JP2016027031A - Medicine or food for neural network reconstruction/activation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide medicines and foods and drinks useful for prevention/treatment of a cognitive dysfunction disease caused by a failure of a neural network.SOLUTION: Drynaria Fortunei or extract thereof has the action of improving memory including object cognitive memory and object place memory, and is useful as a neural network reconstruction agent for cognitive dysfunction. The Drynaria Fortunei or extract thereof is also useful as a component to be blended with foods and drinks for improving the faculty of memory.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a drug and food for reconstructing a neural network containing a crude drug effective as an active ingredient for prevention / treatment of cognitive dysfunction caused by the failure of the neural network, and to activate the neural network Relating to pharmaceuticals and foods.

Although the etiology of neurological or neurodegenerative diseases varies, a direct factor that leads to impaired neurological function is the breakdown of the neural network. In addition to preventive / prophylactic treatment that removes the etiology, it is important to rebuild the neural network.
For drugs that reconstruct neural networks, for example, components of Indian ginseng (Ashwagandha) and its metabolites (Patent Document 1, Non-Patent Document 1), A mixture (Patent Document 2) is known.

On the other hand, the herbal medicine osseous (Kotsusaiho) is the rhizome of Drynaria fortune. The traditional effects of osteoclast are known as osteoclast, hemostasis, blood stasis, anti-inflammation, muscular pain, tinnitus, toothache, etc. (for example, Patent Document 3), but the relationship between osteoclast and neurological diseases Is not known about.
In Non-Patent Document 2, honey is mixed with a high-dose osteoclast extract (120 g of osteoclast supplement, 40 minutes for the first time and 25 minutes for the second time). However, as a result of prescribing Hachimi-jio-maru 9g for a long time, there is a description that improvement in cognitive ability has been confirmed, but only one subject is administered, and the specific aspect of long-term follow-up is also unclear In addition, it cannot be said that the improvement effect of cognitive function by osteoclast replacement was confirmed. Thus, the relationship between osteoclast and cognitive function has not been known so far.

JP2006-176428 WO2006 / 068155 JP2005-330290

YAKUGAKU ZASSHI, 128 (8), 1159-1167, 2008. Journal of Traditional Chinese Medicine, 25 (4), 290-291, 2005.

  Neurodegenerative diseases such as Alzheimer's disease, senile dementia, cerebrovascular dementia, and Parkinson's disease all have different etiologies, but all indicate symptoms that impair memory and cognition due to the breakdown of the neural network. For these diseases for which there is no effective therapeutic method, there is a real need for a treatment that can bring the nerve function closer to normal even when the nerve network has already been impaired. Moreover, considering the patient's standpoint, treatment with less burden is more desirable than nerve cell transplantation and gene therapy that require surgery. Therefore, it is necessary to develop a drug that regenerates the neural network even when the nerve cells are damaged or after the damage.

  Amyotrophic lateral sclerosis is an intractable disease in which the limbs of the cerebral cortex motor area, the spinal cord, and the brain stem lose their limbs, and there is no effective treatment. Furthermore, motor neurons in the cerebral cortex motor cortex are also damaged by cerebral hemorrhage, cerebral infarction, brain tumor, brain trauma, etc., leading to paralysis of the body. Traumatic spinal cord injury also results in limb paralysis due to damage to spinal motor neurons. In addition, multiple cerebral infarction is a demyelinating disease of the central nervous system caused by abnormal activation of autoimmunity, but axonal loss occurs as the disease progresses, which is an irreversible malfunction of the neural network. Leads to. In either case, it is a dysfunction due to the breakdown of the neural network, but if the surviving nerve cells are activated to form a neural network again, recovery of the function can be expected.

In view of the circumstances as described above, the present inventors sought herbal medicines for a component exhibiting excellent activity for improving the atrophy / degeneration of nerve axons in the brain that has been degenerated and dysfunctional, and its action We have been conducting research based on the idea that by exploring the mechanism, it is possible to find new compositions that cannot be predicted from the pathological analysis of conventional neurodegenerative diseases.
As a result, we found that the axon repairing action was found in the extract of herbal medicine. Since its action is effective in improving memory impairment, it has gained new knowledge that it is effective as a neural network restructuring agent that reshapes and constructs a neural network, and as an activator of a neural network, Further studies were made and the present invention was completed.

