JP2019104730A - Composition for raising cognitive function - Google Patents

Abstract

To provide compositions for raising, maintaining, and/or improving cognitive function.SOLUTION: The present invention provides a composition for use in raising, maintaining, and/or improving cognitive function, comprising a black tea aroma component as an active ingredient. The black tea aroma component can be at least one selected from the group consisting of geraniol, linalool oxide, and 2-phenylethanol. The composition of the present invention can be provided in inhalation composition form via the respiratory tract, and in food form and oral preparation form.SELECTED DRAWING: None

Description

  The present invention relates to a composition for use in the improvement, maintenance and / or improvement of cognitive function.

  Improvement, maintenance and improvement of cognitive function are required for a wide range of generations, from young people to elderly people. Improvement, maintenance, and improvement of memory and learning ability are important not only for students and working people studying for examinations and qualification exams, but also for daily work and life. In addition, in the elderly, memory decline and concentration decline are related to the quality of life, so it is required to prevent, reduce, improve, maintain and improve cognitive function.

  Furthermore, in old age, an increase in mental illness causing cognitive decline with aging is becoming a social problem along with an increase in elderly people. Not only dementia such as Alzheimer's disease, but also mental diseases such as depression and delirium have been reported as diseases causing cognitive function decline (Non-patent Document 1).

  By the way, teas are mainly divided into two kinds of infermented tea such as green tea and fermented tea such as black tea and oolong tea according to the manufacturing method. In the fermentation process, volatile aroma components are produced by enzyme reaction such as glycosidase present in fresh tea leaves, so in fermented tea, in addition to the aroma components contained in unfermented tea, new aroma components, The amount is huge. The aroma of tea is important for commercialization because it affects the quality and taste of tea, while the aroma component of tea is known for its health function such as anti-stress (Patent Document 1).

International Publication No. 2005/011718

Tanaka Hisashi, Takeda Masatoshi (2011) Nippon Clinical, Extra Issue, pages 52-56

  The present inventors have now found that black tea aroma components can improve, maintain and / or improve cognitive function. The present inventors have also found that black tea aroma components enhance sympathetic nerve activity to activate the brain and improve, maintain and / or improve cognitive function. The present inventors further found that the combined use of the black tea aroma component and the whey enzyme degradation product can further enhance the improvement, maintenance and / or improvement effect of cognitive function. The present invention is based on these findings.

  That is, the object of the present invention is to provide a composition and agent for improving, maintaining and / or improving cognitive function.

According to the present invention, the following inventions are provided.
[1] A composition for improving, maintaining and / or improving cognitive function (sometimes referred to herein as "the composition of the present invention") and cognitive function comprising a black tea aroma component as an active ingredient Improvers, maintainers and improvers (sometimes referred to herein as the “agent of the present invention”).
[2] The composition and preparation according to the above-mentioned [1], wherein the cognitive function is a memory function.
[3] The composition and preparation according to the above-mentioned [2], wherein the memory function is a long-term memory function.
[4] The composition and preparation according to the above-mentioned [1], wherein the cognitive function is an attention function.
[5] The composition and preparation according to the above-mentioned [1], wherein the cognitive function is a memory acquisition function and / or a memory fixation function.
[6] The composition and preparation according to any one of the above [1] to [5], wherein the black tea aroma component is at least one selected from the group consisting of geraniol, linalool oxide and 2-phenylethanol.
[7] The composition and preparation according to any one of the above [1] to [6], for use in healthy subjects.
[8] The composition according to any one of the above [1] to [7] for use in the treatment, prevention or amelioration of a disease or condition which can be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function Substances and preparations.
[9] The composition and preparation according to any one of the above [1] to [8], which is a composition for inhalation via the respiratory tract.
[10] The composition and preparation according to any one of the above [1] to [9], which is in the form of a food or an oral preparation.
[11] The composition according to [10] above, which further comprises a milk protein enzyme degradation product as an active ingredient.
[12] The composition according to [10] above, for use in combination with a milk protein enzyme degradation product or a composition containing the same.

  According to the present invention, a composition containing a black tea aroma component capable of enhancing, maintaining and / or improving cognitive function is provided. The black tea aroma component which is the active ingredient of the present invention is an ingredient derived from black tea which has been ingested by humans for many years as a food, and the present invention is advantageous in that it can be used as a functional material safe for mammals including humans. .

A: Immunohistochemical staining image of phosphorylated CREB in the hippocampal dentate gyrus area with the control and black tea flavor presentation in Example 1. B: It is a graph showing the ratio of the staining cell area of black tea flavor presentation to control presentation in the same visual field area. Expressed as mean ± standard deviation. *** indicates p <0.001 (t test vs. control group). It is a graph showing monoamine production of brain tissue by black tea flavor addition in Example 2 (A: serotonin, B: norepinephrine, C: dopamine, D: dihydroxyphenylacetic acid, E: 3-methoxytyramine, F: homovanillic acid). Expressed as mean ± standard deviation. *** indicates p <0.001 (ANOVA Dunnett's test vs. control group). FIG. 16 is a diagram showing an outline of a novel object recognition test of a mouse in Example 4. It is a figure showing the time rate (Discrimination index) of the novel object search of the mouse in Example 4 (A: black tea flavor, B: each compound, C: five-component mixture). It is expressed as mean ± standard error. * Indicates p <0.05 (Dunnett's test vs. control group). It is a figure showing the search time ratio of each of the novel object search of a mouse | mouth in Example 4, and a novel object search (A: black tea flavor, B: each compound, C: five-component mixture). It is expressed as mean ± standard error. * Indicates p <0.05 (t test vs. search time ratio of non-new object search). It is a figure showing the time rate (Discrimination index) of the novel object search of the mouse in Example 5 (A: geraniol, B: geraniol, C: linalool oxide, D: 2-phenylethanol). It is expressed as mean ± standard error. *** indicates p <0.001, * indicates p <0.05 (Dunnett's test vs. control group). It is a figure showing the time rate (Discrimination index) of the novel object search of a mouse about the presentation timing of an aroma component (example 6). ** indicates p <0.01 (Dunnett's test vs. control group). It is a figure showing the time rate (Discrimination index) of the novel object search of a mouse about the presentation timing of an aroma component (example 7). It is a figure showing the time rate (Discrimination index) of the novel object search of a mouse about the combined use effect of an aroma component and whey enzyme degradation product on cognitive function (example 8). ** indicates p <0.01, * indicates p <0.05 (Dunnett's test vs. control group). It is a figure showing the time rate (Discrimination index) of the novel object search of a mouse about the combined use effect of an aroma component and whey enzyme degradation product on cognitive function (example 9). ** indicates p <0.01, * indicates p <0.05 (Dunnett's test vs. control group). FIG. 16 is a view showing the flow of test perfume inhalation and pupillary reflex measurement in the test carried out in Example 10. It is a figure showing the measurement result of relative CR value (%) in the autonomic nerve reaction evaluation test carried out in Example 10. Open bars indicate the control perfume, and black bars indicate the test perfume. The control perfume value was displayed as 100. * Indicates p <0.05, ** indicates p <0.01 (1-sample t-test vs. control group). It is a figure showing the measurement result of relative VD value (%) in the autonomic nerve reaction evaluation test carried out in Example 10. Open bars indicate the control perfume, and black bars indicate the test perfume. The control perfume value was displayed as 100. * Indicates p <0.05, ** indicates p <0.01 (1-sample t-test vs. control group).

Detailed Description of the Invention

  In the present invention, "black tea aroma component" means a component having a smell contained in black tea leaves, that is, an aroma component contained in black tea leaves (hereinafter sometimes referred to as "black tea aroma component of the present invention") . As the black tea aroma component of the present invention, for example, even if it is an aroma component contained in black tea leaf itself, it may be an aroma component contained in a black tea extract obtained by subjecting the black tea leaf to an extraction treatment. . The black tea aroma component of the present invention can also be incorporated into the composition in the form of black tea extract. Examples of the black tea extract include extracts from black tea leaves used as raw materials for producing black tea beverages and black tea flavors and processed products thereof (for example, concentrated liquid extracts, powder extracts and the like).

  The black tea aroma component of the present invention may be a single aroma component or a combination of two or more aroma components. As a component which comprises the black tea aroma component of this invention, a linalool, geraniol, a benzyl alcohol, linalol oxide, 2-phenylethanol is mentioned, for example. Further, the composition of the present invention preferably contains at least one selected from the group consisting of geraniol, linalool oxide and 2-phenylethanol as a black tea aroma component.

