Compositions for improving complex attention, cognitive flexibility and / or performance functions

Abstract

The problem addressed by the invention is that of providing a new means for improving complex attention, cognitive flexibility and / or execution functions. There is provided a composition for improving at least one selected from the group consisting of complex attention, cognitive flexibility and executive function, the composition comprising bifidobacteria and a water-soluble dietary fiber.

Description

Compositions for improving complex attention, cognitive flexibility, and / or executive function Technical Field

[0001] The present invention relates to a composition for improving complex attention, cognitive flexibility and / or executive function. Background Technology

[0002] Dementia is an age-related disease characterized by impaired cognitive functions such as memory and judgment due to brain cell death or functional decline, leading to problems in social life and interpersonal relationships. It is estimated that about 15% of people aged 65 and older have dementia, and about 4 million people have mild cognitive impairment (MCI), a precursor to dementia.

[0003] Dementia and MCI have various subtypes, including those related to memory and those related to reaction time. Memory also has various types. Specifically, Cognitrax has been developed and used as a tool to measure these memory and cognitive functions. More specifically, Cognitrax measures the following items and determines which areas of function are declining: overall memory, verbal memory, visual memory, cognitive speed, reaction time, complex attention, sustained attention, simple attention, cognitive flexibility, processing speed, motor speed, executive function, social cognition, logical reasoning, working memory, and neurocognitive indices. Among these functions, complex attention, cognitive flexibility, and executive function are particularly important because they affect various judgments in social life. In these cases, there is an urgent need to develop new means to improve complex attention, cognitive flexibility, and executive function.

[0004] Citation List Non-Patent Literature Non-Patent Literature 1: Jpn Pharmacol Ther. Vol. 48, No. 4, 2020, 685-679 Non-Patent Literature 2: Jpn Pharmacol Ther. Vol. 47, No. 5, 2019, 739-749 Non-Patent Literature 3: Jpn Pharmacol Ther. Vol. 48, No. 3, 2020, 475-486 Non-Patent Literature 4: J Biosci Bioeng. 2012; 113: 587-91 Non-Patent Literature 5: Biosci Microbiota Food Health. 2016; 35(4): 163-171 Non-Patent Literature 6: Arch Clin Neuropsychol. 2006, 7, 623-643 Non-Patent Literature 7: Molecules. 2020, 18, Non-Patent Literature 8: Psychiatry Clin Neurosci. 2007, 1, 112-119 Non-Patent Literature 9: PLoS One. 2014, 4, e95192 Non-Patent Literature 10: Microbiome. 2021, 1, 95 Non-Patent Literature 11: Nutrients. 2019, 4, 839. Summary of the Invention

[0005] Technical Problem: The problem this invention aims to solve is to provide a new means for improving complex attention, cognitive flexibility, and / or executive function.

[0006] In response to the aforementioned situation, the inventors conducted extensive research and discovered that the combination of Bifidobacteria and soluble dietary fiber can improve complex attention, cognitive flexibility, and / or executive function. More specifically, the inventors found that the combination of Bifidobacteria and soluble dietary fiber improves overall cognitive abilities (average scores for memory, information processing speed, reaction time, attention, and judgment), particularly improving attention (the ability to maintain attention and respond accurately), judgment (the accuracy and speed of judgment, and the ability to respond to and process changes in instructions), and / or executive function (the ability to understand rules and concepts and make decisions). This invention is based on these new findings. In particular, the inventors found that even in healthy middle-aged and older adults, ingestion of the composition of this invention can improve complex attention, cognitive flexibility, and / or executive function. Therefore, this invention provides the following item: Item 1. A composition for improving at least one selected from complex attention, cognitive flexibility, and executive function, the composition comprising Bifidobacteria and soluble dietary fiber.

[0007] Project 2. The composition according to Project 1, wherein the water-soluble dietary fiber comprises inulin.

[0008] Project 3. A composition according to Project 1 or 2, which is used to improve neurocognitive indices.

[0009] Project 4. A composition according to any one of Projects 1 to 3, wherein the Bifidobacteria have the ability to proliferate in the intestine.

[0010] Project 5. A composition according to any one of Projects 1 to 4, wherein Bifidobacterium has an anti-metabolic syndrome effect.

[0011] Item 6. A composition according to any one of items 1 to 5 is a food.

[0012] Item 7. A composition according to any one of Items 1 to 6, which is a food for inhibiting cognitive decline, maintaining cognitive function or improving cognitive function.

[0013] Project 8. Use of Bifidobacteria and water-soluble dietary fiber in the manufacture of compositions, wherein the compositions are used to improve at least one selected from complex attention, cognitive flexibility and executive function.

[0014] Item 9. As per the use of Item 8, the water-soluble dietary fiber contains inulin.

[0015] Item 10. As per the use of Item 8 or 9, wherein the composition is used to improve neurocognitive indices.

[0016] Item 11. According to the use of any one of Items 8 to 10, wherein Bifidobacteria have the ability to proliferate in the intestine.

[0017] Item 12. According to any of the uses in Items 8 to 11, Bifidobacterium has an anti-metabolic syndrome effect.

[0018] Item 13. The use of any one of items 8 to 12, wherein the composition is a food.

