Composition for improving cognitive function

Abstract

To provide a novel composition for maintaining, enhancing or improving cognitive functions.SOLUTION: The present invention provides a composition for maintaining, enhancing or improving cognitive functions, comprising a hop oxidation reaction product and a whey protein enzyme degradation product as active ingredients. The cognitive functions include a memory function (for example, a spatial memory function, a working memory function). The composition can be provided as a food composition and can be also provided in a unit package form per meal.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for use in maintaining, enhancing or improving cognitive function. [Background technology]

[0002] The maintenance, improvement, and improvement of cognitive function is required by a wide range of generations, from young people to the elderly. The maintenance, improvement, and improvement of memory and learning ability is important not only for students and working people studying for exams and qualification exams, but also for daily work and life. In addition, for the elderly, a decline in memory and concentration affects the quality of life, so there is a demand to prevent the decline in cognitive function and to maintain, improve, and improve it.

[0003] To date, peptides having specific amino acid sequences (Patent Document 1) and hop oxidation products (Non-Patent Document 1) have been known as ingredients that have an effect of improving cognitive function. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2015 / 194564 [Non-patent literature]

[0005] [Non-Patent Document 1] T. Ayabe et al., Scientific Reports volume 8, Article number: 15372 (2018) Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention aims to provide novel compositions and agents that maintain, enhance or improve cognitive function. [Means for solving the problem]

[0007] The present inventors have now found in an animal model study that co-administration of a whey protein enzymatic hydrolysate and a hop oxidation reaction product results in a significant improvement in cognitive function compared to administration of either of them alone. The present invention is based on this finding.

[0008] According to the present invention, the following inventions are provided. [1] A composition for maintaining, enhancing and / or improving cognitive function (hereinafter sometimes referred to as the "composition of the present invention") and an agent for maintaining, enhancing and improving cognitive function (hereinafter sometimes referred to as the "agent of the present invention"), which comprises a hop oxidation reaction product and an enzymatic hydrolysate of whey protein as active ingredients. [2] The composition and agent described in [1] above, wherein the cognitive function is a memory function. [3] The composition and agent described in [2] above, wherein the memory function is a spatial memory function and / or a working memory function. [4] The composition or agent according to any one of [1] to [3] above, wherein the hop oxidation reaction product is an aqueous medium extract. [5] The composition or agent according to any one of [1] to [4] above, wherein the hop oxidation reaction product is a water extract. [6] The composition or agent according to any one of [1] to [5] above, wherein the hop oxidation reaction products include tricyclooxyisohumulone, tricyclooxyisocohumulone, scorpiofumulinol and scorpiocohumulinol. [7] The composition or agent according to any one of [1] to [6] above, wherein the concentration of bitter acid in the hop oxidation reaction product is 1 to 20 mass%. [8] The composition and agent described in any of [1] to [7] above, wherein the whey protein enzymatic hydrolysate comprises a peptide having the amino acid sequence of GTWY (sequence number 1) and a peptide having the amino acid sequence of WY. [9] The composition and agent described in [8] above, wherein the content of GTWY in the whey protein enzymatic hydrolysate (converted into solid content) is 0.01 to 1 mass% of the total amount, and the content of WY in the whey protein enzymatic hydrolysate (converted into solid content) is 0.005 to 0.5 mass% of the total amount.

[10] The composition or agent according to any one of [1] to [9] above, wherein the ratio of the hop oxidation reaction product to the whey protein enzymatic hydrolysate in the composition (mass ratio converted into solid content) is 0.1 to 2.

[11] The composition according to any one of [1] to

[10] above, which is a food composition.

[12] The composition and preparation according to any one of [1] to

[11] above, which is in a unit-packaged form per meal.

[13] The composition or agent according to any one of [1] to

[12] above, wherein the hop oxidation reaction product is ingested by a human in an amount of 10 to 5,000 mg (in terms of solid content) per meal.

[14] The composition or agent according to any one of [1] to

[13] above, wherein the whey protein enzymatic hydrolysate is ingested by a human in an amount of 1 to 50,000 mg (converted into solid content) per meal.

[15] The composition or agent according to

[13] or

[14] above, which contains 0.01 to 10 mg of GTWY and 1 to 100 mg of a bitter acid.

[0009] The active ingredients of the present invention, the hop oxidation reaction product and the whey protein enzymatic hydrolysate, are both derived from food materials that humans have been ingesting for many years, and therefore the composition and preparation of the present invention are advantageous in that they can be used as functional foods that improve cognitive function and are safe for mammals, including humans. [Brief description of the drawings]

[0010] [Figure 1] FIG. 1 shows the results of HPLC analysis (HPLC chromatogram) of the hop oxidation reaction products in Reference Example 1. [Diagram 2]2 is a diagram showing waveform processing when calculating the area of ​​bitter acids derived from hop oxidation reaction products in Reference Example 2. In the figure, A shows a caffeine analysis chromatogram, B shows an analysis chromatogram for confirming the elution position (a: trans-isocohumulone, b: trans-isohumulone, c: trans-isoadhumulone), and C shows a sample analysis chromatogram. In the sample analysis chromatogram of C, after background correction, a baseline is drawn at an intensity of 0 between caffeine and trans-isocohumulone to define the base of the peak, and the area of ​​that section (gray part) is calculated. [Diagram 3] FIG. 3 is a graph showing the total number of entries into the arms of mice orally administered with each sample in Example 3. [Figure 4] 4 is a graph showing the spontaneous alternation behavior fluctuation rate of mice orally administered with each sample in Example 3. * indicates p<0.05, ** indicates p<0.01 (Tukey-Kramer test). Description of the Invention

[0011] <Compositions and preparations> According to the present invention, there is provided a composition and a preparation comprising a hop oxidation reaction product and a whey protein enzymatic hydrolysate. The hop oxidation reaction product and the whey protein hydrolysate (hereinafter, simply referred to as "whey hydrolysate") will be described below.

[0012] Hop oxidation products In the present invention, the hop oxidation reaction product refers to a product obtained by oxidizing hops or processed products thereof (hop pellets, hop extract, etc.) The hop oxidation reaction product provided by the present invention can be obtained, for example, by contacting hops with oxygen in the air to oxidize them.

[0013] The hop oxidation reaction product can be produced by subjecting hops to an oxidation treatment, for example, according to the method described in International Publication No. 2012 / 081675. The oxidation treatment is preferably carried out by heating hops in air. The heating temperature is not particularly limited, but a preferred upper limit is 100°C, and a more preferred upper limit is 80°C. A heating temperature of 100°C or lower is advantageous in that oxidation proceeds preferentially over isomerization. A preferred lower limit of the heating temperature is 60°C. A heating temperature of 60°C or higher is advantageous in that the oxidation reaction proceeds efficiently. The reaction period is also not particularly limited, and can be appropriately determined depending on the variety of hops and the reaction temperature. For example, 48 to 120 hours at 60°C and 8 to 24 hours at 80°C are preferred. The form of hops when subjected to the oxidation reaction is not particularly limited as long as it can contact oxygen in the air, but the reaction time can be shortened by making the hops into a powder form.

