WO2019225942A1 - Composition for improving memory or composition for preventing and treating alzheimer's disease, comprising cooked processed silkworm product having silk protein - Google Patents

Abstract

The present invention relates to a composition for improving memory, or to a composition for preventing and treating Alzheimer's disease. In particular, the present invention provides a composition for improving memory or a composition for preventing and treating Alzheimer's disease, which comprises a cooked processed silkworm product having a silk protein, containing, as an active ingredient, a variety selected from among silkworm varieties that has the most desirable effects in terms of improving synaptic structure, protecting neuronal cells, and enhancing mitochondrial activity. Accordingly, as various health supplements or drugs using the present invention are anticipated to be developed and commercialized, the present invention can be expected to significantly contribute to the advancement of sericulture by increasing the utility of cooked processed silkworm products having silk glands and also turning same into high value-added products, and furthermore, the present invention can be expected to reduce rising social costs due to population ageing (health care costs, etc.), thereby greatly contributing to the improvement of the quality of life of people.

Description

Memory improving composition comprising cooked silkworm processed material having silk protein or composition for preventing and treating Alzheimer's dementia

The present invention relates to a composition for improving memory or a composition for preventing and treating Alzheimer's dementia comprising a cooked silkworm processed product having a silk protein, and more particularly, a difference in efficacy exhibited by various varieties among silkworm varieties. The composition for improving memory, including a silkworm, which has a silkworm protein, by selecting varieties that enhance memory, improve synaptic structure, protect nerve cells, and enhance mitochondrial activity, To provide a composition for preventing and treating Alzheimer's dementia.

In 2015, the number of school age population reached 887 million, and many people are studying. People of all ages, including the majority of students, including the majority of students, have better memory and learn more. I want to raise. In response to this, various types of memory-improving products have been released, but there are still many products that do not meet the expectations of consumers, and the results of colleges and job seekers, including elementary, middle and high school students, are poor. I am suffering from stress.

In addition, the number of Alzheimer's dementia patients due to an aging population is increasing rapidly, which is emerging as a social solution, but there are few food ingredients or medicines that can effectively prevent or treat it. Dementia is a neurological disorder in which more than 10% of the 65-year-old population suffers. About 70% of dementia has been reported as Alzheimer's dementia. To date, there is no way to prevent or treat Alzheimer's dementia. As of 2018, the elderly population has surpassed 7.3 million, 14% of the population, and is expected to increase to 41.0% by 2060. In addition, the number of patients with dementia reached 750 thousand as of 2018, and as the aging progresses, it is expected to increase to 1.16 million in 2040 and 212.7 million in 2050.

According to the analysis of the Ministry of Health and Welfare and the National Center for Dementia, the national management costs of dementia patients are expected to surge from 10 trillion 300 billion won in 2012 to 78 trillion 400 billion won in 2040 and 134 trillion won in 2050. . According to the result of dementia prevalence survey conducted in 2008 at Seoul National University Hospital, the prevalence of dementia decreased by 20% after 40 years of onset of dementia. It is expected to be alleviated. However, the current treatment of dementia is a treatment for symptomatic relief rather than a solution of the root cause, and it is necessary to develop a substance for suppressing, delaying or treating the onset.

Thus, the following studies have been conducted in connection with the treatment of dementia, and various studies using silkworm-derived materials have been known to show excellent efficacy in preventing dementia while being particularly harmless to the human body. As a technology, Korean Patent No. 10-0575229 (Food composition for preventing or treating brain diseases), Korean Patent No. 10-1430387 (Silk fibroin-derived peptide and memory, learning ability, improvement of cognition or dementia including the same Prophylactic or therapeutic compositions) and the like.

However, the prior art is to use only a silkworm cocoon consisting only of silk protein, not using all the useful active substances contained in silkworms, costly expensive when hydrolyzing the cocoon for ingestion, the loss of useful components In order to solve this problem, it is necessary to use silk protein together with all the ingredients in silkworms. However, in case of silkworms, the silk protein becomes stronger after 3 days of age 5, so it is impossible to consume silkworms. In order to solve this problem, the Rural Development Administration has developed a technology that allows silkworms with high silk protein content, silkworms from 5 to 4 days after sleep.

The present invention is to increase the mitochondrial activity of brain cells, increase the amount of ATP production, improve the synapse, protect neurons, inhibit beta-amyloid production, increase sociality, targeting the cooked silkworm processed material having various components of silkworm and silk protein as above. As a result of improving curiosity through treatment of increased curiosity, increased spatial memory, and increased short-term memory, or treatment of Alzheimer's dementia, the functional effect on memory improvement or Alzheimer's treatment effect using silkworms from the 5th and 4th days from this period Is not known.

An object of the present invention was carried out to improve the memory of cooked silkworms containing a large amount of functional material derived from silk thread and mulberry leaves, and to investigate the prevention and treatment effect of Alzheimer's dementia, especially among various silkworm varieties. Improving the synaptic structure, enhancing the activity of the mitochondria, and selecting a variety excellent in neuronal neuroprotective effect, the composition for improving memory, including a silkworm processed product having a silkworm protein containing the same as an active ingredient; Compositions for preventing and treating Alzheimer's dementia; Treatment methods using the same; And to provide a manufacturing method for manufacturing the same.

Memory improvement, Alzheimer's dementia prevention and improvement of the pharmaceutical composition or food composition of the present invention for achieving the above object, characterized in that the silkworm varieties of the silkworm processing, the silkworm varieties are white dog breeds, yellow dog breeds It is characterized in that it is any one of a soft green dog breed, a red dog breed, and more preferably the silkworm breed is a yellow dog breed.

The cooked silkworm processing is characterized in that the whole silkworm having a silk protein is cooked for 100 to 150 minutes with the heat of steam at 80 ~ 125 ℃ can be ingested for diet in the state containing silk protein.

Technical problems to be achieved by the present invention are not limited to the above-mentioned technical problems, and other technical problems not mentioned above may be clearly understood by those skilled in the art from the following description. There will be.

According to the present invention, it is expected to provide a composition for improving memory and preventing or treating Alzheimer's dementia by cooked silkworm processing containing a large amount of silk protein and mulberry leaf-derived functional substance, and among them, various varieties of silkworm with different color of silk protein Among them, the neurosynthetic structure is improved, mitochondrial activity is enhanced and the neuronal protective effect is selected for the color varieties with the best, and the composition for improving memory including cooked silkworm processing having a silk protein containing this as an active ingredient, or prevent Alzheimer's dementia And as the therapeutic composition is provided, it is expected that various health supplements or medicines using the present invention will be developed and marketed, thereby contributing to the development of the sheep sleep industry by creating utilization and high value-added cooked silkworm processing having a silk gland. It is expected to be effective, The aging is expected to greatly contribute to reducing soaring social costs (health care costs, etc.) and improving the quality of life of the people. In addition, it is expected to greatly contribute to reducing the burden on learning and grades of students and job candidates by improving memory.

1 is a graph showing the protein content of silkworms cooked by the production stage and silkworm varieties (cooked simmer; red yam) of silkworms.

Figure 2 is a graph showing the neurosynaptic improvement effect of the normal Drosophila when ingested silkworm processing by silkworm varieties.

Figure 3 is a graph showing the effect of improving the synaptic structure of Alzheimer's dementia Drosophila model when ingested cooked silkworms (Hwangsae cooked snooze).

Figure 4 is a graph showing the life expectancy increase effect of the Alzheimer's dementia fruit flies model when ingested cooked silkworms (hwangjong breed cooked sleep).

5 is a graph showing the effect of increasing the life span of the Alzheimer's dementia fruit fly model when ingested cooked silkworms (hwang-jong cooked snooze).

Figure 6 is a graph showing the memory enhancement effect upon ingestion of cooked silkworms (hwangjong breed cooked sleep) for the short-term memory loss treated mice.

Figure 7a is a graph comparing the memory enhancement effect of ingested silkworm processing of white dog breed and yellow dog breed for short-term memory loss treated mice. Figure 7b shows the effect of varying the concentration of cooked silkworms (yellow breed cooked silkworms) 0.1g, 0.2g and 0.5g in various ways.

Figure 8a shows the effect of improving the spatial memory according to the treatment of cooked silkworms (hwangsae ripened silk) in the dementia model. Figure 8b shows the effect of improving postural control ability according to the treatment of cooked silkworms (hwangjong breed cooked sleep) in the dementia model. Figure 8c shows the effect of improving the recognition ability for a new object by the treatment of cooked silkworms (hallowed breed silkworm) in the dementia model. Figure 8d shows the effect of improving sociality by the treatment of cooked silkworms (hallowed breed silkworm) in the dementia model. FIG. 8E shows the effect of reducing the production of toxic beta amyloid polymers in the brain following treatment of cooked silkworms (hwang jong-sook).