  The black tea aroma component of the present invention may be a synthetic product or a natural product, and a commercially available product may be used. When the black tea aroma component of the present invention is a natural product, the black tea aroma component is extracted from black tea leaves according to known methods (including those collected by the head space method and dissolved in a solvent), and in some cases desired The purified or crudely purified product, ie, the extract from black tea leaves and its processed products can be used. The extract from black tea leaves and the processed product thereof may use a commercially available raw material for producing a black tea beverage (for example, a black tea extract, a black tea flavor).

In the present invention, the term "black leaf" means that the buds, leaves or stems of a tea tree ( Camellia sinensis var.), Which is an evergreen tree of the camellia family, are withered, strained, fermented and dried. The tea leaf used in the present invention is not particularly limited in tea season, shape of tea leaf, place of production, variety, kind, grade and the like, and dry tea leaf generally marketed for drinking can be used. The types of tea leaves are preferably Sri Lankan tea leaves (eg, Uba, Dimbra, Kandy, Ratnapura, Gal, Ruhuna), Indian tea leaves (eg, Assam, Darjeeling, Nilgiri), Kenya, Keyman, Java, and more preferably. Are Darjeeling and Dimbra. One of these may be used, or two or more may be used in combination.

  In the composition and preparation of the present invention, the black tea aroma component of the present invention can be used alone or in combination with other components. The blending amount of the black tea aroma component of the present invention in the composition and preparation of the present invention can be optionally determined according to the purpose, use, form, dosage form, symptom, age, etc., and the present invention is limited thereto Although not preferred, it can be blended at a content of 0.001 to 10% by mass (preferably 0.01 to 1% by mass) based on the total amount. In the present invention, the preparation of the present invention can be composed of the black tea aroma component of the present invention, and the composition of the present invention can be composed of the black tea aroma component of the present invention and other components.

  The black tea aroma component content in the composition of the present invention can be measured by gas chromatography mass spectrometry, and can be measured, for example, according to the procedure described in the following Example (Example 3).

  As described in the examples below, the black tea aroma component has an effect of enhancing, maintaining and / or improving cognitive function including memory function. Therefore, the black tea aroma component can be used as an active ingredient of a composition for improving, maintaining and / or improving cognitive function, and also a method for improving cognitive function, a method for maintaining cognitive function and a method for improving cognitive function It can be used. In addition, the black tea aroma component can be used as an active ingredient of a cognitive function improver, a cognitive function maintainer and a cognitive function improver. In the present invention, "cognitive function" is a concept encompassing all processes of information processing from perception to judgment, and in this process, memory functions (memory acquisition function, memory fixing function, memory recall function) And various higher brain functions such as memory and reproduction functions, attention functions and judgment functions. That is, in the present invention, "cognitive function" is used in the meaning including these higher brain functions.

  In the present invention, “improvement of cognitive function” includes, for example, enhancing cognitive function from the current state. Also, "maintenance of cognitive function" includes, for example, preventing deterioration of cognitive function. Furthermore, "improvement in cognitive function" includes, for example, recovery of symptoms that have already shown a decline in cognitive function or decline. The improvement, maintenance and / or improvement of cognitive function is used in the sense including enhancement of cognitive function and suppression of decline in cognitive function.

  In the present invention, improvement, maintenance and / or improvement of cognitive function can be evaluated using higher brain function involved in cognitive function as an index. Model animal tests for evaluating higher brain functions involved in cognitive function are known, for example, by novel object recognition tests for evaluating memory function which is one of higher brain functions (see Examples 4 to 9 below), The improvement, maintenance and / or improvement of cognitive function can be evaluated.

  In the present invention, the "memory function" refers to a function including a spatial cognitive function, a working memory function, a working memory function, an episode function, a visual memory function, and a learning function, and is used in a meaning including medium to long-term memory function. Working memory includes working memory of language, working memory of work and working memory of space. In the present invention, the memory function is preferably a long-term memory function.

  In the present invention, "improvement of memory function" includes, for example, enhancing the memory function from the present condition, promoting medium- to long-term memory consolidation, and thus promoting brain development. Further, "maintenance of memory function" includes, for example, preventing deterioration of memory function. Furthermore, "improvement of memory function" includes, for example, recovery of symptoms with memory function or signs of deterioration. The improvement, maintenance and / or improvement of the memory function include enhancement of the memory function and suppression of deterioration of the memory function.

  In the present invention, "attention function" is used in the meaning including directing attention to a part of information, changing an object, and distributing attention to a plurality of objects at the same time. That is, the "attention function" is a selection function (function to turn attention to specific things from various information coming from the outside world), a maintenance function (function to maintain one-time attention for a certain period), conversion It includes functions (function to shift attention once to other things) and allocation function (function to distribute attention when performing multiple tasks simultaneously). Attention function can be reworded as concentration.

  In the present invention, "improvement of the attention function" includes, for example, enhancing the attention function more than the current state. Also, "maintenance of the attention function" includes, for example, preventing the deterioration of the attention function. Furthermore, "improvement of the attention function" includes, for example, recovery of a symptom that has once had a decline in attention function or decline. The improvement, maintenance and / or improvement of the attention function include enhancement of the attention function and suppression of deterioration of the attention function.

  In the present invention, the “decision function” is a function in which several cognitive functions are integrated, space recognition for grasping the state or shape of an object, orientation for judging the current situation of oneself, and things are divided into groups. It means functions that include situation judgment and decision-making that are required in everyday life, such as group thinking.

  In the present invention, “improvement of the judgment function” includes, for example, enhancing the judgment function from the current state. In addition, “maintenance of the determination function” includes, for example, preventing the deterioration of the determination function. Furthermore, "improvement of the judgment function" includes, for example, recovery of a symptom which has once deteriorated and which has a sign of deterioration. The improvement, maintenance and / or improvement of the judgment function include enhancement of the judgment function and suppression of deterioration of the judgment function.

  In the present invention, treatment, prevention or amelioration of diseases and conditions which can be treated, prevented or ameliorated by improvement, maintenance and / or amelioration of cognitive function (preferably treatment by improvement, maintenance and / or amelioration of memory function, Tea aroma components can be used in the treatment, prevention or amelioration of diseases and conditions that may be prevented or ameliorated. Therefore, according to the present invention, it comprises the black tea aroma component as an active ingredient, for use in the treatment, prevention or amelioration of diseases and conditions that can be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function. There are provided a composition, and an agent for treating, preventing and ameliorating diseases and conditions that can be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function, which comprises a black tea aroma component as an active ingredient.

  Diseases and conditions that may be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function include, for example, memory impairment (conditions in which memory can not be recalled or can not remember new things), orientation Disorders (time, place, misplacement of people), cognitive impairment (computational ability, poor judgment, aphasia, agnosia, apraxia, executive dysfunction) may be mentioned.

  The composition for use in the improvement, maintenance and / or improvement of cognitive function of the present invention, and the agent for improving, maintaining and / or improving the cognitive function of the present invention, the therapeutic agent, the preventive agent and the improving agent of the present invention Provided in the form of fragrance, cosmetics, food, medicine, quasi-drugs, feed (including pet food), etc., and can be carried out according to the following description.

  The compositions and compositions of the invention can be inhaled by the subject via the respiratory tract. Here, the inhalation via the airway is inhalation via the nasal passage, in particular, of the upper airway. Also, the subject is a mammal including human, and human includes healthy persons. By allowing the subject to inhale the tea aroma component of the present invention through the nasal cavity, the sense of smell is stimulated, and an effect of improving, maintaining and / or improving cognitive function can be obtained.

  When the composition and agent of the present invention are inhaled through the airway, they may be in a form suitable for inhalation through the airway. For example, since the black tea aroma component of the present invention can be contained in a fragrance and can be inhaled by the use of the fragrance, it can be formulated as a fragrance or a fragrance composition. Examples of the aromatics include aromatic oils, aromatic candles, room fragrances, car fragrances and incense. In the present invention, it is preferable to dilute the black tea aroma component of the present invention with a solvent such as ethanol, oil, etc., atomize it using a diffuser (aroma diffuser), and inhale it into the subject. It can be formulated as a fragrance suitable for use in a diffuser.

