Compositions, foods, drinks, feeds and medicines for maintaining, enhancing or improving cognitive function, preventive and therapeutic drugs for dementia, and compositions, foods, drinks, feeds and medicines for preventing or improving brain fatigue

Abstract

The composition for maintaining, enhancing or improving cognitive function contains cells of Bifidobacterium adolescentis JCM 7045 strain or a processed product thereof.

Description

Technical Field

[0001] The present invention relates to compositions, foods and drinks, feeds and medicines for maintaining, improving or enhancing cognitive functions, preventive and therapeutic agents for dementia, and compositions, foods and drinks, feeds and medicines for preventing or improving brain fatigue, to the drug and the like.

Background Art

[0002] According to the World Health Organization, it is estimated that there are over 55 million people with dementia worldwide, and it is suggested that about 10 million new cases of dementia occur every year (see Non-Patent Document 1). In dementia, it is known that damage to the brain accumulated due to nerve cells destroyed in chronic inflammation that occurs with aging causes a decline in cognitive function and the like.

[0003] In Japan, it is estimated that about 16% of people aged 65 and over have dementia. It is thought that the proportion of dementia patients will continue to rise in the future. In a country with a rapidly aging population, the increase in dementia patients is a concern as it may further exacerbate social problems such as a shortage of caregivers and a burden on insurance medical costs.

[0004] The most typical dementia is Alzheimer's disease (AD). 60 - 70% of dementia patients have AD. As treatment means for dementia and related diseases, for example, administration of drugs such as cholinesterase inhibitors like donepezil, NMDA receptor antagonists like memantine, and selective serotonin inhibitors has become common (see Non-Patent Document 2). In recent years, antibody drugs such as lecanemab have also been actively developed.

Prior Art Documents

Non-Patent Documents

[0005]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0006] The above-mentioned drugs and antibody drugs are difficult to incorporate into daily life for preventing cognitive function decline or improving cognitive function.

[0007] The present invention has been made in view of the above circumstances, and is easy to incorporate into daily life, and an excellent cognitive function improvement effect can be obtained recognized To provide a composition, food, drink, feed, and medicine for maintaining, improving, or enhancing cognitive function, as well as a preventive and therapeutic drug for dementia.

[0008] In addition, the present invention is easy to incorporate into daily life, and an excellent brain fatigue improvement effect can be obtained brain To provide a composition, food, drink, feed, and medicine for preventing or improving fatigue.

Means for Solving the Problems

[0009] The composition for maintaining, improving, or enhancing cognitive function according to the first aspect of the present invention contains cells or processed products thereof of Bifidobacterium adolescentis JCM 7045 strain.

[0010] The cognitive function is a function related to memory, which may also be the case.

[0011] The function related to the memory is a long-term memory function, which may also be the case.

[0012] The composition for preventing or improving brain fatigue according to the second aspect of the present invention is comprising cells of Bifidobacterium adolescentis JCM 7045 strain or a processed product thereof.

[0013] The food or drink for maintaining, improving or enhancing cognitive function according to the third aspect of the present invention is comprising the composition according to the first aspect of the present invention.

[0014] The food or drink for preventing or improving brain fatigue according to the fourth aspect of the present invention is comprising the composition according to the second aspect of the present invention.

[0015] The feed for maintaining, improving or enhancing cognitive function, or the feed for preventing or improving brain fatigue according to the fifth aspect of the present invention is comprising cells of Bifidobacterium adolescentis JCM 7045 strain or a processed product thereof.

[0016] The medicine for maintaining, improving or enhancing cognitive function according to the sixth aspect of the present invention is comprising the composition according to the first aspect of the present invention.

[0017] The medicine for preventing or improving brain fatigue according to the seventh aspect of the present invention is comprising the composition according to the second aspect of the present invention.

[0018] The prophylactic or therapeutic agent for dementia according to the eighth aspect of the present invention is comprising cells of Bifidobacterium adolescentis JCM 7045 strain or a processed product thereof.

[0019] wherein the dementia is Alzheimer's dementia, which is also acceptable.

Advantages of the Invention

[0021] The composition, food and drink, feed, and medicine according to the present invention, as well as the preventive and therapeutic agents for dementia, are easy to incorporate into daily life and can provide excellent cognitive function improvement effects. In addition, the composition, food and drink, feed, and medicine according to the present invention are easy to incorporate into daily life and can provide excellent brain fatigue improvement effects. Further, the composition, food and drink, feed, and medicine according to the present invention, as well as the preventive and therapeutic agents for dementia, also have a high co-aggregation ability against, for example, Fusobacterium nucleatum, which is a periodontal pathogen.