[0019] Item 14. According to the use of Item 13, wherein the composition is a food for inhibiting cognitive decline, maintaining cognitive function or improving cognitive function.

[0020] Project 15. Use of water-soluble dietary fiber in the manufacture of compositions for improving at least one selected from complex attention, cognitive flexibility and executive function, the compositions comprising Bifidobacterium.

[0021] Item 16. As per the use of Item 15, the water-soluble dietary fiber contains inulin.

[0022] Item 17. As per the use of Item 15 or 16, wherein the composition is used to improve neurocognitive indices.

[0023] Item 18. As used in any of Items 15 to 17, wherein Bifidobacteria have the ability to proliferate in the gut.

[0024] Item 19. According to any of the uses in Items 15 to 18, Bifidobacterium has an anti-metabolic syndrome effect.

[0025] Item 20. The use of any one of items 15 to 19, wherein the composition is a food.

[0026] Item 21. According to the use of Item 20, wherein the composition is a food for inhibiting cognitive decline, maintaining cognitive function or improving cognitive function.

[0027] Project 22. Use of Bifidobacterium in the manufacture of compositions for improving at least one selected from complex attention, cognitive flexibility and executive function, the compositions comprising water-soluble dietary fiber.

[0028] Item 23. As per the use of Item 22, the water-soluble dietary fiber contains inulin.

[0029] Item 24. As per the use of Item 22 or 23, wherein the composition is used to improve neurocognitive indices.

[0030] Item 25. As used in any of Items 22 to 24, wherein Bifidobacteria have the ability to proliferate in the gut.

[0031] Item 26. According to any of the uses in Items 22 to 25, Bifidobacterium has an anti-metabolic syndrome effect.

[0032] Item 27. The use of any one of items 22 to 26, wherein the composition is a food.

[0033] Item 28. According to the use of Item 27, wherein the composition is a food for inhibiting cognitive decline, maintaining cognitive function or improving cognitive function.

[0034] Project 29. A method for improving at least one selected from complex attention, cognitive flexibility, and executive function, the method comprising: administering an effective amount of bifidobacteria and water-soluble dietary fiber to a subject in need.

[0035] Project 30. According to the method of Project 29, wherein the water-soluble dietary fiber contains inulin.

[0036] Project 31. Based on the methods of Project 29 or 30, it is used to improve the neurocognitive index.

[0037] Project 32. The method of any one of Projects 29 to 31, wherein Bifidobacteria have the ability to proliferate in the intestine.

[0038] Project 33. According to the method of any one of Projects 29 to 32, Bifidobacterium has an anti-metabolic syndrome effect.

[0039] Project 34. The method of any one of Projects 29 to 33, wherein Bifidobacteria and water-soluble dietary fiber are ingested in the form of food.

[0040] Project 35. A method according to any one of Projects 29 to 34, used to inhibit cognitive decline, maintain cognitive function, or improve cognitive function, wherein bifidobacteria and water-soluble dietary fiber are ingested in the form of food.

[0041] Project 36. A combination of Bifidobacteria and water-soluble dietary fiber for the purpose of improving at least one of complex attention, cognitive flexibility, and executive function.

[0042] Item 37. A combination of uses according to Item 36, wherein the water-soluble dietary fiber contains inulin.

[0043] Item 38. A combination of the uses of Items 36 or 37, which is used to improve neurocognitive indices.

[0044] Item 39. A combination of any of the uses in items 36 to 38, wherein Bifidobacteria have the ability to proliferate in the gut.

[0045] Item 40. A combination of any of the uses in Items 36 to 39, wherein Bifidobacterium has an anti-metabolic syndrome effect.

[0046] Item 41. It is a food product according to any combination of uses from items 36 to 40.

[0047] Item 42. A food for inhibiting cognitive decline, maintaining cognitive function or improving cognitive function, based on any combination of uses in items 36 to 41.

[0048] Project 43. Use of water-soluble dietary fiber in combination with Bifidobacteria to improve at least one selected from complex attention, cognitive flexibility and executive function.

[0049] Item 44. Water-soluble dietary fiber for use according to Item 43, which contains inulin.

[0050] Item 45. Water-soluble dietary fiber used in accordance with the uses in Item 43 or 44, for use in combination with Bifidobacteria to improve neurocognitive indices.

[0051] Item 46. Water-soluble dietary fiber for any of the uses of items 43 to 45, wherein bifidobacteria have the ability to proliferate in the intestine.

[0052] Item 47. Water-soluble dietary fiber for any of the uses in Items 43 to 46, wherein Bifidobacteria have an anti-metabolic syndrome effect.

[0053] Item 48. Water-soluble dietary fiber for any of the uses in Items 43 to 47, in food form.

[0054] Item 49. Water-soluble dietary fiber for any of the uses in Items 43 to 48, in a food form for use in combination with Bifidobacteria to inhibit cognitive decline, maintain cognitive function or improve cognitive function.

[0055] Project 50. Use of Bifidobacteria in combination with water-soluble dietary fiber to improve at least one selected from complex attention, cognitive flexibility and executive function.

[0056] Item 51. Bifidobacteria for use according to Item 50, wherein the water-soluble dietary fiber contains inulin.

[0057] Item 52. Bifidobacteria for use in combination with water-soluble dietary fiber to improve neurocognitive index, as per Item 50 or 51.

