Composition, food or beverage, feed and medicine for maintaining, improving or ameliorating cognitive function; prophylactic agent and therapeutic agent for dementia; composition, food or beverage, feed and medicine for preventing or ameliorating brain fatigue; and composition

Abstract

Provided is a composition for maintaining, improving or ameliorating a cognitive function, the composition containing cells of Bifidobacterium adolescentis JCM 7045 strain or a treated product of the cells.

Description

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

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

[0002] According to the World Health Organization, it is estimated that there are more than 55 million dementia patients worldwide, and it has been suggested that approximately 10 million new cases of dementia may develop each year (see Non-Patent Document 1). It is known that dementia causes cognitive decline and other symptoms due to accumulated brain damage caused by nerve cells destroyed by chronic inflammation that occurs with aging.

[0003] In Japan, it is estimated that approximately 16% of people aged 65 or older suffer from dementia. The proportion of dementia patients is expected to continue to rise. With Japan's super-aging population, there are concerns that the increase in dementia patients will exacerbate social problems, particularly the shortage of caregivers and the pressure on health insurance costs.

[0004] The most common type of dementia is Alzheimer's disease (AD), accounting for 60 to 70% of dementia patients. Common therapeutic approaches for dementia and related diseases include the administration of drugs such as cholinesterase inhibitors such as donepezil, NMDA receptor antagonists such as memantine, and selective serotonin inhibitors (see Non-Patent Document 2). Furthermore, antibody drugs such as lecanemab have also been actively developed in recent years.

[0005] “Dementia”, [online], March 15, 2023, World Health Organization, [Retrieved January 17, 2024], Internet <URL: https: / / www.who.int / news-room / fact-sheets / detail / dementia> Shin Kitamura, “The latest treatments for dementia, particularly Alzheimer’s disease”, Journal of the Japan Medical University Medical Association, Japan Medical University Medical Association, 2012, Vol. 8, No. 4, pp. 291-295

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

[0007] The present invention has been made in view of the above circumstances, and aims to provide a composition that can be easily incorporated into daily life and that has an excellent effect of improving cognitive function, as well as compositions, foods and drinks, feeds and medicines for maintaining, enhancing or improving cognitive function, and preventive and therapeutic drugs for dementia.

[0008] Another object of the present invention is to provide a composition that can be easily incorporated into daily life and has an excellent effect of improving brain fatigue, as well as a composition, food, drink, feed, and medicine for preventing or improving brain fatigue.

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

[0010] The cognitive function may be a memory-related function.

[0011] The memory-related function may be a long-term memory function.

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

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

[0014] A food or drink for preventing or improving brain fatigue according to a fourth aspect of the present invention comprises the composition according to the second aspect of the present invention.

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

[0016] A pharmaceutical for maintaining, enhancing or improving cognitive function according to a sixth aspect of the present invention comprises the composition according to the first aspect of the present invention.

[0017] A pharmaceutical for preventing or ameliorating brain fatigue according to a seventh aspect of the present invention comprises the composition according to the second aspect of the present invention.

[0018] A preventive or therapeutic agent for dementia according to an eighth aspect of the present invention comprises Bifidobacterium adolescentis JCM 7045 strain cells or a processed product thereof.

[0019] The dementia may be Alzheimer's disease.

[0020] A composition according to a ninth aspect of the present invention comprises bacterial cells of Bifidobacterium adolescentis JCM 7045 strain or a processed product thereof, wherein the content of the bacterial cells or the processed product thereof is 1.0 x 10 ８ cells / g~1.0×10 １３ cells / g.

[0021] The compositions, foods, drinks, feeds, and medicines, as well as preventive and therapeutic drugs for dementia, according to the present invention, can be easily incorporated into daily life and provide excellent effects in improving cognitive function. Furthermore, the compositions, foods, drinks, feeds, and medicines according to the present invention can be easily incorporated into daily life and provide excellent effects in improving brain fatigue. Furthermore, the compositions, foods, drinks, feeds, and medicines, as well as preventive and therapeutic drugs for dementia, according to the present invention, also have high coaggregation ability with, for example, Fusobacterium nucleatum, a periodontal disease bacterium.