[0014] In the present invention, hops may be in any form as long as they contain lupulin, and may be harvested and dried before drying, harvested and dried, compressed, crushed, or processed into pellets, but are preferably in the form of hop pellets. Hop pellets may be commercially available products, such as hop cones compressed into pellets (Type 90 pellets), pellets in which the lupulin portion is selectively concentrated (Type 45 pellets), or isomerized hop pellets (e.g., Isomerized Pellets (HopSteiner)).

[0015] A hop extract oxidation reaction product produced by subjecting a hop extract to an oxidation treatment may be provided as the hop oxidation reaction product of the present invention. The hop extract oxidation reaction product can be produced by subjecting a hop extract to an oxidation treatment according to the method described in WO 2012 / 081675, for example.

[0016] The content of the hop oxidation reaction product (converted into solid content) in the compositions and preparations of the present invention can be determined arbitrarily depending on the purpose, use, form, dosage form, etc., and the present invention is not limited thereto. For example, in the case of a liquid composition, the content can be 0.1 to 50,000 mg / 100 mL (preferably 1 to 5,000 mg / 100 mL, more preferably 10 to 600 mg / 100 mL) relative to the total amount, and in the case of a solid composition, the content can be 1 to 99 mass% (preferably 10 to 80 mass%, more preferably 40 to 75 mass%) relative to the total amount.

[0017] Hops contain acidic resin components such as α-acids (humulones), β-acids (lupulones), and iso-α-acids (isohumulones). In the present invention, "humulones" is used to include humulone, adhumulone, cohumulone, posthumulone, and prehumulone. In the present invention, "lupulones" is used to include lupulone, adlupulone, colupulone, postlupulone, and prelupulone. Furthermore, in the present invention, "isohumulones" is used to include isohumulone, isoadhumulone, isocohumulone, isoposthumulone, isoprehumulone, Rho-isohumulone, Rho-isoadhumulone, Rho-isocohumulone, Rho-isoposthumulone, Rho-isoprehumulone, tetrahydroisohumulone, tetrahydroisoadhumulone, tetrahydroisocohumulone, tetrahydroisoprehumulone, tetrahydroisoposthumulone, hexahydroisohumulone, hexahydroisoadhumulone, hexahydroisocohumulone, hexahydroisoposthumulone, and hexahydroisoprehumulone. Isohumulones include cis and trans stereoisomers, and unless otherwise specified, the term is used to include both.

[0018] By subjecting hops to oxidation treatment, the content of α acids, β acids, and iso-α acids is reduced, and the content of other components is increased. Examples of such hop oxidation reaction products include those in which, when HPLC analysis is performed in the same manner as in Example 1, the ratio of the peak area of ​​α acids, β acids, and iso-α acids to the total HPLC peak area of ​​the oxidation reaction products is 20% or less, preferably 10% or less.

[0019] Components other than α acids, β acids, and iso-α acids contained in the oxidation reaction product of the present invention can be easily detected by known analytical means such as HPLC. For example, a hop oxidation reaction product prepared by a procedure similar to that of Example 1 of WO 2012 / 081675 contains components other than α acids, β acids, and iso-α acids, and the peaks corresponding to these components (also referred to as "S-fraction (S-Fr)" in this specification) can exhibit physiological activity. The peaks in the range indicated by the arrows in Figure 1A of Example 1 of WO 2012 / 081675 (excluding the peaks of α acids and β acids) correspond to the S-fraction.

[0020] HPLC analysis of hop oxidation products prepared under the same conditions as those described in WO 2012 / 081675 and the results (HPLC chromatogram) are shown in Reference Example 1 and FIG. 1 of the present specification. The peaks in the range indicated by the arrows A1 and A2 (excluding α-acid and β-acid peaks) correspond to the S-fraction. Here, in FIG. 1, the peak area value in the range fractionated by the arrows A1 and A2 is the sum of the peak area value of A1 from 3 to 25 minutes of retention time and the peak area value of A2 from 32 to 39 minutes of retention time (excluding α-acid and β-acid peaks). Here, "up to 25 minutes of retention time" in A1 means up to the appearance of a peak identified as trans-isocohumulone. In addition, characteristic peaks were observed at around 9.7 minutes of retention time, around 11.8 minutes of retention time, and around 12.3 minutes of retention time in the range fractionated by A1 in FIG. 1. In addition, a shoulder peak was observed in the range fractionated by A2 in Figure 1, with its start point at approximately 32 minutes retention time, its top point (excluding the α acid and β acid peaks) at approximately 35 to 36 minutes retention time, and its end point at approximately 39 minutes retention time.

[0021] The hop oxidation reaction product preferably contains α-acid oxides, iso-α-acid oxides, and β-acid oxides, and examples of such oxides include "tricyclooxyisohumulones." Here, "tricyclooxyisohumulones" include tricyclooxyisocohumulone A (TCOIcoH A: see formula 1 below, IUPAC name: (3aS,5aS,7S,8aS)-3,3a-dihydroxy-7-(1-hydroxy-1-methylethyl)-6,6-dimethyl-2-(2-methylpropanoyl)-5a,6,7,8-tetrahydro-3aH,5H-cyclopenta[c]pentalene-1,4-dione), tricyclooxyisohumulone A (TCOIH A: See formula 2 below, IUPAC name: (3aS,5aS,7S,8aS)-3,3a-dihydroxy-7-(1-hydroxy-1-methylethyl)-6,6-dimethyl-2-(3-methylbutyryl)-5a,6,7,8-tetrahydro-3aH,5H-cyclopenta[c]pentalene-1,4-dione), a group of compounds including tricyclooxyisoadhumulone A (TCOIadH A: See formula 3 below, IUPAC name: (3aS,5aS,7S,8aS)-3,3a-dihydroxy-7-(1-hydroxy-1-methylethyl)-6,6-dimethyl-2-(2-methylbutanoyl)-5a,6,7,8-tetrahydro-3aH,5H-cyclopenta[c]pentalene-1,4-dione). In the present specification, TCOIcoH A, TCOIH A and TCOIadH A may hereinafter be collectively referred to as TCOIHsA. The content of TCOIHsA is measured by the method described in Example 1 below.

[0022] [ka]

[0023] [ka]

[0024] [ka]

[0025] Examples of oxidation products other than "tricyclooxyisohumulones" contained in the hop oxidation reaction products (preferably S-fraction) include scorpiofumulinol A and scorpiocohumulinol A.

[0026] In the present invention, the hop oxidation reaction product may be provided as an aqueous medium extract. The aqueous medium is not particularly limited as long as it is one that is usually used in food production, but is preferably water or ethanol, and more preferably water (an extract using water is specifically referred to as a water extract). The extraction temperature is not particularly limited, but is preferably 60°C or lower, and taking into consideration the extraction efficiency, 50 to 60°C is more preferable.