That is, the present invention relates to a neural network restructuring / activating agent (reconstructing agent and / or activating agent, hereinafter simply referred to as a restructuring agent) comprising a bone fracture complement (Kussaiho) or an extract of bone fracture complement as an active ingredient. , Activator, agent, etc.).
In such an agent, the osteoclastic extract may be extracted with, for example, water and / or alcohol or a mixture thereof.

In the reconstruction / activation agent of the present invention, the reconstruction / activation of the neural network is due to neural circuit formation by axon (nerve axon) repair and / or axon (nerve axon) extension (extension). May be.
In a specific aspect, the reconstruction / activator of the present invention may be one in which the cognitive memory function (cognitive function) is improved and / or enhanced (or improved) by forming a neural circuit. In such an aspect, the recognition memory function may be, for example, any one of object recognition memory, object location memory, and episode memory, or a combination thereof.

In the present invention, an agent for the prevention and / or treatment of a neurological disease (a prophylactic agent and / or a therapeutic agent for a neurological disease, a prophylactic and / or a neurological disease, comprising osteoclastic or an extract of osteoprosthesis Therapeutic drugs). The present invention also includes agents for repair and / or extension of nerve axons, including osteoclastic prostheses or extracts of osteoclastic prostheses. In these agents, the form of the extract of osteoclastic prosthesis may be extracted with water and / or alcohol or a mixture thereof as described above.
In the agent for the prevention and / or treatment of the neurological diseases, the neurological diseases include, for example, Alzheimer's disease, dementia (for example, senile dementia, cerebrovascular dementia, Pick's disease, frontotemporal dementia, Lewy body dementia, vascular dementia, etc.), Parkinson's disease, Huntington's disease, brain contusion, spinal cord injury, amyotrophic lateral sclerosis and multiple cerebral infarction Good.
According to the present invention, since it involves axon repair and / or extension, it is effective for the prevention and / or treatment of a wide range of neurological diseases involving not only cognitive function but also axon degeneration and atrophy.

The present invention further includes a food or drink (bone) containing the agent (reconstruction / activation agent, agent for preventing and / or treating neurological diseases, agent for repairing and / or extending nerve axons). Food or drink containing an extract of crushed or crushed bone). Such food and drink may be, for example, supplements, health foods, functional display foods, nutritional functional foods, or foods for specified health use.
Although the aspect of the food / beverage products of this invention is not specifically limited, For example, a tablet, a powder, a granule, or a capsule may be sufficient. In such a food or drink, the content ratio of the osteoclastic supplement or the osteoclastic extract may be, for example, about 0.01 to 50% by weight. Such a food or drink may contain a food additive.
Examples of typical food and drink products include, for example, tablets or capsule supplements, containing 1 to 50% by weight of osteoclastic or extract of osteoclastic supplement, and containing two or more food additives Etc. are included.

According to the present invention, the neural network can be reconstructed and / or activated by using osteoclastic prosthesis or an extract of osteoclastic prosthesis. Such reconstruction and activation are usually accompanied by axonal repair and / or extension (extension). Therefore, the agent and food of the present invention are effective for the prevention, treatment or improvement of various neurological diseases caused by degeneration or atrophy of axons as well as improvement or improvement (enhancement) of the cognitive memory function.
The present inventors have specifically confirmed this. For example, osteoclast or an extract of osteoclast is extracted from mouse cerebral cortical nerve-derived cultured cell line by Aβ25-35 treatment. It has the action of repairing or extending axons of cells whose density has been significantly reduced. In addition, osteoclast or osteoclast extract has a memory improvement effect such as object recognition memory and object location memory for 5XFAD mouse, which is one of Alzheimer's disease model mice. Useful as an agent. Furthermore, since the extract of osteoclast is recognized to improve memory in the cognitive function of normal mice, it is useful as a compounding ingredient for foods and drinks for the purpose of improving memory.