[0058] Item 53. Bifidobacteria for any of the uses in Items 50 to 52, which have the ability to proliferate in the intestine.

[0059] Item 54. Bifidobacteria for any of the uses in Items 50 to 53, which have anti-metabolic syndrome effects.

[0060] Item 55. Bifidobacteria for any of the uses in Items 50 to 54, in food form.

[0061] Item 56. Bifidobacteria for any of the uses in Items 50 to 55, in a food form intended for use in combination with water-soluble dietary fiber to inhibit cognitive decline, maintain cognitive function, or improve cognitive function.

[0062] Advantages of the Invention: The present invention provides a novel means for improving complex attention, cognitive flexibility, and / or executive function. Each item measured by Cognitrax is typically independent of the others; even if a substance improves some of these items, it may not necessarily improve others. Specifically, ergothioneine is known to be effective for cognitive speed but ineffective for complex attention, cognitive flexibility, or executive function (Non-Patent Literature 1, Tables 2 and 3; inter-group comparison). Phospholipids are known to be effective only for verbal memory and ineffective for complex attention, cognitive flexibility, or executive function (Non-Patent Literature 2, Table 4; inter-group comparison). Furthermore, GABA is known to be effective only for motor speed and ineffective for complex attention, cognitive flexibility, or executive function (Non-Patent Literature 3, Table 1; inter-group comparison). There are currently no reports on the effects of combining Bifidobacteria with water-soluble dietary fiber to improve complex attention, cognitive flexibility, and executive function; therefore, the effects of the present invention cannot be predicted from the prior art. Attached Figure Description

[0063] Figure 1 shows a flowchart from subject recruitment to analysis in an embodiment.

[0064] Figure 2 shows the quantitative results of fecal Bifidobacteria in the examples. Detailed Implementation

[0065] Compositions for improving complex attention, cognitive flexibility, and / or executive functionThis invention provides a composition for improving at least one selected from complex attention, cognitive flexibility, and executive function, the composition comprising Bifidobacteria and water-soluble dietary fiber. Examples of Bifidobacteria in this invention include, but are not particularly limited to, *Bifidobacterium animalis*, *Bifidobacterium adolescentis*, *Bifidobacterium bifidum*, *Bifidobacterium breve*, *Bifidobacterium catenulatum*, *Bifidobacterium longum*, and *Bifidobacterium pseudochain*, with *Bifidobacterium animalis* being preferred. Among *Bifidobacterium animalis*, *Bifidobacterium animalis subsp. lactis* is preferred. The Bifidobacteria are preferably those capable of proliferating in the intestine. More specifically, the Bifidobacteria are preferably those capable of increasing the total number of Bifidobacteria in the intestine. For example, the ability to proliferate in the intestine can be determined by measuring the number of Bifidobacteria in a fecal sample using real-time PCR. More specifically, the measurement and evaluation can be performed according to the method described in Non-Patent Document 4. In this invention, if proliferation in the intestine is confirmed by the above method, Bifidobacteria are selected as the preferred Bifidobacteria of this invention. Preferably, the bacteria are capable of proliferation such that the number of Bifidobacteria becomes two times or more, preferably five times or more, more preferably ten times or more, compared to the number of bacteria ingested as measured by the above method. Furthermore, in this invention, Bifidobacteria are preferably those with anti-metabolic syndrome effects. The anti-metabolic syndrome effect can be confirmed, for example, by measuring visceral fat area. More specifically, the measurement and evaluation can be performed according to the method described in Non-Patent Document 5. In this invention, if a reduction in visceral fat area is confirmed by the above method, Bifidobacteria are selected as the preferred Bifidobacteria of this invention. Bifidobacteria are preferably those that, when measured by the above method, reduce the visceral fat area to 0.98 times or less, more preferably 0.96 times or less. In a preferred embodiment of the invention, an example of Bifidobacterium includes GCL2505, which belongs to the subspecies of Bifidobacterium animalis (Lactobacillus lactis). Strain GCL2505 is deposited with accession number FERM ABP-21918. This strain was deposited on February 17, 2010, under the Treaty of Budapest at the Patent Microbial Collection Center of the National Institute for Technical Evaluation (NITE) of Japan (Room 120, 2-5-8 Wataru, Kisarazu City, Chiba Prefecture, Japan, 292-0818).

[0066] The number of Bifidobacteria in the compositions of the present invention is not limited and can be appropriately set in the range of, for example, 10 million / g, preferably 30 million / g, more preferably 50 million / g, even more preferably 80 million / g or more, and particularly preferably 100 million / g or more. There is also no particular upper limit, for example, 10 trillion / g or less, 100 billion / g or less, or 10 billion / g or less. In a typical embodiment of the present invention, live Bifidobacteria are used.

[0067] Examples of water-soluble dietary fibers include inulin, indigestible dextrin, water-soluble soybean polysaccharides, polydextrose, sodium alginate, psyllium husk, fucoidan, brown algae starch, sodium carboxymethyl cellulose, pullulan, gelatin, and low molecular weight hemicellulose, with inulin being preferred. These water-soluble dietary fibers can be used alone or in combination of various types.

