KR20110057550A - Product fermented by lactic acid bacteria effective to prevention and treatment of dementia, dementia-related disease and improvement of cognition impairment and preparation method and use thereof - Google Patents

Abstract

PURPOSE: A Lactobacillus fermentation product having an effect of preventing/treating dementia and dementia-associated diseases and improving cognitive impairment is provided. CONSTITUTION: A composition for preventing and treating dementia contains Lactobacillus fermentation as an active ingredient, which is obtained by culturing Lactobaicllus in a milk-containing medium and fermenting. The Lactobacillus is Lactobacillus sp., Enterococcus sp., or Bificobacterium sp. A method for producing the Lactobacillus fermentation comprises: a step of inoculating Lactobacillus to the milk-containing medium and culturing for fermentation; a step of removing Lactobacillus from the fermentation; and a step of adding a solvent to the fermentation to precipitate and isolate active ingredients.

Description

Product fermented by lactic acid bacteria effective to prevention and treatment of dementia, dementia-related disease and improvement of cognition impairment and preparation method and use pretty}

The present invention relates to lactic acid bacteria fermentation products that are effective in the prevention / treatment of dementia, dementia-related diseases and cognitive impairment, and a method and use of the same. More specifically, the lactic acid bacteria fermentation obtained by culturing and fermenting lactic acid bacteria in a medium containing milk It relates to water, a method for preparing the same, and a composition for preventing and treating dementia or cognitive impairment comprising the same as an active ingredient.

Dementia is a type of syndrome that causes people's intellectual and social activities to be lost due to a number of causal diseases, resulting in impairments in everyday life. In other words, dementia is a brain disease caused by damage and loss of nerve cells, and severe disorders such as memory, concentration, speech, and cognition are progressively progressed for a long time, leading to death.

The causative diseases are Alzheimer's disease, Vascular dementia, Parkinson's disease, Lewy body dementia, Huntington's disease, Creutzfeldt-Jacob disease And Pick's disease are known.

The highest incidence of these causative diseases is Alzheimer's disease, which accounts for 50-70% of all dementia patients. Alzheimer's disease is classified into sporadic type and familial type depending on the cause. More than 80% of cases are sporadic and the cause of the disease is not known. Most cases occur after age 65 (Physiol Rev 2001, 81 : 741-66). Familiality accounts for less than 20% of the total, and mutations in genes such as amyloid precursor protein (APP), presenilin 1 (PS1), and presenilin 2 (PS2) are the main causes of 100% and early onset. Occurs below. Alzheimer's disease is a pathological feature in which senile plaques and neurofibrillary tangles accumulate outside and inside neurons, respectively. The senile plaques are generally spherical and are mainly found in the limbic system or neocortex, and peptides called beta amyloid (β-amyloid, Aβ) bind and deposit with each other to form nuclei and degenerated nerve fiber axons around them. Axons, dendrites, activated microglia, and astrocytes are clustered. Neuronal fiber concentrates are substances in neurons formed by binding to each other after abnormally high phosphorylation of a protein called tau is observed in various brain tissues of patients with Alzheimer's disease.

The number of dementia patients, including Alzheimer's disease, continues to grow as the population ages. According to a survey by the Korea Institute for Health and Social Affairs, 350,000 people, about 8.3% of the elderly aged 65 or over, are dementia patients, and the number is expected to increase to 530,000 in 2015. have. Although the number of dementia patients is small in the total population, many studies have been conducted for the treatment of dementia due to the enormous socio-economic burden of one year of treatment and care costs for a single dementia patient. In Korea, dementia research is being actively conducted recently.

To date, research has shown that the main cause of Alzheimer's disease is Aβ, a constituent of the senile plaque, which causes neuronal damage and causes Alzheimer's disease. Aβ is a peptide formed by decomposition of β-secretase and γ-secretase, which is one of the main pathogens of familial Alzheimer's disease, consisting of 40 to 42 amino acids. (Nature, 1999, 402: 533-7). Based on these mechanisms, studies on the development of β- and γ-secretase inhibitors and α-secretase activity promoters, which inhibit the production of Aβ, have been conducted. In addition, they inhibit the accumulation of Aβ or promote the accumulation of accumulated Aβ. Drugs are also under development (Nat Rev Neurosci, 2004, 5: 677-85). Recently, efforts to develop Aβ as a vaccine have also been attempted by multinational pharmaceutical companies. However, mechanisms of dementia based on mechanism of action require a long time to be commercialized, and those currently on the market are acetylcholine esterase inhibitors such as Cognex, Donepezyl, and Rivastigmin, which have improved memory and short-term relief. , Reminyl (Galantamine) and NMDA receptor antagonist memantine (Ebixa). In addition, blood circulation improvers such as niceroline, elcarnitine, and ginkgo biloba extract are indirectly confirmed to be effective for vascular dementia.

As discussed above, the current dementia treatment mainly plays a role in alleviating the symptoms of early dementia, but the therapeutic effect is weak and the new drug based on the mechanism of action is still required for development and release. It does not appear and is failing in the clinic (J Alz Dis. 2008, 15; 327-338). For this reason and for the purpose of prevention, new substances have been required that are harmless for long-term use and have the effect of preventing and improving Alzheimer's disease.

Lactobacillus has been used for a long time and it is recognized that Bulgarians who take Lactic Acid Bacteria fermented foods have a high rate of healthy longevity. In addition, Mechinkov's research shows that the harmful substances produced by enteric harmful bacteria enter the blood and pose a threat to health, and many studies have been conducted on the function of lactic acid bacteria from attempts to replace the intestinal flora with beneficial lactic acid bacteria. Many beneficial functions have been reported, including intestinal action, anticancer effect, immune enhancing effect and anti-aging effect (Trends Biotechnol, 1997, 15; 270-274). Currently, lactic acid bacteria and fermented products are widely used as pharmaceuticals or health foods.

Therefore, the present inventors have studied to search for new materials related to dementia, dementia-related diseases and cognitive dysfunction, and inhibited β-secretase action when culturing and fermenting a medium containing milk components with lactic acid bacteria. Fermentation products are produced that are effective in the prevention and treatment of dementia, and confirmed through the animal experiments that the fermentation products are effective in preventing / treating dementia and improving cognitive function to complete the present invention.

Accordingly, it is an object of the present invention to provide a new fermentation product, a method for producing the same, and a use thereof for improving, preventing and treating dementia, dementia-related diseases and cognitive impairment.

In order to achieve the above object, the present invention provides a composition for preventing and treating dementia comprising a lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk as an active ingredient.