The length of pNF-H positive axons between Aβ25-35 untreated cells and Aβ25-35 cells and MAP2 positive neurons in each image are measured, and the comparison of the length of axons per nerve cell is shown. FIG. It is a figure which shows the result of the object recognition memory test of a normal mouse. It is a figure which shows the result of the object location memory test of a normal mouse. It is a figure which shows the result of the object recognition memory test of the wild type mouse group which administered only the 5XFAD mouse group and the solvent. It is a figure which shows the result of the object location memory test of the wild type mouse group which administered only the 5XFAD mouse group and the solvent. It is a figure which shows the result of the episode memory test of the wild type mouse group which administered only the 5XFAD mouse group and the solvent. It is a figure which shows the result of the spontaneous momentum test of a normal mouse. It is a figure which shows the result of the locomotor activity test of the wild type mouse group which administered only the 5XFAD mouse group and the solvent.

The osteoclastic prosthesis used in the present invention is the rhizome of the Drynaria fortune.
Osteoclast may be used as it is, may be used as an extract (extract), or may be used in combination.
Extraction of bone fracture is performed after extracting the bone fracture or its crushed material using water or alcohol (methanol, ethanol, etc.) or a mixture thereof at normal temperature or under heating by a conventional method or a method equivalent thereto. The product can be obtained by room temperature drying or freeze drying.
Although the dried extract may be used as it is, it is preferable to make it into a powder form.

When using osteoclastic or an extract of osteoclastic as a nerve network remodeling agent, etc., the osteoclastic or extract may be used as it is, and tablets, powders, granules, capsules, etc. And may be orally administered.
Furthermore, it may be formulated into a suppository, an injection, an instillation, an eye drop, an external preparation and administered parenterally, but it is desirable to administer as an oral preparation.
For oral preparations, tablets, powders, granules, and capsules can be produced by conventional methods by adding a binder, lubricant, disintegrant, coloring agent, corrigent and the like as necessary.
Moreover, antiseptic | preservative, antioxidant, a stabilizer, etc. can be added as needed.

  The dose of osteoclastic or the extract of osteoclastic extract varies depending on the disease, symptom, age, therapeutic treatment used in combination, and the like, but when administered to humans as an oral preparation, the dosage is 0. 1 to 5000 mg / kg, preferably 1 to 3000 mg / kg, more preferably 5 to 1000 mg / kg (eg 10 to 800 mg / kg), especially 30 to 1000 mg / kg (eg 100 to 800 mg / kg) In this case, 0.1 to 10 mg / kg may be administered once to several times a day.

In this invention, it can also be set as the food / beverage products containing the extract of a bone fracture complement or a bone fracture complement. Here, “food and drink” means food and drink in general, but in addition to general foods including so-called health foods, health foods such as foods for specified health use and nutritional function foods stipulated in the health functional food system of the Ministry of Health, Labor and Welfare. Functional foods are included, and supplements (nutritional supplements), feeds, food additives, and the like are also included in the food and drink of the present invention. In addition, the food and drink may be food or drink for a specific subject [for example, for elderly people, patients or patients (for example, for patients having a neurological disease (such as the above-described neurological diseases)]). Good.
In order to use osteoclastic or extract of osteoclastic for food and drink, it can be produced as it is or mixed with raw materials of food and drink such as processed meat and soft drink together with various nutritional components. .
In addition, when using the extract of osteoclastic as a health food, nutritional supplement, etc., for example, using conventional means, tablets, capsules (soft capsules, hard capsules, etc.), powders, granules, liquids (suspensions, syrups) Agents), emulsions, jellies, sticks and the like. The tablet includes a disintegrating tablet (orally disintegrating tablet).