[0068] The content of water-soluble dietary fiber in the composition of the present invention is not limited. For example, it can be set in the range of 0.5 to 10 wt%, preferably 1 to 5 wt%, more preferably 1.5 to 3 wt%, based on the total mass of the composition.

[0069] In this invention, the combination of Bifidobacterium and water-soluble dietary fiber, as active ingredients, can be used in the form of a composition for improving at least one selected from complex attention, cognitive flexibility, and executive function. Alternatively, the combination can be used in the form of a composition in combination with various pharmaceutically acceptable carriers or carriers that can be added to food (e.g., isotonic agents, chelating agents, stabilizers, pH adjusters, preservatives, antioxidants, solubilizers, thickeners, excipients, and binders). In embodiments containing various carriers, the total content of Bifidobacterium and water-soluble dietary fiber in the composition is not limited and can be suitably set in the range of, for example, 50% or more by mass, 60% or more by mass, 70% or more by mass, 80% or more by mass, 90% or more by mass, 95% or more by mass, 99% or more by mass, etc.

[0070] Examples of isotonic agents include sugars such as glucose, trehalose, lactose, fructose, mannitol, xylitol, and sorbitol; polyols such as glycerol, polyethylene glycol, and propylene glycol; and inorganic salts such as sodium chloride, potassium chloride, and calcium chloride. These isotonic agents can be used alone or in combination of two or more.

[0071] Examples of chelating agents include edetates, such as disodium edetate, sodium calcium edetate, trisodium edetate, tetrasodium edetate, and calcium edetate, as well as ethylenediaminetetraacetic acid, hypozoxytriacetic acid or its salts, sodium hexametaphosphate, and citric acid. These chelating agents can be used alone or in combination of two or more.

[0072] Examples of stabilizers include sodium bisulfite.

[0073] Examples of pH adjusters include acids such as hydrochloric acid, carbonic acid, acetic acid, and citric acid; alkali metal hydroxides such as sodium hydroxide and potassium hydroxide; alkali metal carbonates or bicarbonates such as sodium carbonate; alkali metal acetates such as sodium acetate; alkali metal citrates such as sodium citrate; and bases such as tromethamine. These pH adjusters can be used alone or in combination of two or more.

[0074] Examples of preservatives include sorbic acid, potassium sorbate, parabens (such as methylparaben, ethylparaben, propylparaben, and butylparaben), quaternary ammonium salts (such as chlorhexidine gluconate, benzalkonium chloride, benzyl chloride, and cetylpyridinium chloride), alkyl polyaminoethyl glycine, chlorobutanol, polyquaternary ammonium salts, polyhexamethylene biguanide, and chlorhexidine. These preservatives can be used alone or in combination of two or more.

[0075] Examples of antioxidants include sodium bisulfite, dried sodium sulfite, sodium metabisulfite, and mixed tocopherol concentrates. These antioxidants can be used alone or in combination of two or more.

[0076] Examples of solubilizers include sodium benzoate, glycerin, D-sorbitol, glucose, propylene glycol, hydroxypropyl methylcellulose, polyvinylpyrrolidone, polyethylene glycol, and D-mannitol. These solubilizers can be used alone or in combination of two or more.

[0077] Examples of thickeners include polyethylene glycol, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, xanthan gum, sodium chondroitin sulfate, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, and polyvinyl alcohol. These thickeners can be used alone or in combination of two or more.

[0078] Examples of excipients include lactose, corn starch, L-cysteine, trehalose, maltitol, and sorbitol. These excipients can be used alone or in combination of two or more.

[0079] Examples of adhesives include crystalline cellulose, starch, sucrose, hydroxypropyl cellulose, gelatin, gum arabic, polyvinylpyrrolidone, pullulan, dextrin, cyclodextrin, methylcellulose, ethylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, and polyethylene glycol. These adhesives can be used alone or in combination of two or more.

[0080] The intake of the composition of the present invention is not limited. The daily intake of Bifidobacterium, as the active ingredient, can be appropriately set in the range of, for example, 10 million to 10 trillion, preferably 100 million to 1 trillion, more preferably 1 billion to 100 billion, even more preferably 5 billion to 30 billion, and particularly preferably 10 billion to 20 billion Bifidobacterium. The intake of the composition of the present invention based on the weight of Bifidobacterium is also not limited. The daily intake of Bifidobacterium, as the active ingredient, can be appropriately set in the range of, for example, 1 mg to 10 g, preferably 10 mg to 1000 mg, more preferably 40 mg to 300 mg, even more preferably 200 mg or more.

[0081] In addition to the aforementioned Bifidobacteria, the compositions of the present invention may also contain substances known to improve complex attention, cognitive flexibility, and / or executive function. Examples of substances known to improve complex attention, cognitive flexibility, and / or executive function include propolis and chlorogenic acid. These substances may be used alone or in combination of two or more. The compositions of the present invention may also contain substances known to improve functions other than complex attention, cognitive flexibility, and executive function, i.e., substances known to improve overall memory, verbal memory, visual memory, cognitive speed, reaction time, sustained attention, simple attention, processing speed, motor speed, social cognition, logical reasoning, working memory, neurocognitive index, etc. Examples of substances known to improve such functions include GABA, paraginose, phosphatidylcholine, and ergothioneine. These substances may be used alone or in combination of two or more.