In order to achieve another object of the present invention, the present invention comprises the steps of (a) inoculating, culturing and fermenting lactic acid bacteria in a medium containing milk; (b) removing the lactic acid bacteria from the fermentation product; And (c) provides a method for producing a lactic acid bacteria fermentation product comprising the step of separating the active ingredient with a solvent selected from the group consisting of acetone, alcohol having 1 to 6 carbon atoms to remove the lactic acid bacteria.

In order to achieve the another object of the present invention, the present invention provides a composition for the prevention and treatment of cognitive dysfunction comprising a lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a food composition for improving cognitive function comprising a lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk as an active ingredient.

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

The lactic acid bacteria fermented product of the present invention is characterized in that obtained by culturing and fermenting lactic acid bacteria in a medium containing milk, the purpose of preventing and improving dementia, the purpose of treatment, or the improvement of cognitive function or the improvement, prevention and treatment of cognitive dysfunction. This can be usefully used.

In the present invention, lactic acid bacteria may be used alone or in combination without limitation to those known in the art, preferably Streptococcus genus, Lactococcus genus, Enterococcus genus, Lactobacillus genus Lactobacillus Means one or more lactic acid bacteria selected from the group consisting of the genus, Pediococcus genus, Leukonostoc genus, Weissella genus and Bifidobacterium genus, the lactic acid bacteria are preferred Preferably 10 ¹⁰ to 10 ¹¹ CFU / ml. The lactic acid bacteria are Lactobacillus bacteria into lactic acid bacteria, Enterococcus genus Lactobacillus or Bifidobacterium may be lactic acid, and more preferably Lactobacillus ash FIG filler's (Lactobacillus acidophilus), Lactobacillus Kasei (L. casei), Lactobacillus ramno L. rhamnosus , L. gasseri , L. helveticus , L. helveticus , L. fermentum , L. fermentum , L. paracasei , Lactobacillus plan tareom (L. plantarum), ruteri Lactobacillus (L. reuteri), Bifidobacterium Annie Marlies (Bifidobacterium animalis), Bifidobacterium lactis (B. lactis), Enterococcus paekeol lease (Enterococcus faecalis), Enterococcus Lactococcus passive help (E. faecium), Lactobacillus breather ratio (L. breve), Bifidobacterium ronggeom (B. longum), Bifidobacterium bipyridinium bushes (B. bifidum), Lactobacillus Lee bariwooseu (L. salivarius), Lactobacillus not re kusu (L. bulgaricus) may be alone or a mixture of these strains, most preferably Lactobacillus ash FIG filler's (Lactobacillus acidophilus) ATCC 4356, Lactobacillus Kasei (L . casei) ATCC 393, Lactobacillus ramno suspension (L. rhamnosus) ATCC 7469, Lactobacillus biasing Li (L. gasseri) ATCC 33323, Lactobacillus helveticus (L. helveticus) ATCC 15009, Lactobacillus momentum spread (L. fermentum ) ATCC 14931, L. paracasei ATCC 25302, Lactobacillus plantarum L. plantarum ATCC 14917, L. reuteri ATCC 23272, Bifidobacterium animalis ATCC 25527, B. lactis DSM 10140, Enterococcus faecalis ATCC19433, E. faecium ATCC 19434, Lactobacillus L. breve ATCC 15701, B. doum B. longum ATCC 15707, L. Bifidobacterium B. bifidum ATCC29521, L. salivarius ATCC 11742, Lactobacillus Bacillus bulgaricus ATCC 11842 alone or mixed strains thereof.

In addition, the culturing of lactic acid bacteria according to the present invention may be by the lactic acid bacteria culture method known in the art. In addition, as a culture medium, a medium consisting of a carbon source, a nitrogen source, vitamins and minerals, or whey medium using milk components, skim milk medium, and the like may be used. At least one selected from the group consisting of glucose, sucrose, maltose, fructose, lactose, xylose, galactose, arabinose and combinations thereof may be used as the carbon source in the culture medium for producing the culture of the present invention. , Preferably at least one selected from the group consisting of sucrose, fructose, glucose, galactose, arabinose and lactose, more preferably glucose. The nitrogen source available in the culture medium for producing a culture of the present invention is one selected from the group consisting of yeast extract, soytone, peptone, bee extract, tryptone, cachetone and combinations thereof Or more, preferably one or more selected from the group consisting of yeast extract, peptone, tryptone and soyton, and more preferably yeast extract or soyton.

In particular, the culture medium in the present invention is a medium containing milk (milk), milk can be used in the milk of natural mammals (milk, sheep milk, goat milk, goat oil, camel milk, etc.), or processed. Examples of the processed milk include milk powder, and include both whole milk powder containing the original milk component and skim milk powder made by drying fat remaining after separating fat from milk. As the medium of the present invention, preferably SG medium or milk can be used.

More specifically, the lactic acid bacteria fermented product of the present invention

(a) inoculating, culturing and fermenting lactic acid bacteria in a medium containing milk;

(b) removing the lactic acid bacteria from the fermentation product; And

(C) can be prepared by a method comprising the step of separating the active ingredient with a solvent selected from the group consisting of acetone, alcohol having 1 to 6 carbon atoms from which the lactic acid bacteria have been removed.

In step (a), lactic acid bacteria are inoculated into the medium containing milk, and the culture is fermented. Inoculation of lactic acid bacteria is carried out by adding a suitable amount of pre-cultured lactic acid bacteria to the culture medium so that the lactic acid bacteria can fully grow under culture conditions. The amount of lactic acid bacteria to be inoculated may be added to the culture medium of the pre-cultured lactic acid bacteria culture medium 1 to 5% (v / v). Cultivation and fermentation of lactic acid bacteria can be made according to the milk containing medium and culture conditions known in the art. This process can be easily adjusted and used by those skilled in the art according to the strain selected. Such various methods are disclosed in various documents (eg, James et al., Biochemical Engineering, Prentice-Hall International Editions). Suspension cultures and adherent cultures are classified into batch, fed-batch and continuous cultures according to the culture method.

 In addition, during the culture, antifoaming agents such as fatty acid polyglycol esters can be used to suppress bubble generation. In addition, in order to maintain an aerobic state, oxygen or an oxygen-containing gas (eg, air) may be injected. The temperature at the time of culture is usually 25 ℃ to 40 ℃, preferably 35 ℃ to 40 ℃, can be incubated for 7 hours to 40 hours.

In step (b), the lactic acid bacteria are removed from the fermented product fermented and cultured in the step (a). The state of the fermentation may be different depending on the culture conditions and inoculated lactic acid bacteria, but due to the milk component contained there was a curd (curd) formation and separation from whey appears. After completion of the culture, centrifugation or filtration may be performed to separate and remove the lactic acid bacteria cells from the fermentation product (fermentation product, culture product, culture product), which steps may be performed by those skilled in the art as needed. have. At this time, removal of the bacteria means that the lactic acid bacteria cells have been substantially removed through a method for removing conventional cells.

