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

Abstract

Disclosed is a composition for maintaining, improving or ameliorating cognitive function, which comprises a cell body of Bifidobacterium adolescentis JCM 7045 strain or a processed product thereof.

Description

Technical Field

[0001] This invention relates to a composition for maintaining, improving or enhancing cognitive function, food and beverage, feed and medicine, preventive and therapeutic drugs for dementia, a composition for preventing or improving brain fatigue, food and beverage, feed and medicine, and other compositions. Background Technology

[0002] According to estimates by the World Health Organization, there are over 55 million people with dementia worldwide, and data suggests that approximately 10 million new cases may be added each year (see Non-Patent Literature 1). It is known that in dementia, cognitive decline is caused by the accumulation of brain damage resulting from nerve cells destroyed in chronic inflammation that occurs with age.

[0003] In Japan, it is estimated that approximately 16% of people aged 65 and over suffer from dementia. This proportion is expected to continue rising. With Japan's population rapidly aging, the increase in dementia cases is worrying and may further exacerbate social problems, particularly the shortage of caregivers and the burden of medical insurance costs.

[0004] The most representative form of dementia is Alzheimer's disease (AD). 60-70% of dementia patients have AD. Treatment options for dementia and related diseases include commonly used cholinesterase inhibitors such as donepezil, NMDA receptor antagonists such as memantine, and selective serotonin reuptake inhibitors (SSRIs) (see Non-Patent Literature 2). In addition, antibody drugs such as lencanezumab are under active development in recent years.

[0005] (Existing technical documents) (Non-patent literature) (Non-Patent Document 1) "Dementia", [Online], March 15, 2023, World Health Organization, [Searched January 17, 2024], Internet <URL: https: / / www.who.int / news-room / fact-sheets / detail / dementia> (Non-Patent Literature 2) Shin Kitamura, “The Latest Treatment of Cognitive Disorders Centered on Alzheimer’s Disease”, Journal of the Japan Medical Association, 2012, Vol. 8, No. 4, pp. 291-295 Summary of the Invention Technical problems to be solved The aforementioned drugs and antibody drugs are unlikely to prevent cognitive decline or improve cognitive function through daily intake.

[0006] The present invention was made in view of the above circumstances, and aims to provide a composition that is easy to take daily and can achieve excellent cognitive function improvement, as well as compositions, foods, feeds and medicines for maintaining, improving or enhancing cognitive function, and preventive and therapeutic drugs for dementia.

[0007] Furthermore, the present invention aims to provide a composition that is easy to take daily and can achieve excellent brain fatigue improvement effects, as well as compositions, food and beverage products, feed and medicines for preventing or improving brain fatigue.

[0008] Solution to the problem The composition for maintaining, improving or enhancing cognitive function according to the first aspect of the present invention comprises cells of Bifidobacterium adolescentis strain JCM 7045 or a treatment thereof.

[0009] The cognitive function mentioned can also be a function related to memory.

[0010] The memory-related functions mentioned can also be long-term memory functions.

[0011] The composition for preventing or improving brain fatigue according to the second aspect of the present invention comprises cells of Bifidobacterium adolescentis strain JCM 7045 or a treatment thereof.

[0012] The dietary product for maintaining, improving or enhancing cognitive function according to the third aspect of the present invention comprises the composition according to the first aspect of the present invention.

[0013] The food and beverage for preventing or improving brain fatigue according to the fourth aspect of the present invention comprises the composition according to the second aspect of the present invention.

[0014] The feed for maintaining, improving or enhancing cognitive function, or for preventing or improving brain fatigue, as described in the fifth aspect of this invention, comprises cells of Bifidobacterium adolescentis strain JCM 7045 or a treatment thereof.

[0015] The medicine for maintaining, improving or enhancing cognitive function according to the sixth aspect of the present invention comprises the composition according to the first aspect of the present invention.

[0016] The medicine for preventing or improving brain fatigue according to the seventh aspect of the present invention includes the composition according to the second aspect of the present invention.

[0017] The dementia prevention or treatment medicine according to the eighth aspect of the present invention comprises the cells of Bifidobacterium adolescentis strain JCM 7045 or a treatment thereof.

[0018] The dementia mentioned can also be Alzheimer's dementia.

[0019] The composition according to the ninth aspect of the present invention comprises cells of Bifidobacterium adolescentis strain JCM 7045 or a treatment thereof, wherein the content of said cells or the treatment thereof is 1.0 × 10⁻⁶. 8 cells / g ~ 1.0 × 10 13 cells / g.