  The black tea aroma component of the present invention can also be formulated into a cosmetic or cosmetic composition because it can be contained in a cosmetic and the subject can be inhaled by using the cosmetic. Examples of the cosmetic include perfumes such as parfum, eau de parfum, eau de toilette, eau de cologne, etc .; skin care cosmetics such as milky lotion, cream, lotion, beauty fluid, body lotion, body powder, aroma oil; hair cream, rinse, conditioner, hair styling Hair and other hair cosmetics; Body soaps, shampoos, rinse-in shampoos, facial cleansers, hand soaps, soaps and other detergents; Clothes deodorants, sizing agents, antistatic agents, softeners, etc .; Laundry finish agents; Agents, gluing agents, anti-static agents. The fragrance and cosmetics and the aroma composition and the cosmetic composition containing the black tea aroma component of the present invention can be prepared according to known methods.

  When the composition and preparation of the present invention are provided as a fragrance or a cosmetic, the black tea aroma of the present invention is used in an amount such that the cognitive function is improved, maintained and / or improved through olfactory stimulation by inhalation. The ingredients can be blended. The content of the black tea aroma component of the present invention in the fragrance and the cosmetic can be appropriately determined so as to obtain the improvement, maintenance and / or improvement effect of the cognitive function, in which case the black tea aroma described later The dose of ingredients, types of fragrances and cosmetics, the amount necessary for diffusion for fragrances, and the amount and location of application per application, application temperature, etc. for cosmetics can be considered. For example, when the composition and preparation of the present invention are provided as a fragrance for a diffuser, the content of the black tea aroma component of the present invention in the fragrance is from 0.000001 to 1% by mass (preferably 0.00001) To 0.1% by mass, more preferably 0.0001 to 0.01% by mass). When the composition and preparation of the present invention are provided as a skin care cosmetic, the content of the black tea aroma component of the present invention in the cosmetic is 0.001 to 1.0% by mass (preferably 0.01%). It can be made into-0.5 mass%).

In the present invention, cognitive function can be improved, maintained and / or improved by inhaling a subject with the black tea aroma component of the present invention at a predetermined dose. Although the present invention is not limited by the following description, for example, 0.011 m ³ of room (a black tea aroma component of the present invention dissolved in liquid paraffin at a concentration of 0.00001 to 0.1 mass%) An amount equivalent to the black tea aroma component to be inhaled when volatilized with a diffuser at room temperature (23 ° C, humidity 55 ± 5%) in area of 0.076 m ² ), preferably for 10 seconds or more per subject Can be inhaled for 1 minute or more, more preferably 5 minutes or more. The upper limit of the inhalation time is not particularly limited, and can be, for example, 30 minutes or 1 hour.

  The compositions and preparations of the present invention can also be taken orally or orally administered to a subject. As described above, the black tea aroma component of the present invention exerts an effect of enhancing, maintaining and / or improving cognitive function by olfactory stimulation via the nasal cavity, but the black tea aroma component of the present invention is effective by oral intake or oral administration In the present invention, it is intended to improve, maintain and / or improve cognitive function by oral intake or oral administration of a tea aroma component in the present invention, since part of the substance diffuses into the nasal cavity through the oral cavity and pharynx and stimulates sense of smell. it can. Here, the subject is a mammal including a human, and the human includes a healthy person.

  The compositions and preparations of the present invention can be provided in a form suitable for oral ingestion or oral administration, and examples of such forms include food and oral preparations. Here, "food" means health food, functional food, dietary supplement, supplement, health food (eg, food for specified health, food for nutrition, food for functional indication), food for special use (eg, for infants) It is used in the meaning including food, food for pregnant women, food for sick people). The form of the “food” is not particularly limited, and may be, for example, a form of a beverage, a semi-liquid or gel form, or a solid form. The composition and preparation of the present invention is a food which is chewed in the oral cavity (eg, chewing gum), a preparation which is chewed in the oral cavity so that the tea aroma component of the present invention can be easily diffused into the nasal cavity through the oral cavity and pharynx. (For example, chewable tablets), food for retention in the oral cavity (for example, candy, gummi, confectionery), preparations for retention in the oral cavity (for example, troche), confectionery (for example, jelly) and beverages (for example, black tea beverages) It is preferable to provide in the form of a milk-containing beverage, a milk-fermented beverage, an energy drink, a sports drink, but is not limited thereto.

  When the composition and preparation of the present invention are provided as a food or an oral preparation, the black tea aroma component as the active ingredient of the present invention is diffused into the nasal cavity through the oral cavity and pharynx to stimulate smell and improve cognitive function. The black tea aroma component can be contained (for example, blended) in an amount such that a maintenance and / or improvement effect is exhibited. The content of the black tea aroma component of the present invention is determined in consideration of the type of food and oral preparation, the usual intake per dose, the residence time in the oral cavity, the temperature at the time of intake, the surface area of the preparation, the shape etc. be able to. For example, when the composition and preparation of the present invention are provided as a chewable tablet, the content of the black tea aroma component of the present invention in the chewable tablet scheduled for single administration to adults is 0.01 to 10. It can be 0% by mass (preferably 0.1 to 5% by mass, more preferably 0.5 to 1.5% by mass). When the composition and preparation of the present invention are provided as chewable tablets, in addition to the black tea aroma component, excipients, binders, thickeners, sweeteners, acidulants, flavors, coloring agents, preservatives, stable It can manufacture by a conventional method by mix | blending 1 type (s) or 2 or more types of components selected from an agent, an emulsifier, an antioxidant, a coating agent etc. In addition, when the composition and preparation of the present invention are provided as a beverage, the content of the black tea aroma component of the present invention in a beverage (for example, 500 mL) scheduled for single intake of an adult is 0.001 to 1 It can be made into mass% (preferably 0.01 to 0.5 mass%). When the composition and preparation of the present invention are provided as a beverage, they can be produced according to the usual procedure for producing a beverage except that the black tea aroma component is contained (for example, blended) in the raw materials.

  The composition and the preparation of the present invention can be used to improve, maintain and / or improve memory function in cognitive function. In this case, the composition and preparation of the present invention may be inhaled, ingested or administered at either or both of memory behavior (memory acquisition / fixation) and memory recall behavior (memory recall / regeneration) or both. And preferably can be inhaled, ingested or administered during memory action. The timing of inhalation, intake or administration may be simultaneous with or immediately before the action, preferably immediately before the action, at either or both of the memory action and the memory call action. be able to.

  The compositions and preparations of the present invention are not limited in combination with other ingredients and materials. For example, when used in combination with a component or material that can be expected to improve, maintain and / or improve cognitive function, the effect of improving, maintain and / or improve cognitive function, or improve, maintain and / or improve cognitive function The effects such as treatment, prevention or amelioration of diseases and conditions that can be treated, prevented or ameliorated can be further enhanced. Ingredients that can be expected to have an improvement, maintenance and / or improvement effect on cognitive function include milk protein enzyme degradation products. That is, according to the present invention, a composition for improving, maintaining and / or improving cognitive function and an improvement and maintenance of cognitive function, comprising a black tea aroma component and a milk protein enzyme decomposition product as active ingredients And / or a remedy is provided. According to the present invention, there are also provided compositions and preparations of the present invention for use in combination with a milk protein enzyme degradate or a composition containing the same.

  The milk protein enzyme decomposition product (hereinafter, sometimes referred to as "milk protein decomposition product of the present invention"), which is an active ingredient of the composition and preparation of the present invention, is not particularly limited as long as it is an enzyme decomposition product of milk protein. As milk protein, whole milk, milk powder, casein or whey can be used, preferably whey. Here, "whey" is also referred to as whey, whey or whey, and means an aqueous solution obtained by removing milk fat, casein and the like from milk. Whey is composed of β-lactoglobulin, α-lactalbumin, serum albumin, immunoglobulin and the like. Although the animal origin of the whey used by this invention does not matter, it is preferable to use milk origin whey. The milk protein hydrolyzate according to the present invention is preferably whey protein enzyme hydrolyzate (hereinafter sometimes referred to simply as "whey enzyme hydrolyzate" or "whey hydrolyzate").

  The content (in terms of solid content) of the degraded milk protein of the present invention in the composition and preparation of the present invention can be arbitrarily determined according to the purpose, use, form, dosage form, symptoms, age, etc. Although this invention is not limited to this, For example, it can be set as 0.001-99 mass% (preferably 0.01-95 mass%) with respect to the whole amount.