Figure 9a is a graph showing the effect of improving the activity of the mitochondrial ATP production complex of the control group and dementia model brain when ingested cooked silkworms (hwangsaeng cooked snooze). 9b shows the effect of enhancing the activity of mitochondrial complexes by the treatment of cooked silkworms (hwang-jong cooked silkworms). Figure 9c shows the effect of reducing the amount of reactive oxygen species (ROS) increased with aging by the treatment of cooked silkworms (hwangsae ripening sleep). Figure 9d shows the effect of increasing the concentration of ATP according to the activity of mitochondrial complexes enhanced by the treatment of cooked silkworms (yellow varieties cooked sleep).

10 is a graph showing the cell division promoting effect (neural cell protective effect) of the methanol extract of the silkworm processed varieties of silkworm varieties.

Hereinafter, the present invention will be described in detail, and detailed descriptions of well-known functions and configurations that may unnecessarily obscure the subject matter of the present invention will be omitted.

According to an aspect of the present invention, the present invention provides a pharmaceutical composition or food composition for improving memory, comprising a cooked silkworm processed product containing silk silk protein as an active ingredient; And compositions for preventing or treating Alzheimer's dementia; Or it provides a food composition for preventing or improving Alzheimer's dementia.

That is, the present invention relates to a composition for improving memory, comprising a silkworm protein and a mulberry leaf-derived functional material as an active ingredient, or a composition for preventing and treating Alzheimer's dementia, wherein the silkworm varieties of silkworm processing are white dogs. It is characterized by being any one of a breed, a yellow dog breed, a soft green dog breed, a light red dog breed, and more preferably the silkworm breed is a yellow dog breed.

To describe, the silkworms cooked containing the silk protein, means that all silkworms having a silk protein, such as three-sided nap, four-sided nap is prepared through the process, and is used in the present specification mixed with a snooze or red silk .

That is, silkworms having silk protein may be preferably selected from silkworms after 5 days and 3 days of silkworm silkworms or silkworms to sleep. At this time, the silkworm after 5 days and 3 days is preferably used from the silkworm of 5 days and 4 days, more preferably it is best to use silkworms of the sleeping period. This is because it can be used for silkworms of 5 days and 3 days, but the silkworm has less silk protein than silkworms after 5 days and 4 days, and the content of silkworm dung is higher. On the other hand, silkworm of snooze is a silkworm filled with silk protein in the silkworm body, which means silkworms until the cocoon is produced. Since silkworms of snooze are mostly discharged, pharmaceutical compositions or food compositions ( It is most suitable for use as an active ingredient of health functional food).

In the present invention, in order to be able to eat all the silk protein contained in the silkworm as described above, the silkworm is processed into a processed form to be used as an active ingredient of a pharmaceutical composition or food composition (health functional food), wherein The silkworm is cooked and used as heat of steam, and preferably cooked for 100 to 150 minutes with heat of steam at 80 to 125 ° C. so as to contain all silk protein. In other words, if the production time is over 150 minutes with steam heat of less than 80 ℃, the manufacturing time is too long, rather, the active ingredients that silkworm itself contains silk silk protein may be destroyed, more than 125 minutes less than 100 minutes If manufactured by the temperature is too high, because there is a possibility that adversely affect the active ingredients that silkworm itself contains silk silk protein.

After such a cooked process, it may be dried and used in a conventional manner to prepare a powder form, and preferably, it may be dried and powdered by any one selected from hot air drying or vacuum freeze drying, and loss of active ingredients. In order to minimize the vacuum freeze drying method is most suitable.

The powdering may be performed using any method such as a domestic mixer, a hammer mill, a roller mill, an air jet mill (freeze grinding), but a roller mill method may be used to minimize sample loss, prevent component distortion, and fine powder. good. In general, when grinding with a domestic mixer or hammer mill, it is very difficult to grind the powder particles to about 0.1 mm or less.However, if the powder is repeatedly milled several times with a roller mill, the powdered particles can be micropowdered to an average of about 10㎛. It is expected to work in favor of the product.

In addition, after the powder form, the silkworm processed powder may be prepared in the form of an extract, in which case it may be prepared by extracting with water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. The lower alcohol having 1 to 4 carbon atoms may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol. The water, lower alcohol having 1 to 4 carbon atoms or a mixed solution thereof may use 1 to 40 times the volume (1 to 40 L based on 1 kg) based on the weight of the cooked silkworm powder, and preferably 5 to 40 times the volume. Can be used. Extraction conditions of the extract may be 1 to 48 hours at 20 ~ 100 ℃, the process may be repeated 1 to 4 times.

In another method, the silkworm processed powder obtained by extracting and concentrating the extract obtained by extracting and concentrating with water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof is preferably suspended by adding water, preferably 1 to 1 to the weight of the extract. Suspended by adding 1000 times, more preferably 1 to 500 times, most preferably 1 to 50 times water, and then adding a solvent selected from the group consisting of hexane, chloroform, ethyl acetate, and butanol to the suspension. It can be prepared from the fraction obtained.

The preparation temperature of the extract or a fraction thereof may be 20 to 100 ° C., but is not limited thereto. Extraction or fractionation time is not particularly limited, but is preferably extracted within 10 minutes to 2 days, it is possible to use a conventional extraction device, ultrasonic mill extractor or fractionator as the extraction device. The extract or fraction thus prepared can be removed by hot air drying, reduced pressure drying or lyophilization. In addition, the extract or fractions may be purified using column chromatography. The chromatography may include silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, and medium pressure liquid chromatography. chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

The cooked silkworms have the following effects: Alzheimer's dementia, reduced life expectancy and health life recovery effect, short-term memory loss recovery effect, mitochondrial dysfunction recovery effect in the brain, neurosynaptic recovery effect, neuronal cell protective effect, short-term memory reduction Inhibitory effect, aggression reduction effect, social enhancement effect, curiosity enhancement effect, spatial memory increase effect, postural control ability, beta-amyloid reduction effect is excellent, in the present invention for improving memory or preventing and treating Alzheimer's dementia It is characterized by being used as an active ingredient in pharmaceutical compositions or food compositions (health functional foods), and in particular, silkworm varieties of the silkworm processed material may be used as any type of silkworm, but among them, white dog breed, yellow dog breed and lotus It is characterized by using either the green dog breed or the red dog breed. In addition, more preferably, the recovery effect as described above is most excellent in the yellow dog breed, and for improving memory, including a selected silkworm processed product using the selected variety as an active ingredient, or for preventing or treating Alzheimer's dementia, a pharmaceutical composition or food It is best to provide a composition.

To explain, first of all, as people get older, their memory gradually decreases, which is highly correlated with the structure of the synaptic structures in the brain. The number of neurosynaptic connections between nerve cells in the brain's memory areas decreases gradually as aging progresses. As a result, the number of neuronal synapses decreases. As time passes, the neurosynaptic decreases below a certain level, causing Alzheimer's dementia. Known. In other words, Alzheimer's dementia begins with a decrease in neurosynaptic prior to detecting the onset of the disease, and thus the memory gradually decreases. At some point, severe Alzheimer's dementia develops. To prevent memory loss and prevent Alzheimer's dementia, There is a need for a method that prevents or augments the reduction of junctions. As shown in the following experimental examples, in the present invention, as a result of confirming the number of the neurosynthesis by producing a feed with cooked silkworm powder prepared with a yellow dog breed, a white dog breed, a soft green dog breed, a red dog breed, etc. in a normal fruit fly, The silkworms of the silkworm varieties and the white silkworm varieties showed excellent neurosynaptic number enhancement effects. Among them, the silkworms of the silkworm varieties showed the most excellent synaptic number enhancement effect. Therefore, the two types of normal and Alzheimer's dementia fruit flies were divided into two groups, male and female, using the silkworms of the yellow dog breed to determine whether the neurosynaptic structure is abnormal and the improvement and prevention effect of the synaptic structure before dementia symptoms. As a result of the experiment, the normal model, Alzheimer's dementia model and the male and female model showed excellent neurosynaptic effect. Thus, the processed silkworms were found to be effective in improving memory and preventing Alzheimer's dementia. there was.

In addition, the HT22 cell line, a mouse hippocampal nerve cell line, was treated with alcohol extracts from silkworms of white dog and yellow dog varieties, which showed the best synaptic improvement effect. In the case of the silkworms of the breed (white jade), up to 159.5% increased, showing an excellent cell division promoting effect, and the silkworms of the varieties of the silkworm (Golden silk) showed an amazing cell division promoting effect of 253.0%. It was found that the cooked silkworm processed material had the most excellent effect.

Second, most patients with Alzheimer's dementia have a lower life expectancy. To determine the preventive effect, Alzheimer's dementia using the silkworm-processed silkworms showed the best synaptic improvement and cell division promoting effect on life expectancy. Experiments were conducted on fruit flies and found that life expectancy was increased.

Third, Alzheimer's patients with Alzheimer's dementia experienced a significant decrease in life expectancy and a significant decrease in their lifespan. To determine the preventive effect, Alzheimer's dementia patients tested the effects of increasing the life expectancy on normal fruit flies and Alzheimer's dementia fruit flies. In addition, normal fruit flies and Alzheimer's dementia fruit flies were found to have increased health life.