In step (c), the active ingredient is precipitated with a solvent selected from the group consisting of acetone and an alcohol having 1 to 6 carbon atoms. Precipitation of the active ingredient is carried out to separate and concentrate the active ingredient, and may be carried out by adding acetone or an alcohol having 1 to 6 carbon atoms to the fermented product and obtaining a precipitate. Preferably, the precipitation may be carried out using ethanol, more preferably may be carried out using an edible alcohol. More specifically, the precipitation solvent may be added by mixing 1 to 4 times or less with respect to the volume of the fermentation product, and the active ingredient may be separated by centrifugation. Preferably, the precipitated solvent is added by mixing 1 times the volume of the fermented product (or the ultrafiltration fermented product), centrifuged to remove the precipitate, and then, the precipitated solvent is added to the supernatant by 1 times the volume of the supernatant, mixed and centrifuged A precipitate in which the component is precipitated can be obtained. At this time, the process may be repeated once more to precipitate the active ingredient.

The precipitated active ingredient can be used by itself or in other solvents.

On the other hand, the production method of the lactic acid bacterium fermentation of the present invention may further comprise the step of ultrafiltration (ultrafiltration) of a substance having a molecular weight of 5,000 or less for smooth separation and concentration of the active ingredient. Thus the production method of the present invention

(a) inoculating, incubating and incubating the lactic acid bacteria in a medium containing skim milk or milk;

(b) centrifuging the fermented product to remove lactic acid bacteria;

(c) obtaining a substance having a molecular weight of 5,000 or less from the fermented product from which the lactic acid bacteria have been removed; And

(D) may be a method comprising the step of separating the active ingredient with a solvent selected from the group consisting of acetone, alcohol having 1 to 6 carbon atoms to the obtained material.

Steps (a), (b) and (d) can be performed as described above.

In the above (c) step (b) to obtain a substance with a molecular weight of 5,000 or less from the fermentation product from which the lactic acid bacteria have been removed. This may be preferably performed by a method of ultrafiltration using a filter that filters only a substance having a molecular weight of 5,000 or less, to obtain an ultrafiltration solution, but is not limited thereto.

On the other hand, the present invention provides a lactic acid bacteria fermented product prepared by the production method of the present invention. Lactic acid bacteria fermented product of the present invention contains the active ingredient produced through the fermentation of milk components, which is to improve, prevent and cure dementia, dementia-related diseases and improve brain or cognitive function, improve, prevent and Effective for treatment

On the other hand, the production of Aβ, the causative protein of Alzheimer's disease, is a precursor protein formed by the amyloidogenic pathway (Nature, 1999, 402: 533-7). This process involves proteases called β- and γ-secretase. First, β-secretase acts on the N terminus of Aβ present in APP to produce CTFβ including APPβ and Aβ. Neurons have a amyloid production process that produces Aβ and a non-amyloidogenic pathway that inhibits their production. In this process, a protease called α-secretase acts on the 16th amino acid of Aβ in APP to form CTFα, which includes APPα and 16th amino acid added from the N terminal of Aβ, starting with the 17th amino acid. The action of α-secretase not only inhibits the production of Aβ, but it is known that APPα formed has a neuroprotective effect. The two pathways are balanced in neurons to inhibit the overproduction of Aβ, but the development of amyloid is enhanced by various factors such as external environment, aging, and genetic predisposition, leading to the development of Alzheimer's disease. Cell lines and animal models with enhanced amyloid production have been produced and developed for the development and research of Alzheimer's disease dementia based on these mechanisms.

In one embodiment of the present invention, according to the production method of the present invention, the lactic acid bacteria strains were cultured in a medium containing milk, the lactic acid bacteria cells were removed, and the active ingredients were precipitated to prepare fermented lactic acid bacteria, and the effects on APP metabolism were confirmed. . As a result, Enterococcus fascium ATCC 19434, Lactobacillus brevi ATCC 15701, Bifidobacterium longgum ATCC 15707, Bifidobacterium bifidem ATCC29521 alcoholic deposits did not inhibit or weaken the production of APPβ but In 14 alcoholic liquor sediments, at least 45.2% (Enterococcus facules ATCC19433) up to 85.4% (Bifidobacterium lactis DSM 10140) inhibited the production of APPβ, and enterococcus was treated with 1: 4 alcohol sediment. Alcohol sediment of fascium ATCC 19434 and Lactobacillus Bulgaricus ATCC 11842 showed relatively weak inhibition rate, whereas the remaining 16 strains of alcohol sediment showed excellent inhibitory effect. Lactobacillus ashdophyllus ATCC 4356 alcohol sediment with 93.3% inhibition rate and most of Lactobacillus alcohol sediment It can be seen that at 60% inhibition rate.

In another embodiment of the present invention, to determine whether the fermentation product of the present invention is effective against Alzheimer's disease, which is related to dementia or related diseases, Aβ40 change was measured after oral administration of each alcoholic precipitate in rats. As a result, it was confirmed that Aβ40 was suppressed from 11.7% to 45% at the time of administration of each alcoholic precipitate. As a result, it was found that the lactic acid bacteria fermentation product of the present invention had an effect of improving dementia in Alzheimer's disease dementia animal model.

In another embodiment of the present invention, a water maze test was conducted using 9-week-old dominant ICR mice to confirm the degree of cognitive improvement. As a result, all the lactic acid bacteria precipitates were found to improve the cognitive function, and Lactobacillus ashdophyllus ATCC 4356 (38.0 ± 10.1) and Bifidobacterium lactis DSM 10140 (36.1 ± 10.9) were found to have improved effects. It was found to be the best.

Accordingly, the present invention provides a composition for preventing and treating dementia comprising the lactic acid bacteria fermentation product of the present invention as an active ingredient. In addition, the present invention provides a composition for the prevention and treatment of cognitive impairment comprising the lactic acid bacteria fermentation product of the present invention as an active ingredient. The present invention also provides a brain or cognitive function food composition comprising the lactic acid bacteria fermentation product of the present invention as an active ingredient.

Dementia is a neurodegenerative disorder characterized by learning and cognitive deficits, usually involving behavioral symptoms, mental symptoms and motor symptoms. Alzheimer's disease, Alzheimer's disease, Vascular dementia, Parkinson's disease, Lewy body dementia, Huntington's disease, Creutzfeldt-Jacob disease Jacob disease), Pick's disease, or may be accompanied by the disease. Therefore, the dementia of the present invention is Alzheimer's disease dementia, Vascular dementia, Parkinson's disease dementia, Lewy body dementia, Huntington's disease dementia, and Creutz. Creutzfeldt-Jacob disease dementia, Pix's disease dementia.