[0020] The effects of the invention The compositions, foods, feeds, and pharmaceuticals, as well as preventive and therapeutic drugs for dementia, involved in this invention are easily ingested daily and can achieve excellent cognitive function improvement effects. Furthermore, the compositions, foods, feeds, and pharmaceuticals involved in this invention are easily ingested daily and can achieve excellent brain fatigue improvement effects. In addition, the compositions, foods, feeds, and pharmaceuticals, as well as preventive and therapeutic drugs for dementia, involved in this invention, also exhibit high coagulation ability against Fusobacterium nucleatum, a periodontal bacterium. Attached Figure Description

[0021] Figure 1 This is a graph showing the TNF-α concentration in the culture medium involved in Example 1.

[0022] Figure 2 This is a diagram showing the latency of the response in the passive avoidance test involved in Example 2.

[0023] Figure 3 This is a graph showing the coagulation rate with periodontal bacteria involved in Example 3. Detailed Implementation

[0024] The embodiments of the present invention will be described with reference to the accompanying drawings. However, the present invention is not limited to the embodiments and drawings described below. Furthermore, in the following embodiments, expressions such as "having," "comprising," or "containing" also include the meaning of "composed of" or "constituted by."

[0025] (Effective Method 1: Composition) The composition involved in this embodiment comprises cells of *Bifidobacterium adolescentis* strain JCM 7045 (hereinafter also referred to as "JCM 7045") or a treatment thereof. JCM 7045 can be obtained from the RIKEN Center for Biological Resources Research (RIKEN BRC) in Japan. JCM 7045 is preferably cultured at or near its optimum temperature, for example, 27–45°C, 30–42°C, or 35–40°C. Preferably, the culture temperature is 37°C. JCM 7045 can be cultured in liquid culture medium or agar medium. Preferably, the bifidobacteria can be cultured under anaerobic conditions, for example, while anaerobic gas such as carbon dioxide is introduced. Alternatively, it can be cultured under microaerobic conditions such as liquid static culture. The culture medium for culturing JCM 7045 is not particularly limited, and a culture medium commonly used for culturing bacteria belonging to the genus *Bifidobacterium* can be used.

[0026] As carbon sources for culture, sugars such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolysate, and molasses can be used. As nitrogen sources, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, as well as nitrates, can be used. Furthermore, as inorganic salts, sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate can be used. Additionally, organic components such as peptone, soybean flour, defatted soybean meal, meat extract, and yeast extract can also be used. For example, the medium for JCM 7045 is a BL medium containing nutrient broth.

[0027] Bacterial cells can be obtained as a precipitate by centrifuging the cultured medium. Bacterial cells can be live or dead, wet or dried. Processed products of bacterial cells refer to bacterial cells, compositions derived from or containing a portion of bacterial cells, obtained through some form of treatment. Examples include freeze-dried bacterial cells, acetone-dried bacterial cells, extracts obtained by solvent extraction of bacterial cells, and fragments obtained by disrupting bacterial cells or dried bacterial cells using methods such as ultrasound. Bacterial cells, compositions derived from or containing a portion of bacterial cells, are examples such as culture supernatants or culture media. Therefore, processed products of bacterial cells also include culture supernatants and culture media.

[0028] Solvents used in solvent extraction processes can include water, organic solvents, and mixtures thereof. Examples of organic solvents include diethyl ether, chloroform, benzene, hexane, methanol, ethanol, isopropanol, and mixtures thereof. Extracts obtained using extraction solvents can be used directly in liquid form, or they can be concentrated or diluted and used in liquid, gel, or paste form. Furthermore, they can be used as dried products. Drying can be carried out using known methods such as spray drying, freeze drying, vacuum drying, and fluidized bed drying.

[0029] The bacterial cells can be used directly after culturing in JCM 7045, or they can be used after dilution or concentration. The processed bacterial cells can also be used directly, or after dilution or concentration.

[0030] The composition described in this embodiment contains JCM 7045 bacterial cells or a treatment thereof as the active ingredient, and may also contain other ingredients besides the active ingredient. There are no particular limitations on the ingredients other than the active ingredient; for example, additives used in pharmaceuticals. Furthermore, when culture supernatant or culture medium is used as the treatment for the bacterial cells, the composition may contain, for example, ingredients derived from the culture medium as other than the active ingredient.

[0031] As shown in Example 2 below, the composition according to this embodiment has a cognitive function improvement effect, particularly an improvement effect on long-term memory impairment. Therefore, as for the use of the composition according to this embodiment, examples include maintaining, improving or enhancing cognitive function, and preventing or improving mental fatigue. Preferably, the cognitive function is a function related to memory. More preferably, the memory-related function is long-term memory function.