Fourth, in order to confirm the preventive effect of the short-term memory loss, which is a clinical feature in the early stages of Alzheimer's dementia, the experimental mice were treated with scopolamine, which is a short-term memory loss agent, while ingesting silkworm processed powders of wild dog breeds. It is confirmed that there is a very good effect to save memory. As a result of comparing the preventive effect on the loss of short-term memory of the silkworms from the yellow and white varieties, the silkworms from the silkworms and the silkworms from the yellow varieties were more effective. It was confirmed to appear.

Fifth, the pathological characteristics at the molecular level, which are common in Alzheimer's dementia due to memory deterioration and aging, are the main characteristics of the ability of ATP to be deteriorated due to the deterioration of the complexes that generate the transition between the inner and outer membranes of mitochondria. . This study confirmed the function of mitochondrial complexes I to IV using normal fruit flies and Alzheimer's dementia fruit flies while ingesting silkworms that had been found to have the best synaptic recovery and short-term memory loss. The mitochondrial complexes of normal fruit flies and dementia model fruit flies ingested were significantly increased. At the same time, we found that the amount of reactive oxygen species (ROS) associated with cell aging, disease development, and progression decreased significantly in normal fruit flies fed silkworm-treated silkworms.

Sixth, in order to confirm the reduction effect on the production of beta-amyloid polymer, which is a characteristic of Alzheimer's dementia, ingested silkworm processed powders of wild-breed varieties in dementia mice and confirming the level of beta-amyloid polymer in the brain, the beta produced by Alzheimer's dementia It has been found that there is a very good effect in reducing the production of amyloid polymers.

Seventh, as a result of testing the behavioral characteristics of test mice with Alzheimer's dementia gene, test mice without ingested silkworm processing showed dementia symptoms and changed their aggressiveness, making them very quarreled with each other and making new friends. Test mice that consumed cooked silkworms showed no dementia symptoms, so they made good interests with new friends, showed peaceful societies, such as not fighting and staying peacefully. In contrast, test mice that did not consume much less interest in new items, whereas test mice that received cooked silkworms had the same curiosity as new test mice that did not have dementia genes. In the space memory test to determine whether or not a new destination is well found in a particular space, the test mice that did not consume cooked silkworms were new compared to normal test mice that did not have Alzheimer's dementia gene. Although the frequency of finding the destination was greatly reduced, the test mice fed the cooked silkworms were significantly increased compared to the untreated dementia gene-bearing test mice, and the spatial memory was improved to be close to that of the normal test mice. The dementia test mice that did not consume the cooked silkworms by holding the tails of the test mice in order to lift their hands and feet with little movement due to the damage to the nerve cells, while the test mice ingested cooked silkworms on I could see the same widely observed moving vigorously hanba attitude control ability is also excellent pyeolchimyeonseo hands and feet with normal test mouse.

As described above, it was found that the processed silkworms were excellent in improving memory and preventing Alzheimer's dementia. Among them, the silkworms exhibited the most excellent efficacy in the cooked silkworms of the yellow dog breed. Or to provide a pharmaceutical composition or a food composition comprising a cooked silkworm processed material produced in a variety of varieties of silkworm cocoons of various colors as an active ingredient for preventing and treating Alzheimer's dementia, and among them, a yellow silk protein It was found that it is best to provide a pharmaceutical composition or a food composition comprising a silkworm processed material as an active ingredient.

In the pharmaceutical composition of the present invention, the cooked silkworm processed material may be added as 0.001 to 100% by weight to the pharmaceutical composition of the present invention. The pharmaceutical compositions may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to a conventional method. Carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, saccharin, naphtha extract (mongroside), stevia, starch, acacia Rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil Can be. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid form preparations may contain at least one excipient such as starch, calcium carbonate, sucrose or It is prepared by mixing lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of skill in the art and generally dosages range from 0.01 mg / kg / day to approximately 2000 mg / kg / day. More preferred dosage is 1 mg / kg / day to 500 mg / kg / day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

Subjects to which the pharmaceutical composition of the present invention is administered include all mammals such as rats, livestock, humans, etc., but are not limited thereto as long as the desired effect is exerted from the subjects to be administered.

In addition, the pharmaceutical compositions of the present invention can be administered by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections. Since the compound of the present invention has almost no toxicity and side effects, it is a drug that can be used safely even when taken for long periods of time.

At this time, the cooked silkworm is an effective content for showing the desired efficacy of the present invention, it is preferable to contain 0.01 to 100% by weight relative to the total weight of the composition in terms of powder weight, more preferably 0.1 to 90 weight It is good to contain in%.

In addition, the present invention may be provided as a food composition for improving the memory or preventing and improving Alzheimer's dementia by including the silkworm processed as an active ingredient. At this time, the cooked silkworm processed material may be used as 0.01 to 100% by weight in the food composition of the present invention. The food composition of the present invention includes tablets, capsules, pills, liquids, and the like, and includes various drinks, meat, sausages, breads, candy, snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, and alcoholic beverages. , Gum, tea and vitamin complexes.

In addition, according to another aspect of the invention, the effective amount of the above-described memory composition for improving the memory; And administering a pharmaceutically acceptable carrier to the subject.

Further, according to another aspect of the present invention, an effective amount of the pharmaceutical composition for preventing or treating Alzheimer's dementia described above; And administering a pharmaceutically acceptable carrier to the subject, the method for preventing or treating Alzheimer's dementia.

In addition, according to another aspect of the present invention, the present invention comprises the step of ripening the silkworm whole as a heat of steam, as described above, a memory composition for improving the memory comprising a cooked silkworm processed material containing silk silk protein as an active ingredient Or food compositions; And compositions for preventing or treating Alzheimer's dementia; Or it provides a method for producing a food composition for preventing or improving Alzheimer's dementia.

Since the methods of the present invention utilize the compositions described above, duplicated content is omitted in order to avoid excessive complexity of the present specification.

As such, the present invention is expected to greatly improve the memory function of the brains of all citizens, including the students of various schools suffering from academic stress, and the elderly, and it is expected to greatly improve memory, and also effectively prevent the development of neurological diseases including Alzheimer's dementia. It is expected to contribute greatly to improving the quality of life of the people living in an aging age, and contributing to the reduction of national dementia management costs.

Hereinafter, the present invention will be described in more detail with reference to examples, but these examples are for illustrative purposes only and are not intended to limit the protection scope of the present invention.

<Example 1. Preparation of cooked silkworm processed products according to silkworm varieties>

1) Preparation of cooked silkworm products (cooked simmer; red yam)

Breeding golden silk (cross breeding produced by breeding Japanese breeder Jam 311 and Chinese breeder Jam 312) (yellow breed) with yellow silk protein, it is 110 to 130 minutes with steam at 100 ℃ when it comes to sleep. After the silkworms were cooked to prepare a silkworm processed product, the silkworm-cooked silkworm processed product was vacuum-freeze-dried to prepare a powder form and applied as a sample of the following experiments.

2) Manufacture of silkworm processed products

It is produced in the same manner as the method for producing silkworm processed silkworm varieties of 1) above, but the hybrid breed produced by breeding the light red silk breed (Japanese breed breed Jam 325 and Chinese breed breed Jam 326 instead of the yellow breed). Silkworms were bred to prepare silkworm processed products.

3) Manufacture of silkworm processed products

Manufactured in the same manner as the method of manufacturing silkworm processed silkworm varieties of 1), but instead of the yellow breed, the white jade sleep (white breed cocoon 123 for white cocoon and Chinese breeder jam 124 for white cocoon) Breeding silkworms were produced by breeding breeding silkworms.

4) Manufacture of silkworm processed products

Manufactured by the same method as the method of manufacturing silkworm processed silkworm varieties of 1) above, the soft green silk (Yellow seedling 315 producing green cocoons and Chinese breeder Jam 316 producing white cocoons) Breeding silkworms produced by breeding silkworm dog breeds were prepared.

<Experimental Example 1. Comparison of Component Analysis of Silkworm Processed and Silkworm Processed by Different Types

1) Comparison of ingredients analysis of silkworms and cooked silkworms before cooking

Before the ripening and cooked silkworms were subjected to the main component analysis through a hammer mill mill having a particle size of 0.1mm, respectively, as shown in Table 1 and Table 2, FIG.

TABLE 1

TABLE 2

Through the above results, it was confirmed that the content of various components in the slumber before ripening became variously changed as it became a cooked silkworm, and the pharmacological effect of the cooked silkworm processed by the changes in the components of the cooked silkworm could be changed. It can be confirmed that.

2) Comparison of ingredient analysis by roller mill grinding method for silkworm varieties

The composition analysis of silkworm processed by silkworm varieties was compared through the grinding method of roller mill, which is a grinding method with less material loss and less change in composition.

As a result, it was shown in Figure 1 and Tables 3 to 9.

TABLE 3

TABLE 4

TABLE 5

TABLE 6

TABLE 7

TABLE 8

TABLE 9

As a result of the experiment, there was a difference in the components contained in the silkworm powder cooked by silkworm varieties, and based on this, it was confirmed that the varieties appear differently in the memory improvement and prevention of Alzheimer's dementia as follows.