Prevention, treatment and improvement of dementia prevents, treats and improves symptoms of dementia. Symptoms of dementia include:

a) behavioral symptoms such as sleep disorders, delirium (including agitation), attacks and progression,

b) psychological symptoms such as hallucinations, delusions, anxiety and depression,

c) motility symptoms, meaning impaired ability to perform motor activity despite intact motor function and

d) impaired learning and cognitive impairment, for example impaired ability to learn new information or recall previously learned information (e.g. impaired social memory), aphasia, execution, actual authentication, impairment in executive function, etc. .

Cognitive deficits or cognitive dysfunctions include cognitive function or cognitive domains such as working memory, attention and vigilance, language learning and memory, visual learning and memory, reasoning and problem solving such as executive functions, processing speed or Deterioration of social awareness. In particular, cognitive deficits or cognitive impairments integrate attention deficit, disrupted thinking, slow thinking, difficulty understanding, poor concentration, problem solving disorders, poor memory, difficulty in expressing thoughts and / or thoughts, emotions and behaviors. Difficulty in doing, or the disappearance of irrelevant thoughts. The terms "cognitive defect" and "cognitive impairment" are intended to refer to the same and are used interchangeably.

Diseases classified as cognitive impairment include, but are not limited to, cognitive deficits associated with schizophrenia, cognitive deficiencies associated with age-induced memory impairment or psychosis, cognitive deficiencies associated with diabetes, cognitive deficiencies associated with post-stroke, hypoxia and Associated memory defects, cognitive and attention deficits associated with senile dementia, memory problems associated with mild cognitive impairment, impaired cognitive function associated with dementia, impaired cognitive function associated with Alzheimer's disease, impaired cognitive function associated with Parkinson's disease, impaired cognitive function associated with vascular dementia, Cognitive problems associated with brain tumors, Pick's disease, cognitive deficiencies due to autism, cognitive deficiencies after electroconvulsion therapy, cognitive deficiencies associated with traumatic brain injury, memory loss disorders and delirium.

The lactic acid bacteria fermentation product of the present invention may be provided as a food composition (or food) or a pharmaceutical composition (or medicine).

The food composition includes all forms such as functional foods, nutritional supplements, health foods and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

For example, as a health food, the lactic acid bacteria fermented product of the present invention may be prepared in the form of tea, juice, and drink for drinking, or may be consumed by granulation, encapsulation, and powdering. In addition, the lactic acid bacteria fermentation product of the present invention can be mixed with other active ingredients to prepare in the form of a composition.

In addition, functional foods include beverages (including alcoholic beverages), fruits and processed foods (e.g. canned fruit, canned foods, jams, marmalade, etc.), fish, meat and processed foods (e.g. ham, sausage corned beef) Breads and noodles (e.g. udon, soba noodles, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, syrups, dairy products (e.g. butter, cheese), edible vegetable oils, margarine, vegetable protein , Retort foods, frozen foods, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding the lactic acid bacteria fermentation product of the present invention.

In addition, in order to use the lactic acid bacteria fermented product of the present invention in the form of food additives can be prepared in the form of powder or concentrate.

The preferred content of the lactic acid bacteria fermentation product of the present invention in the food composition of the present invention may include about 0.01 to 10% by weight relative to the total weight of the food.

The pharmaceutical composition according to the present invention may include a pharmaceutically effective amount of the lactic acid bacterium fermentation product of the present invention alone or further include one or more pharmaceutically acceptable carriers. In addition, the composition of the present invention may have a composition of 0.001 to 99.999% by weight of lactic acid bacteria fermentation and 99.999 to 0.001% by weight of carrier. As used herein, the term “pharmaceutically effective amount” refers to an amount that exhibits more response than a negative control, and preferably refers to an amount sufficient to treat or prevent dementia, dementia-related diseases, and cognitive impairment. Pharmaceutically effective amount of the lactic acid bacteria fermentation according to the present invention is 0.01 to 100 mg / day / kg body weight. However, the pharmaceutically effective amount may be appropriately changed depending on various factors such as the disease and its severity, the patient's age, weight, health condition, sex, route of administration and duration of treatment.

As used herein, "pharmaceutically acceptable" is a non-toxic composition that, when administered physiologically to humans, does not inhibit the action of the active ingredient and typically does not cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like. Say The composition of the present invention may be formulated in various ways according to the route of administration by a method known in the art together with the pharmaceutically acceptable carrier. Routes of administration may be administered orally or parenterally, but not limited to these. Parenteral routes of administration include, for example, several routes such as transdermal, nasal, abdominal, muscle, subcutaneous or intravenous.

In the case of oral administration of the pharmaceutical composition of the present invention, the pharmaceutical composition of the present invention is prepared in powder, granule, tablet, pill, dragee, capsule, liquid, gel according to a method known in the art together with a suitable oral carrier. , Syrups, suspensions, wafers and the like. Examples of suitable carriers include sugars, including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol and starch, cellulose, starch including corn starch, wheat starch, rice starch and potato starch, and the like. Fillers such as cellulose, gelatin, polyvinylpyrrolidone, and the like, including methyl cellulose, sodium carboxymethylcellulose, hydroxypropylmethyl-cellulose, and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate and the like may optionally be added as a disintegrant. Furthermore, the pharmaceutical composition may further include an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, and a preservative.

In addition, when administered parenterally, the pharmaceutical compositions of the present invention may be formulated according to methods known in the art in the form of injections, nasal inhalants, together with suitable parenteral carriers. Such injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. Examples of suitable carriers for injection include, but are not limited to, solvents or dispersion media comprising water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols, etc.), mixtures thereof and / or vegetable oils Can be. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanol amine or sterile water for injection, 10% ethanol, 40% propylene glycol and 5%

In addition, functional foods include beverages (including alcoholic beverages), fruits and processed foods (e.g. canned fruit, canned foods, jams, marmalade, etc.), fish, meat and processed foods (e.g. ham, sausage corned beef) Breads and noodles (e.g. udon, soba noodles, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, syrups, dairy products (e.g. butter, cheese), edible vegetable oils, margarine, vegetable protein , Retort foods, frozen foods, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding the lactic acid bacteria fermentation product of the present invention.

In addition, in order to use the lactic acid bacteria fermented product of the present invention in the form of food additives can be prepared in the form of powder or concentrate.

The preferred content of the lactic acid bacteria fermentation product of the present invention in the food composition of the present invention may include about 0.01 to 10% by weight relative to the total weight of the food.