[0032] The composition described in this embodiment can also be used for the prevention or improvement of brain fatigue. Brain fatigue refers to a state in which the brain is subjected to excessive stress, resulting in a decline in brain function. Fatigue is considered to be caused by a decline in original function due to the chronic decrease in immune cytokines or the induction of inflammatory cytokines caused by stress, etc. (Japanese Journal of Biological Psychiatry 24(4):200-210, 2013). To improve brain fatigue, for example, the induction of inflammatory cytokines can be inhibited. As shown in Example 1 below, this composition inhibits the secretion of inflammatory cytokines in central nervous system cells and has excellent anti-inflammatory effects. Therefore, this composition is useful for the prevention or improvement of brain fatigue.

[0033] The composition, for example, contains 0.1–99% by weight, 1–50% by weight, or 1–20% by weight of JCM 7045 cells or its processed form as an active ingredient. The content of JCM 7045 cells or its processed form in the composition according to this embodiment can be appropriately adjusted according to the effective amount of the composition in the stated application. An effective amount refers to the amount required to obtain the desired result; it is the amount required to delay, inhibit, prevent, reverse, or cure the progression of the treated or managed condition. When the active ingredient is JCM 7045 cells, its effective amount is, for example, 1.0 × 10⁻⁶. 2 ~1.0×10 10 Colony forming units (cfu) / kg, 1.0 × 10⁻⁶ 3 ~1.0×10 9 cfu / kg or 1.0×10 4 ~1.0×10 8 cfu / kg. Furthermore, from another perspective, the effective dose could also be 1.0 × 10⁻⁶. 8 cells / kg ~ 1.0 × 10 13 cells / kg, 1.0×10 9 cells / kg ~ 1.0 × 10 12 cells / kg or 1.0×10 10 cells / kg ~ 1.0 × 10 12 Cells / kg. When using a treatment of JCM 7045 cells as the active ingredient, the treatment of cells is, for example, an amount equivalent to the effective amount of cells. Furthermore, the method for counting (determining) the number of cells can be any method commonly used in the field, specifically, examples include, for instance, the DAPI staining method, the plate plating method, etc.

[0034] The composition described in this embodiment is intended for use on humans and animals other than humans. Mammals are preferred, and more specifically, dogs, cats, cattle, pigs, horses, sheep, and deer are examples. The dosage of the composition is appropriately determined based on the sex, age, weight, and symptoms of the recipient. The composition is administered in an effective amount of JCM 7045 bacteria or its processed form. The composition can be administered once daily or in multiple applications. It can be administered at various frequencies, such as daily, every other day, weekly, every other week, or monthly. Furthermore, amounts outside the above ranges may be used as needed. The method of administration is not particularly limited and can be administered via injection, nasal route, transdermal route, pulmonary route, or oral route. Oral administration is particularly preferred.

[0035] The composition described in this embodiment contains the cells of Bifidobacterium JCM 7045 or a processed form thereof. Therefore, it is easily ingested daily and can achieve excellent cognitive function improvement.

[0036] Another aspect of this embodiment involves a composition comprising JCM 7045 strain cells or a treatment thereof, and the content of the strain cells or the treatment thereof is 1.0 × 10⁻⁶. 8 cells (units) / g ~ 1.0 × 10⁻⁶ 13 cells / g or 1.0×10 9 cells / kg ~ 1.0 × 10 12 cells / kg

[0037] (Implementation Method 2: Food and Drink) In another embodiment, a food or beverage comprising the composition described in Embodiment 1 is provided. The purpose of this food or beverage is to maintain, enhance, or improve cognitive function, or to prevent or improve mental fatigue.

[0038] Examples of food products that can be categorized as food supplements, food products, functional foods, and food additives include these. Furthermore, the composition described in Embodiment 1 above can be used as a food product or added to food products and functional foods.

[0039] There are no particular restrictions on the form of supplements; they can be in any form, such as tablets, powders, granules, capsules, sugar-coated tablets, films, lozenges, chewable tablets, solutions, emulsions, or suspensions. Supplements can contain any ingredients commonly used as supplements.

[0040] "Functional foods" refer to foods or beverages consumed for the purpose of maintaining health, including foods for specific health purposes, foods with functional claims, nutritional functional foods, health foods, and nutritional supplements. Foods for specific health purposes or nutritional functional foods are preferred as health foods. Furthermore, when commercialized as functional foods, these products may contain various additives used in food processing, specifically colorants, preservatives, thickeners and stabilizers, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, sweeteners, seasonings, emulsifiers, fortifiers, preparative agents, and flavorings.