[0027] The hop oxidation reaction product (preferably an aqueous medium extract of a hop oxidation reaction product) used in the present invention can be characterized by the ratio (on a dry mass basis) of the total amount of tricyclooxyisohumulone A and tricyclooxyisocohumulone A to the total amount of scorpiofumulinol A and scorpiocohumulinol A; for example, an aqueous medium extract of a hop oxidation reaction product in which this ratio is in the range of 1 to 30, preferably an aqueous medium extract of a hop oxidation reaction product in which this ratio is in the range of 2 to 20, can be used.

[0028] The hop oxidation reaction product used in the present invention can be characterized by the TCOIHsA content (converted into dry mass), and for example, a hop oxidation reaction product (preferably an aqueous medium extract of a hop oxidation reaction product) having a TCOIHsA content of 5 to 15 mass%, preferably 5 to 12 mass% can be used. The content of TCOIHsA can be measured by the same method as that for mature hop bitter acids described in Biosci., Biotechnol., Biochem., 2015 (79):1684-1694.

[0029] The Brix value of the aqueous medium extract of the hop oxidation reaction product is not particularly limited, but is, for example, 3 or less, and preferably 1.5 to 3. The aqueous medium extract of the hop oxidation reaction product may have insoluble components removed by decantation or filter paper, etc. The aqueous medium extract of the hop oxidation reaction product may also be treated with activated carbon.

[0030] The hop oxidation reaction products can also be characterized by the total amount of a group of compounds (sometimes referred to as "bitter acids" in this specification) having a β-tricarbonyl structure represented by formula 4 as a common structure. The bitter acid content is measured by the method described in Reference Example 2. The bitter acid content in the water extract of the hop oxidation reaction products is 0.1 to 90% by mass (preferably 0.5 to 50% by mass, more preferably 1 to 20% by mass). [ka] (In the above formula, R, R1, R2, R3, and R4 may be the same or different from each other, and R represents a hydrogen atom or an alkyl group having 1 to 10 carbon atoms. R1, R2, R3, and R4 each represent any substituent, and each may be independent or may be bonded to each other.)

[0031] Whey protein enzyme hydrolysate In the present invention, "whey" is also called milk serum, whey or whey, and refers to an aqueous solution obtained by removing milk fat, casein, etc. from milk. Whey is composed of β-lactoglobulin, α-lactalbumin, serum albumin, immunoglobulin, etc. The whey used in the present invention can be derived from any animal or plant, but it is preferable to use whey derived from cow's milk. The whey hydrolysate, which is an active ingredient of the composition and agent of the present invention, is not limited as long as it is an enzymatic hydrolysate of whey.

[0032] The content of the whey hydrolysate (converted into solid content) in the compositions and preparations of the present invention can be determined arbitrarily depending on the purpose, use, form, dosage form, etc., and the present invention is not limited thereto. For example, in the case of a liquid composition, the content can be 0.001 to 100 mg / 100 mL (preferably 0.005 to 50 mg / 100 mL, more preferably 0.01 to 10 mg / 100 mL) relative to the total amount, and in the case of a solid composition, the content can be 0.1 to 90 mass% (preferably 0.5 to 80 mass%, more preferably 1 to 70 mass%) relative to the total amount.

[0033] The whey hydrolyzate of the present invention may preferably contain a peptide having the amino acid sequence of GTWY (SEQ ID NO: 1) and a peptide having the amino acid sequence of WY (hereinafter, these may be referred to as "the peptide of the present invention"). Here, "a peptide having an amino acid sequence" means a peptide whose sequence is specified by the amino acid sequence.

[0034] The content of the tetrapeptide GTWY in the whey hydrolysate of the present invention (converted into solid content) is, for example, 0.01 to 1 mass% (preferably 0.05 to 0.5 mass%, more preferably 0.1 to 0.2 mass%) relative to the total amount of the whey hydrolysate, and the content of the dipeptide WY (converted into solid content) is, for example, 0.005 to 0.5 mass% (preferably 0.01 to 0.1 mass%, more preferably 0.03 to 0.09 mass%) relative to the total amount of the whey hydrolysate.

[0035] The content of the peptide of the present invention is measured by liquid chromatography tandem mass spectrometry (LC-MS / MS) according to the method described in Example 2 below. In this case, it goes without saying that a standard peptide having a purity suitable for LC-MS / MS measurement is used during the measurement, and for example, AQUA peptide manufactured by SIGMA ALDRICH can be used.

[0036] The method for producing the whey hydrolysate of the present invention (particularly, the whey hydrolysate containing the peptide of the present invention) is known, and can be produced, for example, according to the description in International Publication No. 2017 / 086303. Alternatively, commercially available whey hydrolysates (e.g., HW-3 (Snow Brand Megmilk), HWP-205 (Tatua), Thermax690 (Glanbia), Protherma (Glanbia)) may be used as the whey hydrolysate.

[0037] The whey hydrolysate containing the peptide of the present invention can be produced, for example, by allowing an enzyme preparation containing a whey degrading enzyme to act on whey protein.

[0038] The concentration of whey protein to be subjected to the enzyme reaction is not limited as long as the protein is soluble; however, from the viewpoint of suppressing gelation and aggregation and eliminating the trouble of concentration, the concentration is preferably 1 to 30 w / v%, more preferably 1 to 20 w / v%, and even more preferably 5 to 15 w / v%.

[0039] Whey may be directly or after being concentrated or diluted for the enzyme reaction, and pH adjustment may be performed as necessary. When the raw material protein is a solid such as a powder, it may be dissolved in any aqueous solvent as long as the enzyme reaction proceeds, but it is preferable to dissolve it in water or a food additive grade buffer in consideration of its use as a food. It is preferable to use a buffer in order to prevent the pH of the reaction solution from changing due to amino acids generated in the enzyme reaction. The type of buffer is arbitrary and may be determined in consideration of the subsequent use, flavor, taste, and mineral content, but a composition that can maintain the pH of the reaction solution at 4 to 9, preferably 5 to 8, and more preferably 7 to 8 is preferable. Potassium phosphate buffer is most preferable. The concentration of the buffer is arbitrary as long as it is within a range in which a buffering effect can be obtained, but in consideration of the flavor, taste, and mineral content, it can be 0.01 to 0.5 M, preferably 0.05 to 0.2 M, and more preferably about 0.1 M.

[0040] Any enzyme preparation containing a protease can be used as the enzyme, but an enzyme preparation containing a neutral protease is preferable, and one or more types can be used in combination. The enzyme preparation can be derived from a microorganism such as Bacillus subtilis, Aspergillus oryzae, or Aspergillus melleus, and among these, an enzyme preparation derived from Aspergillus oryzae and an enzyme preparation derived from Aspergillus melleus are preferable, and an enzyme preparation derived from Aspergillus melleus is more preferable.

[0041] In the present invention, a commercially available enzyme preparation can be used, and for example, enzyme preparations are available from Amano Enzyme, Shin Nippon Chemical Industry, DSM, Danisco, Novozymes, HBI, etc. The amount of the enzyme preparation to be added is arbitrary, but considering an appropriate hydrolysis reaction rate and costs, it can be, for example, 0.01 to 5 w / v%, preferably 0.05 to 4 w / v%, and more preferably 0.1 to 0.5 w / v%.