In other words, as a result of analyzing the components of cooked silkworms, the cooked silkworms contain large amounts of crude proteins, amino acids such as glycine, alanine and serine, linolenic acid, unsaturated fatty acids and vitamins such as beta-carotene, and are harmful to humans. Heavy metals such as trans fat and mercury, and five sugars such as sucrose were not detected. In addition, compared to dried silkworms and uncooked silkworms at 5 days of age, the cooked silkworms were found to have a very high amino acid content of crude protein, glycine, alanine, and serine. By silkworm varieties, all the same components were detected in all varieties such as white jade, golden silk, lotus leaf and red dog varieties, and the content comparison of each component contained different varieties for each silkworm. It became.

Experimental Example 2. Confirmation of Structural Abnormalities of Neural Junctions According to Silkworm Products Treated by Silkworm Types of the Present Invention

1. Experiment Method

The effects of neurosynaptic structure on normal Drosophila were studied on silkworms of silkworms. Normal control fruit flies were cultivated using cooked snooze feed prepared by adding silkworm powder cooked by silkworm type, which added 10% of the total amount of yeast, the main protein source, among the components of the normal feed. In the larval stage, which is the stage of adulthood, the 3rd homeless larvae were dissected in a dissolution solution without Ca ²⁺ , and then fixed using 0.1 M phosphate buffer solution of pH 7.2 containing 4% paraformaldehyde. The immobilized samples were washed three times at 10-minute intervals using 0.01M phosphate buffered saline containing 0.1% Triton X-100, and then stained by neurosynthesis using anti-HRP-FITC. The number of neurosynthesis was confirmed by.

2. Experimental Results

As a result of the experiment, as shown in Figure 2, it was confirmed that there is an effect of improving the synaptic structure in the normal Drosophila test group ingested the feed including the yellow dog breed, white dog breed, and soft green dog breed ripe snooze. In terms of the overall effect on improving memory and the prevention of Alzheimer's dementia, the silkworms of the yellow and white varieties were found to be the best among the silkworm varieties showing various color cocoons. The processed product was judged to be superior to the cooked silkworm processed product produced from white dog breeds.

Specifically, the number of neurosynthesis in the test group ingested cooked silkworms of the yellow dog breed increased to 25% (137.2 + 5.76%) compared to the neurosynthesis in the test group that consumed only normal diet (109.9 + 2.97). It was found that P (0.005) increased, white breeds increased 13% (124.5 + 4.64), and soft green breeds increased 5%. .

Therefore, the experiment was carried out using the silkworms cooked silkworm varieties showed the best synaptic improvement effect shown in the experimental example.

<Experimental Example 3 1) Confirmation of structural abnormality of the neural junction according to the treated silkworm processing of Example 1 of the present invention>

1. Experiment Method

Two types of Drosophila expressing normal and Alzheimer's dementia genes, Amyloid precusor protein (APP) and Beta site amyloid precusor protein cleaving eznyme 1 (BACE1) and Microtubule associated protein tau (MAPT), using the cooked silkworms This study was conducted to investigate whether the neurosynaptic structure abnormalities appeared in the stage before dementia symptoms, and to improve and prevent the synaptic structure.

Two independent Alzheimer's dementia model fruit flies and normal control fruit flies were cultured using cooked silkworms prepared by adding 10% of the total amount of yeast, the main protein source, among the components of normal feed. . In the larval stage, which is the stage of adulthood, the 3rd homeless larvae were dissected in Ca ²⁺ -free anatomical solution and fixed using 0.1M phosphate buffer solution with pH 7.2 containing 4% paraformaldehyde. The immobilized samples were washed three times at 10-minute intervals using 0.01M phosphate buffered saline containing 0.1% Triton X-100, and then stained by neurosynthesis using anti-HRP-FITC. The number of neurosynthesis was confirmed by.

2. Experimental Results

As a result of the experiment, as shown in FIG. 3, the normal model, the Alzheimer's dementia model, and the male and female model confirmed the effect that the synaptic was improved. As a result, the cooked silkworms can effectively improve memory and prevent Alzheimer's dementia. Confirmed.

Specifically, in females, the neurosynaptic of the test group ingested cooked silkworms increased significantly to 42% compared to the test group in which dementia model 1 only consumed normal diet (81.7 + 0.895) (116.2 + 2.34). It was statistically significant (p <0.005), and in the case of dementia model 2, the test group ingested cooked silkworms increased 33% compared to the test group (79.7 + 1.613) who consumed only normal feed ( 113.4 + 1.71) statistically significant (p <0.005) increased, and even in the normal test group, the test group ingested cooked silkworms (114.0) compared to the test group (111 + 2.45) that consumed only normal feed (114.0). + 2.45) showed a synapse growth rate of 2.7%.

In the case of males, dementia model 1 significantly increased (103.2 + 2.342) the synapse of the test group ingested cooked silkworms (63.2 + 2.342) compared to the test group ingesting only normal diet (63.2 + 0.645). (P <0.005), and dementia model 2 also increased by 24% (94.4 + 1.80) in the test group that consumed cooked silkworms compared to the test group (76.0 + 2.39) that consumed only normal feed. It was statistically significant (p <0.005), and even in the normal test group, the test group ingesting cooked silkworms increased 24% (115.7 +) compared to the test group (93.7 + 2.40) that consumed only normal feed. 2.29) showed a statistically significant increase (p <0.005), indicating that cooked silkworms effectively prevent Alzheimer's dementia and significantly increase memory of normal individuals.

<Experimental Example 4 Confirmation of Life Expectancy Increase Effect of Silkworm Processing>

1. Experiment Method

Most patients with Alzheimer's dementia have a lower life expectancy. In order to confirm the preventive effect, Alzheimer's dementia fruit flies were tested using the silkworm-processed silkworm varieties that showed the best synaptic improvement effect on life expectancy. Was

In other words, the Drosophila model expressing both human amyloid precusor protein (APP) and beta site amyloid precusor protein cleaving eznyme 1 (BACE1) and microtubule associated protein tau (MAPT) in humans with Alzheimer's dementia can be normal or 10% cooked silkworm. Cultivation was carried out using a feed containing the processed product. After culturing more than 100 adult fruit flies into 10 vials, the number of living fruit flies was checked until all fruit flies disappeared at three-day intervals. Drosophila were bred at 28 ° C and 60% RH. Statistical analysis was performed by Kaplan-Meyer survival analysis to identify statistical differences.

2. Experimental Results

As shown in FIG. 4, in the case of Alzheimer's dementia fruit flies that consumed feed containing cooked silkworm processing powder, life expectancy increased by 18.3% from 23.97 days to 28.35 days, compared to Alzheimer's dementia fruit flies that consumed only normal feed. Statistical analysis using Meyer survival analysis showed that the risk index was 0.47 (95% confidence interval: 0.35 ~ 0.64), which showed statistically significant ( p value <0.0001) life expectancy increase.

Experimental Example 5. Confirmation of the Effect of Increased Health Life on the Processed Silkworm Processing>

1. Experiment Method

Alzheimer's dementia patients also experienced a decrease in life expectancy and a significant decrease in their life expectancy. To determine the preventive effect, Alzheimer's dementia patients were tested for the effect of increasing the lifespan on Alzheimer's dementia and normal fruit flies. Two normal control fruit flies and two Alzheimer's dementia model fruit flies were bred from feed containing silkworm-derived silkworm powder, which accounted for 10% of the amount of yeasts of protein in normal feed and normal feed. After dividing more than 100 adult fruit flies into 10 vials, behavioral control ability was evaluated at 3 day intervals. To evaluate the behavioral control ability, the vial containing fruit flies was shaken at 2000 rpm for 10 seconds in a vortex mixer, followed by a 20 second rest period. After that, the fruit flies that moved to the wall or entrance of the vial, rather than the bottom of the vial, were counted as being considered fruit fly-controlled fruit flies. Measurements were made every three days until all the fruit flies remained on the ground after vibration. The fruit life of fruit flies is defined as the period during which 50% of fruit flies have behavioral control ability.

2. Experimental Results

As shown in FIG. 5, the Alzheimer's model fruit flies that consumed only normal feed without ingesting cooked silkworms had a shorter health life compared to normal fruit flies (general fruit flies = 17.9 days, dementia model 1 = 15.3 days, Dementia model 2 = 14.7 days), and statistical analysis showed that, in the Alzheimer's dementia fruit fly group, the risk indices of 2.8050 (dementia model 1) and 2.1795 (dementia model 2) were both statistically significant ( p <0.01). Compared to the results of deterioration of health life due to Alzheimer's dementia (Fig. 5A), the normal test group who consumed the feed containing the cooked silkworm powder powder had a healthy lifespan of 17.9 to 20.5 days compared to the normal test group that consumed only normal feed. The risk index was increased by 14.5% and the risk index was 0.6674 (95% CI, 0.5153 ~ 0.8643), which was statistically significant ( p <0.01).