[0041] Functional foods can be either food or beverages; there are no particular limitations as long as they can be taken orally. Examples of functional food forms include beverages, snacks, processed cereals, minced / fish paste products, dairy products, and seasonings. Examples of beverages include nutritional drinks, soft drinks, black tea, and green tea. Examples of snacks include candy, biscuits, candy tablets, chewing gum, and jelly. Examples of processed cereals include bread, rice, and biscuits. Examples of minced / fish paste products include sausages, ham, and fish cakes. Examples of dairy products include butter and yogurt.

[0042] Food additives can be in the form of pastes, gels, powders, liquid preparations, suspending agents, emulsions, granules, etc., to facilitate their addition to food.

[0043] The food and beverage products involved in this embodiment may contain water, vitamins, minerals, organic acids, organic bases, fruit juice, flavorings, functional ingredients, food additives, etc., within the range that does not affect the maintenance of cognitive function and improves or enhances the effectiveness of preventing or improving brain fatigue. The food and beverage products may be prepared from the composition involved in Embodiment 1 above, and other ingredients as needed, using known methods.

[0044] The content of the composition described in Embodiment 1 in the food and beverage products involved in this embodiment is appropriately adjusted according to the effective amount of the food and beverage products in the stated use. When the composition contains JCM 7045 cells, the cell content of the food and beverage products is, for example, 1.0 × 10⁻⁶. 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. Furthermore, from another perspective, the bacterial count could also be 1.0 × 10⁻⁶. 8 cells (units) / g ~ 1.0 × 10⁻⁶ 13 cells / g, 1.0×10 9 cells (units) / g ~ 1.0 × 10⁻⁶ 12 cells / g or 1.0×10 10 cells / kg ~ 1.0 × 10 12 Cells / kg. When the composition contains a treatment of JCM 7045 bacteria, the amount of the treatment of bacteria in the food or beverage is, for example, an amount equivalent to the above-mentioned bacterial content. Furthermore, the above-mentioned bacterial content can also be used as a reference when discussing the effective amount of the feed, preventative medicine, therapeutic medicine, or oral environment improvement composition involved in this embodiment described later.

[0045] The food and beverage can be packaged in one or more containers in a manner that constitutes a one-day intake, in which case it is preferable to contain a one-day intake of the food and beverage in one container.

[0046] The food and beverage products are provided in a manner that distinguishes them from other products in that they contain the composition described in Embodiment 1 above and are used to maintain, improve or enhance cognitive function or prevent or improve mental fatigue. For example, at least one of the packaging, instructions, and promotional materials of the food and beverage products is labeled as having the effect of maintaining, improving or enhancing cognitive function or preventing or improving mental fatigue.

[0047] Food and beverage products may also be provided or sold with the indication that they are intended to maintain, improve or enhance cognitive function, or prevent or improve mental fatigue (including health care purposes). "Indication" includes all actions taken to inform consumers of these uses. Any statement that leads to the association or inference of these uses falls under the category of "indication," regardless of the purpose, content, object, or medium of the indication.

[0048] "Labeling" is preferably carried out in a way that allows the consumer to directly identify the aforementioned uses. Specifically, examples include: recording the aforementioned uses on goods or packaging involving food and beverages and transferring, delivering, displaying for transfer or delivery, or importing them; recording the aforementioned uses on advertisements, price lists, or transaction documents related to the goods and displaying or distributing them; or recording the aforementioned uses in information containing these uses and providing them through electromagnetic (Internet, etc.) methods.

[0049] "Labeling" includes labels for health foods, functional foods, enteral nutrition foods, foods for special purposes, health-functional foods, foods for specific health uses, nutritional functional foods, foods with functional claims, and quasi-drugs. Specifically, examples include labels recognized under regulations concerning foods for specific health uses, nutritional functional foods, or foods with functional claims, or similar regulations. More specifically, examples include labels for foods for specific health uses, labels for conditionally designated foods for specific health uses, labels indicating effects on body structure or function, labels indicating reduced disease risk, and scientifically based functional labels. More concretely, typical examples include labels for foods for specific health uses (especially labels for health purposes) and similar labels.

[0050] (Implementation method 3: Feed) In another embodiment, a feed comprising the composition described in Embodiment 1 is provided. Regarding the feed described in this embodiment, the main differences from the diet and food described in Embodiment 2 will be explained. For aspects of this feed not specifically mentioned, please refer to the description of the diet and food described in Embodiment 2. Furthermore, the composition described in Embodiment 1 can also be used as feed.