[0082] The compositions of the present invention are preferably oral compositions. Oral compositions include food or beverage compositions and pharmaceutical compositions. In the present invention, food or beverage compositions include health functional foods (foods with nutritional function claims, foods for specific health purposes, and foods with functional claims).

[0083] Examples of food or beverage compositions include beverages such as vegetable juice beverages, fruit juice beverages, mixed vegetable and fruit juice beverages, fermented milk beverages, and almond-containing beverages; and foods such as ice cream, frozen fruit syrups, candies, gummies, almond-containing foods, biscuits (e.g., cream-filled biscuits), chocolate (including quasi-chocolate), and fermented dairy products (yogurt and cheese). Ice cream is preferably lactic acid ice cream (e.g., lactic acid ice cream containing fermented milk). Quasi-chocolate includes corn-containing quasi-chocolate. Dairy products include fermented milk, ice cream, and milk beverages. The food or beverage compositions of the present invention also include supplements, etc.

[0084] In another embodiment, the present invention provides the use of Bifidobacterium and water-soluble dietary fiber in the manufacture of a composition for improving at least one selected from complex attention, cognitive flexibility, and executive function; the use of water-soluble dietary fiber in the manufacture of a composition for improving at least one selected from complex attention, cognitive flexibility, and executive function, the composition comprising Bifidobacterium; the use of Bifidobacterium in the manufacture of a composition for improving at least one selected from complex attention, cognitive flexibility, and executive function, the composition comprising water-soluble dietary fiber; a method for improving at least one selected from complex attention, cognitive flexibility, and executive function, the method comprising administering (ingesting) an effective amount of Bifidobacterium and water-soluble dietary fiber to a subject in need; the use of a combination of Bifidobacterium and water-soluble dietary fiber for improving at least one selected from complex attention, cognitive flexibility, and executive function; the use of water-soluble dietary fiber in combination with Bifidobacterium to improve at least one selected from complex attention, cognitive flexibility, and executive function; and the use of Bifidobacterium in combination with water-soluble dietary fiber to improve at least one selected from complex attention, cognitive flexibility, and executive function. In these embodiments, the detailed information (type, amount, etc.) regarding Bifidobacterium, soluble dietary fiber, and optional other ingredients, the method of use (pharmaceuticals, food, and beverages, etc.), and the purpose of use (improving at least one selected from complex attention, cognitive flexibility, and executive function) are the same as described above. Examples of subjects suitable for administering (or ingesting) Bifidobacterium and soluble dietary fiber in this invention include mammals, such as humans, mice, rats, and guinea pigs (preferably humans). In the case of human subjects, examples include, but are not particularly limited to, humans who do not have dementia or mild cognitive impairment. The age of the subjects is also not particularly limited, and examples include age groups where complex attention, cognitive flexibility, executive function, etc., are generally considered to decline with age. In a preferred embodiment, the invention can improve such complex attention, cognitive flexibility, and / or executive function.

[0085] According to the present invention, the combination of Bifidobacteria and water-soluble dietary fiber can improve at least one (preferably two, more preferably three) selected from complex attention, cognitive flexibility, and executive function. According to the present invention, judgment can be improved by improving these functions (especially cognitive flexibility, etc.). Complex attention and executive function can also be referred to as "attention" and "executive ability," respectively. Therefore, in one embodiment, the present invention provides a composition for improving at least one selected from attention, judgment, and executive function, the composition comprising Bifidobacterium and water-soluble dietary fiber; use of Bifidobacterium and water-soluble dietary fiber in manufacturing a composition for improving at least one selected from attention, judgment, and executive function; use of water-soluble dietary fiber in manufacturing a composition for improving at least one selected from attention, judgment, and executive function, the composition comprising Bifidobacterium; use of Bifidobacterium in manufacturing a composition for improving at least one selected from attention, judgment, and executive function, the composition comprising water-soluble dietary fiber; a method for improving at least one selected from attention, judgment, and executive function, the method comprising administering (ingesting) an effective amount of Bifidobacterium and water-soluble dietary fiber to a subject in need; use of a combination of Bifidobacterium and water-soluble dietary fiber for improving at least one selected from attention, judgment, and executive function; use of water-soluble dietary fiber in combination with Bifidobacterium to improve at least one selected from attention, judgment, and executive function; and use of Bifidobacterium in combination with water-soluble dietary fiber to improve at least one selected from attention, judgment, and executive function.

[0086] The specific embodiments of the present invention will now be described in more detail with reference to the examples. However, the present invention is not limited to these examples.

[0087] The examples were randomized, double-blind, placebo-controlled, parallel group comparison studies to evaluate the potential of GCL2505 and inulin to improve cognitive function.

[0088] Test methods Subjects (target group for analysis)This study was approved by the Ethics Committee of Koyukai Kofuku Plastic Surgery Hospital on May 12, 2022 (Approval date: March 12, 2022; Approval number: MK-2205-01). All participants signed written informed consent forms based on the Declaration of Helsinki. Participants were Japanese men and women aged 50 to 80 years or younger, meeting the inclusion criteria and exclusion criteria, and were deemed eligible to participate by the principal investigator. The inclusion criteria are as follows: (1) a score of 24 or higher on the Mini Mental State Examination-Japanese (MMSE-J), a score of 17 or higher on the Japanese version of the Montreal Cognitive Assessment (MoCA-J), or a score of 5 or lower on the Geriatric Depression Scale-short Version-Japanese (GDS-SJ) in the screening test; (2) having symptoms of subjective memory loss, or having been reported by close relatives or acquaintances to have symptoms of objective memory loss; and (3) having fully understood the purpose and content of the study, having the capacity to consent, and having voluntarily participated in the study based on a full understanding of the purpose and content of the study, and having provided a written informed consent form for participation in the study.