In addition, the Alzheimer's dementia model test group that consumed the feed containing the cooked silkworm powder powder increased the life expectancy of Alzheimer's dementia compared to the Alzheimer's dementia model test group that consumed only normal feed. The Alzheimer's dementia model test group 1, who received feed containing powder, had a 14.4% increase in health life from 15.3 days to 17.5 days, compared to the Alzheimer's dementia model test group 1 who consumed only normal feed. The index was 0.7513 (95% CI, 0.5801-0.973) was found to be statistically significant ( p <0.05) lowered (Fig. 5C).

In addition, the Alzheimer's dementia model test group 2 showed a 14.3% increase in health life from 14.7 days to 16.8 days and a risk index of 0.7532 (95% CI, 0.582 ~ 0.9746), which was statistically significant ( p <0.05). It is believed that this is because ingestion of cooked silkworms effectively prevents the development of Alzheimer's dementia (FIG. 5D).

Experimental Example 6. Confirmation of Preventive Effect on Short-term Memory Loss by Processing Silkworm Processing>

1. Experiment Method

In order to confirm the preventive effect of short-term memory loss that is characteristic of early Alzheimer's dementia, experiments were performed by treating scopolamine, which is a short-term memory loss agent, while ingesting silkworm processed powders in experimental mice. Silkworms cooked in male mice 4 weeks old were administered for 3 weeks at a ratio of 1.0 g / Kg body weight. After three weeks of administration, the mice were transported to the laboratory 30 minutes before the experiment on the first day of the experiment, and then acclimated for 2 minutes in a bright room of a passive avoidance apparatus. On the second day of the experiment, mice were injected with scopolamine dissolved in saline at a rate of 0.75 mg / kg body weight 30 minutes before starting the experiment. The control group was injected with the same amount of saline. After placing the mouse in the bright room of the passive evacuation device, 30 seconds later, the passage to the dark room was opened, and when the mouse enters the tail of the mouse, an electric shock of 0.4 mA was given for 4 seconds. After that, the period of entry into the dark room was measured. On the third day of the experiment, the mice were transferred to the bright room, the passage to the dark room was opened, and the period of entry to the tail was measured. After the experiment, the difference between the control group and the experimental group was performed by ANOVA and T-test for statistical analysis.

2. Experimental Results

As a result of the experiment, as shown in FIG. 6, in the case of a mouse fed only a general feed, short-term memory loss was observed when scopolamine was administered, but in a test group fed a feed containing cooked silkworm powder, It was confirmed that memory loss was greatly reduced.

Specifically, in the case of the normal mouse experiment group that consumed only general feed, the time required to enter the dark room of his favorite room was 52.3 + 5.0 seconds on the first day before the electric shock, but it was 222.7 on the second day after the electric shock. + Increasing the time to 41.8 seconds to suppress the instincts because of the previous scared memory, but the long time to restrained, but in the case of mice that lost short-term memory with scopolamine administration, the first day of the experiment before the electric shock Even though it took 50.9 + 4.1 seconds to enter the dark room, on the second day, despite the terrible electric shock, I could not remember it at all and confirmed entering the dark room in 40.0 + 8.18 seconds.

On the other hand, in the normal test group that consumed the cooked silkworm processed powder, it took 46.4 + 6.82 seconds to enter the dark room on the first day of the experiment before the electric shock, but after the electric shock, the duration was increased to 300.0 seconds, more than when not ingested. It appeared to remember that shock for a long time.

In addition, in the test group administered scopolamine, it took 49.5 + 10.9 seconds on the first day before the electric shock, and the time to enter the dark box was increased by 176% to 136.4 + 22.7 seconds on the 2nd day after the electric shock. Due to the intake of cooked silkworm processed powder, it was found that the treatment of short-term memory as well as normal mice was very effective in saving short-term memory.

Experimental Example 7. Comparison of the Inhibitory Effect of Short-term Memory Loss on the Treatment of Cooked Silkworms from Yellow and White Breeds

1. Experiment Method

In order to compare the preventive effect of the short-term memory loss characteristic of early Alzheimer's dementia, the silkworm powder and the silkworm processed powder of silkworm varieties were studied. Ingestion was performed by treating scopolamine, a short-term memory loss agent. In addition, at this time, the concentration of Hwangsam varieties of red silk was variously applied to 0.1g, 0.2g and 0.5g / Kg body weight, confirming the effect according to the concentration.

Four-week-old male mice were ingested with yellow or white breed silkworms at a rate of 0.5 g / kg body weight for three weeks. After 3 weeks of administration, the mice were transported to the laboratory 30 minutes before the experiment on the first day of the experiment, and then acclimated for 2 minutes in a bright room of a passive avoidance apparatus. On the second day of the experiment, mice were injected with scopolamine dissolved in saline at a rate of 0.75 mg / kg body weight 30 minutes before starting the experiment. The control group was injected with the same amount of saline. After the mouse was placed in the bright room of the passive evacuation apparatus, 30 seconds later, the passage into the dark room was opened, and when the mouse enters the tail of the mouse, the electric shock of 0.4 mA was given for 4 seconds, and the duration of entering the dark room was measured at the same time. On the third day of the experiment, the mice were moved to a bright room, the passage to the dark room was opened, and the period of entry to the tail was measured. After the experiment, statistical analysis was performed by performing ANOVA and T-test on the difference between the control group and the experimental group.

2. Experimental Results

As a result of the experiment, as shown in FIG. 7a, when the rats ingested the white silkworm cooked silkworms and the mice ingested the yellow silkworm cooked silkworms were treated with saline-treated white silkworm cooked silkworms, the experimental group entered the dark room on the first day. The time taken was 15.0 + 6.99 seconds. However, the time it took to move to a dark room after receiving an electric shock was 205.75 + 64.4 seconds, and he had a memory of electric shock. In comparison, the experimental group consumed saline-treated silkworm cooked silkworms took 34.0 + 21.17 seconds to enter the dark room on Day 1, but did not go to 300.0 seconds after the electric shock. This result shows that both white dog and yellow dog varieties are effective even in experiments that do not induce short-term memory loss.

In the experimental group that induced short-term memory loss by the administration of scopolamine, mice fed the white silkworm cooked silkworms were allowed to enter the dark room on the first day at 20.5 + 7.61 seconds, and then dark room on the second day. While the time taken to enter was 42.0 + 14.16 seconds, mice fed the silkworm cooked silkworms were increased to 81.75 + 37.98 seconds after being shocked at 38.0 + 14.79 seconds on the first day. These results also show that both the white and yellow dog breeds have excellent short-term memory loss suppression effects, and that the short-term memory loss effect of cooked silkworms made from the yellow dog breeds is higher when inducing short-term memory loss.

In addition, as shown in Figure 7b, it was confirmed that as the concentration of cooked silkworm processing increases, the effect of inhibiting short-term memory loss is significantly increased. Mice fed the cooked silkworms at 0.1 g / kg body weight had increased short-term memory to 33.3% (66.8 seconds) compared to mice fed normal diets. Mice fed the cooked silkworms at 0.2 g / kg body weight had increased short-term memory by 88.1 seconds to 75.8% compared to mice fed normal diets. Mice fed the cooked silkworms at 0.5 g / kg body weight had increased short-term memory by 136.8 seconds to 173% compared to mice fed normal diets.

Experimental Example 8. Comparison of effects on dementia mouse model by treatment of cooked silkworms

1) Comparison of Spatial Memory Enhancement Effect by Processed Silkworm Processing in Dementia Mouse Model

1. Experiment Method

In order to compare the effects on the loss of spatial memory characteristic of Alzheimer's dementia, using the silkworms silkworms which showed excellent effects in the experimental examples, the silkworms were cooked in 5xFAD dementia mice, a dementia-induced transgenic mouse model. Spatial memory was tested after ingestion of feed containing snooze or red yam) powder. Dementia mice received cooked slums at a ratio of 1 g / kg body weight. The Y-maze test is a method of evaluating spatial cognition and the Y-maze consists of three channels. The passage is 34 cm long, 15 cm high, 3 cm wide at the bottom, and 9 cm wide at the top. The three passages are each connected at an angle of 120 degrees, allowing the experimental mouse to move freely through the three passages. After placing the experiment mouse in the middle of the Y-maze, the movement path was recorded for 10 minutes. When the experimental mouse moved to another passage, it was considered to have moved only if it completely entered the tail. The number and order of entry into each passage shall be measured. If entering three passages sequentially, one point shall be regarded as one point. The ratio of spontaneous alteration in the new direction was converted to% by dividing the number of consecutive passages in all three passages by subtracting 2 from the total passage entry times.

2. Experimental Results

As a result of the experiment, as shown in Figure 8a, the 23-week-old dementia mice ingested cooked silkworm processed feed had a spatial memory at a level similar to the normal control mice ingested the general feed. The average mouse had 66.77 + 2.43% of spatial memory, but the normal feed demented mouse had 50.78 + 2.54% of statistically significant decrease ( p <0.05). Dementia mice fed cooked silkworm feed at 1g / Kg body weight ratio were 60.29 + 3.38, which was not statistically different from that of normal mice. ( p <0.05) was confirmed. This result shows that even when Alzheimer's dementia develops, the effect of improving spatial memory of cooked silkworms is excellent.