[0042] The enzyme reaction temperature and reaction time can be set so that the hydrolysis of the raw material protein is sufficiently performed and the quality of the enzymatic hydrolyzate 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 can be 1 to 12 hours, preferably 2 to 10 hours, and more preferably 4 to 5 hours. The reaction temperature and reaction time can be appropriately adjusted while checking the production amount of the peptide of the present invention.

[0043] The enzyme reaction can also be carried out while increasing the temperature. For example, the reaction can be carried out while increasing the temperature from 30°C to 75°C over 4 to 10 hours. Preferably, the reaction can be carried out while increasing the temperature from 35°C to 75°C over 5 to 8 hours, and more preferably, the reaction can be carried out while increasing the temperature from 35°C to 75°C over 6 to 8 hours. The temperature can be increased at any speed, but it is preferable to hold the temperature between 45°C and 55°C for a longer period (5 to 7 hours), then quickly increase the temperature to 60°C, and then hold the temperature between 60°C and 75°C for a longer period (for example, 1 to 3 hours). The most preferred method is to add the enzyme at 50°C, hold the temperature for 5 to 7 hours, then raise the temperature at any speed, and hold the temperature at the target temperature of 60 to 65°C or 65 to 75°C for 1 to 3 hours.

[0044] From the viewpoint of reaction efficiency, it is preferable to stir the reaction solution during the reaction. The faster the stirring speed, the better so that the substrate comes into contact with the enzyme. However, if the stirring speed is too fast, the reaction solution may be splashed. For example, the stirring speed may be set to 100 to 500 rpm, preferably 200 to 400 rpm, and more preferably about 250 rpm.

[0045] Once the desired peptide is obtained, the reaction solution is preferably subjected to a step of terminating the enzyme reaction. In the step of terminating the enzyme reaction, a method of increasing the temperature of the reaction solution, or adding a chelating agent to change the chemical structure of the enzyme, or a method of removing the enzyme by membrane treatment can be adopted. A preferred method is inactivation treatment at high temperature. This method can be carried out by holding the reaction solution at 80 to 90°C for 5 to 30 minutes, preferably at 80 to 90°C for 20 to 30 minutes. In addition, when the temperature is increased in the concentration step described below, this can also be carried out as the concentration step.

[0046] The reaction liquid (milk protein hydrolysate) that has been subjected to the enzyme reaction step and the enzyme reaction stopping step may be further subjected to a sterilization step. Examples of the sterilization step include a membrane treatment step and a high-temperature sterilization step described below. The heat sterilization step can also serve as the enzyme reaction stopping step, which is advantageous in terms of simplifying the production process.

[0047] The reaction solution (milk protein hydrolysate) that has been subjected to the enzyme reaction step and the enzyme reaction stopping step may be further subjected to a purification step. An example of the purification step is a membrane treatment step, and a preferred membrane treatment is ultrafiltration. The molecular weight cutoff for ultrafiltration is preferably 3 to 100 kDa, and more preferably 5 to 50 kDa. The purification step is advantageous in that the flavor of the peptide composition can be improved compared to when it is not performed. In addition, the purification step can also serve as the enzyme reaction stopping step and the sterilization step, which is advantageous in terms of simplifying the production process.

[0048] The reaction liquid (whey hydrolyzate) that has been subjected to the enzyme reaction step and the enzyme reaction stopping step may be further subjected to a concentration step from the viewpoint of storage and transportation. Any method may be selected for the concentration step, but vacuum concentration, freeze-drying, spray-drying, and concentration by membrane processing (e.g., a method using a reverse osmosis membrane) are preferred, and freeze-drying and spray-drying are more preferred. Spray-drying is particularly preferred from the viewpoint of performing concentration in large quantities and efficiently.

[0049] The composition and preparation of the present invention may contain a hop oxidation reaction product and a whey protein enzymatic hydrolysate in a specific ratio. In order to better maintain, improve or ameliorate cognitive function, the ratio of the hop oxidation reaction product to the whey protein enzymatic hydrolysate in the composition and preparation (mass ratio converted into solid content) may be 0.1 to 2, and preferably 0.5 to 1.5.

[0050] When the composition and agent of the present invention contain a hop oxidation reaction product, bitter acids can be used as an indicator, and 0.1 to 1000 mg (preferably 0.5 to 500 mg, more preferably 1 to 100 mg) of bitter acids can be contained per meal. When the composition and agent of the present invention contain an enzymatic hydrolyzate of whey protein, the peptide of the present invention can be used as an indicator, and 0.001 to 100 mg (preferably 0.005 to 50 mg, more preferably 0.01 to 10 mg) of GTWY and 0.0001 to 30 mg (preferably 0.005 to 15 mg, more preferably 0.001 to 3 mg) of WY can be contained per meal.

[0051] <Application> As described in the Examples below, hop oxidation reaction products and whey hydrolysates (hereinafter, these may be referred to as "active ingredients of the present invention") have the effect of maintaining, enhancing and / or improving cognitive functions including memory function. Therefore, the combination of hop oxidation reaction products and whey hydrolysates can be used as an active ingredient for maintaining, enhancing and / or improving cognitive function, and can be used as an active ingredient in a method for maintaining, improving and improving cognitive function. The combination of hop oxidation reaction products and whey hydrolysates can also be used as an active ingredient in an agent for maintaining, improving and improving cognitive function. In the present invention, "cognitive function" is a concept that encompasses all information processing processes from perception to judgment, and various higher brain functions such as memory function (including memory acquisition function, memory consolidation function, memory retrieval function and memory reproduction function), attention function and judgment function are involved in this process. That is, in the present invention, "cognitive function" is used in a sense that includes these higher brain functions.

[0052] In the present invention, "maintaining cognitive function" includes, for example, suppressing or preventing a decline in cognitive function. Furthermore, "improving cognitive function" includes, for example, increasing cognitive function from its current state. Furthermore, "improving cognitive function" includes, for example, restoring cognitive function that has declined or symptoms that show signs of decline. The terms "maintaining, enhancing, and / or improving cognitive function" are used to mean, for example, enhancing cognitive function and suppressing a decline in cognitive function.

[0053] In the present invention, the maintenance, enhancement and / or improvement of cognitive function can be evaluated using higher brain functions involved in cognitive function as an index. Model animal tests for evaluating higher brain functions involved in cognitive function are known, and for example, the maintenance, enhancement and / or improvement of cognitive function can be evaluated by a Y-maze test (see Example 3 below) for evaluating short-term memory and spatial memory / working memory.

[0054] In the present invention, the term "memory function" refers to functions including spatial cognitive function, working memory function, working memory function, episodic memory function, visual memory function, and learning function, and is used to mean medium- to long-term memory function. Working memory includes linguistic working memory, task working memory, and spatial working memory. In the present invention, the memory function is preferably a long-term memory function.