[0022] Fig. 1 is a diagram showing the concentration of TNF-α in a medium according to Example 1. Fig. 2 is a diagram showing the reaction latency of a retention trial in a passive avoidance test according to Example 2. Fig. 3 is a diagram showing the coaggregation rate with periodontal disease bacteria according to Example 3.

[0023] Embodiments of the present invention will be described with reference to the drawings. Note that the present invention is not limited to the following embodiments and drawings. Note that in the following embodiments, the expressions "have," "include," or "contain" also include the meaning of "consisting of" or "consisting of."

[0024] (Embodiment 1: Composition) The composition according to this embodiment contains cells of Bifidobacterium adolescentis JCM 7045 strain (hereinafter simply referred to as "JCM 7045") or a processed product thereof. JCM 7045 is available from the RIKEN BioResource Research Center (RIKEN BRC). JCM 7045 is preferably cultured at or near the optimum temperature, 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 either a liquid medium or an agar medium. Preferably, bifidobacteria can be cultured under anaerobic conditions, for example, by aerating with anaerobic gas such as carbon dioxide. Alternatively, they may be cultured under microaerobic conditions, such as liquid static culture. The medium for culturing JCM 7045 is not particularly limited, and any medium commonly used for culturing bacteria belonging to the genus Bifidobacterium can be used.

[0025] Examples of carbon sources that can be used in the culture include sugars such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolysates, and blackstrap molasses. Examples of nitrogen sources that can be used include ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, and nitrates. Examples of inorganic salts that can be used include sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate. Organic components such as peptone, soybean flour, defatted soybean meal, meat extract, and yeast extract may also be used. For example, the JCM 7045 medium is a BL medium containing nutrient broth.

[0026] The bacterial cells can be obtained as a precipitate by centrifugation of the culture medium after culture. The bacterial cells may be live or dead, and may be wet or dried. The processed bacterial cells are, for example, freeze-dried bacterial cells, bacterial cells dried with acetone, extracts obtained by subjecting bacterial cells to solvent extraction, and crushed products obtained by crushing bacterial cells or dried bacterial cells with ultrasound or the like, bacterial cells obtained by subjecting bacterial cells to some treatment, or compositions derived from bacterial cells or containing parts of bacterial cells. The bacterial cells, compositions derived from bacterial cells, or containing parts of bacterial cells are, for example, culture supernatants or culture media. Therefore, processed bacterial cells also include culture supernatants and culture media.

[0027] The solvent used in the solvent extraction treatment can be water, an organic solvent, or a mixture thereof. Examples of organic solvents include ether, chloroform, benzene, hexane, methanol, ethanol, isopropanol, and mixtures thereof. The extract extracted using the extraction solvent can be used as is, or after concentration or dilution, in the form of a liquid, gel, or paste. Furthermore, this can be dried and used as a dried product. Drying can be carried out by known methods such as spray drying, freeze drying, vacuum drying, and fluidized bed drying.

[0028] After culturing JCM 7045, the resulting bacterial cells may be used as they are, or may be diluted or concentrated before use. The treated bacterial cells may also be used as they are, or may be diluted or concentrated before use.

[0029] The composition according to this embodiment contains JCM 7045 cells or a processed product thereof as an active ingredient, and may contain other ingredients in addition to the active ingredient. The other ingredients are not particularly limited and may be additives contained in pharmaceuticals. Furthermore, when a culture supernatant or a medium is used as the processed product of the cells, the composition may contain, for example, a component derived from the medium as an ingredient in addition to the active ingredient.