2) Comparison of Postural Control Enhancement Effect by Processing Silkworm Processing in Dementia Mouse Model

1. Experiment Method

5xFAD dementia mouse, a dementia-induced transgenic mouse model, to compare the effects on the loss of postural control, which is one of the abnormal behaviors seen in Alzheimer's dementia mice, by using a silkworm-derived silkworm processed product that showed excellent effects in the experimental examples. A rat clasping test was performed on rats 32 and 36 weeks old after the feed containing the silkworm processed powder at a ratio of 1 g / kg body weight.

In the posture control test, the video was recorded for 10 seconds after catching the tail of the subject, and the video was scored from 0 to 4 depending on how many legs each of the two researchers observed. If all four legs could not be extended, four points were given. If all four legs could be extended, zero points were given. The results of the two analyzes were averaged. The better the posture control, the lower the score near zero.

2. Experimental Results

As a result of the experiment, as shown in Figure 8b, the control of the 32-year-old control mice fed the cooked silkworm feed did not show a problem as the posture control ability is 0.1 + 0.1. In contrast, dementia mice fed the normal diet did not have one or two legs stretched to 1.36 + 0.41. However, dementia mice fed cooked silkworm feed had a postural control ability of 0.16 + 0.16, similar to that of control mice.

Similarly, the postural control ability after 4 weeks of the same mice showed that the postural control ability was 0.45 + 0.16 and 1.61 + 0.09, respectively. However, the demented mice fed cooked silkworms were not significantly different from the normal control group, with postural control ability of 0.18 + 0.13, similar to 32 weeks. The results show that the improved silkworm postural control ability improved even when dementia developed.

3 ) Comparison of Cognition Enhancement Effects of New Silkworms Treated with Processed Silkworms in a Dementia Mouse Model

1. Experiment Method

The study was conducted to investigate whether the cooked silkworms treated with yellow varieties, which had excellent effects in the experimental examples, could suppress the deterioration of cognition deterioration of new objects, which is one of the abnormal behaviors of Alzheimer's dementia mice. After that, it was confirmed by conducting a Novel objective recognition assay for new objects.

The first and second days allowed the experimental mouse to move freely for 10 minutes in an empty 50 cm wide x 50 cm wide x 50 cm high. On the third day, two identical objects were put in, and the movement of the mouse was recorded for 10 minutes, and the time of staying in each object was measured. On the last day, one old and one new item was put in, and after 10 minutes of recording, the time spent on the old and new items was measured.

2. Experimental Results

As a result of the experiment, as shown in Figure 8c, demented mice ingested cooked silkworms had a recognition ability for a new object to a level similar to the normal control mice ingested the general feed.

In the control rats of 22-week-old diets, the time to stay in the vicinity of two identical objects was 47.4 + 1.1% and 52.6 + 1.1%, respectively. However, if you change one to a new shape, it was 60.2 + 2.2% for the new one, which was statistically significant ( p <0.05) compared to 39.8 + 2.22% of the old one. However, there was no statistically significant difference in the degree of interest in new or old items in the diet fed demented mice, 48.1 + 3.0% and 51.7 + 1.9%, respectively. On the other hand, demented mice with cooked silkworm feed were significantly interested in new and old items with 67.7 + 3.4% and 32.3 + 3.4%, respectively ( p <0.05). . These levels were higher than normal mice.

This trend was similar in behavioral analysis conducted after breeding the same mice for 8 weeks. Normal feed intake was 63.8 + 2.2% and 36.2 + 2.2% for new and old items, respectively, but the interest in new items was statistically significant ( p <0.05). In the case of 37.0 + 4.6%, the percentage of the time of interest in new items was significantly lower than that of 57.8 + 5.2% of the time of interest in old items ( p <0.05). In demented mice with cooked silkworm feed, 71.2 + 2.7% of the time spent on new items and 28.8 + 2.7% of the old items showed statistically significant interest in new items than normal mice ( p. <0.05). These results show that ingestion of cooked silkworms can suppress the reduction of curiosity about new objects in demented mice.

4) Comparison of Sociability Enhancement Effect by Processed Silkworm Processing in Dementia Mouse Model

1. Experiment Method

In order to compare the effects on the loss of social characteristics of Alzheimer's dementia, using the silkworms cooked silkworm varieties showed excellent effects in the experimental examples, the silkworms cooked in dementia mice were ingested at a ratio of 1g / Kg body weight. The change of interaction assay) was tested.

Social analysis was performed using a mouse social analysis device divided into three spaces (width 60cm x height 40cm x height 40cm). The three spaces were divided into two transparent acrylic plates with holes for the mouse to pass through. On both sides, there is a cage with a wire mesh of 10 cm in diameter and 6 cm in height. Initially, the mice were placed in both cages with no mouse, so that they could adapt freely for 5 minutes. When the adaptation was finished, the mouse was placed in one cage, which had never met, and the other was in an empty cage, and the mouse was placed in the middle space, and the action was recorded for 10 minutes. Finally, the new mouse was placed in an empty cage and the action was recorded for 10 minutes. The recorded behaviors were analyzed to determine how long each experimental mouse stays in a space with an empty cage or with a mouse and in a space with a mouse and a new mouse.

2. Experimental Results

As a result of the experiment, as shown in Fig. 8d, demented mice ingested cooked silkworms had sociality at a level similar to that of normal control mice ingested with general feed.

The 23-week-old mouse was 65.1 + 2.12% of the time spent in the space with the new mouse, which was statistically significantly higher ( p <0.05) than the 34.9 + 2.12% of the time spent in the empty cage. The percentage of time to stay in the space with the new mouse was 59.7 + 1.73%, which was statistically significantly higher than the ratio of time to stay in the space with the mouse once met (40.3 + 1.73%) ( p <0.05). However, in the case of normal feed mice, the percentage of time spent in the space with an empty cage was 49.2 + 2.37%, which was no different from 50.7 + 2.37% of time spent in the space with a new mouse. The retention time ratio was 44.4 + 2.57%, which was statistically significantly lower than that of the 55.6 + 2.57% time spent in the space where the mice had already met ( p <0.05). However, for cooked silkworm feed-induced dementia mice, the time spent in an empty cage (36.5 + 3.06%) was statistically greater than the percentage of time spent in a new mouse (63.5 + 3.06%), as in normal mice. Was significantly lower ( p <0.05), and the percentage of time spent in the space where the mouse was already met (40.1 + 2.49%) was statistically significantly lower than the ratio of time spent in the space with the new mouse (55.6 + 2.57%). ( p <0.05).

This trend was similar for the experiment at 31 weeks of the same mice. In the case of normal mice, the percentage of time spent in empty space (43.9 + 2.09%) was statistically significantly lower than that of new mice (56.1 + 2.09%) ( p <0.05). The percentage of time spent in a new mouse (53.1 + 2.24%) was statistically significantly higher than that of a new mouse (46.9 + 2.24%) ( p <0.05). However, the ratio of time spent in empty cages (42.9 + 4.14%) was significantly lower than that of new mice (57.1 + 4.15%) ( p <0.05). However, the time ratio of staying in the space where the new mouse is located (38.4 + 3.76%) shows that the socialization rate is deteriorated because the time rate of staying in the space where the mouse is once met (61.6 + 3.76%) is very high ( p <0.05). In contrast, cooked silkworm ingested dementia mice preferred space with new mice (63.4 + 2.82%) over empty cages (36.6 + 2.82%), and new mice rather than space with mice once met (39.0 + 3.02%). Statistically significant ( p <0.05) stayed in the space with (60.9 + 3.02%). These results show that the ingestion of silkworms processed by social deprivation in demented mice is corrected.

In addition to these social enhancement effects, the dementia mice fed cooked slums had significantly fewer aggressions compared to the normal feed demented mice, indicating that their aggressiveness was significantly reduced.

5) Comparison of the effects of reduced silkworm beta amyloid polymer production on the brains of treated silkworms in dementia mouse models

1. Experiment Method

Toxicity in the brain that is characteristic of Alzheimer's dementia using the silkworm-treated silkworms that showed excellent effects in the experimental examples. Beta amyloid polymer production level was confirmed. After 50 weeks of dementia mice were painlessly sacrificed using CO ² gas, the mice were perfused for 1 hour with a pH 7.2 buffer containing pH 4.0 containing cold 4.0% Paraformaldehyde. The extracted brain is fixed with the same fixative for at least 24 hours, and then sections are made to a thickness of 1 mm using a mouse brain matrix. The prepared sections were clarified in CUBIC-L solution for 24 hours, and then stained beta amyloid polymer using ethanol containing 0.05% Thioflvin-S. This is followed by clearing again in CUBIC-R solution for 24 hours. Stained brain sections were determined for distribution of beta amyloid polymer using LSM700 confocal laser microscopy.

2. Experimental Results

As a result of the experiment, as shown in Figure 8e, dementia mice fed 50-week-old cooked silkworm feed was significantly lower production level of toxic beta amyloid polymer in the brain than dementia mice fed the normal feed. Beta-amyloid polymers were observed in all parts of the brain in general feed-ingested mice, and dementia mice ingested cooked silkworms were almost free of beta-amyloid polymers bound to Thioflavin-S. These results show that silkworms cooked in demented mice have an excellent effect on reducing beta amyloid polymer production.