[0055] In the present invention, "maintaining memory function" includes, for example, suppressing or preventing a decline in memory function. Furthermore, "improving memory function" includes, for example, increasing memory function from the current level, promoting the consolidation of medium- to long-term memory, and ultimately promoting brain development. Furthermore, "improving memory function" includes, for example, recovering a memory function that has declined or a symptom that shows signs of decline. Examples of maintaining, enhancing, and / or improving memory function include enhancing memory function and suppressing a decline in memory function.

[0056] In the present invention, the "spatial memory function" is a function for storing spatial information required for searching for an object, heading to a destination, returning home, and the like.

[0057] In the present invention, "maintaining spatial memory function" includes, for example, suppressing or preventing a decline in spatial memory function. Furthermore, "improving spatial memory function" includes, for example, enhancing spatial memory function from its current state. Furthermore, "improving spatial memory function" includes, for example, recovering a spatial memory function that has declined or a symptom showing signs of decline. Examples of maintaining, enhancing and / or improving spatial memory function include enhancing spatial memory function and suppressing a decline in spatial memory function.

[0058] In the present invention, the term "working memory function" refers to the function of working while temporarily holding information.

[0059] In the present invention, "maintaining working memory function" includes, for example, suppressing or preventing a decline in working memory function. Furthermore, "improving working memory function" includes, for example, increasing working memory function from its current level. Furthermore, "improving working memory function" includes, for example, recovering a working memory function that has declined or a symptom showing signs of decline. Examples of maintaining, enhancing and / or improving working memory function include enhancing working memory function and suppressing a decline in working memory function.

[0060] The compositions and agents of the present invention can be provided in the form of pharmaceuticals (e.g., pharmaceutical compositions), quasi-drugs, foods (e.g., food compositions), feeds (including pet foods), etc., and can be implemented according to the following description.

[0061] The active ingredient of the present invention can be orally administered to humans and non-human animals. Oral preparations include granules, powders, tablets (including sugar-coated tablets), pills, capsules, syrups, emulsions, and suspensions. These preparations can be formulated using pharmaceutically acceptable carriers by methods commonly used in the art. Pharmaceutically acceptable carriers include excipients, binders, diluents, additives, flavors, buffers, thickeners, colorants, stabilizers, emulsifiers, dispersants, suspending agents, and preservatives.

[0062] The active ingredient of the present invention can be orally ingested by humans and non-human animals, and a typical form of ingestion is food. When the active ingredient of the present invention is provided as food, it can be provided by being contained in food. The food provided in this way is a food containing an effective amount of the active ingredient of the present invention. In this specification, "containing an effective amount" of the active ingredient of the present invention refers to a content such that the active ingredient of the present invention is ingested in a range described below when the amount normally consumed in each food is ingested. In addition, "food" is used to mean health food, functional food, nutritional supplement, health functional food (e.g., specific health food, nutritional functional food, functional food), special purpose food (e.g., food for infants, food for pregnant women, food for sick people) and supplements. It goes without saying that when the active ingredient of the present invention is ingested by animals other than humans, the food referred to in the present invention is used as feed.

[0063] Since the active ingredient of the present invention has the effect of improving cognitive function as described above, it can be contained in foods that are taken daily, or can be provided as a supplement. That is, the composition and the agent of the present invention can be provided in the form of food. In this case, the composition and the agent of the present invention can be provided in a unit package form in which the amount to be taken per meal is predetermined. Examples of the unit package form per meal include a form in which a certain amount is specified by a pack, a package, a can, a bottle, etc. In order to better exert the various effects of the composition and the agent of the present invention, the intake amount per meal can be determined according to the daily intake amount of the active ingredient of the present invention, which will be described later. The food of the present invention may be provided with an explanatory matter regarding the intake amount displayed on the package, or with a document or the like in which the explanatory matter is described.

[0064] The predetermined intake amount per meal in the unit package form may be the effective intake amount per day, or the effective intake amount per day divided into two or more times (preferably two or three times). Therefore, the unit package form of the composition and preparation of the present invention may contain the active ingredient of the present invention at the daily intake amount per human described below, or may contain the active ingredient of the present invention in an amount of 1 / 2 to 1 / 20 of the daily intake amount per human described below. For convenience of intake, the composition and preparation of the present invention are preferably provided in a "unit package form per meal" in which the intake amount per meal is the effective intake amount per day.

[0065] The form of the "food" is not particularly limited, and may be, for example, a beverage, a semi-liquid or gel form, or a solid or powder form. In addition, examples of the "supplement" include tablets produced by adding an excipient, a binder, etc. to the active ingredient of the present invention, kneading the mixture, and then tableting, and capsules encapsulated in capsules. When providing the supplement, in addition to the above-mentioned unit package form per meal, it is also suitable to provide it in a unit package form per day, per week, or per month.

[0066] The foods provided by the present invention are not particularly limited as long as they contain the active ingredient of the present invention, and examples thereof include non-alcoholic beverages such as soft drinks, carbonated drinks, drinks containing fruit juice, drinks containing vegetable juice, drinks containing fruit juice and vegetable juice, livestock milk such as cow's milk, soy milk, dairy drinks, drink-type yogurt, drink-type or stick-type jellies, coffee, cocoa, tea drinks, nutritional drinks, energy drinks, sports drinks, mineral water, near-water, and non-alcoholic beer-flavored drinks; carbohydrate-containing foods and beverages such as rice, noodles, bread, and pasta; cheeses, hard or soft yogurt, fresh cream made from livestock milk or other fats and oils, ice cream, etc. dairy products; Western confectionery such as cookies, cakes, and chocolates, Japanese confectionery such as manju and yokan, tablet confectionery (refreshing confectionery) such as ramune, candies, gums, cold confectionery such as jelly and pudding, ice confectionery, snacks, and other confectionery; alcoholic beverages such as whiskey, bourbon, spirits, liqueur, wine, fruit wine, sake, Chinese wine, shochu, beer, non-alcoholic beer with an alcohol content of 1% or less, sparkling wine, other miscellaneous alcohol, and chuhai; processed foods such as processed foods using eggs, processed foods (including delicacies) of seafood and meat (including liver and other organ meats), soups such as miso soup, seasonings such as miso, soy sauce, furikake, and other seasoning seasonings, and liquid foods such as thick liquid foods. Mineral water includes both sparkling and non-sparkling mineral water. The food provided by the present invention also includes both food manufacturing raw materials and food additives.

[0067] Tea beverages include all of fermented, semi-fermented, and non-fermented teas, such as black tea, green tea, barley tea, brown rice tea, sencha, gyokuro tea, roasted green tea, oolong tea, turmeric tea, pu-erh tea, rooibos tea, rose tea, chrysanthemum tea, ginkgo leaf tea, and herbal teas (e.g., mint tea, jasmine tea).

[0068] Examples of fruits used in fruit juice beverages and beverages containing fruit and vegetable juice include apples, mandarin oranges, grapes, bananas, pears, peaches, mangoes, acai, blueberries, and plums, while examples of vegetables used in vegetable juice beverages and beverages containing fruit and vegetable juice include tomatoes, carrots, celery, pumpkins, cucumbers, and watermelons.