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

[0031] The composition according to this embodiment may be used to prevent or improve brain fatigue. Brain fatigue refers to a state in which the brain is continuously subjected to excessive stress, resulting in impaired brain function. Fatigue is thought to occur when the brain's natural functions are impaired as a result of a chronic decrease in immune cytokines or the induction of inflammatory cytokines due to stress or other factors (Japanese Journal of Biological Psychiatry 24(4):200-210, 2013). Brain fatigue can be improved, for example, by suppressing the induction of inflammatory cytokines. 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 preventing or improving 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 JCM 7045 cells or a processed product thereof as an active ingredient. The content of JCM 7045 cells or a processed product thereof in the composition according to this embodiment may be adjusted appropriately depending on the effective amount of the composition for the above-mentioned use. An effective amount is the amount necessary to obtain the desired result, i.e., the amount necessary to delay, inhibit, prevent, reverse, or cure the progression of the condition being treated or treated. When the active ingredient is JCM 7045 cells, the effective amount is, for example, 1.0 x 10 ２ ~1.0 x 10 １０ Colony forming units (cfu) / kg, 1.0 x 10 ３ ~1.0 x 10 ９ cfu / kg or 1.0 x 10 ４ ~1.0 x 10 ８ cfu / kg. From another perspective, the effective amount is 1.0 x 10 ８ cells (pieces) / kg ~ 1.0×10 １３ cells / kg, 1.0×10 ９ cells (pieces) / kg ~ 1.0×10 １２ cells / kg or 1.0 x 10 １０ cells (pieces) / kg ~ 1.0×10 １２ The effective amount of the bacterial cell count may be 1000 cells / kg. When a processed product of JCM 7045 bacterial cells is used as the active ingredient, the processed product of the bacterial cells is, for example, an amount equivalent to an effective amount of the bacterial cells. The method for measuring (measuring) the number of bacteria may be any method commonly used in this field, and specific examples include fluorescent (DAPI) staining and plate smear methods.

[0033] The composition of this embodiment is administered to humans and non-human animals. The animal is preferably a mammal, more specifically, dogs, cats, cows, pigs, horses, sheep, deer, etc. The dosage of the composition is determined appropriately based on the gender, age, weight, symptoms, etc. of the recipient. The composition is administered so that an effective amount of JCM 7045 cells or a processed product thereof is obtained. The composition can be administered once a day or in divided doses. The composition may be administered at various frequencies, such as daily, every other day, once a week, every other week, or once a month. If necessary, the composition may be used in an amount outside the above range. The method of administration of the composition is not particularly limited, but may be via injection, nasal administration, transdermal administration, pulmonary administration, oral administration, etc. Oral administration is particularly preferred.

[0034] The composition according to the present embodiment contains cells of the bifidobacterium JCM 7045 or a processed product thereof, making it easy to incorporate into daily life and providing excellent cognitive function improvement effects.

[0035] A composition according to another aspect of the present embodiment includes JCM 7045 strain cells or a processed product thereof, and the content of the cells or the processed product thereof is 1.0 × 10 ８ cells / g~1.0×10 １３ cells / g or 1.0 x 10 ９ cells (pieces) / kg ~ 1.0×10 １２ cells / kg.

[0036] (Embodiment 2: Food and Drink) In another embodiment, there is provided a food and drink containing the composition according to the above-mentioned embodiment 1. The food and drink is used for maintaining, enhancing, or improving cognitive function, or for preventing or improving brain fatigue.

[0037] Examples of foods and beverages include supplements, foods and beverages, functional foods, food additives, etc. The composition according to the first embodiment may be used as foods and beverages, or may be added to foods and beverages and functional foods.

[0038] The form of the supplement is not particularly limited and may be any form such as tablets, powders, granules, capsules, sugar-coated tablets, films, lozenges, chewable tablets, solutions, emulsions, suspensions, etc. The supplement may contain any ingredient typically used as a supplement.

[0039] "Functional foods" refer to foods or beverages consumed for the purpose of maintaining health, and include foods with health claims, foods with specified health uses, foods with functional claims, foods with nutrient functions, health foods, and nutritional supplements. Functional foods are preferably foods with health claims or foods with nutrient functions. When commercialized as functional foods, the foods and beverages may contain various additives used in foods, specifically colorants, preservatives, thickening agents, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, sweeteners, seasonings, emulsifiers, strengthening agents, manufacturing agents, flavorings, etc.