[0089] Exclusion criteria are as follows: (1) Those with a current or past history of mental disorders (including depressive symptoms), cerebrovascular diseases, and sleep disorders; (2) Those with serious liver, kidney, heart, respiratory, endocrine, or metabolic diseases; (3) Smokers; (4) Frequent drinkers (daily intake of 60 mg / L). (5) Individuals who cannot comply with restrictions on food and supplements that affect the gut environment during the study; (6) Individuals who have taken antibiotics within one month prior to the start of the study; (7) Individuals who have undergone digestive system surgery (excluding appendicitis); (8) Individuals with a history of allergy to any food component in the study; (9) Individuals currently taking medications that may affect cognitive function (e.g., antipsychotics, anxiolytics, antidepressants, anti-Parkinson's drugs, antimanic drugs, antiepileptics, and anticoagulants); (10) Individuals with visual or hearing impairments that may interfere with cognitive function testing; (11) Individuals who frequently consume foods or supplements that may affect cognitive function; (12) Individuals with a current or past history of drug or alcohol dependence; (13) Individuals who participate in studies involving the intake of other foods or the use of drugs, or studies involving the application of cosmetics or drugs, or who participate in other studies during the period of participation in this study; and (14) Any other disqualifications identified by the principal investigator.

[0090] Test foodThe tested foods included a dairy beverage (test beverage) containing inulin (Orafti GR; BENEO GmbH, Mannheim, Germany) and GCL2505, as well as a placebo. The test beverage was designed to contain 2.0 g of inulin per 100 g and 1×10⁻⁶ ppm per 100 g. 10 GCL2505 with colony-forming units (cfu). The placebo was prepared using the same ingredients as the test product, with the addition of food-grade acetic acid and lactic acid to adjust its flavor and pH. The base ingredients were skim milk powder, high-fructose corn syrup, sucrose, yeast extract, acidifier, stabilizer, and flavoring agent. Table 1 shows the nutritional details of the test food and placebo.

[0091] [Table 1] Experimental Design This test was conducted as a randomized, placebo-controlled, double-blind, parallel-group comparative study. Participants were randomly assigned to two subject groups using stratified permutation randomization based on age, sex, and Cognitrax (short version), MMSE-J, and MoCa-J scores at the time of screening. Block size was set at 4. A controller (assignment administrator) assigned the two groups to the experimental food intake group and the control food intake group. Participants in both the active and placebo groups consumed 100g of the dairy beverage once daily for 12 weeks. Throughout the trial, neither participants nor observers were aware of their group assignments. The double-blinding method was achieved by labeling the experimental food with only an identification number. The primary outcome was Cognitrax (long version) at 12 weeks after the start of the trial. Secondary outcomes were Cognitrax (long version), fecal Bifidobacterium count, and SF-36v2 (registered trademark) at 8 weeks after the start of the trial.

[0092] Cognitrax test (PC-based cognitive function test) Participants' cognitive functions were assessed using the Cognitrax measurement, developed as a Japanese version of the CNS Vital Signs (CNSVS) (Non-Patent Document 6). The Cognitrax cognitive function tests were conducted in the following order: Verbal Memory Test (VBM), Visual Memory Test (VIM), Finger Tapping Test (FTT), Symbolic Digit Coding Test (SDC), Stroop Test (ST), Attention Shift Test (SAT), Continuous Performance Test (CPT), Affective Perception Test (POET), Nonverbal Reasoning Test (NVRT), and Four-Part Continuous Performance Test (FPCPT). The tests were conducted according to a prior procedure (Non-Patent Document 7).

[0093] Mental health status At weeks 0, 8, and 12, the World Health Organization (WHO) Five-in-One Well-being Index (WHO-5-J) (Non-Patent Literature 8) was used to confirm that the subjects had no abnormal mental health status during the study period.

[0094] Biochemical parameters Blood pressure, pulse, and weight were measured at weeks 0, 8, and 12. Biochemical parameters were also measured at weeks 0 and 12. Blood samples were collected from each subject 4 hours after fasting or consuming beverages (except water) prior to testing. Biochemical parameters included hematological tests (white blood cell count, red blood cell count, hemoglobin, hematocrit, MCV, MCH, MCHC, platelet count, and white blood cell count), biochemical tests (total protein, albumin, total bilirubin, AST, ALT, LD (IFCC), ALP (IFCC), γ-GT, blood urea nitrogen (UN), creatinine, uric acid, sodium, chloride, potassium, calcium, total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, phospholipids, glucose, HbA1c (NGSP), and insulin), and urinalysis (protein, glucose, bilirubin, urinary ketones, occult blood reaction, urobilinogen, pH, and specific gravity). Serum BDNF was also quantified. All these tests were performed by LSI Medience Corporation (Tokyo, Japan).