Experimental Example 9 Confirmation of Mitochondrial Function Increase Effect According to Processed Cooked Silkworm Processing>

1) Comparison of Mitochondrial Complex Activities According to the Processing of Cooked Silkworms in Alzheimer's Dementia Drosophila Model

1. Experiment Method

The effects of mitochondrial complex I to IV were verified using normal fruit flies and Alzheimer's dementia fruit flies while ingesting silkworms processed by the yellow dog breed, which showed the best synaptic improvement and neuronal protection among the silkworm varieties. The main function of mitochondria is to produce ATP, the cell's main energy substance, which is produced by five respiratory complexes in the inner membrane of the mitochondria. Complex I to IV produce potential differences in the mitochondrial lining by oxidative phosphorylation and Complex V actually produces ATP. Therefore, a study comparing the activity of mitochondrial complex 1 to 4 was conducted.

To test the activity of mitochondrial complex I, 10 ml of mitochondria extracted from Drosophila adult brains were mixed with 182 ml of activity assay buffer (25 mM PB, 0.35% BSA, 0.06 mM DCIP, 0.07 mM decylubiquinone, 1 mM AA), and 5 mM. After mixing with 8 ml of NADH and reacted at 37 ℃ for 30 minutes, the absorbance change was measured at 600 nm for 4 minutes 30 seconds, the activity was measured in five replicate samples. For assaying mitochondrial complex II, 1 ml in 184 ml activity assay buffer (80 mM PB pH 7.8, 0.1% BSA, 2 mM EDTA, 0.2 mM ATP, 80 mM DCIP, 50 mM decylubiquinone, 1 mM AA, 3 mM rotenone) After adding 10 ml of malonate and mitochondrial extract, the reaction was carried out at 37 ° C. for 30 minutes, and then the absorbance change was measured at 600 nm for 4 minutes at 30 second intervals. For assaying mitochondrial complex III, mitochondrial extract 10 in 80 ml of activity assay buffer (50 mM Tris-HCl pH 7.5, 4 mM NaN3, 0.1 mM decylubiquinone, 80 mM Succinate, 2 mM KCN, 3 mM rotenone, 40 mM Cytochrome C) After adding ml and 10 ml of enzyme reaction solution, the mixture was measured at 25 ° C. and 550 nm for 30 minutes at 30 second intervals. For the assay of mitochondrial complex IV, 1 ml of activity assay buffer (10 mM Tris-HCl pH 7.0, 120 mM KCl), 10 ml Ferrocytochrome C substrate (0.22 mM Ferrocytochrome and 0.5 mM DTT), 2 ml of enzyme reaction and mitocone After adding 2 ml of Delia extract, measure for 1 minute at 25 ℃ and 550nm at 10 second intervals. It was measured on five replicates. Tumor-Kramer post hoc analysis was performed after confirming the differences between clusters by ANOVA.

2. Experimental Results

As a result of the experiment, as shown in Fig. 9a, the function of the mitochondrial complex I to IV using normal fruit flies and Alzheimer's dementia fruit flies while ingesting the cooked silkworm processed food flies, the fruit flies ingested only normal feed both normal fruit flies and Alzheimer's dementia flies. The mitochondrial complex of fruit flies fed feed containing cooked silkworms was significantly increased ( p <0.05).

Specifically, the consumption of cooked silkworms increased 50.9% in normal fruit flies, 70.2% in dementia models, and 23.1 in normal fruit flies. The dementia model fruit flies increased by 106.9%. In the case of Complex III, normal Drosophila increased 57.7%, dementia model Drosophila increased 263%, the activity of Complex IV increased 43.7% in normal Drosophila, and 126.6% increased in Dementia model Drosophila experimental group and Alzheimer's dementia Drosophila In the experimental group, it was confirmed that the mitochondrial complex increased significantly. These results indicate that ingestion of cooked silkworms activates mitochondrial function in both normal and Alzheimer's dementia.

2) Comparison of ATP Concentration According to the Processing of Cooked Silkworms in Alzheimer's Dementia Drosophila Model

1. Experiment Method

In order to confirm whether the amount of ATP was increased in the brain cells in accordance with the mitochondrial activity enhanced by the treatment of cooked silkworms of the yellow varieties, which showed the best synaptic improvement and neuronal protection among silkworm varieties. Inspected. Normal control fruit flies and dementia model fruit flies expressing the human Alzheimer's disease genes APP, BACE1, MAPT were cultured for 20 days in a general feed or cooked silkworm feed, and 10 animals were prepared for ATP measurement buffer (250mM Tricine, 50mM MgSO4). , 1mM EDTA, 1mM Sodium Azide) was homogenized, and then refrigerated centrifugation at 13,000rpm to secure the supernatant. The amount of ATP present in the supernatant was measured in 83.5 μl of ATP activity measurement solution (20 mM Tris-HCl pH8.0, 1 mM DTT, 10% glycerol, 0.1% BSA). 5 μl 1 mM Luciferin, 0.5 μl 250 μg / ml Luciferase, 1 After adding 10 μl of the supernatant to 100 μl DTT, the change in fluorescence was measured using a Luminometer.

2. Experimental Results

As a result of the experiment, as shown in Fig. 9b, while confirming the amount of ATP using a normal Drosophila and Alzheimer's dementia Drosophila model while ingesting the cooked silkworms, both the normal Drosophila and the Alzheimer's dementia Drosophila were cooked silkworms compared to Drosophila ingested only normal feed. It was confirmed that the ATP concentration of Drosophila fed the included feed was significantly increased ( p <0.05).

Specifically, in normal feed cultured fruit flies, the amount of ATP decreased by 31.0% on the 20th day compared to the 7th day, whereas the cooked silkworm cultured fruit flies decreased by only 3.9%. These results show that the yellow silkworm cooked silkworm suppresses the decrease in the amount of ATP produced by normal aging.

Comparing the ATP levels of normal flies and Alzheimer's dementia model Drosophila cultures for 20 days with normal feed or cooked silkworm product medium, the normal Drosophila cultured silkworm product culture normal fruit flies showed 26.2% more ATP than normal cultured fruit flies. . In the case of normal cultured dementia Drosophila model, it had 48.9% of ATP compared to normal Drosophila. In the case of cooked silkworm processed food-growing Alzheimer's dementia model Drosophila, the ATP amount was 45.0% higher than that of the normal-feeding Alzheimer's dementia model Drosophila. These results show that silkworms that have learned the reduction of ATP caused by Alzheimer's dementia are inhibited.

Therefore, this result suggests that the consumption of cooked silkworms increases the life expectancy and health life, greatly improves memory, and effectively prevents Alzheimer's dementia by activating mitochondrial function in both normal and Alzheimer's dementia to increase the production of ATP. It is noted that.

3) Reduction of Reactive Oxygen Species Production in Aged Normal Drosophila by Ingestion of Cooked Silkworm Processing

1. Experiment Method

Consumption of 10 Drosophila flocked silkworm processed foods with the best synaptic improvement and neuronal protection among normal or various silkworm varieties for 5 or 20 days Buffered Reactive Oxygen Species (ROS) After homogenizing with a solution (0.04 M Tris-HCl buffer, pH 7.4), the supernatant was obtained by centrifugation at 13,000 rpm. 20 μl of supernatant was mixed with 200 μl of ROS buffer solution and 1 μl of 10 mM 2, 7, dichlorofluorescein diacetate. After reacting at 37 ° C. for 30 minutes, the change in fluorescence was measured. Statistical analysis was performed after three repeated experiments.

2. Experimental Results

As aging occurs, the amount of reactive oxygen species increases in normal fruit flies, but the amount of active oxygen species decreased in normal fruit flies fed cooked silkworms.

Specifically, as shown in Figure 9C, 5 days old normal feed intake Drosophila has an active oxygen species of 1.23 x 10 ⁹ . Intake of free radicals of normal fruit flies increased to 3.58 x 10 ⁹ at 20 days. In comparison, the normal Drosophila on day 5 of fed silkworms had a statistically reduced free radical species ( p <0.05) of 1.05 x 10 ⁹ , and the Drosophila Drosophila on day 20 had a statistically decreased activity of 3.05 x 10 ⁹ . Oxygen species (p <0.05). These results indicate that cooked silkworms increase the production of ATP and at the same time reduce the amount of free radical species that is increased by enhancing the activity of mitochondria.

4) Comparison of ATP Concentration According to Treated Silkworm Processing in Short-term Memory Loss Mouse Model

1. Experiment Method

In order to confirm whether the amount of ATP was increased in the brain tissue of a real short-term memory loss mouse model according to the mitochondrial function enhanced by the treatment of cooked silkworms of the yellow varieties showing excellent effects in the previous examples, the concentration of ATP was examined.

In this case, the short-term memory loss mouse model was subjected to a short-term memory change behavior study after treating scopolamine, which is a short-term memory loss agent, while ingesting silkworm (powder) powder cooked in the experimental mouse, as described in Experimental Example 7a. Thereafter, the concentration of ATP in the brain tissue of the experimental mouse was examined. In addition, by applying various concentrations of silkworms cooked, the effect of the concentration was confirmed.