[0069] The intake amount of the active ingredient of the present invention can be determined depending on the recipient's sex, age, and body weight, symptoms, intake time, dosage form, intake route, combined drugs, etc. The daily intake amount (converted into solid content) of a hop oxidation reaction product for the purpose of maintaining, enhancing, or improving cognitive function for an adult is, for example, 10 to 5000 mg (preferably 50 to 2000 mg, more preferably 100 to 1000 mg) as a water extract, which is, for example, 0.1 to 1000 mg (preferably 1 to 100 mg, more preferably 10 to 60 mg) when converted into bitter acid. In addition, the daily intake (in terms of solid content) of whey hydrolysates for adults for the purpose of maintaining, improving or improving cognitive function is, for example, 1 to 50,000 mg (preferably 10 to 10,000 mg, more preferably 100 to 5,000 mg), which is converted to GTWY, for example, 0.1 to 5,000 mg (preferably 1 to 1,000 mg, more preferably 10 to 500 mg), and converted to WY, for example, 0.01 to 1,000 mg (preferably 0.1 to 300 mg, more preferably 3 to 100 mg). The above-mentioned intake amount of the active ingredient of the present invention and the following intake timing and intake period are applicable to both non-therapeutic and therapeutic uses of the active ingredient of the present invention, and in the case of therapeutic purposes, intake can be read as administration. The active ingredient of the present invention can also be administered to mammals other than humans (e.g., mice, rats, rabbits, dogs, cats, cows, horses, pigs, monkeys, dolphins, sea lions, etc.), and the intake amount, intake timing and intake period can be determined with reference to the descriptions for humans.

[0070] It is preferable to continue taking the active ingredient of the present invention during the period in which the maintenance, improvement or improvement of cognitive function is expected. The period of taking the active ingredient of the present invention can be, for example, at least one day (preferably three days, more preferably five days) at the above-mentioned daily dose, and can be one week or more, preferably one month or more, particularly when the maintenance of cognitive function is expected. In addition, when the maintenance, improvement or improvement of memory function, particularly among cognitive functions, is expected, it can be at least one day. The interval of taking the active ingredient of the present invention can be once every three days, once every two days, or once a day at the above-mentioned daily dose.

[0071] The active ingredient of the present invention can also be started to be taken before the date and time when the maintenance, improvement or improving effect of cognitive function is expected. Examples of the date and time before the expected maintenance, improvement or improving effect of cognitive function include before studying for an exam when the cognitive function (particularly memory function) is expected to be improved, and before the cognitive function starts to decline due to age.

[0072] The composition, agent, and food of the present invention may be labeled with a statement that it has an effect of maintaining, enhancing, or improving cognitive function. In this case, the labeling can be easily understood by consumers.

[0073] According to another aspect of the present invention, there is provided a combination of a hop oxidation reaction product and a whey protein enzymatic hydrolysate for use in maintaining, enhancing and / or improving cognitive function. The combination of the present invention includes, for example, a kit for use in maintaining, enhancing and / or improving cognitive function, comprising a hop oxidation reaction product and a whey protein enzymatic hydrolysate in separate forms (as independent entities). The combination of the present invention can be carried out according to the description of the composition, agent and food of the present invention.

[0074] According to another aspect of the present invention, there is provided a composition for maintaining, enhancing and / or improving cognitive function, and an agent for maintaining, enhancing and improving cognitive function, comprising a hop oxidation reaction product, which is a composition and preparation for use in combination with a whey protein enzymatic hydrolysate. According to another aspect of the present invention, there is also provided a composition for maintaining, enhancing and / or improving cognitive function, and an agent for maintaining, enhancing and improving cognitive function, comprising a whey protein enzymatic hydrolysate, which is a composition and preparation for use in combination with a hop oxidation reaction product. The above-mentioned compositions and preparations can be carried out according to the description of the compositions, preparations and foods of the present invention.

[0075] According to another aspect of the present invention, there is provided a method for maintaining, enhancing or improving cognitive function, comprising ingesting or administering to a subject in need thereof a combination of a hop oxidation reaction product and a whey protein enzymatic hydrolysate or a composition containing the same. The method of the present invention can be carried out according to the description of the composition and preparation of the present invention.

[0076] According to yet another aspect of the present invention, there is provided use of a combination of a hop oxidation reaction product and a whey protein enzymatic hydrolysate, or a composition containing the same, for the manufacture of an agent for maintaining, improving or improving cognitive function, as an agent for maintaining, improving or improving cognitive function, or in a method for maintaining, improving or improving cognitive function of the present invention. The use of the present invention can be carried out according to the description of the composition and agent of the present invention.

[0077] According to yet another aspect of the present invention, there is provided a combination of a hop oxidation reaction product and a whey protein enzymatic hydrolysate for use in maintaining, enhancing or improving cognitive function. The combination of the hop oxidation reaction product and the whey protein enzymatic hydrolysate can be carried out according to the description of the composition and preparation of the present invention.

[0078] The method and use of the present invention may be used in mammals, including humans, and both therapeutic and non-therapeutic uses are intended. As used herein, "non-therapeutic" does not include surgical, therapeutic or diagnostic procedures for humans (i.e., medical procedures for humans), and specifically does not include methods in which a physician or a person under the supervision of a physician performs surgical, therapeutic or diagnostic procedures on humans. EXAMPLES

[0079] The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

[0080] Reference Example 1: Preparation of hop pellet oxidation reaction product The hops used were pellet-shaped Hallertauperle (HPE). The hops were ground in a mill and heated at 80°C for up to 24 hours. The resulting product was pretreated as follows and then subjected to HPLC analysis.

[0081] [Reactant analysis pretreatment] The collected product was added to ethanol to a concentration of 10% w / v, and extraction was carried out for 1 hour at 50° C. The resulting extract was diluted 10-fold with ethanol. [HPLC configuration equipment] Pump: LC-10ADvp x 3 (SHIMADZU) Degasser: DGU-20A5 (SHIMADZU) System controller: CBM-20A (SHIMADZU) Autosampler: SIL-20ACHT (SHIMADZU) Column oven: CTO-20AC (SHIMADZU) Photodiode array detector: SPD-M20A (SHIMADZU) Waveform analysis software: LCSolution (SHIMADZU)

[0082] [HPLC conditions] Column: Alltima C18 2.1mm ID x 100mm Particle size 3μm Flow rate: 0.6mL / min Dissolution medium A: Water / phosphoric acid, 1000 / 0.2, (v / v) + EDTA(free) 0.02%(w / v) Elution solvent B: Acetonitrile Dissolution medium C: Water Injection volume: 3μL Column temperature: 40℃ Detection wavelength: 270 nm (oxidation reaction products, iso-α acids, α acids, β acids) Gradient conditions: Table 1

[0083] [Table 1]

[0084] Under the above analytical conditions, the ratio (%) of the area values ​​of the α-acid, β-acid, and iso-α-acid peaks to the total area value (mAU·min) of all peaks detected at a detection wavelength of 270 nm was calculated. In the waveform analysis, the regions where solvent peaks and negative peaks due to injection shock occurred were excluded from the analysis. The HPLC chromatogram of the above product analysis is shown in Figure 1.