[0051] The feed is given to non-human animals. Non-human animals are preferably mammals. Examples of mammals include primates such as chimpanzees, laboratory animals such as rats, mice, and rabbits, livestock such as pigs, cattle, horses, sheep, and goats, and pet animals such as dogs and cats. In addition to providing nutrients for the non-human animals, the feed also includes supplements for their consumption as a treat.

[0052] (Implementation method 4: drugs, dementia prevention drugs, dementia treatment drugs) In another embodiment, a medicine, a dementia prevention drug, and a dementia treatment drug comprising the composition described in Embodiment 1 are provided (hereinafter, sometimes collectively referred to as "medicines, etc."). Regarding the medicines, etc., described in this embodiment, aspects differing from those described in Embodiment 1 will be primarily explained. For aspects not specifically mentioned in this medicine, please refer to the description of the composition described in Embodiment 1. Furthermore, the composition described in Embodiment 1 can also be used as a medicine, etc.

[0053] The pharmaceuticals involved in this embodiment may contain ingredients other than those involved in Embodiment 1 above, such as excipients, lubricants, binders, disintegrants, solvents, solubilizers, suspending agents, isotonic modifiers, buffers, analgesics, etc. Furthermore, pharmaceuticals may include additives such as preservatives, antioxidants, colorants, and sweeteners as needed.

[0054] Excipients include, for example, lactose, white sugar, D-mannitol, starch, crystalline cellulose, and light anhydrous silica. Lubricants include, for example, magnesium stearate, calcium stearate, talc, and colloidal silica. Binders include, for example, crystalline cellulose, white sugar, D-mannitol, dextrin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Disintegrants include, for example, starch, carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, and sodium carboxymethyl starch.

[0055] Solvents include, for example, water for injection, ethanol, propylene glycol, and polyethylene glycol. Cosolvents include, for example, polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, tris(hydroxymethyl)aminomethane, cholesterol, triethanolamine, sodium carbonate, and sodium citrate. Suspending agents are surfactants and hydrophilic polymers, such as stearyltriethanolamine, sodium lauryl sulfate, dodecylaminopropionic acid, lecithin, benzalkonium chloride, benzyl chloride, glyceryl monostearate, polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethyl cellulose, methylcellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose.

[0056] Isotonic adjusters include, for example, sodium chloride, glycerol, and D-mannitol. Buffers include, for example, buffers containing phosphates, acetates, carbonates, and citrates. Analgesics include, for example, benzyl alcohol. Preservatives include, for example, parabens, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, and sorbic acid. Antioxidants include, for example, sulfites and ascorbic acid.

[0057] Since the medicine involved in this embodiment can maintain, improve or enhance cognitive function, or prevent or improve brain fatigue, it can be used, for example, to treat or prevent diseases caused by decreased brain function, such as dementia, depression, schizophrenia, and delirium.

[0058] In the dementia prevention and treatment drugs involved in this embodiment, dementia is not particularly limited, and can include Alzheimer's dementia, Lewy body dementia, frontotemporal dementia, cerebrovascular dementia, etc. Preferably, the dementia is Alzheimer's dementia.

[0059] (Effective 5: Composition for improving oral environment) As shown in Example 3 below, JCM 7045 exhibits high co-aggregation ability against *Fusobacterium nucleatum*, a periodontal pathogen. Therefore, the cells of JCM 7045 or their treatments also possess antibacterial activity against periodontal pathogens, i.e., the effect of improving the oral environment. Therefore, in another embodiment, a composition comprising the cells of JCM 7045 or their treatments for the prevention, maintenance, or improvement of oral function is provided. A composition comprising the cells of JCM 7045 or their treatments for the prevention or improvement of periodontal disease is provided. Furthermore, a composition comprising the cells of JCM 7045 or their treatments for inhibiting biofilm formation is provided. Further, a composition comprising the cells of JCM 7045 or their treatments and possessing co-aggregation ability with *Fusobacterium nucleatum* is provided.

[0060] The composition described in this embodiment, in addition to JCM 7045 bacterial cells or their processed form as the active ingredient, may further contain ingredients (raw materials) commonly used for improving the oral environment. 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, colorings, etc. From the viewpoint of adjusting taste or sweetness, various sugars, fruit juices, dairy products, etc., may be incorporated into the compound.

[0061] The composition described in this embodiment can be formulated into various forms as needed, such as paste, gel, liquid, and solid. Specific examples of these forms include toothpaste, dental gel, dental paste, mouthwash, oral spray, tooth powder, gel, candy, gummies, chewing gum, and tablets. These forms can be prepared by combining the aforementioned ingredients (raw materials).