  The degraded milk protein (in particular, degraded whey) of the present invention is preferably a peptide having the amino acid sequence of GTWY (SEQ ID NO: 1) and a peptide having the amino acid sequence of WY (hereinafter referred to as "the peptide of the present invention" Can be used. Here, "a peptide having an amino acid sequence" means a peptide whose sequence is specified by the amino acid sequence.

  The content (in terms of solid content) of the tetrapeptide GTWY in the degraded milk protein (in particular, the degraded whey) of the present invention is, for example, 0.001 to 10% by mass (preferably 0.01 to 5% by mass) The content (in terms of solid content) of the dipeptide WY is preferably, for example, 0.0005 to 5% by mass (preferably 0.005 to 1% by mass, more preferably 0. 1 to 5% by mass). 03 to 0.3% by mass).

  The content of the degraded milk protein (in particular, whey degradation product; converted to solid content) in the composition and preparation of the present invention may be optionally determined according to the purpose, use, form, dosage form, condition, body weight etc. Although the present invention is not limited thereto, it can be, for example, 0.001 to 50% by mass (preferably 0.01 to 10% by mass) with respect to the total amount.

  The method for producing the milk protein hydrolyzate of the present invention (in particular, a milk protein hydrolyzate containing the peptide of the present invention) is known and can be produced, for example, according to the description of WO 2017/086303. Alternatively, commercially available whey degradation products (for example, HW-3 (Snowmark Meg Milk Co., Ltd.)) may be used as the milk protein degradation products of the present invention.

  A milk protein hydrolyzate containing the peptide of the present invention can be produced, for example, by reacting a raw material protein with an enzyme preparation containing a proteolytic enzyme.

  When whey protein is used as a raw material, the concentration of whey protein to be subjected to the enzyme reaction is not limited as long as the protein can be dissolved, but from the viewpoint of suppressing gelation and aggregation and omitting time and effort for concentration It is preferable to set it as v%, More preferably, it is 1-20 w / v%, More preferably, it is 5-15 w / v%.

  When the raw material protein is an aqueous solution such as whey or whey, it may be subjected to an enzyme reaction as it is, or after concentration or dilution, and the pH may be adjusted as necessary. When the raw material protein is a solid such as powder, it may be dissolved in any aqueous solvent as long as the enzyme reaction proceeds, but it is dissolved in water or a food additive grade buffer considering its use as a food. Is preferred. It is preferable to use a buffer to prevent the pH of the reaction solution from being changed by the amino acid generated by the enzyme reaction. The type of buffer solution is optional, and may be determined in consideration of the subsequent use, taste, taste and mineral content, but the pH of the reaction solution is 4 to 9, preferably 5 to 8, more preferably 7 to 8. A composition that can be maintained at is preferred. Most preferably, it is potassium phosphate buffer. The concentration of the buffer solution is optional as long as a buffer effect can be obtained, but in consideration of taste, taste and mineral content, it can be 0.01 to 0.5 M, preferably 0.05 to 0. It is 2 M, more preferably about 0.1 M.

  Any enzyme may be used as the enzyme as long as it contains a proteolytic enzyme, but it is preferably an enzyme preparation containing a neutral protease, and one or more may be used in combination. As the enzyme preparation, for example, those derived from microorganisms such as Bacillus subtilis, Aspergillus oryzae, Aspergillus oryzae, etc. can be used, among which, an enzyme preparation derived from Aspergillus oryzae and an enzyme preparation derived from Aspergillus mereus are used. Preferably, it is an enzyme preparation derived from Aspergillus mereus, more preferably.

  In the present invention, commercially available enzyme preparations can be used. For example, enzyme preparations can be obtained from Amano Enzyme, Shin Nippon Chemical Industries, DSM, Danisco, Novozyme, HBI and the like. The addition amount of the enzyme preparation is optional, but in consideration of the appropriate hydrolysis reaction rate and cost, for example, 0.01 to 5 w / v%, preferably 0.05 to 4 w / v%, more preferably 0. .1 to 0.5 w / v%.

  The enzyme reaction temperature and the enzyme reaction time can be set so that the raw material protein is sufficiently hydrolyzed and the quality of the enzyme degradation product is maintained. That is, the enzyme reaction temperature can be, for example, 30 to 70 ° C., preferably 40 to 70 ° C., and more preferably 45 to 65 ° C. The enzyme reaction time may be 1 to 12 hours, preferably 2 to 10 hours, and more preferably 4 to 5 hours. In addition, reaction temperature and reaction time can be suitably adjusted, confirming the production amount of the peptide of this invention.

  The enzymatic reaction can also be carried out with increasing temperature. For example, the reaction can be performed while raising from 30 ° C. to 75 ° C. over 4 to 10 hours. Preferably, the reaction can be performed while increasing from 35 ° C. to 75 ° C. over 5 to 8 hours, more preferably from 35 ° C. to 75 ° C. for 6 to 8 hours. The temperature rise speed is arbitrary, but the holding time between 45 ° C and 55 ° C is extended (5 to 7 hours) and then raised rapidly to 60 ° C and then extended between 60 ° C and 75 ° C. It is preferable to hold (for example, 1 to 3 hours). Most preferably, the enzyme is introduced at 50 ° C., held for 5 to 7 hours, heated at an arbitrary rate, and held at a target temperature of 60 to 65 ° C. or 65 to 75 ° C. for 1 to 3 hours.

  At the time of reaction, it is preferable to stir the reaction solution from the viewpoint of reaction efficiency. The solution agitation speed should be fast so that the substrate is in good contact with the enzyme, but if it is too fast, the reaction solution may be splattered, so it can be 100-500 rpm, preferably 200-400 rpm, More preferably, it is about 250 rpm.

  When the desired peptide is obtained, the reaction solution is preferably subjected to an enzyme reaction termination step. In the enzyme reaction termination step, a method of raising the temperature of the reaction solution, adding a chelating agent to change the chemical structure of the enzyme, or a method of removing the enzyme by membrane treatment can be employed. The preferred method is high temperature deactivation treatment. The method can be carried out at 80 to 90 ° C. for 5 to 30 minutes, preferably at 80 to 90 ° C. for 20 to 30 minutes. Moreover, when it becomes high temperature by the below-mentioned concentration process, it can carry out combining a concentration process.

  The reaction solution (milk protein degradation product) subjected to the enzyme reaction step and the enzyme reaction termination step may be further subjected to a sterilization step. As a sterilization process, the below-mentioned membrane treatment process and a high temperature sterilization process are mentioned, for example. The heat sterilization step can also serve as the enzyme reaction termination step, which is advantageous in simplifying the production process.

  The reaction solution (milk protein hydrolyzate) subjected to the enzyme reaction step and the enzyme reaction termination step may be further subjected to a purification step. The purification step may, for example, be a membrane treatment step, and a preferred membrane treatment is ultrafiltration. The molecular weight cut off of ultrafiltration is preferably 3 to 100 kDa, more preferably 5 to 50 kDa. The implementation of the purification step is advantageous in that the flavor of the peptide composition can be improved as compared to the case where it is not performed. In addition, the purification step can be combined with the enzyme reaction termination step and the sterilization step, which is also advantageous in terms of simplification of the production step.

  The reaction solution (milk protein degradation product) subjected to the enzyme reaction step and the enzyme reaction termination step may be further subjected to a concentration step from the viewpoint of storage and transportation. The concentration step can be selected from any method, preferably by vacuum concentration, lyophilization and spray drying (spray drying), concentration by membrane treatment (for example, a method using a reverse osmosis membrane), more preferably freezing Drying and spray drying. Spray drying is particularly preferred from the viewpoint of mass and efficient concentration.

  The measurement of the content of the peptide of the present invention can be carried out by liquid chromatography tandem mass spectrometry (LC / MS / MS). Those skilled in the art can easily set the conditions, and it goes without saying that, for the measurement, one having a purity for LC / MS / MS measurement is used as a standard peptide, for example, SIGMA ALDRICH The AQUA peptide manufactured by Japan can be used.

  The daily intake and dose (in terms of solid content) of the degraded milk protein of the present invention for the purpose of improvement, maintenance and improvement of cognitive function in the present invention are, for example, 0.01 to 100 g (preferably 0). 1 to 10 g). The daily intake and dose (in terms of solid content) of the peptide of the present invention for the purpose of improvement, maintenance and improvement of cognitive function are, for example, 0.01 to 100 mg (preferably 0.1 to 10 mg) is there.