Brief Description of the Drawings

[0022]

Figure 1

Figure 2

Figure 3

Modes for Carrying Out the Invention

[0023] Embodiments according to the present invention will be described with reference to the drawings. It should be noted that the present invention is not limited by the following embodiments and drawings. In the following embodiments, expressions such as "having", "including", or "containing" also include the meaning of "consisting of" or "composed of".

[0024] (Embodiment 1: Composition) The composition according to this embodiment contains cells of Bifidobacterium adolescentis JCM 7045 strain (hereinafter also simply referred to as "JCM 7045") or a processed product thereof. JCM 7045 can be obtained from the RIKEN BRC. JCM 7045 is preferably cultured at or near the optimum temperature, and the culture temperature is, for example, 27 to 45 °C, 30 to 42 °C, or 35 to 40 °C. Preferably, the culture temperature is 37 °C. JCM 7045 can be cultured in a liquid medium or an agar medium. Preferably, the bifidobacteria can be cultured under anaerobic conditions, for example, while aerating an anaerobic gas such as carbon dioxide gas. Also, it may be cultured under microaerophilic conditions such as liquid static culture. The medium for culturing JCM 7045 is not particularly limited, and a medium commonly used for culturing bacteria belonging to the genus Bifidobacterium can be used.

[0025] As the carbon source in the culture, for example, sugars such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolyzate, and molasses can be used. As the nitrogen source, for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, ammonium nitrate, and nitrates can be used. Also, as inorganic salts, for example, sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, ferrous sulfate, etc. can be used. Also, organic components such as peptone, soybean powder, defatted soybean meal, meat extract, and yeast extract may be used. For example, the medium for JCM 7045 is a BL medium containing Nutrient Broth.

[0026] The cells can be obtained as a precipitate by centrifuging the culture medium after cultivation. The cells may be viable or dead, and may be wet or dry. The processed product of the cells is, for example, freeze-dried cells, cells dried with acetone, an extract obtained by subjecting the cells to solvent extraction treatment, a disrupted product obtained by disrupting the cells or dry cells with ultrasonic waves or the like, cells obtained by applying some treatment to the cells, a composition derived from the cells or containing a part of the cells. The cells, a composition derived from the cells, or a composition containing a part of the cells is, for example, a culture supernatant or a culture medium. Therefore, the processed product of the cells also includes the culture supernatant and the culture medium.

[0027] As the solvent used for the solvent extraction treatment, water, an organic solvent, and a mixture thereof can be used. Examples of the organic solvent include ether, chloroform, benzene, hexane, methanol, ethanol, isopropanol, and a mixed solution thereof. The extract extracted using the extraction solvent can be used in a liquid state, or can be concentrated or diluted and used in a liquid, gel, or paste form. Further, it can be used as a dried product obtained by drying it. Drying can be performed by known methods such as spray drying, freeze drying, vacuum drying, and fluidized drying.

[0028] After culturing JCM 7045, the obtained cells may be used as they are, or may be diluted or concentrated before use. The same applies to the processed product of the cells, which may be used as it is, or may be diluted or concentrated before use.

[0029] The composition according to this embodiment contains, as an active ingredient, the cells of JCM 7045 or a processed product thereof, and components other than the active ingredient may be blended. The components other than the active ingredient are not particularly limited and are additives or the like formulated in pharmaceuticals. Further, when a culture supernatant or a culture medium is used as the processed product of the cells, components derived from the culture medium, for example, may be included in the composition as components other than the active ingredient.

[0030] As shown in Example 2 below, the composition according to this embodiment has an effect of improving cognitive function, particularly an effect of improving long-term memory impairment. Therefore, the uses of the composition according to this embodiment include maintenance, improvement or enhancement of cognitive function, and prevention or improvement of brain fatigue. Preferably, the cognitive function is a function related to memory. More preferably, the function related to memory is the long-term memory function.

[0031] The use of the composition according to this embodiment may be prevention or improvement of brain fatigue. Brain fatigue refers to a state in which the brain continues to be subjected to excessive stress and its function deteriorates. Fatigue is considered to occur as a result of a chronic decrease in immune cytokines or induction of inflammatory cytokines due to stress or the like, resulting in a decrease in the original function (Japanese Journal of Biological Psychiatry 24(4):200-210, 2013). In order to improve brain fatigue, for example, induction of inflammatory cytokines may be suppressed. As shown in Example 1 below, the composition suppresses the secretion of inflammatory cytokines in central nervous system cells and has an excellent anti-inflammatory effect. Therefore, the composition is useful for prevention or improvement of brain fatigue.