The pharmaceutical composition according to the present invention may include a pharmaceutically effective amount of the lactic acid bacterium fermentation product of the present invention alone or further include one or more pharmaceutically acceptable carriers. In addition, the composition of the present invention may have a composition of 0.001 to 99.999% by weight of lactic acid bacteria fermentation and 99.999 to 0.001% by weight of carrier. As used herein, the term “pharmaceutically effective amount” refers to an amount that exhibits more response than a negative control, and preferably refers to an amount sufficient to treat or prevent dementia, dementia-related diseases, and cognitive impairment. Pharmaceutically effective amount of the lactic acid bacteria fermentation according to the present invention is 0.01 to 100 mg / day / kg body weight. However, the pharmaceutically effective amount may be appropriately changed depending on various factors such as the disease and its severity, the patient's age, weight, health condition, sex, route of administration and duration of treatment.

As used herein, "pharmaceutically acceptable" is a non-toxic composition that, when administered physiologically to humans, does not inhibit the action of the active ingredient and typically does not cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like. Say The composition of the present invention may be formulated in various ways according to the route of administration by a method known in the art together with the pharmaceutically acceptable carrier. Routes of administration may be administered orally or parenterally, but not limited to these. Parenteral routes of administration include, for example, several routes such as transdermal, nasal, abdominal, muscle, subcutaneous or intravenous.

In the case of oral administration of the pharmaceutical composition of the present invention, the pharmaceutical composition of the present invention is prepared in powder, granule, tablet, pill, dragee, capsule, liquid, gel according to a method known in the art together with a suitable oral carrier. , Syrups, suspensions, wafers and the like. Examples of suitable carriers include sugars, including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol and starch, cellulose, starch including corn starch, wheat starch, rice starch and potato starch, and the like. Fillers such as cellulose, gelatin, polyvinylpyrrolidone, and the like, including methyl cellulose, sodium carboxymethylcellulose, hydroxypropylmethyl-cellulose, and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate and the like may optionally be added as a disintegrant. Furthermore, the pharmaceutical composition may further include an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, and a preservative.

In addition, when administered parenterally, the pharmaceutical compositions of the present invention may be formulated according to methods known in the art in the form of injections, nasal inhalants, together with suitable parenteral carriers. Such injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. Examples of suitable carriers for injections include, but are not limited to, solvents or dispersion media comprising water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols, etc.), mixtures thereof and / or vegetable oils Can be. More preferably, suitable carriers may include Hanks' solution, Ringer's solution, isotonic solution such as phosphate buffered saline (PBS) containing triethanol amine or injection dextrose, and the like. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like may be further included. In addition, the injection may in most cases further comprise an isotonic agent such as sugar or sodium chloride. In the case of transdermal administrations, ointments, creams, lotions, gels, external preparations, pasta preparations, linen preparations, air rolls and the like are included. These formulations are described in Remington's Pharmaceutical Science , 15th Edition, 1975, Mack Publishing Company, Easton, Pennsylvania, a prescription generally known in pharmaceutical chemistry.

In the case of inhaled dosages, the compounds used according to the invention may be pressurized packs or by means of suitable propellants, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be delivered conveniently from the nebulizer in the form of an aerosol spray. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in inhalers or blowers can be formulated to contain a mixture of the compound and a suitable powder base such as lactose or starch.

Other pharmaceutically acceptable carriers can be found in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, Pa., 1995).

On the other hand, the experiments used in the present invention are well known in the art and described in the following literatures: Molecular Cloning, 3rd ed., Cold Spring Harbor Laboratory Press, New York, 2001.

Accordingly, the present invention provides a lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk, a method for producing the same and a use thereof. The fermented product contains an active ingredient that selectively inhibits the production of APPβ by inhibiting the β-secretase activity of the amyloid production process that induces the production of Aβ40, thereby preventing and treating dementia and dementia-related diseases, or improving cognitive function or recognition. It can be usefully used for the purpose of preventing and treating dysfunction.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

≪ Example 1 >

Preparation of Lactic Acid Bacteria Fermentation and Selection of Lactic Acid Bacteria with β-Secretase Inhibition

<1-1> Preparation of Lactic Acid Bacteria Fermentation

Eighteen human-derived lactic acid strains used in the experiment were distributed from a depository institution such as Korea Microorganism Conservation Center, and the fermentation products cultured with these strains were examined for APPβ inhibition activity. Lactic acid strains used were: Lactobacillus acidophilus ATCC 4356, L. casei ATCC 393, L. rhamnosus ATCC 7469, Lactobacillus gaseri ( L. gasseri ) ATCC 33323, L. helveticus ATCC 15009, L. fermentum ATCC 14931, Lactobacillus paracasei ATCC 25302, Lactobacillus plantarum ) ATCC 14917, L. reuteri ATCC 23272, Bifidobacterium animalis ATCC 25527, B. lactis DSM 10140, Enterococcus faecalis ATCC19433, E. faecium ATCC 19434, L. breve ATCC 15701, Bifidobacterium longgum ( B. longum ) ATCC 15707, Bifidobacterium bifidom ( B . Bifidum) ATCC29521, Lactobacillus salivarius (L. salivarius) ATCC 11742, Lactobacillus Bulgaria kusu (L. bulgaricus) ATCC 11842.

As lactic acid bacteria, subcultured or precultured in MRS medium (de Man-Rogosa-Sharpe media) was used. In order to prepare the lactic acid bacterium fermentation product of the present invention, the preculture liquid containing each lactic acid bacterium was 3% (v / v) in SG (10% skim milk powder, 0.5% glucose, pH 6.8) medium or milk containing skim milk powder as a main component. The culture solution was obtained by inoculating with a method of incubating at 37 ° C. for 24 hours. All cultured in SG medium or milk showed curd formation and separated from whey, just like lactic acid bacteria fermented milk. As a control, a medium inoculated at 3% (v / v) in MRS medium was cultured under the same conditions.

Each culture was centrifuged at 15,000 rpm for 10 minutes to recover the supernatant, which was first filtered with a 0.22 μm syringe filter. Next, ultrafiltration was performed with a filter membrane for filtering a molecular weight of 5,000 or less. Subsequently, the alcohol was mixed with 1 times the volume of the ultrafiltrate first, followed by centrifugation at 15,000 rpm for 10 minutes to separate the precipitate and the supernatant. This precipitate was referred to as 1-time precipitate, and 1-fold volume of the ultrafiltrate was added to the remaining supernatant and centrifuged in the same manner to separate the precipitate and the supernatant. This precipitate was called a 2-fold precipitate, and a 4-fold precipitate was obtained in the same manner. Each precipitate was recovered and dried and weighed by concentration in 0.5% trifluoroacetic acid (TFA). Suspension was used for the test. In the cognitive function improvement test, the alcoholic sediment containing 1: 2 and 1: 4 alcoholic sediments is added to the test by adding 4 times the alcohol directly to the ultrafiltration volume without recovering the alcoholic sediment as described above. It was.