The food of the present invention may be labeled as having an effect of enhancing, maintaining and / or improving cognitive function. In this case, in order to make it easy for the consumer to understand, the food of the present invention may be marked as part or all of the following. In the present invention, it is needless to say that "improvement, maintenance and / or improvement of cognitive function" is used in the meaning including the following indications.
Maintain, enhance, improve, improve, enhance, support, maintain memory Maintain, improve, improve, improve, enhance, support cognitive, maintain, maintain memory Help prevent deterioration · Maintain, enhance, improve, improve, enhance, support, maintain, prevent · prevent forgetting, improve retention of memory · Accuracy of memory Increase the age ・ Reduce the decline in memory with age

  According to another aspect of the invention, the method includes inhaling the mammal into the effective amount of the black tea aroma component through the respiratory tract or ingesting or administering the effective amount of the black tea aroma component to the mammal. And providing a method of enhancing, maintaining and / or improving cognitive function. The method of the present invention can be practiced according to the description of the composition and agent of the present invention.

  According to another aspect of the present invention, it is also provided that the mammal inhales an effective amount of the tea aroma component through the airway or that the mammal ingests or administers an effective amount of the tea aroma component. There is provided a method for treating, preventing or ameliorating a disease or condition which can be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function. The treatment, prevention or amelioration method of the present invention can be practiced according to the description of the composition and agent of the present invention.

  The methods of the invention may be for use in mammals, including humans, and are intended for both therapeutic and non-therapeutic use. As used herein, "non-therapeutic" means that it does not include the act of surgery, treatment or diagnosis of a human (ie, a medical act on a human), specifically, a doctor or a doctor's instruction Means that the person does not include methods for surgery, treatment or diagnosis on humans.

  According to yet another aspect of the present invention, the use of black tea aroma components for the preparation of cognitive function enhancers, maintainers and / or improvers, and black tea as a cognitive function enhancer, maintainer and / or improver The use of aroma components is provided. Also according to the present invention, use of a black tea aroma component for the manufacture of a therapeutic, prophylactic or ameliorating agent for diseases or conditions that can be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function. The use of the tea aroma component is provided as a therapeutic, preventive or ameliorating agent for diseases or conditions which can be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function. The use of the present invention can be carried out according to the description of the composition and preparation of the present invention.

  According to still another aspect of the present invention, a black tea aroma component for use in the improvement, maintenance and / or improvement of cognitive function and a disease which can be treated, prevented or improved by the improvement, maintenance and / or improvement of cognitive function Alternatively, a tea aroma component is provided for use in the treatment, prevention or amelioration of symptoms. The above-mentioned black tea aroma component can be implemented according to the description regarding the composition and preparation of this invention.

  The present invention will be more specifically described based on the following examples, but the present invention is not limited to these examples.

Example 1: Effect of black tea flavor on memory enhancement (1) Preparation of test ingredients A black tea flavor extracts aroma components from Dinbra black leaves according to a conventional method to prepare a black tea flavor (solvent: ethanol), which is used as a test ingredient (Same below in Examples 2 to 5). Also, MilliQ water was used as a control component. 100 .mu.L of a test component or a control component was absorbed per filter paper (Whatman Co., Ltd., qualitative filter paper grade 1, 2 cm.times.2 cm, the same applies hereinafter), and six each were prepared.

(2) Test method A 10-week-old male C57BL / 6 mouse (Oriental Yeast Co., Ltd.) is bred in one cage (length 18.2 cm, width 26 cm, height 12.8 cm) one by one and acclimated for 3 hours, The experimental group and the control group (3 animals in each group) were divided. Two pieces of filter paper imbibed with the test component (black tea flavor solution) prepared in the above (1) per mouse in the experimental group, and filter paper imbibed with the control component (MilliQ water) per mouse in the control group Two were presented for 30 minutes each on the floor of the cage. Reflux fixation was performed with 4% paraformaldehyde 1.5 hours after the start of presentation of the filter paper, and the brain was removed. Then, after replacing with 30% sucrose, the brain was embedded in OTC compound, and a 30 μm thick slice per one brain was prepared using a microtome (Leica CM3050, manufactured by Leica).

  In order to examine the memory enhancing effect of the test component (black tea flavor), phosphorylated CREB (pCREB) antibody (manufactured by cell signaling technology) as a primary antibody and Histofine simple stain mouse MAX-PO (R) rabbit as a secondary antibody The obtained sections were subjected to immunohistochemical staining using an antibody (manufactured by Nichirei Bio). Subsequently, the sections were developed with 3,3'-diaminobenzidine (DAB) (manufactured by SIGMA), and then sealed on a slide glass. The tissue image of the section was taken, the phosphorylated CREB stained cell area in the same visual field area was calculated using image analysis software ImageJ, and the ratio of the phosphorylated CREB expressing cell area of the test component presentation to the control component presentation was calculated. . Phosphorylated CREB functions as a transcription factor for memory-related genes and is known to regulate long-term memory and short-term memory (Suzuki A et al., J Neurosci., 31 (24): 8786). -802 (2011)).

(3) Results The results are shown in FIG.

  From the results of FIG. 1A, in the experimental group presenting the black tea flavor, the expression (shown by black arrowhead) of phosphorylated CREB in the hippocampal dentate gyrus region was confirmed. On the other hand, in the control group, expression of phosphorylated CREB (the same region is indicated by white arrowheads) was hardly observed. Also, from the results shown in FIG. 1B, in the experimental group that presented the black tea flavor (Dimbra black tea leaf extract flavor), the expression area of phosphorylated CREB in the hippocampal dentate gyrus region was approximately twice that in the control group and significantly The rise indicates that the black tea flavor has a memory enhancing effect by olfactory stimulation.

Example 2: Effect of black tea flavor on monoamine production of brain tissue (1) Test method Five 11-week-old male ICR mice (Japan Charles River Co., Ltd.) were used. The mouse is decapitated, the brain is rapidly removed, and Hank's balanced salt solution (HBSS) is cooled by adding 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid (HEPES) (GIBCO) to 2 mM and cooling. ) (Manufactured by GIBCO). A hippocampal-containing brain slice was prepared using Neo-Linear Slicer-MT (manufactured by Dohan EM Co., Ltd.). A 300 μm thick sagittal brain slice was prepared from five mice, and data of four replicates was acquired using four brain slices per test area. Moreover, the black tea flavor solution prepared in the said Example 1 (1) was used as a test component. Specifically, by adding to the HBSS containing 2 mM HEPES so that the concentration of the black tea solution is 0% (not added), 0.001%, 0.01% or 0.1%, the control (not added) ) And 3 test sections were prepared and incubated at 37 ° C. 500 μL of black tea perfume sample was added per brain slice, and after 30 minutes, 100 μL of black tea perfume sample was collected (collected sample). Each collected sample is deproteinized and then treated with serotonin (5-HT), dopamine (DA), norepinephrine (NE) synthesized from dopamine, and dopamine metabolites using HPLC-ECD system (700 series, manufactured by ACOM). The concentrations of certain dihydroxyphenylacetic acid (DOPAC), 3-methoxytyramine (3-MT) and homovanillic acid (HVA) were measured. Serotonin, dopamine and norepinephrine are neurotransmitters of the monoamine nervous system, and serotonin is involved in various functions in brain function (Adv. Pharmacol., 68: 167-197 (2013)), and dopamine is cognitive function, Norepinephrine is involved in higher brain functions such as memory function (Nichishikan, 65 (4-5): 113-129 (2003)), and attention function, memory and learning function etc. (total rehabilitation, 25 (11) : 1229-1236 (1997)).

(2) Results The results are shown in FIG.

  It was confirmed from the results in FIG. 2 that the addition (stimulation) of the black tea flavor sample increased serotonin, dopamine, 3-methoxytyramine and homovanillic acid produced from the brain slice in a manner dependent on the black tea flavor concentration. As well as their production, their metabolism has been enhanced, and it has been shown that the produced dopamine acts functionally.