[0032] The composition contains, for example, 0.1 to 99% by weight, 1 to 50% by weight, or 1 to 20% by weight of the cells of JCM 7045 or a processed product thereof as an active ingredient. The content of the cells of JCM 7045 or a processed product thereof in the composition according to this embodiment may be appropriately adjusted according to the effective amount in the above uses of the composition. The effective amount is the amount necessary to obtain a desired result and is the amount necessary to bring about a delay, inhibition, prevention, reversal or cure of the progression of the condition to be treated or disposed of. When the active ingredient is the cells of JCM 7045, the effective amount is, for example, 1.0×10 2 ~1.0×10 10 colony forming units (cfu) / kg, 1.0×10 3 ~1.0×10 9 cfu / kg or 1.0×10 4 ~1.0×10 8 cfu / kg. Also, from another perspective, the effective amount is 1.0×10 8cells (pcs) / kg ~ 1.0×10 13 cells (pcs) / kg, 1.0×10 9 cells (pcs) / kg ~ 1.0×10 12 cells (pcs) / kg or 1.0×10 10 cells (pcs) / kg ~ 1.0×10 12 cells (pcs) / kg may also be used. When using the treated product of the cells of JCM 7045 as the active ingredient, the treated product of the cells is, for example, an amount corresponding to the effective amount of the cells. In addition, any method for measuring (determining) the number of bacteria may be used as long as it is a method commonly used in this field. Specifically, for example, fluorescence (DAPI) staining method, plate smearing method, etc. may be mentioned.

[0033] The administration targets of the composition according to this embodiment are humans and animals other than humans. As animals, mammals are preferred, and more specifically, dogs, cats, cows, pigs, horses, sheep, deer, etc. may be mentioned. The dosage of the composition is appropriately determined according to the sex, age, weight, symptoms, etc. of the administration target. The composition is administered so that the cells of JCM 7045 or its treated product is in an effective amount. The composition can be administered once a day or divided into multiple times. The composition may be administered at various dosing frequencies such as daily, every other day, once a week, every other week, once a month, etc. In addition, if necessary, the composition may be used in an amount outside the above range. The administration method of the composition is not particularly limited, but it is administered by injection, nasal, transdermal, pulmonary, oral, etc. As the administration method, oral administration is particularly preferred.

[0034] The composition according to this embodiment contains the cells of Bifidobacterium JCM 7045 or its treated product. Therefore, it is easy to incorporate it into daily life, and an excellent cognitive function improvement effect can be obtained.

[0035] The composition according to another aspect in this embodiment contains the cells of JCM 7045 strain or its treated product, and the content of the cells or its treated product is 1.0×10 8 cells (pcs) / g ~ 1.0×10 13 cells (pcs) / g or 1.0×10 9cells (pcs) / kg ~ 1.0×10 12 is cells (pcs) / kg.

[0036] (Embodiment 2: Food and Drink) In another embodiment, a food or drink containing the composition according to Embodiment 1 is provided. The use of the food or drink is to maintain, improve or enhance cognitive function, or to prevent or improve brain fatigue.

[0037] Examples of the food or drink include supplements, food and drinks, functional foods, food additives, etc. Note that the composition according to Embodiment 1 may be used as a food or drink, or may be added to food and drinks and functional foods.

[0038] The form of the supplement is not particularly limited and may be any form such as tablets, powders, granules, capsules, dragees, films, lozenges, chewables, solutions, emulsions, suspensions, etc. The supplement may contain any component usually used as a supplement.

[0039] "Functional food" means a food or beverage ingested for the purpose of maintaining health, and includes foods for specified health uses which are health functional foods, foods with function claims, nutritional functional foods, health foods, dietary supplements, etc. As the functional food, a food for specified health use or a nutritional functional food which is a health functional food is preferable. Note that when commercialized as a functional food, the food or drink may contain various additives used in foods, specifically, coloring agents, preservatives, thickening and stabilizing agents, antioxidants, bleaching agents, antibacterial and antifungal agents, souring agents, sweetening agents, seasonings, emulsifiers, fortifying agents, manufacturing agents, flavors, etc.

[0040] The functional food can be either a food or a beverage, and is not particularly limited as long as it can be ingested orally. Examples of the forms of functional foods include beverages, confectioneries, processed grains, kneaded products, dairy products, seasonings, and the like. Examples of beverages include nutritional drinks, soft drinks, black tea, green tea, and the like. Examples of confectioneries include candies, cookies, tablets, chewing gums, jelly, and the like. Examples of processed grains include bread, cooked rice, biscuits, and the like. Examples of kneaded products include sausages, hams, kamaboko, and the like. Examples of dairy products include butter, yogurt, and the like.