The food and drink may contain a food additive (food additive) as long as it contains osteoclastic or an extract of osteoclastic. The food additive is not particularly limited. Calcium silicate, etc.), binder (eg, pregelatinized starch, hydroxypropylmethylcellulose, polyvinylpyrrolidone, etc.), disintegrant (eg, cellulose, hydroxypropylcellulose, corn starch, etc.), fluidizing agent (eg, light anhydrous silicic acid) Sucrose fatty acid esters, etc.), oils (eg, vegetable oils such as soybean oil, sesame oil, olive oil, linseed oil, sesame oil, rapeseed oil, coconut oil, corn oil, etc.), nutrients (eg, various Minera , Various vitamins, amino acids), flavorings, sweeteners, flavoring agents, coloring agents, solvents (ethanol), salts, surfactants, pH regulators, buffers, antioxidants, stabilizers, gelling agents, thickening Agents, lubricants, encapsulating agents, suspending agents, coating agents, preservatives and the like.
The food additives may be used alone or in combination of two or more, and in particular, two or more may be used. When two or more kinds of additives are used, two or more components (for example, two or more components selected from the components classified as excipients) may be selected from the same type of additives. May be combined (for example, a combination of an excipient and a binder), or both of them may be selected. In particular, the additive may be a combination of different additives.
In addition, among food and drinks in the form of tablets or the like, at least two or more additives selected from excipients, binders, and disintegrants may be used.

  In the case of using osteoclastic or an extract of osteoclastic as a food additive, the food and drink is not particularly limited. For example, food [eg, noodles (eg, soba noodles, Chinese noodles, instant noodles), confectionery, etc. (Salmon, candy, gum, chocolate, snacks (such as potato chips), biscuits, cookies, gummi, jelly, jam, butter, cream (such as cream puffs), cakes, etc.), breads, marine or livestock processed foods (kamaboko, ham , Sausages, etc.), dairy products (processed milk, fermented milk, etc.), fats and oils and processed foods (salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing, etc.), seasonings (sauce, sauce, etc.), retort Food (curry, stew, rice cake, rice cake, miscellaneous cooking, etc.), frozen dessert (ice cream, sherbet) Etc. shaved ice), fried (croquettes, fries, fried chicken, etc.), etc.], drinks (tea drinks, soft drinks, carbonated drinks, energy drinks, fruit drinks, such as lactic acid beverage), and the like.

The blending amount of the osteoclastic supplement or the osteoclastic extract in the above food and drink varies depending on the addition form and the dosage form, and can be selected from a wide range, for example, 0.001 to 80% by weight, preferably 0.01 It is desirable to blend in an amount of -50% by weight, more preferably 1-50% by weight (for example, 5-20% by weight), usually 0.01-50% by weight (for example, 0.01-10% by weight).
In addition, in food and drink, the amount of bone fracture or the dose (or amount of intake) of bone fracture can be selected from the same range as described above.

  Hereinafter, the present invention will be described in more detail with reference to comparative examples and examples, but these do not limit the present invention. A person skilled in the art can implement the present invention to the maximum extent by making various modifications to the claims according to the present specification as well as the following reference examples and examples. It is included in the scope of claims.

<Water extract of bone fracture supplement> 20 times the amount of water was added to bone fracture complement (Tochimoto Tenkaido, Osaka), and extraction was performed using a decoction device (Uchida Wakayaku, Tokyo). Heated over high heat until the water added to the herbal medicine boiled (15-20 minutes), and then continued to heat for 1 hour. The filtrate was filtered and then filtered with a cotton plug while hot. The filtrate was allowed to cool at room temperature and then frozen with liquid nitrogen. Furthermore, it was made to dry with a freeze dryer, and the extract of the powdered osteoclastic supplement was obtained (yield 7.8%). This was dissolved in physiological saline and used as a crushed rehydration extract for the following tests.

<Statistical processing> In this example, the obtained results were subjected to the following statistical processing:
One-way analysis of variance (one-way ANOVA), post-hoc Dunnett test, and paired t-test were performed using Graphpad Prism 5 (Graphpad Software, La Jolla, (California, USA). * P <0.05,
** P <0.01, *** P <0.001 is statistically significant, and mean values are shown with standard error.