[0040] Functional foods may be either foods or beverages, and are not particularly limited as long as they can be taken orally. Examples of functional foods include beverages, confectioneries, processed grain products, paste products, dairy products, seasonings, etc. Examples of beverages include nutritional drinks, soft drinks, black tea, green tea, etc. Examples of confectioneries include candy, cookies, tablet candy, chewing gum, jelly, etc. Examples of processed grain products include bread, cooked rice, biscuits, etc. Examples of paste products include sausages, ham, kamaboko, etc. Examples of dairy products include butter, yogurt, etc.

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

[0042] The food and drink according to this embodiment may contain water, vitamins, minerals, organic acids, organic bases, fruit juice, flavors, functional ingredients, food additives, etc., within the range that maintains the effectiveness in maintaining, improving, or ameliorating cognitive function, or preventing or ameliorating brain fatigue. The food and drink can be produced by a known method from the composition according to the first embodiment above and, if necessary, other ingredients other than the composition.

[0043] The content of the composition according to the first embodiment in the food or drink according to the present embodiment is adjusted appropriately depending on the effective amount for the intended use of the food or drink. When the composition contains JCM 7045 bacteria, the bacterial content of the food or drink is, for example, 1.0 × 10 ２ ~1.0 x 10 １０ cfu / g, 1.0×10 ３ ~1.0 x 10 ９ cfu / g or 1.0 x 10 ４ ~1.0 x 10 ８ From another perspective, the bacterial cell content is 1.0 × 10 ８ cells / g~1.0×10 １３ cells / g, 1.0×10 ９ cells / g~1.0×10 １２ cells / g or 1.0 x 10 １０ cells (pieces) / kg ~ 1.0×10 １２ The content of the processed bacterial cells in the food or drink may be, for example, an amount equivalent to the above bacterial cell content. The above bacterial cell content can also be used as a reference when determining the effective amount of a feed, a preventive drug, a therapeutic drug, or a composition for improving oral environment according to the present embodiment, which will be described later.

[0044] The food and drink may be divided and stored in one or more containers so that the daily intake is the amount mentioned above, and in this case, it is preferable that one container stores one day's worth of food and drink.

[0045] The food and drink contains the composition according to the first embodiment and is provided in a form that can be distinguished from other products as a product in that it is used for maintaining, enhancing, or improving cognitive function, or for preventing or improving brain fatigue. For example, at least one of the packaging, instructions, and promotional materials for the food and drink product indicates that it has the effect of maintaining, enhancing, or improving cognitive function, or for preventing or improving brain fatigue.

[0046] Food and beverages may be offered or sold with a label indicating their intended use (including health uses) for maintaining, enhancing, or improving cognitive function, or for preventing or alleviating brain fatigue. "Labeling" includes all acts intended to inform consumers of the above-mentioned uses. Any expression that can evoke or infer the above-mentioned uses is considered a "labeling" act, regardless of the purpose, content, object, or medium of the labeling.

[0047] It is preferable that the "labeling" be done in an expression that allows consumers to directly recognize the intended use. Specifically, this includes acts of transferring, delivering, displaying for the purpose of transferring or delivering, or importing food and drink products or product packaging that lists the intended use, displaying or distributing advertisements, price lists, or transaction documents that list the intended use, or providing information containing the above-mentioned content by electromagnetic means (such as the Internet).

[0048] "Labeling" includes labeling as health food, functional food, enteral nutritional food, special dietary food, health functional food, food for specified health uses, food with nutrient functions, food with functional claims, quasi-drugs, etc. Among these, labeling approved under systems related to foods for specified health uses, foods with nutrient functions, or foods with functional claims, or similar systems, is particularly included. Specific examples include labeling as a food for specified health uses, labeling as a conditional food for specified health uses, labeling that indicates an effect on the structure or function of the body, labeling that reduces disease risk, and labeling of functionality based on scientific evidence. More specifically, labeling as a food for specified health uses (especially labeling of health uses) and similar labeling are typical examples.