[0041] The food additive may be in the form of a paste, gel, powder, liquid, suspension, emulsion, granule, etc. so as to be easily added to the food.

[0042] The food or beverage according to the present embodiment may contain water, vitamins, minerals, organic acids, organic bases, fruit juices, flavors, functional components, food additives, etc. within the range of maintaining the effectiveness related to the maintenance, improvement or enhancement of cognitive function, or the prevention or improvement of brain fatigue. The food or beverage can be produced by a known method from the composition according to the above-described Embodiment 1 and, if necessary, other components other than the composition.

[0043] The content of the composition according to the above-described Embodiment 1 in the food or beverage according to the present embodiment is appropriately adjusted according to the effective amount of the food or beverage for the above-described use. When the composition contains the cells of JCM 7045, the cell content of the food or beverage is, for example, 1.0×10 2 ~1.0×10 10 cfu / g, 1.0×10 3 ~1.0×10 9 cfu / g or 1.0×10 4 ~1.0×10 8 cfu / g. From another perspective, the cell content is 1.0×10 8 cells (pcs) / g~1.0×10 13 cells (pcs) / g, 1.0×10 9 cells (pcs) / g~1.0×10 12 cells (pcs) / g or 1.0×10 10cells (pcs) / kg~1.0×10 12 It may be cells (pcs) / kg. When the composition contains a processed product of the cells of JCM 7045, the content of the processed product of the cells in the food or drink is, for example, an amount corresponding to the above cell content. Regarding the above cell content, it can also be used as a reference when considering the effective amount of the feed, preventive drug, therapeutic drug or composition for improving oral environment according to the present embodiment described later.

[0044] The food or drink may be divided and stored in one or more containers so that the daily intake amount is the above-mentioned intake amount. In this case, preferably, one container contains the food or drink for one day.

[0045] The food or drink contains the composition according to Embodiment 1 above, and is provided in a manner that can be distinguished from other products as a product in terms of being used for maintaining, improving or enhancing cognitive function, or preventing or improving brain fatigue. For example, at least one of the packaging, instruction manual and promotional materials of the product related to the food or drink indicates that it has the effect of maintaining, improving or enhancing cognitive function, or preventing or improving brain fatigue.

[0046] The food or drink may be provided or sold with the indication of the use (including health care use) for maintaining, improving or enhancing cognitive function, or preventing or improving brain fatigue. The "indication" act includes all acts for informing the consumer of the above uses. As long as it is an expression that can remind or analogize the above uses, regardless of the purpose of the indication, the content of the indication, the object to be indicated, the medium, etc., it all falls within the "indication" act.

[0047] The "indication" is preferably made by an expression that allows the consumer to directly recognize the above uses. Specifically, it includes the act of transferring, delivering, displaying for transfer or delivery, importing a product related to the food or drink or a product package with the above uses described thereon, displaying, distributing, or publishing an advertisement, price list or transaction document related to the product with the above uses described thereon, or providing information containing these by an electromagnetic (Internet, etc.) method with the above uses described thereon.

[0048] Examples of "representation" include representation as health foods, functional foods, enteral nutritional foods, special-purpose foods, health functional foods, foods for specified health use, nutritional functional foods, foods with functional claims, quasi drugs, etc. Among these, in particular, representations approved under systems related to foods for specified health use, nutritional functional foods, or foods with functional claims, or similar systems, are included. Specifically, examples include representation as a food for specified health use, representation as a conditional food for specified health use, representation indicating an impact on the structure or function of the body, disease risk reduction representation, and representation of functionality based on scientific evidence. More specifically, representation as a food for specified health use (especially representation of health uses) and similar representations are typical examples.

[0049] (Embodiment 3: Feed) In another embodiment, a feed containing the composition according to Embodiment 1 is provided. Regarding the feed according to this embodiment, the points different from the food and drink according to Embodiment 2 will be mainly described. For points not specifically mentioned about the feed, the description regarding the food and drink according to Embodiment 2 can be referred to. Note that the composition according to Embodiment 1 may also be used as a feed.

[0050] The feed is given to non-human animals. The non-human animals are preferably mammals. Mammals include, for example, primates such as chimpanzees, experimental animals such as rats, mice, and rabbits, livestock animals such as pigs, cows, horses, sheep, and goats, and pet animals such as dogs and cats. In addition to being ingested as a nutrient source necessary for non-human animals, the feed also includes supplements for non-human animals given as treats for non-human animals.