(3) Discussion It is shown from this example that the black tea flavor or its constituent components may be transferred into the brain and exert their pharmacological effects. Here, there is no report that the black tea component or its constituent compound exerts a pharmacological effect in the brain, but in other compounds, there is a report that the compound exerts a pharmacological effect in the brain through olfactory stimulation (for example, , Clinical Neurophysiology, 42 (4): 87-99 (2014)). Therefore, the pharmacological effects (cognitive function improvement and the like) by the black tea aroma component of the present invention are considered to include the mechanism exhibited by the black tea perfume and the compounds constituting the same being transferred into the brain. Assuming that the mechanism of the pharmacological effect of the black tea aroma component of the present invention is attributable to the direct migration into the brain of the compound, the pharmacological effect of the black tea aroma component of the present invention is effective in the body by oral intake etc. It is considered to be more effective than the pharmacological action exerted through bloodstream or through some kind of signal transduction after being taken up. Therefore, it can be said that the black tea aroma component of the present invention is advantageous in that it has immediate effect.

Example 3: Component analysis of black tea flavor (1) Analysis method The black tea flavor solution prepared in the above Example 1 (1) is subjected to gas chromatography mass spectrometry (GC-MS) under the following conditions to be contained in the black tea flavor The contents of various components were measured. The standard products used for analysis are linalool (made by Tokyo Chemical Industry Co., Ltd., the same below), geraniol (made by Sigma Aldrich, the same below), benzyl alcohol (made by Sigma Aldrich, the same below), linalool oxide (Tokyo Chemical Industry Co., Ltd.) Made of pure products of 2-phenylethanol (made by Tokyo Kasei Kogyo Co., Ltd., the same below).

[GC-MS conditions]
Instrument (analysis except linalool oxide): 7890A / 5875C inertXL (manufactured by Agilent Technologies)
Device (analysis of linalool oxide): 7890A / 5975C inertXL (manufactured by Agilent Technologies)
Column (analysis except linalool oxide): DB-WAX UI (length 30 m, inner diameter 0.25 mm, film thickness 0.25 μm, manufactured by Agilent Technologies)
Column (analysis of linalool oxide): DB-WAX (length 30 m, internal diameter 0.25 mm,
Film thickness 0.25 μm, manufactured by Agilent Technologies
Injection volume: 1 μL
Inlet temperature: 150 ° C
Column bath temperature: 60 ° C (1 minute) → 15 ° C / minute → 220 ° C
Helium gas flow rate: 1 mL / min

(2) Results The results are shown in Table 1.

  From the obtained chromatogram (data not shown), the detected fragment ions (m / z) were 136, 121 for linalool, 123, 93 for geraniol, 108, 79 for benzyl alcohol, 111, 94 for linalool oxide, The 2-phenylethanol was 122, 91. Further, from the results in Table 1, it is found that the black tea flavor prepared in the above Example 1 (1) contains particularly large amounts of 2-phenylethanol, benzyl alcohol and linalool oxide which are known as main aroma components of black tea. confirmed.

Example 4: Effects of aroma components on cognitive function (1)
(1) Evaluation method of cognitive function (new object recognition test)
When rodents such as mice are recognized as novel objects, they approach from search desires and perform search actions such as confirmation of shape and smell. At this time, the search action is not taken for the stored object, or the time (search time) of the search action is shortened as compared with the novel object. The novel object recognition test utilizes this property and can evaluate the memory state. In the present embodiment, the influence on the memory of various components was evaluated by the novel object recognition test.

  Here, the novel object recognition test is a test that determines how much the shape of an object is stored after 24 hours and examines the memory state (storage retention). The outline of the test performed in this example is as shown in FIG.

  The test was performed as follows. That is, two blocks of the same shape, about the size of a golf ball, were placed at a distance of 4 cm from the adjacent corners of the container (38.5 cm long, 38.5 cm wide, 40 cm high) . The mice were placed in this container, allowed to free explore for 10 minutes, and returned to the rearing cage after completion. This is called "acquisition attempt".

  The same mouse is put into a container in which one of the two blocks X presented 24 hours ago is replaced with a golf ball Y (a novel object) 24 hours after an attempt to acquire it, and a free search is performed for 5 minutes. The search times for the novel object X and the novel object Y were measured. This is called "test trial".

  When the distance between the acquisition trial and the test trial is short, the search time for the novel object (golf ball Y) is longer than that of the non-new object (block X) at the test trial when the interval between the acquisition trial and the test trial is short. It is known to diminish as the distance between test trials increases. Therefore, it is generally considered that the change in the search behavior for the novel object reflects the "memory of the shape of the object at the time of acquisition trial".

The retention of memory was evaluated based on the difference between the search time N for the novel object Y and the search time F for the non-new object X. Specifically, Discrimination index (DI) which is a time rate of the novel object search was calculated by the following equation (1).

  As the Discrimination index is higher, the shape of the object at the time of acquisition trial is stored, and it can be evaluated that long-term memory is maintained.

  Further, the search time ratio (%) of the search time F for the non-new object at the time of the test trial or the search time N for the new object was calculated from the following formulas (2) and (3).

  The higher the search time ratio to the non-new object (equation (2)), the lower the cognitive function, ie, the lower the memory acquisition / fixation and recall / reproduction power, and the search time ratio to the novel object (equation As (3) is higher, it can be evaluated that the cognitive function is higher, that is, the ability to acquire and fix a memory and recall and regeneration is higher.

(2) Presentation Method of Aroma Component Preparation A 6-week-old male CD-1 mouse (Nippon SLC Co., Ltd.) was acclimated for 24 hours and divided into an experimental group and a control group (10 mice in each group). Black tea flavor solution (test ingredient) prepared in the same manner as in Example 1 (1) as an aroma component, geraniol (Wako Pure Chemical Industries, the same below), linalool oxide (Tokyo Chemical Industry Co., Ltd., the same below), 2 -Phenyl ethanol (manufactured by Wako Pure Chemical Industries, Ltd., the same as the following), or a five-component mixture (geraniol, linalool oxide, 2-phenyl ethanol, benzyl alcohol (manufactured by Wako Pure Chemical Industries, the same as the following), linalool (Wako Pure Chemical Industries, Ltd. Made by diluting liquid paraffin (a product containing the same product below) with liquid paraffin (made by Wako Pure Chemical Industries, Ltd., the same as the following) so that the concentration shown in Table 2 is obtained. It was a solution. The solution was absorbed by immersing filter paper (Whatman, glass fiber filter paper, grade GF / B, φ 25 mm) in a test tube filled with the test component solution or the control solution. Next, the filter paper is attached to a diffuser (Clear Forest Air Leaf, manufactured by Mikuni Co., Ltd.), and a cage (22.5 cm in length, 33.8 cm in width, 14 cm in height, and so on) used for aroma component presentation May be set in). The five-component mixture was prepared by mixing the five components such that the concentration of each component in liquid paraffin was as shown in Table 2.
B. Presentation of aroma components to each individual The presentation and trial of aroma components to each individual were conducted in a room adjusted to a room temperature of approximately 23 ° C. and a humidity of approximately 55.5 ± 5%. Just before the acquisition trial, one mouse was placed in the aroma component display cage (test start), and the aroma component was presented (first aroma component presentation) by freely searching for 5 minutes in the cage. Immediately after, I tried to get out of the cage. Twenty-four hours after the start of the test, one mouse was again placed in the aroma component presentation cage, and the aroma component was presented by freely searching for 5 minutes in the cage (second aroma component presentation). Immediately after, a test trial was performed outside the cage. The test according to the procedure of the first aroma component presentation to test trial was performed on each individual. In addition, in 2 times of aroma component presentation, the operation of the diffuser was started simultaneously with the entrance of the mouse to the aroma component display cage and ended simultaneously with the exit of the mouse.

(3) Results The results are shown in FIG. 4 and FIG.

  From the results shown in FIG. 4, it was observed that the discrimination index tended to be high in the groups respectively presenting the black tea flavor, geraniol, linalool oxide, 2-phenylethanol and the five-component mixture. The presentation of these aroma components showed that the memory of the shape of the object at the time of acquisition trial was maintained, that is, the long-term memory was enhanced. In addition, from the results shown in FIG. 5, in the group presenting the black tea flavor, geraniol, linalool oxide, 2-phenylethanol or the five-component mixture, the search time ratio to the novel object tends to increase as compared to the control group. These results are the results of improving memory acquisition / fixation and recall / regeneration ability by inhalation of these aroma components in the group presenting the black tea flavor, geraniol, linalool oxide, 2-phenylethanol and 5-component mixture respectively At the same time, it was thought that this was a result of improvement in the ability to exercise attention regarding object recognition and judgment for novel objects.