[0085] Reference Example 2: Measurement of bitter acids (1) Measurement of bitter acids contained in hop oxidation products Measure using high performance liquid chromatography. For details, follow the method published in the Consumer Affairs Agency's functional food labeling notification (https: / / www.fld.caa.go.jp / caaks / cssc04 / bunsekihoho?bunsekihohoFile=E522%255CE522_bunsekihoho.pdf).

[0086] (i) Preparation of measurement samples The sample is water with an appropriate amount of hop oxidation products dissolved in it. Dichloromethane and 1N hydrochloric acid are added to the sample, and after stirring, the mixture is centrifuged. The bottom layer is collected and evaporated to dryness under a nitrogen stream at room temperature. This is then redissolved in ethanol and centrifuged to obtain the measurement sample.

[0087] (ii) High-performance liquid chromatography conditions Column: Alltima C18 2.1 mm ID x 100 mm, particle size 3 μm Mobile phase A: water / phosphoric acid 1000 / 0.2(v / v), 0.02%w / vEDTA Mobile phase B: Acetonitrile Mobile phase C: water Injection volume: 3μL Column temperature: 40℃ Autosampler temperature: 4℃ Detection: 270nm Gradient conditions: Table 2

[0088] [Table 2]

[0089] (iii) Waveform processing The elution time of the end point of the caffeine peak (Figure 2A) is the start time of the calculation of the area of ​​bitter acids, and the end time is the peak of trans-isocohumulone (peak a in Figure 2B). Perform background processing as necessary, and calculate the peak area of ​​bitter acids (Figure 2C).

[0090] (iv) Calculation of the amount of bitter acid Using the calibration curve prepared with the TCOIHA standard solution, the concentration of bitter acid in the sample is calculated as a TCOIHA equivalent value according to the following formula. Concentration of bitter acid in sample (mg / 100mL) = (peak area of ​​bitter acid in hop oxidation products - intercept of calibration curve prepared with TCOIHA standard solution) / slope of calibration curve prepared with TCOIHA standard solution × 1 / 5 × 100 / 1000

[0091] (2) Preparation of samples for measuring bitter acids in compositions containing hop oxidation products The solid composition of the specimen in (1) above is prepared by the same procedure as in (1), and the liquid composition is prepared as is or by appropriate dilution. It has been concluded that all compounds detected in the bitter acid elution region have a β-tricarbonyl skeleton (Biosci., Biotechnol., Biochem., 2015 (79): 1684-1694, J., Agric., Food Chem., 2015: 63: 10181-10191).

[0092] Example 1: Preparation and component analysis of hop oxidation products (1) Hop oxidation process Hallertauperle (HPE) hops were ground in a pellet mill, and the resulting ground hops were heated at 60° C. for about 120 hours while stirring in the air. Water was added to the resulting heated hops (matured hop pellets: one embodiment of a hop oxidation reaction product) so that the solid content concentration was 5 w / v%, and extraction treatment was carried out for 30 minutes at 50° C. The resulting extract was subjected to solid-liquid separation by decantation to obtain a solid-liquid separated liquid (Brix approximately 2).

[0093] (2) Activated carbon treatment process Activated carbon (Y180C, manufactured by Ajinomoto Fine Chemicals Co., Ltd.; 0.5 w / v% relative to the solid-liquid separated liquid) and polyvinyl polypyrrolidone (Polyclar 10, manufactured by ISP Japan Co., Ltd.; 0.4 w / v% relative to the solid-liquid separated liquid) were added to the solid-liquid separated liquid obtained in (1) above, and the mixture was allowed to stand for 2 hours. A filter aid (diatomaceous earth) was added to the resulting mixture, and the mixture was filtered to obtain a filtrate (Brix value of about 1.5). The resulting filtrate was used in the following examples as a water extract of a hop oxidation reaction product.

[0094] (3) Analysis of hop oxidation products The filtrate obtained in (2) above (water extract of hop oxidation reaction products) was analyzed by HPLC-MS / MS under the following conditions to measure the contents of various components contained in the hop oxidation reaction products. It is known that hop oxidation reaction products contain scorpiofumulinol A, scorpiocohumulinol A, tricyclooxyisohumulone A, and tricyclooxyisocohumulone A as α-acid oxides, iso-α-acid oxides, or β-acid oxides (Biosci., Biotechnol., Biochem., 2015 (79): 1684-1694, J., Agric., Food Chem., 2015: 63: 10181-10191). In addition, the standards used in the analysis were prepared according to the methods described in J., Agric., Food Chem., 2015:63:10181-10191 and J. Nat. Prod. 2014, 77, 1252-1261.

[0095] [HPLC conditions] Column: Unison UK-C18 100 x 2 mm id, particle size 3 μm Flow rate: 0.25mL / min Column temperature: 40℃ Mobile phase A: 1% formic acid in water Mobile phase B: acetonitrile containing 1% formic acid Injection volume: 3μL Gradient conditions: 0 → 30 min, 15 → 31% B 30→40 minutes, 31→80%B 40→43 minutes, 80%B After that, washing and equilibration steps

[0096] [MSMS conditions] Mass spectrometer: AB SCIEX 4000Q Trap Ion source: ESI - negative ion mode Ion spray voltage: -4500V Analysis parameters:

[0097] [Table 3]

[0098] The above (1) and (2) were carried out three times to obtain a filtrate, and the contents of scorpiofumulinol A, scorpiocohumulinol A, tricyclooxyisohumulone A and tricyclooxyisocohumulone A were measured. As a result, the ratio of the total amount of tricyclooxyisohumulone A and tricyclooxyisocohumulone A to the total amount of scorpiofumulinol A and scorpiocohumulinol A ((tricyclooxyisohumulone A + tricyclooxyisocohumulone A) / (scorpiofumulinol A + scorpiocohumulinol A)) was as follows. Lot 1:3.2 Lot 2: 6.8 Lot 3: 12.1

[0099] From the above results, it was found that in the hop oxidation reaction products, the ratio of the total amount of tricyclooxyisohumulone and tricyclooxyisocohumulone to the total amount of scorpiofumulinol and scorpiocohumulinol was approximately 2 to 20.

[0100] Example 2: Determination of the content of tetrapeptide GTWY and dipeptide WY in whey hydrolysates (1) Preparation of analytical samples The whey hydrolyzate (HW-3, Megmilk Snow Brand) was diluted appropriately with sterilized water and filtered to prepare a measurement sample. The whey hydrolyzate (HW-3) was a product obtained by treating whey protein with an enzyme preparation containing a protease, then removing undigested matter by membrane processing, and drying, and contains the tetrapeptide GTWY and dipeptide WY as described below.