[0051] (Embodiment 4: Medicine, Medicine for Preventing Dementia, Medicine for Treating Dementia) In another embodiment, there are provided a medicament containing the composition according to Embodiment 1 above, a preventive agent for dementia, and a therapeutic agent for dementia (hereinafter sometimes collectively referred to as "medicaments, etc."). Regarding the medicaments, etc. according to this embodiment, differences from the composition according to Embodiment 1 above will mainly be described. For points not specifically mentioned regarding the medicaments, etc., the description regarding the composition according to Embodiment 1 above can be referred to. Note that the composition according to Embodiment 1 above may also be used as medicaments, etc.

[0052] The medicaments, etc. according to this embodiment may contain components other than those according to Embodiment 1 above, for example, excipients, lubricants, binders, disintegrants, solvents, solubilizing agents, suspending agents, isotonic agents, buffers, soothing agents, etc. Further, the medicament may contain additives such as preservatives, antioxidants, coloring agents, sweeteners, etc. as necessary.

[0053] Examples of the excipient include lactose, sucrose, D-mannitol, starch, crystalline cellulose, light anhydrous silicic acid, etc. Examples of the lubricant include magnesium stearate, calcium stearate, talc, colloidal silica, etc. Examples of the binder include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, etc. Examples of the disintegrant include starch, carboxymethyl cellulose, carboxymethyl cellulose calcium, croscarmellose sodium, sodium carboxymethyl starch, etc.

[0054] The solvent is, for example, water for injection, alcohol, propylene glycol, macrogol, etc. The solubilizing agent is, for example, polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate, etc. The suspending agent is a surfactant, a hydrophilic polymer, etc., and examples thereof include stearyl triethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate, polyvinyl alcohol, polyvinyl pyrrolidone, sodium carboxymethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, etc.

[0055] The isotonic agent is, for example, sodium chloride, glycerin, D-mannitol, etc. The buffer is, for example, a buffer solution of phosphate, acetate, carbonate, citrate, etc. The soothing agent is, for example, benzyl alcohol, etc. The preservative is, for example, paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, etc. The antioxidant is, for example, sulfite, ascorbic acid, etc.

[0056] The pharmaceutical according to this embodiment can be used for the treatment or prevention of diseases caused by the decline of brain function such as dementia, depression, schizophrenia, delirium, etc., in order to bring about the maintenance, improvement or amelioration of cognitive function, or the prevention or amelioration of brain fatigue.

[0057] In the preventive and therapeutic agents for dementia according to this embodiment, dementia is not particularly limited, and examples thereof include Alzheimer's disease, Lewy body dementia, frontotemporal dementia, vascular dementia, etc. Preferably, dementia is Alzheimer's disease.

[0058] (Embodiment 5: Composition for improving oral environment) JCM 7045 exhibits high co-aggregation ability against Fusobacterium nucleatum, which is a periodontal pathogen, as shown in Example 3 below. Therefore, the cells of JCM 7045 or its processed products also have an antibacterial effect against periodontal pathogens, that is, an effect of improving the oral environment. Thus, in an embodiment according to another aspect, there is provided a composition for preventing, maintaining, or improving oral functions, which contains the cells of JCM 7045 or its processed products. There is provided a composition for preventing or improving periodontal disease, which contains the cells of JCM 7045 or its processed products. In addition, there is provided a composition for inhibiting biofilm formation, which contains the cells of JCM 7045 or its processed products. Furthermore, there is provided a composition having co-aggregation ability with Fusobacterium nucleatum, which contains the cells of JCM 7045 or its processed products.

[0059] The composition according to this embodiment may further contain, in addition to the cells of JCM 7045 or its processed products as an active ingredient, components (materials) that can be used for normal oral environment improvement. For example, the compound may contain excipients, lubricants, binders, disintegrants, and the like. Furthermore, the compound may contain vitamins, minerals, dietary fibers, and the like, and may also contain fragrances, acidulants, seasonings, colorants, and the like. From the viewpoint of adjusting taste or sweetness, various saccharides may be blended in the compound, or fruit juices, dairy products, and the like may also be blended.

[0060] The composition according to this embodiment can be made into various forms as needed, such as paste, gel, liquid, solid, and the like. Specifically as forms, for example, tooth cream, tooth gel, toothpaste, mouthwash, mouth spray, tooth powder, gel, candy, gummy, gum, tablet, and the like can be mentioned. These forms can be prepared by blending the aforementioned components (materials) and the like.

[0061] In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a composition for maintaining, enhancing or improving cognitive function. In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a medicament for maintaining, enhancing or improving cognitive function. In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a composition for preventing or improving brain fatigue. In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a medicament for preventing or improving brain fatigue.