Example 5: Effects of aroma components on cognitive function (2)
(1) Evaluation method of cognitive function (new object recognition test)
Cognitive function was evaluated according to the method described in Example 4 (1) above.

(2) Method of Presenting Aroma Components Five-week-old ICR mice (Charles Reaver Japan, Inc.) were bred in 5 cages in one cage (same size as in Example 5) and acclimated for 1 week. Geraniol as an aroma component is 0% (V / V) (control), 0.0001% (V / V), 0.001% (V / V), 0.01% (V / V) 0% (V / V) (control), 0.01% (V / V), 0.03% (V / V), 0.1% (V / V), 0. 3% (V / V), linalool oxide or 2-phenylethanol 0% (V / V) (control), 0.01% (V / V), 0.03% (V / V), 0. It was prepared to be 1% (V / V) and 0.3% (V / V). The size of the aroma component display cage is 13.6 cm, width 20.8 cm, height 11.5 cm, and the presentation time of the aroma component before the acquisition trial is 10 minutes as described in Example 4 (2) above. It went according to the method. The geraniol test group was 15 animals in each group, and the low concentration geraniol test, linalool oxide test group and 2-phenylethanol test group were 10 animals in each group.

(3) Results The results are shown in FIG.

  From the results shown in FIG. 6, it was observed that the Discrimination index tended to be higher in the group presenting geraniol, linalool oxide or 2-phenylethanol than in the group presenting control. It was shown that inhalation of these aroma components maintained the memory of the shape of the object at the time of acquisition trial, that is, the long-term memory was enhanced.

Example 6: Effect on presentation timing of aroma components and cognitive function (1)
In this example, the relationship between the presentation timing of the aroma component and the memory function was evaluated by the novel object recognition test.

(1) Evaluation method of cognitive function (new object recognition test)
Cognitive function was evaluated according to the method described in Example 4 (1) above.

(2) Presentation Method of Aroma Component The presentation method of the aroma component was performed according to the method described in Example 5 (2) above. Geraniol was prepared to be 0% (V / V) (control) and 0.001% (V / V) as an aroma component. The presentation timing of the aroma component was made into the following 4 patterns, and was 10 animals in each group. In addition, what showed control as an aromatic component was described as "aromatic component presentation: none" (following, the same).
・Control group acquisition trial aroma component presentation before trial: None, test trial aroma component presentation: None ・Test group 1 acquisition trial aroma component before trial presentation: None, test trial aroma component presentation: yes ・Test group 2 Presentation: Yes, test trial aroma component presentation: None · Test group 3 acquisition trial pre-trial aroma component presentation: Yes, test trial aroma component presentation: yes

(3) Results The results are shown in FIG. From the results shown in FIG. 7, in the test group 2 where the aroma component was presented only before the acquisition trial, the Discrimination index was significantly higher than in the control group, and comparable to the test group 3 in which both the acquisition trial and the test trial were presented. Was confirmed. There was no significant difference in the test group 1 which presented the aroma component only before the test trial as compared with the control group.

Example 7: Effects on the presentation timing of aroma components and cognitive function (2)
In this example, the relationship between the presentation timing of the aroma component and the memory function was evaluated by the novel object recognition test.

(1) Evaluation method of cognitive function (new object recognition test)
Cognitive function was evaluated according to the method described in Example 4 (1) above.

(2) Presentation Method of Aroma Component The aroma component presentation method was performed according to the method described in Example 5 (2) except that the presentation time of the aroma component before the test was 10 minutes. Geraniol was prepared to be 0% (V / V) (control) and 0.001% (V / V) as an aroma component. The presentation timing of the aroma component was made into the following 2 patterns, and was 10 animals in each group.
・Control group acquisition trial pre-trial aroma component presentation: None, pre-trial trial aroma component presentation: None ・Test group acquisition pre-trial aroma component presentation: None, pre-trial trial aroma component presentation: Yes

(3) Results The results are shown in FIG. From the results shown in FIG. 8, the discrimination index of the test group which presented the aroma component only before the test and the control group was similar.

(4) Discussion From the results of Example 6 and Example 7 above, the mechanism by which the Discrimination index is increased by inhalation of the aroma component is the Proust effect (a phenomenon in which past related memories are induced by sniffing a specific odor) Instead, it was suggested that inhalation of aroma components prior to acquisition acquisition established the memory of the object's shape and that it was attributed to enhanced long-term memory (ie, memory acquisition and / or memory fixation). . Therefore, it can be said that the black tea aroma component can exert its pharmacological effect by inhalation only during memory action in improving cognitive function.

Example 8: Combined effect of aroma components and whey enzyme degradation product on cognitive function (1)
In the present test, the effect on memory function when using a combination of presentation of a specific aroma component and intake of a composition for improving cognitive function was evaluated.

(1) Effects of Whey Enzyme Degradation Products on Cognitive Function The cognitive function improvement effect is on a peptide having a specific amino acid sequence produced by treating a protein contained in whey (whey) with a specific microorganism-derived enzyme Some have been reported (Ano et al., Neurobiology of Aging, 72 (2018) 23-31 and WO 2017/086303).

(2) Evaluation method of cognitive function (new object recognition test)
Cognitive function was evaluated according to the method described in Example 4 (1) above.

(3) Administration method of whey enzyme degradation product The whey enzyme degradation product (HW-3, Snow Mark Meg Milk Co., Ltd.) was suspended in distilled water so as to be 0.5 mg / mL. Gavage of distilled water (control) or whey enzyme lysate suspension into mice at 10 mL / kg once a day for 3 days (the day before the test, 1 hour before the trial on the day of the test, and on the day of the test) Administered. In addition, when this whey enzyme degradation product was analyzed by the LC / MS / MS method described in WO 2017/086303, the tetrapeptide GTWY contained 1.61 mg / g and the dipeptide WY contained 0.65 mg / g. The

(4) Presentation Method of Aroma Component The presentation method of the aroma component was performed according to the method described in Example 5 (2) above. Geraniol was prepared to be 0% (V / V) (control) and 0.0001% (V / V) as an aroma component.

In this test example, administration of whey enzyme degradation product and presentation of aroma components were made to the following four patterns, and the test group was 10 animals in each group.
・Control group aroma component presentation: None, Whey enzyme decomposition product administration: None ・Test group 1 aroma component presentation: None, whey enzyme decomposition product administration: Yes ・Test group 2 aroma component presentation: Yes, Whey enzyme decomposition product administration: None Test group 3 aroma component presentation: Yes, whey enzyme decomposition product administration: Yes

(5) Results The results are shown in FIG. From the results shown in FIG. 9, P <0.05 was significant in Test Group 1 where only oral administration of whey enzyme degradation product was performed and Test Group 2 where only presentation of aroma components was performed, as compared to the control group. It was confirmed that the Discrimination index is high, and in the test group 3 used in combination, the Discrimination index is further high. Therefore, it was shown that the long-term memory enhancing effect can be obtained only by whey enzyme decomposition product intake or by inhalation of an aroma component. In addition, it has been shown that the combination of inhalation of aroma components and whey enzyme decomposition product intake has a combined effect on the enhancement of long-term memory.

Example 9: Combined effect of aroma components and whey enzyme degradation product on cognitive function (2)
In the present test, the effect on memory function when using a combination of presentation of a specific aroma component and intake of a composition for improving cognitive function was evaluated.

(1) Evaluation method of cognitive function (new object recognition test)
Cognitive function was evaluated according to the method described in Example 4 (1) above.

(2) Administration Method of Whey Enzyme Degraded Product The administration method of whey enzyme decomposed product was carried out according to the method described in Example 8 (3) above.

(3) Presentation Method of Aroma Component The presentation method of the aroma component was performed according to the method described in Example 5 (2) above. Geraniol was prepared to be 0% (V / V) (control), 0.00001% (V / V) and 0.0001% (V / V) as an aroma component.