<1-2> Screening of lactic acid bacteria inhibiting β-secretase activity

Lactobacillus strains having β-secretase activity inhibitory effect were selected using the alcohol precipitates of the fermented lactic acid bacteria fermentations obtained in Example <1-1> as follows. As a control group, the precipitate precipitated by alcohol in the same manner was used only in the SG medium culture medium not inoculated with lactic acid bacteria.

In order to investigate the effects of each lactic acid bacteria fermentation product sample of the present invention on the metabolism of amyloid precursor protein (hereinafter referred to as APP), HEK293 cell line in which they were overexpressed by injecting β-secretase and wild-type APP695 genes , BA-3; Yeon et al, Peptides, 2007, 28: 838-844). BA-3 is useful for the exploration of active ingredients that directly or indirectly affect β-secretase activity by generating a lot of APPβ by overexpression of β-secretase.

BA-3 was inoculated in a 6-well plate at a density of 80% or more using DMEM medium containing 10% FBS (fetal bovine serum) and incubated for 1 day. After removing the medium, 0.9 ml of DMEM (free DMEM) without FBS warmed to 37 ° C. was added, and the ethanol precipitate sample suspended in 0.1 ml free DMEM was treated at 100 μg / ml and incubated for 24 hours.

0.2 ml of each sample-treated medium was recovered and subjected to electrophoresis on 8% SDS-PAGE, and then APPβ in culture was measured by Western blot using Rb53 (polyclonal, Cephalon, USA) antibody, an antibody of APPβ.

As a result, as shown in Fig. 1 and Table 1, enterococcus fascidium ATCC 19434, Lactobacillus brevi ATCC 15701, Bifidobacterium long gum ATCC when 18 kinds of lactic acid bacteria culture liquor precipitates 1: 2 were treated with BA-3 cell line. 15707, Bifidobacterium bifidum ATCC29521 alcoholic sediments did not inhibit or weakly inhibit the production of APPβ, but at least 45.2% in 14 other strains of Lactobacillus strains (Enterococcus faculus ATCC19433), up to 85.4% ( Bifidobacterium lactis DSM 10140) inhibited the production of APPβ. In the treatment of 1: 4 alcoholic sediment, the alcoholic sediments of Enterococcus fascium ATCC 19434 and Lactobacillus vulgaris ATCC 11842 showed relatively weak inhibition, whereas the other 16 strains of alcohol sediment showed excellent inhibitory effect. Lactobacillus ashdophyllus ATCC 4356 alcohol sediment showed 93.3% inhibition and most of the lactic acid strain alcohol sediment showed more than 60% inhibition.

In order to confirm whether the sAPPβ inhibitory effect appears selectively in the alcoholic precipitate obtained by culturing each lactic acid bacteria in the milk medium, the culture filtrate obtained by culturing each lactic acid bacteria in the MRS medium, the lactic acid bacteria culture medium, was ultrafiltration after ultrafiltration. Four times the volume of the filtrate was added to obtain a precipitate, which was treated with 100 μg / ml in the BA-3 cell line. It was confirmed that sAPPβ inhibition of milk medium ethanol sediment was selective as all MRS culture alcohol sediments of 18 lactic acid bacteria did not inhibit sAPPβ at all.

In conclusion, 18 kinds of alcoholic precipitates tested were strongly inhibited in the production of APPβ and were able to inhibit the production of beta amyloid, which is the cause protein of Alzheimer's dementia, although there were differences depending on the method and strain obtained.

In vitro β-secretase Inhibition and sAPPβ Inhibitory Effect of Lactobacillus Alcohol Sediment Strain SAPPβ inhibition rate of each alcoholic sediment (%) SG Badge MRS Badge 1: 2 1: 4 1: 4 L. acidophilus ATCC 4356 74.0 93.3 0.0 L. casei ATCC 393 72.4 87.6 0.0 L. rhamnosus ATCC 7469 83.1 73.0 0.0 L. gasseri ATCC 33323 74.1 86.3 0.0 L. helveticus ATCC 15009 73.6 64.0 0.0 L. fermentum ATCC 14931 39.9 42.4 0.0 L. paracasei ATCC 25302 72.1 83.0 0.0 L. plantarum ATCC 14917 45.6 63.3 0.0 L. reuteri ATCC 23272 68.3 60.2 0.0 B. animalis ATCC 25527 68.5 76.9 0.0 B. lactis DSM 10140 85.4 80.8 0.0 E. faecalis ATCC19433 45.2 67.6 0.0 E. faecium ATCC 19434 8.5 19.0 0.0 L. breve ATCC 15701 0.0 71.6 0.0 B. longum ATCC 15707 0.0 80.2 0.0 B. bifidum ATCC29521 0.0 32.8 0.0 L. salivarius ATCC 11742 72.7 65.4 0.0 L. bulgaricus ATCC 11842 61.7 14.5 0.0

<Example 2>

Confirmation of In vivo Effect of Fermented Products of the Present Invention

250 mg of 1: 4 alcohol precipitates suspended in 1% carboxymethylcellulose in 200-250 g of Sprague Dawley rats to determine if each lactic acid bacteria sediment inhibits the production of Aβ, the cause of dementia. Blood was collected 4 hours after oral administration at / kg, the plasma was separated after centrifugation, and the change in plasma Aβ40 was quantitatively compared using the ELISA kit (the GENETICS Company, Switzerland).

This method was performed according to the methods of Moudden et al. (Current Pharmaceutical Design, 2006, 12: 671-676) and Best et al. (JPET, 2005, 313: 902-908). In general, the efficacy evaluation of a dementia drug is analyzed by using a dementia animal model rat, which improves the shortness of the animal model due to the problems of whether the animal model is expensive, the evaluation period is very long, and the non-genetic dementia can be evaluated. The need for animal model testing has been raised to identify the dementia-improving effects of many drugs over time, and one of these methods has been developed to determine the reduction of Aβ in rat blood after drug administration. Drugs that reduce A [beta] in existing animal models of dementia are also useful in the search for therapeutic drugs by reducing A [beta] by this method.