[0062] In another embodiment, use is provided of JCM 7045 cells or a treatment thereof in the manufacture of compositions for maintaining, improving, or enhancing cognitive function. In another embodiment, use is provided of JCM 7045 cells or a treatment thereof in the manufacture of medicines for maintaining, improving, or enhancing cognitive function. In another embodiment, use is provided of JCM 7045 cells or a treatment thereof in the manufacture of compositions for preventing or improving mental fatigue. In another embodiment, use is provided of JCM 7045 cells or a treatment thereof in the manufacture of medicines for preventing or improving mental fatigue.

[0063] Furthermore, in another embodiment, the use of JCM 7045 microbial cells or processed products thereof in the manufacture of dietary supplements for maintaining, improving, or enhancing cognitive function is provided. In another embodiment, the use of JCM 7045 microbial cells or processed products thereof in the manufacture of dietary supplements for preventing or improving mental fatigue is provided. In another embodiment, the use of JCM 7045 microbial cells or processed products thereof in the manufacture of feed for maintaining, improving, or enhancing cognitive function is provided. In another embodiment, the use of JCM 7045 microbial cells or processed products thereof in the manufacture of feed for preventing or improving mental fatigue is provided. In another embodiment, the use of JCM 7045 microbial cells or processed products thereof in the manufacture of preventative or therapeutic drugs for dementia is provided.

[0064] Furthermore, in another embodiment, a method for maintaining, improving, or enhancing cognitive function is provided, comprising administering JCM 7045 bacterial cells or a treatment thereof to a subject. In another embodiment, a method for preventing or improving brain fatigue is provided, comprising administering JCM 7045 bacterial cells or a treatment thereof to a subject. In yet another embodiment, a method for preventing or treating dementia is provided, comprising administering JCM 7045 bacterial cells or a treatment thereof to a subject.

[0065] Furthermore, in another embodiment, a bacterial cell or a processed form of JCM7045 is provided for maintaining, improving, or enhancing cognitive function. In another embodiment, a bacterial cell or a processed form of JCM7045 is provided for preventing or improving brain fatigue. In yet another embodiment, a bacterial cell or a processed form of JCM7045 is provided for preventing or treating dementia.

[0066] The present invention will be described in more detail through the following embodiments, but the present invention is not limited to these embodiments.

[0067] Example [Example 1: In vitro anti-inflammatory screening] Neuroinflammation caused by aging and other factors is known to be a contributing factor to cognitive decline. To efficiently select Bifidobacteria for improving cognitive function, an anti-inflammatory assessment was conducted. As Bifidobacteria for improving cognitive function (samples), JCM 7045, JCM 1251, and JCM 1275, all Bifidobacterium adolescentis, were used. T (Purchased from the RIKEN Center for Biological Resources Research, Japan; RIKEN BRC).

[0068] Each sample used for anti-inflammatory assessment was cultured for 5 days at 37°C using an anaerobic gas-generating bag (trademark) (manufactured by Mitsubishi Gas Chemical Co., Ltd.). The culture medium shown in Table 1 was used. Afterwards, the collected bacterial cells were washed twice with physiological saline (manufactured by Otsuka Pharmaceutical Co., Ltd.) and then freeze-dried to prepare a dried bacterial powder.

[0069] Table 1

[0070] MG6 cells (RCB2403, purchased from RIKEN Bioresource Research Center, RIKEN BRC) were cultured at 37°C and 5% CO2 using Dulbecco's modified Eagle medium (D-MEM) (high glucose) (manufactured by Fujifilm and Hikari Pure Chemical Industries Co., Ltd.) containing 10% fetal bovine serum (manufactured by MP Bio Japan) and 1% penicillin-streptomycin solution (×100) (manufactured by Fujifilm and Hikari Pure Chemical Industries Co., Ltd.). MG6 cells, intended as a mouse microglial cell line, were prepared, for example, according to Neurosci Lett 2006 407(3):205-10 or Biochim Biophys Acta 2005 1726(2):177-86.

[0071] After detaching the cultured MG6 cells with 0.05% trypsin (Gibco), they were injected with 1×10⁻⁶... 4 Cells were seeded at a concentration of [insert concentration here] in 96-well plates (Corning Corporation) and cultured overnight at 37°C with 5% CO2. Cells that had been passaged less than 10 times were used in the experiment.