[0095] Inflammatory proteome For blood inflammatory markers, proteins in frozen serum were detected using the Olink (registered trademark) Target 96 Inflammatory Protein Detection Plate (Olink Proteomics AB, Uppsala, Sweden) according to the manufacturer's instructions. This plate employs proximity amplification (LAA), a high-throughput multiplex proteomic immunoassay method (see Non-Patent Literature 9). The plate contains 92 immune-related proteins, primarily cytokines and chemokines. In this assay, epitope-specific binding and hybridization were performed using a set of paired oligonucleotide antibody probes, followed by amplification using quantitative PCR. The results were then log-normalized to base 2 to obtain Olink's own relative abundance units (normalized protein expression (NPX) values). Quality control was performed on each sample plate, including both internal and external controls with added internal standards. This testing was outsourced to Pharma Foods International Co., Ltd. (Tokyo, Japan).

[0096] Fecal sample collection Stool samples were submitted in week 0 and week 12. Stool samples were collected at home 7 to 2 days prior to the scheduled visit date. The collected stool samples were immediately transported to the testing company under refrigeration.

[0097] Fecal DNA ExtractionAccording to Tourlousse et al. (Non-Patent Document 10), bacterial DNA was extracted from fecal samples using the ISOSPIN Fecal DNA Kit (Nippon Gene Co., Ltd., Tokyo, Japan). Specifically, the sample (0.2 g fecal sample in this case), 700 μL of FE1 buffer, and 10 μL of RNase were added to a test tube containing magnetic beads. Cells were lysed using a FastPrep-24 (MP Biomedicals, Irvine, CA) at 6 m / s for 1 minute. This was repeated three times, with the sample held at room temperature for 5 minutes during this period. Subsequently, 90 μL of FE2 buffer was added, and the sample was centrifuged at 12,000 × g for 15 minutes. The supernatant (up to 500 μL) was collected and mixed with FB buffer and isopropanol, each at 0.4 times the volume of the resulting supernatant. Finally, the sample was loaded onto a centrifuge column and washed according to the manufacturer's instructions. The purified DNA was eluted with 50 μL Tris-EDTA buffer (pH 8.0).

[0098] fecal bifidobacteria According to Tanaka et al., real-time polymerase chain reaction (PCR) was performed using genus-specific primers capable of detecting Bifidobacterium (including GCL2505). The primer sequences are as follows: Bifidobacterium positive primer: 50-GATTCTGGCTCAGGATGAACGC-30; Bifidobacterium antisense primer: 50-CTGATAGGACGCGACCCCAT-30. Each PCR reaction mixture consisted of 20 pmol of each primer (total volume 10 µL), 5 µL of SYBR (trademarked) premix ExTaq (Takara Bio Inc., Shiga, Japan), and 1 µL of DNA solution (Non-Patent Document 11). This assay was outsourced to the Kyoto Institute of Nutritional Pathology (Kyoto, Japan).

[0099] Statistical analysisAll measurements are expressed as mean and standard deviation (SD). All statistical analyses were performed using IBM SPSS Statistics 27 (IBM, Armonk, NY, USA). A p-value < 0.05 was used as the threshold for significance. As basic statistics, mean and SD are expressed to one decimal place, and percentages (%) are expressed to one decimal place, with rounding. Missing data were treated as missing values, and no substitutes were used. Statistical analyses of Cognitrax (long version) were performed using paired t-tests to compare results at the start of intake (week 0) with those at weeks 8 and 12. For other items, Dunnett's test (two-tailed test) was used to compare results at the start of intake (week 0) with those at weeks 8 and 12, and Wilcoxon's signed-rank test was used for qualitative evaluation items. Comparisons between the active group and placebo group during each test period were performed using unpaired t-tests (two-tailed tests), and Wilcoxon's rank-sum test was used for qualitative evaluation items.

[0100] result Subjects (target group for analysis) An overview of the trial period is shown in the flowchart in Figure 1. In this study, 255 participants were screened. As a result of the screening, 80 participants were eligible: 40 were assigned to the active group and 40 to the placebo group. At the start of the trial, a significant difference in alkaline phosphatase levels was observed between the active and placebo groups; however, this difference was considered acceptable because these values ​​were all within the reference range. For other items, there were no differences in baseline characteristics of the participants' data (Table 2).

[0101] [Table 2] At the end of the trial, one participant from the active group withdrew for personal reasons. After the entire trial concluded, one participant from the placebo group was discontinued due to adherence issues. Additionally, one participant from the active group was excluded due to a diagnosed illness unrelated to the trial (potentially affecting the outcome). Furthermore, nine participants were excluded because they took medications or foods during the trial that could affect the results (n=5 from the active group, n=4 from the placebo group). Additionally, one participant from the active group was excluded due to partially missing data for the primary endpoint. Therefore, a total of 67 participants (32 from the active group, 35 from the placebo group) were included in the target population for the analysis. Furthermore, four participants were excluded from the Cognitrax analysis due to a lower WHO-5-J score (n=1 from the active group, n=3 from the placebo group) because their scores were ≤50, a score used to specify the cutoff for the WHO-5 global version of depression screening diagnosis. A lower WHO-5-J score indicates the likelihood of depression in older adults due to an indoor lifestyle caused by the COVID-19 pandemic; therefore, the Cognitrax test may not have been performed correctly on these participants.