[0062] Also, in another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a food or drink for maintaining, enhancing or improving cognitive function. In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a food or drink for preventing or improving brain fatigue. In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a feed for maintaining, enhancing or improving cognitive function. In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a feed for preventing or improving brain fatigue. In another embodiment, there is provided the use of the cells of JCM 7045 or a processed product thereof for the manufacture of a prophylactic or therapeutic agent for dementia.

[0063] Also, in another embodiment, there is provided a method for maintaining, enhancing or improving cognitive function, which includes administering the cells of JCM 7045 or a processed product thereof to a subject. In another embodiment, there is provided a method for preventing or improving brain fatigue, which includes administering the cells of JCM 7045 or a processed product thereof to a subject. In another embodiment, there is provided a method for preventing or treating dementia, which includes administering the cells of JCM 7045 or a processed product thereof to a subject.

[0064] In another embodiment, the cells of JCM 7045 or a processed product thereof for use in maintaining, enhancing or improving cognitive function are provided. In another embodiment, the cells of JCM 7045 or a processed product thereof for use in preventing or improving brain fatigue are provided. In another embodiment, the cells of JCM 7045 or a processed product thereof for use in preventing or treating dementia are provided.

[0065] The present invention will be described in more detail by the following examples, but the present invention is not limited by the examples.

Example

[0066] [Example 1: in vitro anti-inflammatory screening] Neuroinflammation due to aging etc. is known to be a factor in the decline of cognitive function. In order to efficiently select Bifidobacterium that improves cognitive function, an anti-inflammatory evaluation was carried out. As Bifidobacterium that improves cognitive function (test subjects), JCM 7045, JCM 1251 and JCM 1275, which are Bifidobacterium adolescentis T (obtained from RIKEN BRC, RIKEN BioResource Center) were used.

[0067] Each test subject used for the anti-inflammatory evaluation was cultured at 37°C using AnaeroPack (trademark) (manufactured by Mitsubishi Gas Chemical Company) for a total of 5 days including one subculture. The medium shown in Table 1 was used for the culture. Thereafter, the collected cells were washed twice with physiological saline (manufactured by Otsuka Pharmaceutical Factory, Inc.), and dry bacterial powder was prepared using a freeze dryer.

[0068]

Table 1

[0069] The MG6 cells, a mouse microglia cell line (RCB2403, obtained from the RIKEN BioResource Center; it can also be produced with reference to, for example, Neurosci Lett 2006 407(3):205-10 or Biochim Biophys Acta 2005 1726(2):177-86), were cultured at 37°C under 5% CO 2 conditions using Dulbecco’s Modified Eagle Medium (D-MEM) (High Glucose) (manufactured by Fujifilm Wako Pure Chemical Corporation) containing 10% fetal bovine serum (manufactured by MP Biomedicals Japan) and 1% penicillin-streptomycin solution (×100) (manufactured by Fujifilm Wako Pure Chemical Corporation).

[0070] After detaching the cultured MG6 cells with 0.05% trypsin (manufactured by Gibco), they were seeded in a 96-well plate (manufactured by Corning) at a concentration of 1×10 4 cells / well and cultured overnight at 37°C under 5% CO 2 conditions. For the tests, cells with passage numbers of 10 or less were used.

[0071] After removing the medium from each well, D-MEM (High Glucose) containing 2% fetal bovine serum and 1% penicillin-streptomycin solution (×100) was added to the control group and the inflammation induction group. To the positive control group, D-MEM (High Glucose) containing SB239063 (manufactured by Tokyo Chemical Industry), a p38 MAPK inhibitor dissolved in dimethyl sulfoxide (manufactured by Sigma-Aldrich), at a final concentration of 1.25 μM, along with 2% fetal bovine serum and 1% penicillin-streptomycin solution (×100), was added. To the test substance addition group, each test substance was suspended in D-PBS(-) (manufactured by Fujifilm Wako Pure Chemical Corporation), irradiated with ultraviolet (UV) light for 15 minutes, and D-MEM (High Glucose) containing the irradiated test substance at a final concentration of 1 μg / mL, along with 2% fetal bovine serum and 1% penicillin-streptomycin solution (×100), was added. All groups were cultured for 24 hours.

[0072] After culturing, as an inflammation-inducing component, lipopolysaccharide (LPS, manufactured by Sigma-Aldrich) dissolved in D-PBS(-) was added to groups other than the control group at a final concentration of 1 μg / mL, and the cells were cultured for 6 hours. A part of the culture medium was collected, and the concentration of TNF-α in the culture medium was measured using Mouse TNF-alpha DuoSet ELISA (manufactured by R&D systems).