<Test animals> (1) Normal mice: ddY mice were obtained from Japan SLC (Hamamatsu, Japan). In this example, male and 6-week-old ddY mice were used as normal mice. All mice received food and water ad libitum and were housed in a controlled environment with 22 ± 2 ° C., 50 ± 5% humidity and a 12 hour light-dark cycle starting at 7 am.

(2) Alzheimer's disease model mouse: Transgenic mouse (5XFAD) is considered an animal model of Alzheimer's disease and was obtained from Jackson Laboratory (Bar Harbor, Maine, USA). 5XFAD mice are human APP695 cDNA with mutations in Sweden (K670N and M671L), Florida (I716V) and London (V717I) under the transcriptional control of neuron-specific mouse Thy-1 promoter, And human PS1 cDNA (M146L and L286V mutations) are overexpressed (Oakley, H. et al., J Neurosci, 26, 10129-10140, 2006.). They were maintained by crossing B6 / SJL F1 breeders with hemizygous transgenic mice.

  In this example, 5XFAD mice were male and female, 6-8 months old, with free access to food and water, 22 ± 2 ° C, 50 ± 5% humidity, from 7 am Breeding in a controlled environment with a 12-hour light-dark cycle starting.

<Spontaneous momentum measurement> In this reference test 1, the spontaneous momentum measurement was performed as follows:
For each mouse to be tested, the movement path of the mouse when acclimated for 10 minutes in an open field box was followed using a digital camera system. The distance traveled in 10 minutes was analyzed as mobile activity by EthoVision 3.0 (Noldus, Wageningen, The Netherlands).

<Object recognition memory test>
The object recognition memory test is a test using a habit of showing interest in new things by animals. That is, in the test stage, it is a test for confirming whether or not the object seen in the training stage is remembered. The test was carried out according to the description of the literature (Int J Neurosci, 121, pp.181-190, 2011., and Int J Neurosci, 121, pp.641-648, 2011.) the day after the measurement of the spontaneous momentum. Specifically, it was performed as follows.
The test was conducted in a relatively well-lit room (approximately 100 lux). Appropriate time intervals between the training phase and the test phase were determined in advance by testing with different groups of mice. There are no landmarks on the inner wall of the open field box under test. In the training phase, place two identical objects in the field and let them search for 10 minutes. During the test phase, one of the objects is replaced with a new object, but the place is not changed, and a 10-minute exploratory action is performed. The increase in the number of times the mouse performs an exploratory action with interest in the replaced new object is used as an index of the object memory ability.
In this embodiment, the ratio (%) of the number of searches for a new object with respect to the total search time is calculated as a search index (Preference index).

<Object location memory test (spatial memory test)>
The object location memory test (spatial memory test) is a test using a behavior that an animal is interested in a new place. That is, in the test stage, it is a test for confirming whether or not the location (space) of the object seen in the training stage is remembered. The test was conducted with reference to the description in British Journal of Pharmacology (2009) 157, 1427-1440. Specifically, it was performed as follows.
The test was conducted in a relatively well-lit room (approximately 100 lux). Appropriate time intervals between the training phase and the test phase were determined in advance by testing with different groups of mice. A wallpaper with a characteristic pattern that serves as a mark on the two walls facing each other of the four walls inside the open field box to be tested is attached. In the training phase, two identical objects seen for the first time for the mouse are placed in the field and allowed to search for 10 minutes. In the test stage, change the location of one of the objects and let it search for 10 minutes.
The increase in the number of times the mouse performs an exploration action with interest in an object that has the same place but the same place is used as an index of spatial memory ability.
In this embodiment, the ratio (%) of the number of searches for an object whose location is changed with respect to the total search time is calculated as a search index (Preference index).