After 4 hours of oral administration of SG medium alcohol sediment 1: 4, the concentration of Aβ40 in plasma and the concentration of Aβ40 after oral administration of Lactobacillus ethanol sediment 1: 4 were measured to determine the Aβ40 inhibition rate of Lactic acid bacteria ethanol sediment. Lactic acid bacteria inhibited Aβ40 by more than 10%, and L. acidophilus ATCC 4356 showed the highest inhibition rate of 45.0 ± 7.1%. Bifidobacterium lactis ) DSM 10140 was followed by 38.3 ± 6.9%.

Inhibitory Effect of Lactobacillus Alcoholic Sediment on In vivo Aβ40 Strain Aβ0 Inhibition of 1: 4 Alcohol Sediment (%) L. acidophilus ATCC 4356 45.0 ± 7.1 L. casei ATCC 393 18.3 ± 3.8 L. rhamnosus ATCC 7469 21.5 ± 4.5 L. gasseri ATCC 33323 23.2 ± 3.5 L. helveticus ATCC 15009 14.6 ± 2.8 L. fermentum ATCC 14931 17.7 ± 3.2 L. paracasei ATCC 25302 15.5 ± 2.9 L. plantarum ATCC 14917 15.9 ± 5.3 L. reuteri ATCC 23272 18.5 ± 5.7 B. animalis ATCC 25527 14.1 ± 3.9 B. lactis DSM 10140 38.3 ± 6.9 E. faecalis ATCC19433 19.8 ± 2.3 E. faecium ATCC 19434 16.0 ± 4.9 L. breve ATCC 15701 21.8 ± 2.7 B. longum ATCC 15707 17.0 ± 3.2 B. bifidum ATCC29521 24.4 ± 5.1 L. salivarius ATCC 11742 11.7 ± 1.1 L. bulgaricus ATCC 11842 19.5 ± 3.5

<Example 3>

Cognitive Function Improvement Efficacy Test of Lactic Acid Bacteria Fermentation

Nine-week-old dominant ICR mice were used in each treatment group for cognitive improvement tests. Each type strain lactic acid bacterium fermented product alcohol precipitate was suspended in 1% carboxymethyl cellulose to prepare a administered sample. The alcoholic sediment of each strain was repeatedly orally administered twice a week for 2 weeks at 500 mg / kg dose, and 60 minutes before the test, oral administration of scopolamine at 1 m / kg was used to cause memory damage. maze) test was carried out. As a vehicle, a group treated with scopolamine after administration of saline solution for 2 weeks instead of the drug was used.

During the underwater maze test, the reference memory test was performed after adapting by allowing the swimmer to swim freely for 60 seconds in a platformless tank one day before the start of the test. The test was performed by measuring the time to find the platform submerged 4-5 times daily for 5 days (escape latency, in seconds), and the maximum allowable time was limited to 60 seconds. By the second day of the test, if the platform could not be located, it was guided to find the platform within the 60-second time limit.

24 hours after the end of the reference memory test, the probe test was performed by removing the platform in the tank and allowing the user to swim freely for 60 seconds to measure the time to stay at the platform.

After the administration of 1: 4 alcoholic precipitates obtained by ultrafiltration of each lactic acid bacteria culture filtrate, the reference memory test was performed for 5 days, and the average time for finding the platform of the scopolamine-treated group on the 5th day and the time for finding the platform of each lactic acid bacteria precipitate. The time to find the platform compared to the scopolamine-administered group was expressed as an improvement rate.

Reference memory improvement rate = (t _s -t _{lactic acid bacteria} ) / t _s × 100 (%)

(t _s : mean time to find the platform of scopolamine group; t _lactic acid _bacteria : mean time to find the platform of lactic _acid bacteria alcohol sediment)

As a result, it was found that all strains precipitated the cognitive function, and Lactobacillus ashdophyllus ATCC 4356 (38.0 ± 10.1) and Bifidobacterium lactis DSM 10140 (36.1 ± 10.9) had the best improvement effect. Appeared.

In the probe test conducted after the completion of the reference memory test, the average time spent on the platform in the scopolamine-administered group was compared with the average time spent on the Lactobacillus ethanol sediment-treated group.

Probe test improvement rate = (t _{lactic acid bacteria} -t _s ) / t _s × 100 (%)

(t _s : mean time spent on the platform where the scopolamine-administered group stayed; t mean _lactic acid _bacteria : mean time spent on the platform where the lactobacillus alcoholic sediment was administered)

In the probe test, Lactobacillus ashdophilus ATCC 4356 and Bifidobacterium lactis DSM 10140 alcoholic sediment treated group had the longest time on the platform and the improvement rate was 138.3 ± 9.9% and 85.2 ± 11.1, respectively. Non-phidum ATCC29521 (76.3 ± 9.2), Lactobacillus Bulgaricus ATCC 11842 (56.3 ± 7.0), and Lactobacillus rhamnosus ATCC 7469 (53.1 ± 3.0) also showed excellent improvement.

Improvement of Cognitive Function of Lactobacillus Alcohol Sediment Strain Day 5 improvement of reference memory (% improvement compared to scopolamine group) Probe inspection
(% improvement compared to scopolamine group) L. acidophilus ATCC 4356 38.0 ± 10.1 138.3 ± 9.9 L. casei ATCC 393 17.3 ± 4.8 13.5 ± 2.7 L. rhamnosus ATCC 7469 27.5 ± 6.8 53.1 ± 3.0 L. gasseri ATCC 33323 27.2 ± 7.5 45.0 ± 2.5 L. helveticus ATCC 15009 24.8 ± 3.8 35.1 ± 1.9 L. fermentum ATCC 14931 18.4 ± 4.3 29.8 ± 5.5 L. paracasei ATCC 25302 13.5 ± 4.9 27.6 ± 6.4 L. plantarum ATCC 14917 16.4 ± 8.3 18.0 ± 2.5 L. reuteri ATCC 23272 16.9 ± 3.7 18.7 ± 4.7 B. animalis ATCC 25527 15.4 ± 3.4 16.6 ± 4.1 B. lactis DSM 10140 36.1 ± 10.9 85.2 ± 11.1 E. faecalis ATCC19433 23.7 ± 5.1 32.8 ± 6.0 E. faecium ATCC 19434 13.5 ± 5.3 12.0 ± 5.3 L. breve ATCC 15701 13.9 ± 8.8 17.4 ± 6.9 B. longum ATCC 15707 17.8 ± 5.1 39.0 ± 1.3 B. bifidum ATCC29521 25.5 ± 4.1 76.3 ± 9.2 L. salivarius ATCC 11742 16.3 ± 6.5 17.5 ± 3.0 L. bulgaricus ATCC 11842 21.0 ± 6.7 56.3 ± 7.0

Hereinafter, an example of the preparation of a pharmaceutical composition comprising the compound of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Preparation Example 1 Preparation of Powder