[0072] After removing the culture medium from each well, D-MEM (high glucose) containing 2% fetal bovine serum and 1% penicillin-streptomycin solution (×100) was added to the control and inflammation-inducing groups. The positive control group was added to D-MEM (high glucose) containing SB239063 (manufactured by Tokyo Chemical Industry Co., Ltd.), a p38 MAPK inhibitor, dissolved in dimethyl sulfoxide (manufactured by Sigma-Aldrich), at a final concentration of 1.25 μM, and containing 2% fetal bovine serum and 1% penicillin-streptomycin solution (×100). The sample addition group was added to D-MEM (high glucose) containing each sample at a final concentration of 1 μg / mL, suspended in D-PBS(-) (manufactured by Fujifilm and Hikari Pure Chemicals Co., Ltd.) and irradiated with ultraviolet light (UV) for 15 minutes, and containing 2% fetal bovine serum and 1% penicillin-streptomycin solution (×100). All groups were cultured for 24 hours.

[0073] After culture, as an inflammation-inducing component, lipopolysaccharide (LPS, Sigma-Aldrich) dissolved in D-PBS(-) was added to groups other than the control group at a final concentration of 1 μg / mL, and cultured for 6 hours. A portion of the culture medium was collected, and the concentration of TNF-α in the culture medium was measured using the Mouse TNF-alpha DuoSet ELISA kit (R&D Systems).

[0074] (result) like Figure 1 As shown, in the sample, JCM 7045 statistically significantly inhibited LPS-induced TNF-α secretion, exhibiting the highest anti-inflammatory effect. Furthermore, the statistical significance was calculated using the Dunnett method relative to the inflammation-inducing group.

[0075] [Example 2: In vivo trial] The cognitive function improvement effect of JCM 7045, with its confirmed anti-inflammatory effect, was evaluated in Alzheimer's disease model mice prepared according to the following steps. Male Slc:ddY mice (manufactured by SLC Corporation, Japan) were used. Administration was performed orally once daily for 15 days using a syringe (manufactured by Terumo Corporation) fitted with a mouse oral gastric tube (manufactured by Fuchigami Instruments Co., Ltd.). For the positive control group, the dosage was 10 mL / kg based on the body weight on the day of administration; for other groups, it was 200 μL / head. Here, "head" refers to the body weight of the model mice, approximately 30 g to 45 g. The test group received JCM 7045 at a dose of 1.2 × 10⁻⁶. 8 CFU / animal (total bacterial count 3.8 × 10⁻⁶) 10 (cells / animal). The control group was treated with Bifidobacterium obtained from a known cognitive-improving supplement (trade name: MemoryBifidus, manufactured by Morinaga Milk Industry Co., Ltd.) using well-known methods commonly employed in the field, at a dose of 1.0 × 10⁻⁶ cells / animal. 9 CFU / animal (total bacterial count 1.2 × 10⁻⁶) 11 Cells / animal). The positive control group was given donepezil hydrochloride (trade name: Aricept, manufactured by Eisai Co., Ltd.), an existing dementia treatment, at a dose of 0.5 mg / kg. In addition, the sham-operated group and the media control group were given JCM 7045 media (cryoprotectant: physiological saline = 1:9).

[0076] On day 3, β-amyloid protein 1-42 (manufactured by the Peptide Research Institute) was administered as a single intraventricular injection to all groups except the sham-operated group, while physiological saline was administered as a single intraventricular injection to the sham-operated group. During the anesthesia procedure for injecting β-amyloid protein 1-42 into the ventricles, sodium pentobarbital (manufactured by Tokyo Chemical Industry Co., Ltd.) 40 mg / kg was first administered intraperitoneally, followed by subcutaneous administration of levobupivacaine hydrochloride (manufactured by Maruishi Pharmaceutical Co., Ltd.). Afterwards, the hair on the top of the mice's heads was shaved, and the heads were fixed to a stereotactic brain imaging device. The top of the head was disinfected with povidone-iodine, then incised to expose the skull. Connective tissue on the skull was removed with cotton swabs, and the skull was dried with a hairdryer to facilitate observation of the anterior fontanelle. A small hole was drilled in the skull 1 mm lateral to the anterior fontanelle (right side) and 0.2 mm posterior to it using a dental drill. The stainless steel tube, connected to a 0.5 mm outer diameter silicone tube and a micro-injector, was vertically inserted to a depth of 2.5 mm from the bone surface. Using a micro-injector pump, 3 μL (200 pmol / 3 μL) of β-amyloid protein solution (physiological saline for the sham-operated group) was injected into the ventricle over 3 minutes. After injection, the stainless steel tube was left inserted for 3 minutes, and then slowly withdrawn. The skull hole was sealed with a non-absorbable bone marrow hemostatic agent (Nestop, manufactured by Alfresa Pharma), the scalp was sutured, the mouse was removed from the stereotaxic device, and returned to its cage.