<Episode memory test>
Episodic memory test is a test that examines whether animals retain complex memory for objects and space, using the habit of showing interest in new things and new places. That is, in the test stage, it is a test to confirm whether or not the object and the place seen in the two training stages are remembered. The phenomenon that increases the number is used as an index of the episode memory ability. Specifically, it was performed as follows.
The test was conducted in a relatively well-lit room (approximately 100 lux). An appropriate time interval between the training phase and the test phase was pre-tested with another group of mice and set to 10 minutes. Paste the wallpaper with a characteristic pattern that serves as a mark on the two walls facing each other of the four walls inside the open field box to be tested. In Training 1, place the same four objects that the mouse sees for the first time in the field and let them search for 10 minutes. In training 2, another four objects are placed, two of which are in the same location as training 1 and the other two are in different locations. In the test stage, two of the four places set in training 2 are placed with training 2 objects as they are, and the remaining two places the objects used in training 1. Both allow 10 minutes of exploratory behavior.
In this embodiment, the ratio (%) of the number of times of searching for an object with different types and placement locations relative to the total search time is calculated as a search index (Preference index).

Example 1
The bone replenishing water extract was dissolved in sterilized purified water. Mouse cerebral cortex neurons were obtained from embryonic 14-day-old ddY mice (Japan SLC, Shizuoka), washed with PBS, decapitated, and prepared for primary culture [Neurobasal media (Invitrogen, Carlsbad, CA, USA) ) 12% horse serum (Invitrogen), 2 mM L-glutamic acid, 0.6% glucose dissolved]. In the medium, only the cerebral cortex was isolated under a stereomicroscope (SZ-61, Olympus, Tokyo).

  Cerebral cortex was cut into approximately 1 mm square in a safety cabinet. After centrifugation at 700 rpm for 3 minutes, the supernatant was removed, and 2 mL of 0.05% trypsin-0.53 mM EDTA solution (Life Technologies) was added to the precipitate and suspended. Incubate with stirring at 37 ° C for 15 minutes every 5 minutes, add 4 mL of medium, centrifuge at 700 rpm for 3 minutes to remove the supernatant, and precipitate 600 U / mL DNase I (Life Technologies) -0.03% trypsin 2 mL of inhibitor (Life Technologies) -PBS solution was added and suspended.

  Furthermore, incubation was performed at 37 ° C. for 15 minutes with stirring every 5 minutes, 4 mL of the medium was added, and the mixture was centrifuged at 700 rpm for 3 minutes. 4 mL of calcium / magnesium-free Hank's balanced salt solution (HBSS) was added to the sediment and centrifuged at 700 rpm for 3 minutes. 4 mL of medium was added to the sediment.

  The suspension was gently suspended with a Pasteur pipette with a blunt tip until the cell mass was not visible, and then filtered with a 70 μm nylon cell strainer (Falcon, New Jersey, USA). Cells were cultured on 8-well chamber slides (Corning). The day before coating [Poly-D-Lysine (PDL, Sigma-Aldrich) diluted with PBS to 0.005 mg / mL, and incubated at 37 ° C. Wash twice with sterile purified water on the day of culture. Cells were seeded. Cultivation was started at 37 ° C., 10% CO 2 and saturated steam. After 3 days, the whole medium was replaced with a new medium containing B-27 supplement (Invitrogen) instead of horse serum. At this time, the amyloid beta partial sequence (Aβ25-35) that had been agglutinated in advance was added to 10 μM, and 3 days later, when changing the whole medium to a new one, a bone replenishing water extract or solvent was added, Further, immunostaining was performed 4 days later.

  In the immunostaining, after completion of the drug treatment period, the medium was removed, the plate was washed with PBS, 4% paraformaldehyde (PFA, Wako Pure Chemicals) -PBS solution was added, and the mixture was allowed to stand at room temperature for 2 hours to fix the cells. Washing with a 0.3% triton X-100 (Wako Pure Chemicals) -PBS solution for 5 minutes was performed twice. Thereafter, a primary antibody solution [0.3% triton X-100-PBS solution, 1% normal goat serum (NGS, Wako Pure Chemical Industries, Ltd., primary antibody)] was added and reacted overnight at 4 ° C.