[0049] (Embodiment 3: Feed) In another embodiment, a feed containing the composition according to the above-mentioned embodiment 1 is provided. The feed according to this embodiment will be described mainly in terms of the differences from the food and drink according to the above-mentioned embodiment 2. For points not specifically mentioned about this feed, the description of the food and drink according to the above-mentioned embodiment 2 can be referred to. Note that the composition according to the above-mentioned embodiment 1 may also be used as 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, laboratory 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 necessary nutritional source by non-human animals, feed also includes non-human animal supplements that are given to non-human animals as a luxury item.

[0051] (Embodiment 4: Medicine, dementia preventive medicine, dementia therapeutic medicine) In another embodiment, a medicine, dementia preventive medicine, and dementia therapeutic medicine (hereinafter sometimes collectively referred to as "medicine, etc.") containing the composition according to embodiment 1 above are provided. The medicine, etc. according to this embodiment will be mainly described in terms of differences from the composition according to embodiment 1 above. For points not specifically mentioned about the medicine, etc., the description of the composition according to embodiment 1 above can be referenced. Note that the composition according to embodiment 1 above may be used as a medicine, etc.

[0052] The medicament etc. according to this embodiment may contain components other than those according to the above-mentioned embodiment 1, such as excipients, lubricants, binders, disintegrants, solvents, solubilizers, suspending agents, isotonicity agents, buffers, soothing agents, etc. Furthermore, the medicament may contain additives such as preservatives, antioxidants, colorants, sweeteners, etc., as needed.

[0053] Examples of excipients include lactose, sucrose, D-mannitol, starch, crystalline cellulose, light anhydrous silicic acid, etc. Examples of lubricants include magnesium stearate, calcium stearate, talc, colloidal silica, etc. Examples of binders include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, etc. Examples of disintegrants include starch, carboxymethylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, carboxymethylstarch sodium, etc.

[0054] Examples of solvents include water for injection, alcohol, propylene glycol, macrogol, etc. Examples of solubilizing agents include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate, etc. Examples of suspending agents include surfactants, hydrophilic polymers, etc., such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glycerin monostearate, polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, etc.

[0055] Examples of isotonic agents include sodium chloride, glycerin, D-mannitol, etc. Examples of buffers include phosphate, acetate, carbonate, citrate buffer solutions, etc. Examples of soothing agents include benzyl alcohol, etc. Examples of preservatives include paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, etc. Examples of antioxidants include sulfites, ascorbic acid, etc.

[0056] The medicine of this embodiment maintains, enhances, or improves cognitive function, or prevents or improves brain fatigue, and can therefore be used to treat or prevent diseases caused by decreased brain function, such as dementia, depression, schizophrenia, and delirium.

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

[0058] (Embodiment 5: Composition for Improving Oral Environment) As shown in Example 3 below, JCM 7045 exhibits high coaggregation ability with the periodontal disease bacterium Fusobacterium nucleatum. Therefore, JCM 7045 cells or a processed product thereof have antibacterial activity against periodontal disease bacteria, i.e., the effect of improving the oral environment. Therefore, in another embodiment, a composition for preventing, maintaining, or improving oral function is provided, which comprises JCM 7045 cells or a processed product thereof. A composition for preventing or improving periodontal disease is provided, which comprises JCM 7045 cells or a processed product thereof. Also provided is a composition for inhibiting biofilm formation, which comprises JCM 7045 cells or a processed product thereof. Furthermore, a composition having coaggregation ability with Fusobacterium nucleatum, which comprises JCM 7045 cells or a processed product thereof, is provided.