20 mg of lactic acid bacteria fermented product (powdered) of the present invention

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

&Lt; Formulation Example 2 > Preparation of tablet

10 mg of lactic acid bacteria fermented product of the present invention (powdered)

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Preparation Example 3 Preparation of Capsule

10 mg of lactic acid bacteria fermented product of the present invention (powdered)

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Preparation Example 4 Preparation of Injection

1 mg of lactic acid bacteria fermented product of the present invention (powdered)

Mannitol 180 mg

Sterile distilled water for injection 2793 mg

_{Na 2 HPO 4 · 12H 2 O} 26 ㎎

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Preparation Example 5 Preparation of Liquid

Lactic acid bacteria fermented product of the present invention (powdered) 2 mg

10 g per isomer

5 g mannitol

Purified water quantity

After dissolving each component in purified water according to the usual method of preparing a liquid solution, adding lemon flavor appropriately, mixing the above components, adding purified water, adjusting the whole to 100 ml by adding purified water, and then filling into a brown bottle. The solution is prepared by sterilization.

Preparation Example 6 Preparation of Health Food

Lactic acid bacteria fermented product of the present invention (powdered) 500 mg

Vitamin mixture quantity

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

0.13 mg of vitamin B ₁

0.15 mg of vitamin B ₂

0.5 mg of vitamin B ₆

Vitamin B ₁₂ 0.2 g

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Preparation Example 7 Preparation of Healthy Drinks

100 mg of lactic acid bacteria fermented product of the present invention (powdered)

15 g of vitamin C

100 g of vitamin E (powder)

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B ₁ 0.25 g

Vitamin B ₂ 0.3 g

Water quantification

After mixing the above components according to the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated Used to prepare the healthy beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

As described above, the present invention provides a lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk, a method for preparing the same and use thereof. The fermented product contains an active ingredient that selectively inhibits the production of APPβ by inhibiting the β-secretase activity of the amyloid production process that induces the production of Aβ40, thereby preventing and treating dementia and dementia-related diseases, or improving cognitive function or recognition. It can be usefully used for the purpose of preventing and treating dysfunction.

1 is a result of confirming the β-secretase inhibitory effect of alcohol sediment obtained by culturing each strain strain lactic acid strain in SG medium by ultrafiltration (Media: using only culture medium without administration of precipitate) Band of APPβ in the fermentation of BA-3 cell line; each strain fermentation: in fermentation product of BA-3 cell line by administering alcoholic precipitate of each type strain lactic acid bacteria fermentation to the culture medium at a concentration of 100 μg / ml Band of APPβ). The result is a thin 1: 2 alcoholic sediment treatment and I am treated with a 1: 4 alcoholic precipitate.

2 is a result of confirming the β-secretase inhibitory effect of 1: 4 alcoholic precipitate obtained after ultrafiltration of the culture filtrate obtained by culturing each type strain lactic acid strain in MRS medium by Western blot (Media: Band of APPβ in the fermentation cultured BA-3 cell line using; each strain fermentation product: fermentation of BA-3 cell line by administering the alcoholic precipitate of each type strain lactic acid bacteria fermentation to the culture medium at a concentration of 100 μg / ml Band of APPβ in water).

Figure 3 is a graph of the results of comparing the time to find the platform in order to find out the memory improvement effect of each strain strain lactic acid bacteria in the underwater maze test of scopolamine induced memory damage model mice (Escape latency time ( s): Time taken to find the platform (seconds); Scopolamine: Scopolamine treated group after 2 weeks of physiological saline instead of drug; Scopolamine treated group after oral administration for 2 weeks in kg; Scopolamine + B. lactis DSM10140: Bifidobacterium lactis DSM10140 culture solution after iv administration of alcoholic precipitate at 500 mg / kg for 2 weeks; Amine + L. acidophillus ATCC4356: Scopolamine treated group after oral administration of Lactobacillus ashdophilus ATCC4356 culture liquor precipitate at 500 mg / kg for 2 weeks; Scopolamine + L. plantarum ATC C14917: Scopolamine treated group after oral administration of Lactobacillus plantarum ATCC14917 culture liquor sediment at 500 mg / kg for 2 weeks.

Claims (8)

A composition for preventing and treating dementia comprising a lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk as an active ingredient. The composition of claim 1, wherein the lactic acid bacteria are selected from the group consisting of Lactobacillus sp., Enterococcus spp. And Bifidobacterium sp. According to claim 1, The lactic acid bacteria Lactobacillus acidophilus ( Lactobacillus acidophilus ), Lactobacillus casei ( L. casei ), Lactobacillus rhamnosus ( L. rhamnosus ), Lactobacillus gasery ( L. gasseri ), Lactobacillus L. helveticus , L. fermentum , L. fermentum , L. paracasei , L. plantarum , L. plantarum , L. reuteri , BP Bifidobacterium animalis , Bifidobacterium lactis , Enterococcus faecalis , E. faecium , L. breve , B. longum , B. bifidum , L. salivarius and L. bulgaricus from the group consisting of L. bulgaricus and Bifidobacterium longgum ( B. longum ), B. bifidum , L. salivarius and L. bulgaricus Composition, characterized in that. According to claim 1, wherein the dementia is Alzheimer's disease dementia, Vascular dementia, Parkinson's disease dementia, Lewy body dementia, Huntington's disease dementia , Dementia selected from the group consisting of Creutzfeldt-Jacob disease type dementia and Pick's disease type dementia. (a) inoculating, culturing and fermenting lactic acid bacteria in a medium containing milk; (b) removing the lactic acid bacteria from the fermentation product; (C) a method for producing a lactic acid bacteria fermentation product having a dementia prevention and treatment effect comprising the step of precipitating and separating the active ingredient by adding a solvent selected from the group consisting of acetone, alcohol having 1 to 6 carbon atoms to remove the fermented lactic acid bacteria. The method of claim 5, wherein the manufacturing method (a) inoculating, culturing and fermenting lactic acid bacteria in a medium containing milk; (b) centrifuging the fermented product to remove lactic acid bacteria; (c) obtaining a substance having a molecular weight of 5,000 or less from the fermented product from which the lactic acid bacteria have been removed; And (d) adding a solvent selected from the group consisting of acetone and an alcohol having 1 to 6 carbon atoms to the obtained material to precipitate and separate the active ingredient, the method of producing a lactic acid bacterium fermentation product having a dementia prevention and treatment effect. A composition for the prevention and treatment of cognitive dysfunction comprising the lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk as an active ingredient. A food composition for enhancing brain or cognitive function comprising a lactic acid bacteria fermented product obtained by culturing and fermenting lactic acid bacteria in a medium containing milk as an active ingredient.

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