, and 43 cm high, and was measured in one box for 5 minutes each.

As a result of the above experiment, the total movement distance of the experimental group consuming the negative control group and the nongreen extract was 2172.84 ± 104.2 ㎝ and 2212.31 ± 98.2 ㎝, respectively, which showed no significant difference. Therefore, the results of the above-mentioned 'measurement of anxiety-reducing efficacy according to the measurement of the staying time change' and 'measurement of the change in the number of access to the open space' can be confirmed not by merely improving the athletic performance but by improving the mental health to be.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (9)

A pharmaceutical composition for improving, preventing or treating a neurological disease comprising an extract of Brassica oleracea var. Botrytis aut italiana L. as an active ingredient, wherein the neurological disease is a neurodegenerative disease or a psychiatric disorder, Parkinson's disease, Alzheimer's disease, mild cognitive impairment, senile dementia, Lewy Body dementia and frontotemporal dementia, wherein the mental disorder is selected from the group consisting of anxiety, depression, insomnia, , Memory disorders, and cognitive disorders. The pharmaceutical composition for improving, preventing or treating a neurological disorder.
delete delete delete delete The composition according to claim 1, wherein the nogrin extract inhibits nerve cell death and protects nerve cells.
2. The composition of claim 1, wherein the nogrin extract improves cognitive function and memory.
2. The composition of claim 1, wherein the nogrin extract alleviates anxiety or stress.
A food composition or a functional food composition for improving or preventing a neurological disease comprising an extract of Brassica oleracea var. Botrytis aut italiana L. as an active ingredient, wherein the neurological disease is a neurodegenerative disease or mental disorder, Is selected from the group consisting of Parkinson's disease, Alzheimer's disease, mild cognitive impairment, senile dementia, Lewy Body dementia, and frontotemporal dementia, wherein the mental disorder is selected from the group consisting of anxiety, depression, insomnia, , Stress, memory impairment, and cognitive disorders. &Lt; RTI ID = 0.0 &gt; 18. &lt; / RTI &gt;

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