[0073] (Results) As shown in Fig. 1, among the test subjects, JCM 7045 significantly suppressed the secretion of TNF-α induced by LPS statistically, showing the highest anti-inflammatory effect. The significant difference was calculated by the Dunnett method against the inflammation-inducing group.

[0074] [Example 2: In vivo test] The effect of JCM 7045 on improving cognitive function, for which an anti-inflammatory effect had been confirmed, was evaluated using Alzheimer's disease model mice prepared by the following procedure. Slc:ddY male mice (manufactured by Japan SLC) were used as the animals. For administration, a syringe (manufactured by Terumo) equipped with an oral sonde for mice (manufactured by Fuchigami Kikai) was used, and oral administration was performed once a day for 15 days. The dose volume was calculated at 10 mL / kg based on the body weight on the administration day for the positive control group, and at 200 μL / head for the other groups. Here, "head" refers to the body weight of the above model mice, which is approximately 30 g to 45 g. JCM 7045 was administered to the test subject group at 1.2×10 8 cfu / head (total number of bacteria 3.8×10 10 cells / head). Bifidobacterium obtained by a known method commonly used in this field from a supplement known to have an effect on improving cognitive function (product name: Memory Bifidus, manufactured by Morinaga Milk Industry Co., Ltd.) was administered to the comparative control group at 1.0×10 9 cfu / head (total number of bacteria 1.2×10 11It was administered by intracerebroventricular injection (cells / head). Donepezil hydrochloride (trade name: Aricept, manufactured by Eisai Co., Ltd.), an existing therapeutic drug for dementia, was administered to the positive control group at a dose of 0.5 mg / kg. In addition, the sham operation group and the vehicle control group were administered the vehicle of JCM 7045 (cryoprotectant: physiological saline = 1:9).

[0075] On the third day after administration, β-amyloid 1-42 (manufactured by Peptide Institute, Inc.) was administered once intracerebroventricularly to the groups other than the sham operation group, and physiological saline was administered once intracerebroventricularly to the sham operation group. For anesthesia during intracerebroventricular injection of β-amyloid 1-42, first, sodium pentobarbital (manufactured by Tokyo Chemical Industry Co., Ltd.) 40 mg / kg was administered intraperitoneally, and then, levobupivacaine hydrochloride (manufactured by Maruishi Pharmaceutical Co., Ltd.) 0.1 mL was administered subcutaneously. Thereafter, the hair on the top of the mouse's head was shaved, and the head was fixed to a stereotaxic apparatus. After disinfecting the top of the head with isodine and making an incision to expose the skull, the connective tissue on the skull was removed with a cotton swab and then dried with a blower to make the position of bregma visible. Using a dental drill, a hole for inserting a stainless steel pipe was made in the skull 1 mm lateral (right side) and 0.2 mm posterior to bregma, and a stainless steel pipe connected to a silicon tube with an outer diameter of 0.5 mm and a microsyringe was vertically inserted to a depth of 2.5 mm from the bone surface. 3 μL (200 pmol / 3 μL) of β-amyloid solution (physiological saline for the sham operation group) was injected into the cerebral ventricle over 3 minutes using a microsyringe pump. After injection, it was left standing for 3 minutes with the stainless steel pipe inserted, and the stainless steel pipe was slowly removed. The cranial hole was closed with a non-absorbable bone marrow hemostatic agent (Nestop (trademark), manufactured by Alpharesa Pharma Co., Ltd.), the scalp was sutured, then removed from the stereotaxic apparatus, and the mouse was returned to the breeding cage.

[0076] As a test for learning and memory impairment, the passive avoidance test was conducted on the 10th to 11th days after administration. For the passive avoidance test, a step-through type passive avoidance response apparatus (light-dark box: manufactured by Nippon Bio-Research Center, SHOCK SCRAMBLER, manufactured by Unicom) equipped with a light chamber partitioned by a guillotine door in the center and a dark chamber that applies an electric stimulus from the grit on the floor was used. The mouse was placed in the light chamber, and after 10 seconds, the guillotine door was gently opened, and the time until the mouse entered the dark chamber (latency) was measured. In the acquisition trial (10th day after administration), when the mouse entered the dark chamber, the guillotine door was closed immediately, and an electric stimulus (0.2 mA, 2 seconds, scrambled method) was applied, and the presence or absence of the animal's vocalization during the electric stimulus load was confirmed. The measurement of the latency was up to 300 seconds at most. In the retention trial (11th day after administration), the trial ended when the animal entered the dark chamber or 300 seconds had elapsed in the light chamber.