[0102] Cognitrax test The change in scores in the neurocognitive index domain from week 0 to week 12 in the active group (5.5±7.1) was greater than that in the placebo group (2.3±4.0), and the difference between the two groups was statistically significant (p=0.027 by unpaired t-test). Furthermore, the change in scores in the complex attention domain from week 0 to week 12 in the active group (8.3±11.8) was significantly higher than that in the placebo group (3.2±6.9) (p=0.041 by unpaired t-test); the change in scores in the cognitive flexibility domain in the active group (9.8±11.5) was significantly higher than that in the placebo group (4.8±6.7) (p=0.038 by unpaired t-test); and the change in scores in the executive function domain in the active group (9.5±11.8) was significantly higher than that in the placebo group (4.5±6.9) (p=0.044 by unpaired t-test) (Table 3).

[0103] [Table 3] Table 3. Changes in each parameter of cognitive function due to intervention (placebo group: n=32; active group: n=31). All data are represented as the average Cognitrax score (SD).

[0104] Paired t-tests were used to compare data from weeks 8 and 12 with those from week 0. Changes between the placebo and active groups were compared using unpaired t-tests.

[0105] fecal bifidobacteria count The number of Bifidobacteria in feces was quantified (Figure 2). At week 12, the number of Bifidobacteria in the active group (7.71 ± 0.56 log cells / g feces) was significantly increased compared with week 0 (7.39 ± 0.74 log cells / g feces) (p = 0.005 by Dunnett's multiple comparison test), while no change was observed in the placebo group. Furthermore, at week 12, the total number of Bifidobacteria in the active group was significantly increased compared with the placebo group (7.31 ± 0.90 log cells / g feces) (p = 0.031 by unpaired t-test).

[0106] Blood inflammatory markers The expression levels of a total of 92 inflammatory biomarkers were measured at weeks 0 and 12 using the Olink (registered trademark) Target 96 inflammation detection panel. Seventy-five inflammatory biomarkers were identified as quantifiable. Results showed statistically significant differences between the active and placebo groups in the changes in scores for leukemia inhibitory factor receptor (LIF-R), sulfotransferase 1A1 (ST1A1), CC motif chemokine 23 (CCL23), and tumor necrosis factor (ligand) superfamily member 12 (TWEAK) during weeks 0 to 12. During weeks 0 to 12, there were no statistically significant differences between the active group and the placebo group in the scores for adenosine deaminase (ADA), osteoprotegerin (OPG), eosinophil activation chemokine (CCL11), glial cell line-derived neurotrophic factor (GDNF), fractal chemokine (CX3CL1), interleukin-8 (IL-8), CC motif chemokine 28 (CCL28), interleukin-18 (IL-18), interleukin-10 (IL-10), CC motif chemokine 19 (CCL19), and T cell surface glycoprotein CD5 (CD5); however, the trend was considered significant due to p < 0.1 (Table 4).

[0107] [Table 4] Table 5. Changes in each inflammatory marker due to the intervention (those with intergroup differences P-value <0.1 were selected). All data were obtained using the Olink® Target 96 Inflammation Biomarker Detection Plate and are expressed as the mean (SD) of log-baseline 2 normalized protein expression (NPX).

[0108] Paired t-tests were used to compare the data from week 12 with those from week 0.

[0109] The changes between the placebo group and the active group were compared using an unpaired t-test.

[0110] The study results indicate that consuming a probiotic beverage containing GCL2505 and inulin improved complex attention, cognitive flexibility, executive function, and neurocognitive index domain scores. These cognitive improvements contribute to quality of life, and the results of this study are considered clinically significant. (PCT / RO / 134)

Claims

1. A composition for improving at least one selected from complex attention, cognitive flexibility, and executive function, said composition comprising bifidobacteria and water-soluble dietary fiber.

2. The composition according to claim 1, wherein the water-soluble dietary fiber comprises inulin.

3. The composition according to claim 1 or 2, used to improve neurocognitive index.

4. The composition according to claim 1 or 2, wherein the Bifidobacterium has the ability to proliferate in the intestine.

5. The composition according to claim 1 or 2, wherein the Bifidobacterium has an anti-metabolic syndrome effect.

6. The composition according to claim 1 or 2, wherein it is a food product.

7. The composition according to claim 1 or 2, which is a food for inhibiting cognitive decline, maintaining cognitive function, or improving cognitive function.

8. Use of Bifidobacteria and water-soluble dietary fiber in the manufacture of a composition, wherein the composition is used to improve at least one selected from complex attention, cognitive flexibility and executive function.

9. The use according to claim 8, wherein the water-soluble dietary fiber comprises inulin.

10. The use according to claim 8 or 9, wherein the composition is used to improve neurocognitive index.

11. The use according to claim 8 or 9, wherein the Bifidobacterium has the ability to proliferate in the intestine.

12. The use according to claim 8 or 9, wherein the Bifidobacterium has an anti-metabolic syndrome effect.

13. The use according to claim 8 or 9, wherein the composition is a food product.

14. The use according to claim 8 or 9, wherein the composition is a food for inhibiting cognitive decline, maintaining cognitive function, or improving cognitive function.

Images