[0077] As an examination for learning and memory impairment, a passive avoidance test was performed on days 10–11 after administration. The passive avoidance test used a step-through passive avoidance response device (light-dark chamber: Bio Research Center, Japan; shock scrambler: Unicom) consisting of a bright chamber separated by a central gate and a dark chamber through which electrical stimulation was applied via a floor grid. Mice were placed in the bright chamber, and the gate was gently opened after 10 seconds. The time required for the mouse to enter the dark chamber was measured (response latency). In the acquisition test (day 10 after administration), the gate was closed as the mouse entered the dark chamber, and electrical stimulation (0.2 mA, 2 seconds, scramble mode) was applied. The presence or absence of vocalization during the application of electrical stimulation was monitored. The maximum response latency was set at 300 seconds. In the maintenance test (day 11 after administration), the test ended when the animal entered the dark chamber or after 300 seconds in the bright chamber.

[0078] (result) Figure 2The results show the response latency of the retention test in the passive avoidance test. Compared with the sham-operated group, the media control group showed a statistically significant shorter response latency in the retention test, indicating that a single intraventricular administration of β-amyloid 1-42 leads to long-term memory impairment. On the other hand, the sample group showed a statistically significant longer response latency in the retention test, indicating improvement in long-term memory impairment. Furthermore, the response latency of the sample group in the retention test was comparable to that of the comparative control group, which is known to have cognitive function improvement effects. These results indicate that JCM 7045 has a cognitive function improvement effect, particularly in improving long-term memory impairment. Furthermore, the statistical significance was calculated using the Dunnett method relative to the media control group.

[0079] [Example 3: Coagulation Test] To explore the further effects of JCM 7045, its co-aggregation ability with periodontal bacteria was studied. Fusobacterium nucleatum JCM 8532, purchased from RIKEN BRC, was used as the periodontal bacteria. T Periodontal bacteria were cultured for 6 days at 37°C using modified GAM broth (manufactured by Nissui Pharmaceutical Co., Ltd.) and an anaerobic gas-generating bag, AnaeroPack (trademark). JCM 7045, JCM 7046, and JCM1275 (Bifidobacterium adolescentis) were used as samples. T Samples were cultured overnight at 37°C using GAM broth (manufactured by Nissui Pharmaceutical Co., Ltd.) and an anaerobic gas-generating bag, AnaeroPack (trademark).

[0080] After collection, each bacterial cell was washed once with the buffer solution prepared according to Table 2, and then resuspended in the buffer solution until it reached a certain absorbance (OD). 600 ).

[0081] Table 2

[0082] Suspensions of periodontal bacteria or individual samples (single samples) and suspensions of equal volumes of periodontal bacteria and individual samples (mixed samples) were prepared. After stirring with a vortex mixer, the suspensions were incubated at 110 rpm and 37°C for 30 minutes. Subsequently, the upper layer of the suspension was collected after standing at room temperature for at least 3 minutes, and the absorbance (OD) was measured. 600 ). The OD of a single sample of periodontal bacteria and the target sample. 600 Let the sum be St, and the OD of the mixed sample. 600 Let Sm be the coagulation rate, and use Equation 1 below to calculate the coagulation rate.

[0083]

[0084] (result) Figure 3 The coagulation rates of periodontal bacteria with each sample are shown. JCM 7045 exhibited high coagulation capacity in the samples, indicating its potential to effectively remove periodontal bacteria from the oral cavity. This result suggests that JCM 7045 not only improves cognitive function but also has coagulation capacity with periodontal bacteria, thus improving the oral environment.

[0085] The above embodiments are for illustrative purposes only and do not limit the scope of the invention. That is, the scope of the invention is shown by the claims rather than the embodiments. Furthermore, various modifications implemented within the scope of the claims and their equivalents are considered to be within the scope of the invention.

[0086] 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.

[0087] Industrial availability This invention can be used as a composition, food and beverage, feed and medicine.

Claims

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

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

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

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

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

6. A dietary product for preventing or improving brain fatigue, comprising the composition of claim 4.

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

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

9. A medicament for preventing or improving brain fatigue, comprising the composition of claim 4.

10. A dementia prevention or treatment drug comprising cells of Bifidobacterium adolescentis strain JCM 7045 or a treatment thereof.

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

12. A composition comprising cells of Bifidobacterium adolescentis strain JCM 7045 or a treatment thereof, wherein the amount of said cells or the treatment thereof is 1.0 × 10⁻⁶. 8 cells / g ~ 1.0 × 10 13 cells / g.

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