[0077] (Results) Figure 2 shows the latency of the retention trial in the passive avoidance test. The vehicle control group showed a statistically significant shortening of the latency in the retention trial compared to the sham-operated group, and long-term memory impairment was observed due to a single intracerebroventricular administration of β-amyloid 1-42. On the other hand, the latency of the retention trial in the test substance group was statistically significantly prolonged, and improvement of long-term memory impairment was observed. In addition, the latency of the retention trial in the test substance group was comparable to that of the comparative control group, which is already known to have a cognitive function improving effect. From these results, it was shown that JCM 7045 has a cognitive function improving effect, particularly an improving effect on long-term memory impairment, for example. The significant difference was calculated by the Dunnett method with respect to the vehicle control group.

[0078] [Example 3: Coaggregation test] To explore further effects of JCM 7045, the coaggregation ability with periodontal pathogens was examined. As periodontal pathogens, Fusobacterium nucleatum JCM 8532 obtained from RIKEN BRC T was used. Anaeropack (trademark) was used with modified GAM broth (manufactured by Nissui Pharmaceutical Co., Ltd.), and the periodontal pathogens were cultured at 37°C for 6 days. The test substances used were JCM 7045, JCM 7046, and JCM 1275, which are Bifidobacterium adolescentisT An Aneropack (trademark) was used with GAM broth (manufactured by Nissui Pharmaceutical Co., Ltd.), and the test specimens were cultured overnight at 37°C.

[0079] After collecting each bacterial cell, it was washed once with the buffer solution prepared as shown in Table 2, and then resuspended in the same buffer solution so that the absorbance (OD 600 ) reached a certain value.

[0080]

Table 2

[0081] A suspension of only the periodontal pathogen or each test specimen (single sample) and a suspension in which each test specimen was mixed in equal amounts with the periodontal pathogen (mixed sample) were prepared. After stirring with a vortex mixer, they were incubated at 110 rpm and 37°C for 30 minutes. Then, the upper layer of the suspension that had been allowed to stand at room temperature for 3 minutes or more was collected, and the absorbance (OD 600 ) was measured. Let the sum of the OD 600 of the single sample of the periodontal pathogen and the target test specimen be St, and the OD 600 of the mixed sample be Sm. The co-aggregation rate was calculated using the following formula 1.

[0082]

Equation

[0083] (Results) Figure 3 shows the co-aggregation rate between the periodontal pathogen and each test specimen. Among the test specimens, JCM 7045 showed a high co-aggregation ability, indicating the possibility of efficiently removing the periodontal pathogen from the oral cavity. From this result, it was shown that JCM 7045 has not only an effect of improving cognitive function but also a co-aggregation ability with the periodontal pathogen, that is, an effect of improving the oral environment.

[0084] The above-described embodiments are for explaining the present invention and do not limit the scope of the present invention. That is, the scope of the present invention is indicated not by the embodiments but by the claims. And various modifications made within the scope of the claims and within the scope of the meaning of the invention equivalent thereto are considered to be within the scope of the present invention.

[0085] This application is based on Japanese Patent Application No. 2024-055911, filed on March 29, 2024. The entire specification, claims, and drawings of Japanese Patent Application No. 2024-055911 are incorporated herein by reference.

Industrial Applicability

[0086] The present invention is useful as a composition, food or drink, feed, and medicine.

Claims

1. Bifidobacterium adolescentis JCM 7045 strain cells or a processed product thereof. A composition for maintaining, enhancing or improving cognitive function.

2. The cognitive function is It is a function related to memory, The composition of claim 1.

3. The memory-related function is Long-term memory function, The composition of claim 2.

4. Bifidobacterium adolescentis JCM 7045 strain cells or a processed product thereof. A composition for preventing or improving brain fatigue.

5. 4. A composition comprising the composition of any one of claims 1 to 3. Food and drink for maintaining, enhancing or improving cognitive function.

6. 5. The composition of claim 4, Food and drink for preventing or improving brain fatigue.

7. Bifidobacterium adolescentis JCM 7045 strain cells or a processed product thereof. Feed for maintaining, enhancing or improving cognitive function, or feed for preventing or improving brain fatigue.

8. 4. A composition comprising the composition of any one of claims 1 to 3. A medicine for maintaining, enhancing or improving cognitive function.

9. 5. The composition of claim 4, A medicine for preventing or improving brain fatigue.

10. Bifidobacterium adolescentis JCM 7045 strain cells or a processed product thereof. A preventive or therapeutic drug for dementia.

11. The dementia is Alzheimer's disease, The prophylactic or therapeutic agent according to claim 10.

Images