[0059] The composition according to the present embodiment may further contain, in addition to the JCM 7045 bacteria or a processed product thereof as an active ingredient, other ingredients (materials) that can be used for general oral environment improvement. For example, the compound may contain excipients, lubricants, binders, disintegrants, etc. Furthermore, the compound may contain vitamins, minerals, dietary fiber, etc., and may also contain flavorings, acidulants, seasonings, colorants, etc. From the viewpoint of adjusting taste or sweetness, the compound may be blended with various sugars, fruit juice, dairy products, etc.

[0060] The composition according to the present embodiment can be in various forms, such as paste, gel, liquid, solid, etc., as needed. Specific examples of the form include tooth cream, tooth gel, tooth paste, mouthwash, mouth spray, tooth powder, gel, candy, gummy, chewing gum, tablet, etc. These forms can be prepared by blending the above-mentioned components (materials), etc.

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

[0062] In another embodiment, there is provided the use of JCM 7045 fungus cells or a processed product thereof for the production of food and beverages for maintaining, improving, or ameliorating cognitive function. In another embodiment, there is provided the use of JCM 7045 fungus cells or a processed product thereof for the production of food and beverages for preventing or ameliorating brain fatigue. In another embodiment, there is provided the use of JCM 7045 fungus cells or a processed product thereof for the production of feed for maintaining, improving, or ameliorating cognitive function. In another embodiment, there is provided the use of JCM 7045 fungus cells or a processed product thereof for the production of feed for preventing or ameliorating brain fatigue. In another embodiment, there is provided the use of JCM 7045 fungus cells or a processed product thereof for the production of a preventive or therapeutic drug for dementia.

[0063] In another embodiment, there is provided a method for maintaining, improving, or ameliorating cognitive function, which comprises administering JCM 7045 bacterial cells or a processed product thereof to a subject. In another embodiment, there is provided a method for preventing or ameliorating brain fatigue, which comprises administering JCM 7045 bacterial cells or a processed product thereof to a subject. In another embodiment, there is provided a method for preventing or treating dementia, which comprises administering JCM 7045 bacterial cells or a processed product thereof to a subject.

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

[0065] The present invention will be explained in more detail with reference to the following examples, but the present invention is not limited to these examples.

[0066] Example 1: In vitro anti-inflammatory screening Neuroinflammation due to aging and other factors is known to be a factor in the decline of cognitive function. To efficiently select bifidobacteria that improve cognitive function, anti-inflammatory evaluation was performed. Bifidobacterium adolescentis JCM 7045, JCM 1251, and JCM 1275 were used as bifidobacteria (subjects) that improve cognitive function. Ｔ (obtained from RIKEN BioResource Research Center; RIKEN BRC) was used.

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

[0068]

[0069] MG6 cells (RCB2403, obtained from RIKEN BioResource Research Center; RIKEN BRC; MG6 cells can also be produced by referring to, for example, Neurosci Lett 2006 407(3):205-10 or Biochim Biophys Acta 2005 1726(2):177-86), a mouse microglial cell line, were cultured in Dulbecco's Modified Eagle Medium (D-MEM) (High Glucose) (Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum (MP Bio Japan) and 1% penicillin-streptomycin solution (x100) (Fujifilm Wako Pure Chemical Industries, Ltd.) at 37°C and 5% CO ２ The cells were cultured under the conditions.

[0070] The cultured MG6 cells were detached with 0.05% trypsin (Gibco) and then cultured at 1 × 10 ４ The cells were seeded in a 96-well plate (Corning) at a density of 1000 cells / well and incubated at 37°C, 5% CO ２ The cells were cultured overnight under the conditions described above. The cells used in the test were passaged 10 times or less.

[0071] After removing the medium from each well, D-MEM (High Glucose) supplemented with 2% fetal bovine serum and 1% penicillin-streptomycin solution (100x) was added to the control and inflammation-induced groups. To the positive control group, D-MEM (High Glucose) containing 2% fetal bovine serum and 1% penicillin-streptomycin solution (100x) and containing the p38 MAPK inhibitor SB239063 (Tokyo Chemical Industry Co., Ltd.) dissolved in dimethyl sulfoxide (Sigma-Aldrich) at a final concentration of 1.25 μM was added. For the test sample addition group, each test sample was suspended in D-PBS(-) (Fujifilm Wako Pure Chemical Industries, Ltd.) and irradiated with ultraviolet (UV) light for 15 minutes. Then, D-MEM (High Glucose) containing 2% fetal bovine serum and 1% penicillin-streptomycin solution (x100) containing each test sample at a final concentration of 1 μg / mL was added. All groups were cultured for 24 hours.