  As the primary antibody, mouse anti-phosphorylated NF-H monoclonal antibody (1: 500) (COVANCE, Emeryville, CA, USA) and rabbit anti-MAP2 polyclonal antibody (1: 2000) (abcam, Cambridge, UK) are used. It was. The next day, after removing the primary antibody solution and washing twice with 0.3% triton X-100-PBS solution for 5 minutes, secondary antibody solution [0.3% triton X-100-PBS solution, Alexa Fluor 594 labeled goat Anti-mouse IgG antibody (1: 200) (Molecular Probes, Eugene, OR, USA), Alexa Fluor 488-labeled goat anti-rabbit IgG antibody (1: 200) (Molecular Probes, Eugene, OR, USA)] For 2 hours at room temperature. After the reaction, the secondary antibody solution was removed, washed twice with PBS for 5 minutes, stained with DAPI staining, and then encapsulated with Aqua Poly Mount (Polysciences, Warrington, USA).

  Using a fluorescence microscope (BX-61, Olympus), 10 images of 648 μm × 860 μm were acquired from a well subjected to each drug treatment with a digital image capturing device DP70 (Olympus). Using the image analysis software MetaMorph (Molecular Device, Tokyo) for these images, the length of pNF-H positive axons on the entire screen and the MAP2 positive neurons in each image were measured. Axon length was calculated.

  The results are shown in FIG. Aβ25-35 cells had a significant decrease in axon density compared to Aβ25-35 untreated cells, but significant post-treatment axonal elongation was obtained by post-treatment with a bone rehydration extract (1, 10 μg / mL) Was recognized.

Example 2
The bone replenishment extract was dissolved in physiological saline and orally administered to normal mice once a day at a dose of 500 mg / kg / day. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 7 days. An object recognition memory test was performed on these mice. The interval between the training stage and the test stage was 48 hours. Compared with the control group to which only the solvent was administered, the object recognition memory was significantly enhanced in the bone replenishing water extract administration group (FIG. 2).

Example 3
The bone replenishment extract was dissolved in physiological saline and orally administered to normal mice once a day at a dose of 500 mg / kg / day. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 5 days. An object location memory (spatial memory) test was performed on this mouse. The interval between training and testing was 48 hours. Compared with the control group to which only the solvent was administered, the object location memory was significantly enhanced in the bone replenishing water extract administration group (FIG. 3).

Example 4
The bone replenishment extract was dissolved in physiological saline and orally administered to 5XFAD mice once a day at doses of 50 and 500 mg / kg / day. A 5XFAD mouse group to which only the solvent was administered and a wild type mouse group to which only the solvent was administered were also provided. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 21 days. An object recognition memory test was performed on these mice. The interval between the training phase and the test phase was 1 hour. Object recognition memory is maintained in wild-type mice, whereas memory impairment occurs in the solvent-administered group of 5XFAD mice. On the other hand, improvement of memory impairment was shown in a dose-dependent manner in the crushed rehydration extract administration group, and a significant effect was observed in the 50 and 500 mg / kg / day administration groups (FIG. 4).

Example 5
The bone replenishment extract was dissolved in physiological saline and orally administered to 5XFAD mice once a day at a dose of 500 mg / kg / day. A 5XFAD mouse group to which only the solvent was administered and a wild type mouse group to which only the solvent was administered were also provided. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 24 days. An object location memory (spatial memory) test was performed on this mouse. The interval between the training phase and the test phase was 1 hour. In wild-type mice, object location memory is maintained, whereas in the 5XFAD mouse solvent-administered group, memory impairment occurs. On the other hand, improvement in memory impairment was shown in a dose-dependent manner in the osteoclast rehydration extract administration group, and a significant effect was observed in the 500 mg / kg / day administration group (FIG. 5).

Example 6
The bone replenishment extract was dissolved in physiological saline and orally administered to 5XFAD mice once a day at doses of 50 and 500 mg / kg / day. A 5XFAD mouse group to which only the solvent was administered and a wild type mouse group to which only the solvent was administered were also provided. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 30 days. An episodic memory test was performed on this mouse. The interval between training 1 and training 2 and between training 2 and test were 10 minutes each. Episodic memory is maintained in wild-type mice, whereas memory impairment occurs in the solvent-administered group of 5XFAD mice. On the other hand, improvement of memory impairment was shown in a dose-dependent manner in the crushed rehydration extract administration group, and a significant effect was observed in the 50 and 500 mg / kg / day administration groups (FIG. 6).