In this test example, administration of whey enzyme degradation product and presentation of aroma components were made to the following six patterns, and the test group was eight animals in each group.
・Control group aroma component presentation: None, whey enzyme decomposition product administration: None ・Test group 1 aroma component presentation: None, whey enzyme decomposition product administration: yes ・Test group 2 aroma component presentation: presence (0.00001%), whey Enzyme degradation product administration: None ・Test group 3 aroma component presentation: Yes (0.00001%), whey enzyme decomposition product administration: presence ・test group 4 aroma component presentation: presence (0.0001%), whey enzyme decomposition product administration : None ・Test group 5 Aroma component presentation: Yes (0.0001%), Whey enzyme decomposition product administration: Yes

(4) Results The results are shown in FIG. From the results of FIG. 10, compared with the control group, the test group 1 to which only the oral administration of whey enzyme degradation product was administered and the test group 2 to which only geraniol 0.00001% (V / V) was presented were compared It was confirmed that in the test group 3 in which the whey enzyme degradation product and geraniol 0.00001% (V / V) were used in combination, the Discrimination index was high. Similar results were obtained in Test Groups 4 and 5 for Geraniol 0.0001% (V / V). Therefore, it was confirmed that the long-term memory is enhanced by combining the inhalation of the aroma component and the intake of the whey enzyme decomposition product.

(5) Discussion The results of Example 8 and Example 9 show that inhalation of the aroma component and intake of the whey enzyme degradation product have a combined effect on the enhancement of long-term memory. From this result, the memory function improvement effect is further enhanced by perfuming geraniol to foods containing whey enzyme degradation products and oral preparations, and by combining geraniol-containing foods and foods and whey preparations containing whey enzyme degradation products in combination with oral preparations. It can be said.

Example 10 Verification of Effect of Black Tea Flavor on Cognitive Function (1) Outline of Test This test was a single-blind test to evaluate cognitive function when a test perfume or a control perfume was inhaled from the nose. Here, since it is known that enhancement of sympathetic nervous function affects cognitive function (Journal of Physiology, 454: 373-87, 1992, Applied Psychophysiology Biofeedback, 37 (4): 219-27, 2017 The evaluation of cognitive function was performed by evaluating the autonomic nervous system. More specifically, healthy men and women between 20 and 65 years of age are allowed to inhale autonomic nervous responses by inhaling a perfume containing one kind of compound that is a black tea perfume or its component as a test perfume and distilled water as a control perfume. evaluated.

(2) Subjects The subjects were 7 healthy men and women (5 men and 2 women) aged 25 to 51 years old, and all subjects were tested for all test perfumes according to the schedule described later.

(3) Test method As a summary of the test method, the test schedule is shown in Table 3 (o indicates that the test was performed). On the test day 1, the autonomic nervous response evaluation at the time of control fragrance inhalation and the autonomic nervous reaction evaluation at the time of any test perfume inhalation were performed. The test perfume was changed and the same evaluation was performed after the test day 2 so that all the test perfume could be evaluated by the test day 4. The test was conducted at intervals of one day or more, and the test days were not consecutive. The autonomic nervous system was evaluated by measuring the pupillary reaction as described later.

(4) Inhalation Method of Test Fragrance Inhalation Method of Test Fragrance 0.05 mL of the test fragrance was impregnated into a filter paper (wiper 33330 manufactured by Kimberly-Clark) cut into 2 cm × 2 cm and set in an aroma diffuser (FAD01 manufactured by Mikuni Co., Ltd.). The aroma diffuser was placed about 10 cm away from the subject's nose, and the scent component to volatilize was inhaled from the nose. The inhalation time for each test perfume was 1 minute. The filter paper was replaced for each subject.

B. Composition of test perfume The formulation of the test perfume was as shown in Table 4. Except for the control perfume, it is a perfume that uses 50% aqueous ethanol as a solvent. The black tea flavor is produced by extraction from Dimbra black tea leaf according to a conventional method, and the content of the component known as the main aroma component in the black tea flavor is quantified by the method described in Example 3 and is shown in the table. Described.

(5) Measurement item Measurement item With regard to the pupil's response to the stimulus of predetermined visible light, the miosis phase and the subsequent mydriasis phase are automatically measured and analyzed, and the miosis rate CR (contraction rate of pupils: pupil lateral diameter The amount of change / maximum pupil lateral diameter × 100) and the maximum mydriasis velocity VD (maximum velocity of mydriasis) were determined. The former is widely used as an index of parasympathetic activity and the latter is generally used as an index of sympathetic activity. In any case, an increase in value is evaluated as an increase in neural activity.

B. Measurement method of light pupil reaction Measurement was performed at a fixed time (from 9 am to 12 am) in order to avoid the influence of the daily fluctuation of the pupil. In addition, in order to avoid the influence of other factors on the autonomic nervous system, eating and drinking, especially taking of food and drink including caffeine was prohibited for 2 hours before the measurement. The measurement was performed in a dark room, and a pupil recorder (Iris Coder Dual C10641, manufactured by Hamamatsu Photonics K.K.) was used for measurement and analysis.

The subjects were seated and light pupil responses were measured after 5 minutes of dark adaptation. The flow of daily measurement is shown in FIG.
(I) Blank measurement 2 times, 2 times during control perfume inhalation, 5 minutes rest, 2 measurements during inhalation of control perfume. The blank value was measured in order to calculate the relative value by correcting the individual difference and daily difference of the pupillary reflex (see “(6) Evaluation and analysis” described later).
(Ii) After the measurement with the control fragrance, a 5-minute rest was given, and the same measurement was performed for any of the test fragrances.
(Iii) As described in (3) above, by performing the measurements (i) and (ii) from test day 1 to test day 4, all subjects were evaluated for all test perfumes. According to (i) and (ii), 12 measurements were made per subject per day.

(6) Evaluation and Analysis As described above, the pupil dilation rate CR obtained by the light reflex measurement was evaluated as an index of parasympathetic nerve activity, and the maximum mydriasis velocity VD was evaluated as an index of sympathetic nerve activity. The relative value (relative CR or relative VD) was calculated from the measurement value of the blank and the measurement value of the test perfume. Calculation was performed by following formula (4) and Formula (5). Then, the relative values of the control perfume and the test perfume on the same test day were obtained for each subject, and the effects of the test perfume were evaluated by comparing the values of all the subjects by a one-sample t-test.

(7) Results The results are shown in Table 5 and FIGS.

  Since the relative CR (%) which is an index of parasympathetic nerve activity decreased and the relative VD (%) which is an index of sympathetic nervous activity increased in the black tea perfume test area, inhalation of black tea aroma components is different from autonomic nervous activity. It has been shown to act on sympathetic nerves. The relative CR (%) decreased also in the test group that presented 2-phenylethanol or geraniol, suggesting the possibility of acting on sympathetic nerves. 2-phenylethanol and geraniol are the main components of black tea flavors, and their action on autonomic nervous activity showed a tendency similar to that of black tea flavors, so they were considered to be the components contributing to the sympathetic nerve enhancing action of black tea aroma .

(8) Conclusion It was confirmed that acting on autonomic nervous system activity and enhancing sympathetic nervous system activity by inhaling the black tea flavor, 2-phenylethanol and geraniol singly or in combination through the nose. It has been reported that the scent that makes sympathetic activity dominant activates the brain and contributes to the improvement of cognitive function (Niijima, autonomic nerve, 45 (5), 178-184 (2008), Nagai, Japan Nursing Research Association) From the journal, 23 (4), 11-17 (2000), Jimbo D, et al .; Psychogeriatrics, 9: 173-179 (2009)), it is about black tea flavor, 2-phenylethanol, geraniol singly or in combination Also, it was thought that sympathetic nervous activity was enhanced to activate the brain and improve cognitive function.

Claims (12)

  A composition for improving, maintaining and / or improving cognitive function, comprising a black tea aroma component as an active ingredient.   The composition according to claim 1, wherein the cognitive function is a memory function.   The composition according to claim 2, wherein the memory function is a long-term memory function.   The composition according to claim 1, wherein the cognitive function is an attention function.   The composition according to claim 1, wherein the cognitive function is a memory acquisition function and / or a memory fixation function.   The composition according to any one of claims 1 to 5, wherein the black tea aroma component is at least one selected from the group consisting of geraniol, linalool oxide and 2-phenylethanol.   The composition according to any one of claims 1 to 6, for use in healthy individuals.   The composition according to any one of claims 1 to 7 for use in the treatment, prevention or amelioration of a disease or condition which can be treated, prevented or ameliorated by improvement, maintenance and / or improvement of cognitive function.   The composition according to any one of claims 1 to 8, which is a composition for inhalation via the respiratory tract.   10. A composition according to any one of the preceding claims which is in the form of a food or an oral preparation.   The composition according to claim 10, further comprising a milk protein enzyme degradant as an active ingredient.   11. A composition according to claim 10 for use in combination with a milk protein enzyme digest or a composition containing it.

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