In order to check the amount of ATP in the mouse brain tissue, the brain was divided in half, homogenized in 0.5 ml, 2.5% HClO ₄ , and 0.1 ml of 2.5 M KOH was added thereto. The supernatant was secured by refrigeration centrifugation at 13,000 rpm. The amount of ATP present in the supernatant was measured in 83.5 μl of ATP activity measurement solution (20 mM Tris-HCl pH8.0, 1 mM DTT, 10% glycerol, 0.1% BSA). 5 μl 1 mM Luciferin, 0.5 μl 250 μg / ml Luciferase, 1 After adding 10 μl of the supernatant to 100 μl DTT, the change in fluorescence was measured using a Luminometer.

2. Experimental Results

As a result of the experiment, normal mice ingested cooked silkworm processing as shown in Figure 9d had a statistically significant ATP in brain tissue compared to normal feed-ingested normal mice.

More specifically, the saline-treated control mice that received silkworm powder powder cooked at 0.1 g, 0.2 g, or 0.5 g / kg body weight increased the amount of ATP from 33.1 to 51.1% compared to the normal feed-ingested mice. The brains of short-term memory loss mice had only 52.9% of ATP compared to the normal-fed control mice. However, the brains of short-term memory loss model mice fed silkworms cooked at 0.1 g, 0.2 g, or 0.5 g / kg body weight contained 122.1 to 166.9% more ATP than the brains of normal-feeding short-term memory mice. Was doing.

Thus, these results show that cooked silkworm ingestion increases the production of ATP by activating mitochondrial function in both normal and short-term memory loss models.

<Experimental Example 10. Comparison of accelerated neuronal division of silkworm processed methanol extract of silkworm varieties according to Example 2 of the present invention>

1. Experiment Method

To investigate the neuroprotective effects of silkworms of silkworm varieties and silkworm varieties of silkworm varieties, which showed the best synaptic effect, and to compare the cultivars, methanol extracts of silkworms of different silkworm varieties were prepared and HT22 cell lines were used. The neuronal cell division promoting effect was tested.

First, 10.0 g of cooked silkworms prepared from white and yellow dog varieties were extracted using 2.0 L of 80.0% methanol, and then dried using a vacuum evaporator. After confirming the dry weight, the extract was recovered using distilled water, and the concentration was adjusted at a rate of 10.0 mg / ml. In order to confirm the protective effect on brain neurons, the mouse hippocampal neuronal HT22 cell line was used for MTT cell division experiments, and the number of cells after 24 and 72 hours was confirmed.

2. Experimental Results

As a result of the experiment, as shown in Figure 10, when the silkworm processed methanol extract prepared in HT22 cells, the brain hippocampal cell line, it was confirmed that there is an effect that promotes cell division and cell proliferation of hippocampal cells. In particular, the cooked silkworm extract extract prepared with a yellow dog breed was found to be superior to the methanol extract extracted from a cooked silkworm manufactured with a white dog breed.

More specifically, up to 24 hours, the silkworm processing extract of 0.1% / ml of silkworm or yellow varieties of silkworms did not have a cell proliferation effect. However, when treated with 1.0mg / ml, HT22 cells were treated with whiteworm varieties. It was confirmed that 40,7% and 91.2% of yellow varieties were extracted. In addition, after 72 hours, 0.1 mg / ml treatment resulted in cell proliferation of 7.20%, yellow dog extract 98.2%, and 1.0mg / ml treatment of white dog extract 159.9% Was increased by 253.0%, and the silkworm processed extracts of H. dog varieties showed better neuroprotective effect than that of B. dog varieties.

The results show that the silkworms cooked in white dog breeds have excellent effects in exerting the proliferation of hippocampal cells, which play an important role in brain function, but the silkworms cooked in yellow dog varieties are better. Along with the inhibitory effect of short-term memory loss, it is expected that the silkworms of the breed of the silkworm varieties will be the most effective in improving memory and preventing Alzheimer's dementia.

As described above, the present invention provides a composition for improving memory or a composition for preventing and treating Alzheimer's dementia comprising a cooked silkworm processed product containing silk silk protein and a cooked silkworm processed product having a silk protein containing the same as an active ingredient. It is expected that various health supplements or medicines will be developed and marketed. Therefore, it is expected that the utilization of cooked silkworms with silk lines and the high value added will contribute to the development of the sleep industry. It is expected to contribute greatly to reducing social costs (health care costs, etc.) and improving the quality of life of the people. In addition, improving memory is expected to contribute greatly to reducing the burden on students' learning and grades.

Preparation Example 1. Pharmaceutical Preparation

200 g of cooked silkworm processed powder was mixed with 175.9 g of lactose, 180 g of potato starch, and 32 g of colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.

Preparation Example 2 Food Preparation

Formulation Example 2-1. Preparation of Cooking Seasonings

1% by weight of cooked silkworms were added to the cooking seasoning to prepare a cooking sauce for health promotion.

Formulation Example 2-2. Manufacture of dairy products

0.1% by weight of cooked silkworms were added to milk, and the milk was used to prepare various dairy products such as butter and ice cream.

Formulation Example 2-3. Vegetable Juice Manufacturing

0.5% by weight of cooked silkworm processed products were added to tomato juice or carrot juice to prepare vegetable juice for health promotion.

The present invention has been described with reference to the preferred embodiments, and those skilled in the art to which the present invention pertains to the detailed description of the present invention and other forms of embodiments within the essential technical scope of the present invention. Could be. Here, the essential technical scope of the present invention is shown in the claims, and all differences within the equivalent range will be construed as being included in the present invention.

Claims (20)

Characterized in that it comprises cooked silkworm processed material containing silk silk protein as an active ingredient, Pharmaceutical composition for improving memory. The method of claim 1, Silkworm varieties of the silkworm processing, characterized in that any one of white dog breed, yellow dog breed, soft green dog breed, red dog breed, Pharmaceutical composition for improving memory. The method of claim 2, The silkworm varieties are characterized in that the yellow dog breed, Pharmaceutical composition for improving memory. The method according to any one of claims 1 to 3, The cooked silkworm processed material is produced through the step of ripening the silkworm whole for 100 to 150 minutes with heat of steam of 80 ~ 125 ℃, Pharmaceutical composition for improving memory, Characterized in that it comprises cooked silkworm processed material containing silk silk protein as an active ingredient, Pharmaceutical composition for preventing or treating Alzheimer's dementia. The method of claim 5, Silkworm varieties of the silkworm processing, characterized in that any one of white dog breed, yellow dog breed, soft green dog breed, red dog breed, Pharmaceutical composition for preventing or treating Alzheimer's dementia. The method of claim 6, The silkworm varieties are characterized in that the yellow dog breed, Pharmaceutical composition for preventing or treating Alzheimer's dementia. The method according to any one of claims 5 to 7, The cooked silkworm processed material is produced through the step of ripening the silkworm whole for 100 to 150 minutes with heat of steam of 80 ~ 125 ℃, Pharmaceutical composition for preventing or treating Alzheimer's dementia. Characterized in that it comprises cooked silkworm processed material containing silk silk protein as an active ingredient, Food composition for improving memory. The method of claim 9, Silkworm varieties of the silkworm processing, characterized in that any one of white dog breed, yellow dog breed, soft green dog breed, red dog breed, Food composition for improving memory. The method of claim 10, The silkworm varieties are characterized in that the yellow dog breed, Food composition for improving memory. The method according to any one of claims 9 to 11, The cooked silkworm processed material is produced through the step of ripening the silkworm whole for 100 to 150 minutes with heat of steam of 80 ~ 125 ℃, Food composition for improving memory. Characterized in that it comprises cooked silkworm processed material containing silk silk protein as an active ingredient, Food composition for preventing or improving Alzheimer's dementia. The method of claim 13, Silkworm varieties of the silkworm processing is characterized in that any one of white dog breed, yellow dog breed, soft green dog breed, red dog breed, Food composition for preventing or improving Alzheimer's dementia. The method of claim 14, The silkworm varieties are characterized in that the yellow dog breed, Food composition for preventing or improving Alzheimer's dementia. The method according to any one of claims 13 to 15, The cooked silkworm processed material is produced through the step of ripening the silkworm whole for 100 to 150 minutes with a heat of steam of 80 ~ 125 ℃, Food composition for preventing or improving Alzheimer's dementia. Effective amount of the pharmaceutical composition for improving memory of any one of claims 1 to 4; And administering a pharmaceutically acceptable carrier to the subject. An effective amount of the pharmaceutical composition for preventing or treating Alzheimer's dementia according to any one of claims 5 to 8; And administering a pharmaceutically acceptable carrier to the subject. A method for producing a composition for improving memory, comprising a cooked silkworm processed product comprising silk silk protein, the step of ripening the silkworm whole with heat of steam. A method for producing a composition for preventing or treating Alzheimer's dementia comprising a silkworm, which comprises a silkworm protein, comprising the step of ripening the whole silkworm as an active ingredient.

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