Reference example 1
The bone replenishment extract was dissolved in physiological saline and orally administered to normal mice once a day at a dose of 500 mg / kg / day. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 6 days. When the spontaneous exercise amount of this mouse was measured, there was no significant difference between the control group to which only the solvent was administered and the crushed rehydration extract administration group (FIG. 7).

Reference example 2
The bone replenishment extract was dissolved in physiological saline and orally administered to 5XFAD mice once a day at doses of 50 and 500 mg / kg / day. A 5XFAD mouse group to which only the solvent was administered and a wild type mouse group to which only the solvent was administered were also provided. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 27 days. When the spontaneous exercise amount was measured for these mice, there was no significant difference between the groups (FIG. 8).

Example 7
Add 5g of lactose as an excipient, 1g of corn starch (corn starch) as an excipient, 1g of hydroxypropylcellulose (L-HPC) as a disintegrant, and 1g of polyvinylpyrrolidone as a binder, Granules were prepared by sizing.

Example 8
Add 5 g of lactose and 1.5 g of corn starch (corn starch) as excipients to 5 g of crushed rehydration extract and mix well. Add 0.05 g of light anhydrous silicic acid as a fluidizing agent and mix well to obtain a powder. It was.

Example 9
0.3 g of the powder obtained in Example 2 was packed in a commercially available gelatin capsule (obtained from Hill Half Research Institute, product number “No. 1 capsule”) to obtain a capsule.

Example 10
5 g of the granules obtained in Example 1 were sieved with a 32 mesh sieve, 0.05 g of lubricant magnesium stearate was added, mixed lightly for about 30 seconds, and then tableted to obtain tablets.

  The osteoclastic prosthesis or the extract of osteoclastic prosthesis has a memory improving effect such as object recognition memory and object location memory, and is useful as a neural network restructuring agent. Furthermore, osteoclastic prosthesis or osteoclastic extract is useful as a compounding ingredient for foods and drinks for the purpose of improving memory ability.

Claims (14)

  A neural network restructuring / activating agent comprising bone crush prosthesis or a bone crush extract as an active ingredient.   The neural network reconstructing / activating agent according to claim 1, wherein the extract of bone fracture is extracted with water and / or alcohol or a mixture thereof.   The neural network restructuring / activating agent according to claim 1 or 2, wherein the neural network restructuring / activation is based on neural circuit formation by axonal repair and / or axonal extension.   The neural network restructuring / activating agent according to claim 3, wherein the cognitive memory function is improved and / or enhanced by neural circuit formation.   The neural network reconstruction / activation agent according to claim 4, wherein the recognition memory is any one of object recognition memory, object location memory, and episode memory, or a combination thereof.   An agent for the prevention and / or treatment of neurological diseases, comprising osteoclastic or an extract of osteoclastic.   An agent for repair and / or extension of nerve axons, comprising osteoclastic or an extract of osteoclastic.   The neurological disease is at least one neurological disease selected from Alzheimer's disease, dementia, Parkinson's disease, Huntington's disease, cerebral contusion, spinal cord injury, amyotrophic lateral sclerosis, and multiple cerebral infarction. The agent described in 1.   Food-drinks containing the agent of Claims 1-8.   The food or drink according to claim 9, which is a supplement, health food, functional labeling food, nutritional functional food, or food for specified health use.   The food or drink according to claim 9 or 10, which is a tablet, powder, granule or capsule.   The food or drink according to any one of claims 9 to 11, comprising 0.01 to 50% by weight of osteoclastic or an extract of osteoclastic.   The food-drinks in any one of Claims 9-12 containing the additive for foodstuffs.   The food or drink according to any one of claims 9 to 13, which is a tablet or capsule supplement, comprising 1 to 50% by weight of a bone crush or a bone crush extract, and two or more food additives.