[0101] (2)Analysis method The concentrations of the tetrapeptide GTWY and dipeptide WY in the measurement samples obtained in (1) above were quantified by LC-MS / MS under the following analytical conditions. The concentrations of GTWY and WY in the measurement samples were calculated by the calibration curve method using AQUA Peptide (Sigma Aldrich) as the standard sample.

[0102] <Analysis conditions> Mass spectrometer: 4000Q TRAP (AB Sciex) HPLC equipment: Agilent 1200 Series (Agilent Technologies) Column: TSK gel ODS-100V 3μm 2.0mm ID×150mm (Tosoh) Column temperature: 70℃ Mobile phase A: 0.1% formic acid in water Mobile phase B: 0.1% formic acid in acetonitrile Gradient conditions: The gradient conditions shown in Table 4 were applied.

[0103] [Table 4]

[0104] Flow rate: 0.2mL / min Sample injection volume: 2μL Ionization method: ESI (positive ion detection mode) Curtain gas: 40psi Nebulizer gas: 50 psi Dry gas: 80psi Drying gas temperature: 600℃ Collision gas: Nitrogen Ionization voltage: 5000V

[0105] <Analysis conditions for tetrapeptide GTWY> Set mass number (m / z) / collision energy (eV): 526.4→159.2 / 47, 526.4→368.3 / 23 DP voltage(V):36

[0106] <Analysis conditions for dipeptide WY> Set mass number (m / z) / collision energy (eV): 368.2 → 351.1 / 19, 368.2 → 159.2 / 33 DP voltage(V):51

[0107] (3)Analysis results It was confirmed that 1.62 mg of tetrapeptide GTWY and 0.60 mg of dipeptide WY were contained per gram of whey hydrolyzate (HW-3).

[0108] (4) Measurement of the peptide of the present invention in a composition containing whey hydrolysate When the composition is solid, the analysis sample is prepared from the composition instead of the whey hydrolyzate of (1). When the composition is liquid, the analysis sample is prepared by appropriate dilution and filtration.

[0109] Example 3: Effects of hop oxidation products and whey hydrolysates on cognitive function (particularly short-term memory, spatial memory, and working memory) (1) Method A. Experimental Procedure Six-week-old male CD-1 mice (Japan Oriental Yeast Co., Ltd.) were acclimated for one week, raised on crudely purified feed CE-2 (Japan Clea), and divided into five groups without bias. The samples shown in Table 5 were administered intragastrically for each group. The samples were prepared by adding and dissolving the dried hop oxidation reaction product extract (prepared in Example 1), whey hydrolyzate (prepared in Example 2), and donepezil hydrochloride (Fujifilm Wako Pure Chemical Industries, Ltd.) in distilled water immediately before administration. The concentrations of the samples were 0.03 mg / mL for the whey hydrolyzate and hop oxidation reaction product, and 0.1 mg / mL for donepezil hydrochloride. 40 minutes after administration, scopolamine hydrochloride (SCP) (Sigma) was administered intraperitoneally at 0.85 mg / kg body weight to induce memory impairment, and amnesic model mice were created. 20 minutes after intraperitoneal administration of SCP, the Y-maze test described below was performed. The experiment was performed with 10 animals per group, and the average and standard error of each group were calculated. The dose of donepezil hydrochloride was, for example, the amount described in Behav Brain Res. 2009 Dec 1;204(1):217-25.

[0110] [Table 5]

[0111] B. Cognitive function evaluation When a mouse is placed in a space, if it remembers the route it previously chose, it will choose a different route from the previous one due to its desire to explore novelty. Therefore, when a mouse is placed in a Y-maze with three arms of equal width and length, it will usually enter an arm different from the one it previously entered. The Y-maze test utilizes this property of mice to evaluate short-term memory, spatial memory, and working memory, which are indicators of cognitive function.

[0112] The Y-maze apparatus used had three arms, each 25 cm long, with walls 20 cm high and a floor 5 cm wide, connected at a 120 degree angle. Mice were placed at the end of one of the arms of the Y-maze and allowed to explore freely for 8 minutes, and the order of the arms they moved through was recorded. Spontaneous alternation behavior was defined as a case in which a mouse selected and entered a different arm three times in a row. The total number of entries into the arms and the number of spontaneous alternation behaviors within the time period were counted, and the spontaneous alternation behavior variability (%) was calculated using the following formula (1). A higher spontaneous alternation behavior variability indicates that short-term memory, spatial memory, and working memory are better retained.

[0113]

number

[0114] (2) Results The results are shown in Figure 3 (total number of entries) and Figure 4 (rate of spontaneous alternation behavior fluctuation). Although cognitive function was improved when hop oxidation reaction products or whey hydrolysates were administered alone, it was confirmed that the combined administration of hop oxidation reaction products and whey hydrolysates had a more significant effect on improving cognitive function than administration of either one alone.

Claims

1. A composition for maintaining, enhancing and / or improving cognitive function, comprising a hop oxidation reaction product and a whey protein enzymatic hydrolysate as active ingredients, wherein the hop oxidation reaction product includes tricyclooxyisocohumulone A, tricyclooxyisohumulone A, scorpiofumulinol A and scorpiocohumulinol A, the whey protein enzymatic hydrolysate includes a peptide having the amino acid sequence of GTWY (sequence number 1) and a peptide having the amino acid sequence of WY, the cognitive function is a memory function, and the memory function is a spatial memory function and / or a working memory function.

2. The composition of claim 1 , wherein the hop oxidation reaction product is an aqueous medium extract.

3. 3. The composition of claim 1 or 2, wherein the hop oxidation reaction product is a water extract.

4. The composition according to any one of claims 1 to 3, wherein the ratio (in terms of dry mass) of the total amount of tricyclooxyisohumulone A and tricyclooxyisocohumulone A to the total amount of scorpiofumulinol A and scorpiocohumulinol A is in the range of 2 to 20.

5. The composition according to any one of claims 1 to 4, wherein the concentration of bitter acid in the hop oxidation reaction product is 1 to 20 mass %.

6. The composition according to claim 5, wherein the content of GTWY in the enzymatic hydrolyzate of whey protein (converted into solid content) is 0.01 to 1 mass% based on the total amount, and the content of WY in the enzymatic hydrolyzate of whey protein (converted into solid content) is 0.005 to 0.5 mass% based on the total amount.

7. The composition according to any one of claims 1 to 6, wherein the ratio of the hop oxidation reaction product to the whey protein enzymatic hydrolysate in the composition (mass ratio converted into solid content) is 0.1 to 2.

8. The composition according to any one of claims 1 to 7, which is a food composition.

9. The composition according to any one of claims 1 to 8, which is in a unit package form per serving.

10. The composition according to any one of claims 1 to 9, wherein the hop oxidation reaction product is ingested by a human in an amount of 10 to 5,000 mg (in terms of solid content) per meal.

11. The composition according to any one of claims 1 to 10, wherein the whey protein enzymatic hydrolysate is ingested by a human in an amount of 1 to 50,000 mg (in terms of solid content) per meal.

12. The composition according to claim 10 or 11, comprising 0.01 to 10 mg of GTWY and 1 to 100 mg of a bitter acid.

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