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

[0073] (Results) As shown in Figure 1, among the subjects, JCM 7045 statistically significantly inhibited LPS-induced TNF-α secretion and exhibited the highest anti-inflammatory effect. The significance of the difference was calculated using the Dunnett method relative to the inflammation-induced group.

[0074] [Example 2: In Vivo Test] The cognitive function-improving effect of JCM 7045, whose anti-inflammatory effect was confirmed, was evaluated in Alzheimer's disease model mice prepared according to the following procedure. Male Slc:ddY mice (manufactured by Japan SLC) were used as animals. Administration was performed orally once daily for 15 days using a syringe (manufactured by Terumo Corporation) equipped with a mouse oral probe (manufactured by Fuchigami Kikai). The volume of the administered solution was calculated at 10 mL / kg based on the body weight on the day of administration for the positive control group, and 200 μL / head for the other groups. Here, "head" refers to the body weight of the model mouse, approximately 30 g to 45 g. The subject group received 1.2 x 10 ８ cfu / head (total number of bacteria 3.8 x 10 １０ The control group was administered 1.0 × 10 bifidobacteria obtained by a known method commonly used in this field from a supplement known to have an effect of improving cognitive function (trade name: Memory Bifidus, manufactured by Morinaga Milk Industry Co., Ltd.). ９ cfu / head (total number of bacteria 1.2 x 10 １１ The positive control group received 0.5 mg / kg of donepezil hydrochloride (trade name: Aricept, manufactured by Eisai Co., Ltd.), an existing dementia treatment drug. The sham-operated group and vehicle control group received JCM 7045 vehicle (cryoprotectant: physiological saline = 1:9).

[0075] On the third day of administration, β-amyloid 1-42 (Peptide Institute) was administered intracerebroventricularly to all groups except the sham-operated group, and saline was administered intracerebroventricularly to the sham-operated group. For anesthesia during intracerebroventricular injection of β-amyloid 1-42, 40 mg / kg of pentobarbidal sodium (Tokyo Chemical Industry Co., Ltd.) was administered intraperitoneally, followed by 0.1 mL of levobupivacaine hydrochloride (Maruishi Pharmaceutical Co., Ltd.). The hair on the top of the mouse's head was then shaved, and the head was fixed in a stereotaxic apparatus. The top of the head was disinfected with isodine and then incised 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 bregma easier to visualize. A hole for inserting a stainless steel pipe was drilled into the skull 1 mm lateral (right side) and 0.2 mm posterior to bregma using a dental drill. A 0.5 mm silicone tube and a stainless steel pipe connected to a microsyringe were inserted vertically 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-operated group) was injected into the ventricle over 3 minutes using a microsyringe pump. After injection, the stainless steel pipe was left in place for 3 minutes and then slowly removed. The skull hole was sealed with a non-absorbable bone marrow hemostatic agent (Nestop™, Alfresa Pharma). The scalp was sutured, and the mouse was removed from the stereotaxic apparatus and returned to its cage.

[0076] A passive avoidance test was performed on days 10 and 11 of administration to examine learning and memory impairment. For the passive avoidance test, a step-through passive avoidance reaction apparatus (light-dark box: manufactured by Nihon Bioresearch Center Co., Ltd.; SHOCK SCRAMBLER, manufactured by Unicom Co., Ltd.) was used. The mouse was placed in the light compartment, and 10 seconds later, the guillotine door was gently opened. The time it took for the mouse to enter the dark compartment (response latency) was measured. In the acquisition trial (day 10 of administration), the guillotine door was closed immediately after the mouse entered the dark compartment, and an electrical stimulus (0.2 mA, 2 seconds, scrambled) was administered. The presence or absence of vocalizations from the animal during the electrical stimulus challenge was confirmed. The response latency was measured up to 300 seconds. In the retention trial (day 11 of administration), the test was terminated when the animal entered the dark compartment or 300 seconds had elapsed in the light compartment.

[0077] (Results) Figure 2 shows the response latency in the retention trial in the passive avoidance test. Compared to the sham-operated group, the vehicle control group showed a statistically significant shortening of the response latency in the retention trial, indicating long-term memory impairment due to a single intracerebroventricular administration of β-amyloid 1-42. On the other hand, the response latency in the retention trial in the subject group was statistically significantly prolonged, indicating improvement in long-term memory impairment. Furthermore, the response latency in the retention trial in the subject group was comparable to that of the control group, which is already known to have an effect of improving cognitive function. These results indicate that JCM 7045 has an effect of improving cognitive function, particularly, for example, an effect of improving long-term memory impairment. Note that significant differences were calculated using the Dunnett method relative to the vehicle control group.

[0078] [Example 3: Coaggregation test] To explore further effects of JCM 7045, the coaggregation ability with periodontal disease bacteria was examined. Fusobacterium nucleatum JCM 8532 obtained from RIKEN BRC was used as the periodontal disease bacteria. Ｔ The periodontal disease bacteria were cultured in modified GAM bouillon (manufactured by Nissui Pharmaceutical Co., Ltd.) using Anaeropack (trademark) at 37°C for 6 days. The specimens were Bifidobacterium adolescentis strains JCM 7045, JCM 7046, and JCM 1275. Ｔ The test specimen was cultured overnight at 37°C in GAM bouillon (manufactured by Nissui Pharmaceutical Co., Ltd.) using Anaeropack (trademark).

[0079] After harvesting each bacterial cell, it was washed once with a buffer solution prepared as shown in Table 2, and the absorbance (OD ６００ The suspension was resuspended in the buffer solution so that the total volume of the suspension was 100 ml.

[0080]

[0081] Suspensions of periodontal disease bacteria or each analyte alone (single samples) and suspensions of periodontal disease bacteria mixed with equal amounts of each analyte (mixed samples) were prepared, stirred using a vortex mixer, and then incubated at 110 rpm and 37°C for 30 minutes. After that, the upper layer of the suspension was left to stand at room temperature for 3 minutes or more, and the absorbance (OD ６００ ) was measured. The OD of a single sample of periodontal bacteria and the target subject was６００ The sum of St and the OD of the mixed sample ６００ The coaggregation rate was calculated using the following formula 1, where Sm is the particle size.

[0082]

[0083] (Results) Figure 3 shows the coaggregation rate between periodontal disease bacteria and each test specimen. Among the test specimens, JCM 7045 showed high coaggregation ability, indicating the possibility of efficiently removing periodontal disease bacteria from the oral cavity. These results indicate that JCM 7045 not only has the effect of improving cognitive function, but also has the ability to coaggregate with periodontal disease bacteria, i.e., has the effect of improving the oral environment.

[0084] The above-described embodiments are intended to explain the present invention and are not intended to limit the scope of the present invention. That is, the scope of the present invention is defined by the claims, not by the embodiments. 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.

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

Claims

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

2. The composition according to claim 1, wherein the cognitive function is a memory-related function.

3. The composition according to claim 2, wherein the memory-related function is a long-term memory function.

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

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

6. A food or drink for preventing or improving brain fatigue, comprising the composition according to claim 4.

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

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

9. A medicine for preventing or improving brain fatigue, comprising the composition according to claim 4.

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

11. The preventive or therapeutic drug according to claim 10, wherein the dementia is Alzheimer's dementia.

12. A food product containing Bifidobacterium adolescentis JCM 7045 strain cells or a processed product thereof, wherein the content of the cells or the processed product thereof is 1.0 x 10 ８ cells / g~1.0×10 １３ The composition,

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