JP7344722B2 - Composition for improving constitutive behavior - Google Patents

Description

The present invention relates to a composition for improving constitutive behavior that contains a specific peptide.

According to the International Alzheimer's Association, 9.9 million people were newly diagnosed with dementia in 2015, increasing at a rate of one person every three seconds (Non-Patent Document 1). Furthermore, the number of dementia patients in the world was 46.8 million as of 2015, but it is predicted that this number will reach 131.5 million by 2050. It is known that approximately 70% of dementia cases are caused by Alzheimer's disease, and countermeasures against Alzheimer's disease and other dementias have become an urgent issue around the world. Although there are some drugs that can slow the progression of dementia, there are currently no known cures. Therefore, it has been pointed out that it is important to preventively improve brain function before brain dysfunction occurs, that is, before dementia develops (Non-Patent Documents 2 and 3).
Since it is a preventive method that targets the stage before brain dysfunction occurs, it is desirable that it be a method that is highly safe and can be used continuously.

The act of composition refers to the ability to form a single coherent object by combining multiple constituent elements. Compositional apraxia, in which the ability to perform compositional acts is reduced, is known to be one of the characteristic symptoms of Alzheimer's disease (Non-Patent Document 4). Specific symptoms of compositional apraxia include, for example, getting lost and apraxia (not knowing how to use objects or using them incorrectly).
Therefore, improving constituent behavior in people without brain dysfunction is thought to not only help them lead a healthy life in the present, but also prevent the onset of dementia such as Alzheimer's disease in the future. .

Up to now, an Alzheimer's type dementia improving agent for the elderly is known which contains a casein hydrolyzate and a peptide consisting of Met-Lys-Pro contained therein as an active ingredient (Patent Document 1). Furthermore, it has been found that casein hydrolyzate and a peptide consisting of Met-Lys-Pro contained therein have an ameliorating effect on brain dysfunction caused by amyloid beta protein (Patent Document 2). It has been confirmed that peptides composed of casein hydrolyzate and Met-Lys-Pro have few side effects, are excellent in safety, and do not cause fluctuations in blood pressure.
However, as an effect of casein hydrolyzate and the peptide consisting of Met-Lys-Pro contained therein, it is not known that it improves composition behavior, especially in people without brain dysfunction.

JP2016-69343A Japanese Patent Application Publication No. 2018-154640

World Alzheimer report 2015: the global impact of dementia. An analysis of prevalence, incidence, cost and trends. London: Alzheimer’s Disease International; (2015). Stop Alzheimer's before it starts. Nature. 2017, 547(7662):153-155. Dementia prevention, intervention, and care. Lancet. 2017, 390(10113):2673-2734. Dementia disease treatment guidelines 2017 Japanese Society of Neurology

An object of the present invention is to provide a technique for improving configuration operations that is highly safe and can be continued.

As a result of intensive research to solve the above problems, the present inventors discovered that a peptide having Met-Lys-Pro as an amino acid sequence improves the composition behavior, and completed the present invention. .

That is, the present invention is a composition for improving constitutive behavior, which contains a peptide consisting of Met-Lys-Pro or a peptide containing the above amino acid sequence.
The compositions of the invention are preferably ingested or administered to subjects who do not have dementia.
The compositions of the invention are preferably used to improve the ability to remember and recall directions and/or how to use an article.
The composition of the present invention is preferably a food or drink.
The composition of the invention is preferably a pharmaceutical.

According to the present invention, it is possible to improve the composition behavior of the person taking the medicine (administrator). Moreover, the composition of the present invention has few side effects and is excellent in safety, so it can be taken continuously. The composition of the present invention can be preferably applied to people who do not have dementia, and more preferably to people who do not have brain dysfunction. Furthermore, the composition of the present invention is preferably administered to elderly people.

Next, the present invention will be explained in detail. However, the present invention is not limited to the following embodiments, and can be freely modified within the scope of the present invention.

The composition of the present invention contains a peptide consisting of Met-Lys-Pro or a peptide containing the above amino acid sequence (collectively referred to as "MKP peptide").
Here, Met (M) represents a methionine residue, Lys (K) represents a lysine residue, and Pro (P) represents a proline residue. Preferably, both amino acids are L-type amino acids.

The MKP peptide may be in its salt form. Examples of the salt include alkali metals such as potassium and sodium; alkaline earth metals such as calcium and magnesium.

In the present invention, the term "peptide containing" a predetermined amino acid sequence refers to a peptide in which one or more arbitrary amino acids are added to the N-terminus or C-terminus of the predetermined amino acid sequence. Specifically, a peptide with 1 to 10 residues, preferably 1 to 5 residues, and more preferably 1 to 3 residues added, and a sequence that has an effect of improving compositional behavior is desirable.

The composition of the present invention may contain other peptides as long as they do not interfere with the effects of the present invention.

MKP peptide can be obtained, for example, by (1) decomposing a protein or peptide containing the Met-Lys-Pro amino acid sequence with a hydrolase, etc., and separating and purifying the resulting decomposed product; (2) obtaining the peptide by separating and purifying the resulting decomposed product; A method of synthesizing an MKP peptide by a synthetic method, and then separating and purifying the desired MKP peptide from the resulting synthetic product; (3) a method of obtaining a desired MKP peptide from a plant, animal, or microorganism that produces a peptide, etc. containing the MKP peptide according to the present invention; It can be obtained by a method of extraction and separation and purification from the obtained extract.
The methods (1) and (2) will be specifically explained below.

(1) Method for obtaining MKP peptide by hydrolysis MKP peptide is obtained by hydrolyzing a protein or peptide containing the amino acid sequence of Met-Lys-Pro with a protein hydrolase, acid, alkali, etc. It can be obtained by separating and purifying. Examples of raw materials containing proteins and peptides include proteins derived from milk, soybeans, eggs, wheat, barley, rice, potatoes, sweet potatoes, peas, corn, meat, fish, and seafood. Of these, casein, which is a milk protein, is particularly preferred.

The following describes a method for obtaining MKP peptide by treatment with a hydrolase, using casein as a raw material as an example.
Casein protein is a protein that contains MKP peptide in its primary structure, and can generate MKP peptide when digested with an appropriate hydrolase.
First, before hydrolysis with an enzyme, raw protein is dispersed and dissolved in water or hot water to prepare an aqueous protein solution. The concentration of the aqueous protein solution is not particularly limited, but it is generally preferable to set the protein concentration to a concentration range of 2% by mass or more, more preferably about 5 to 15% by mass.
Furthermore, the protein aqueous solution is subjected to an ion exchange method using a sodium type or potassium type cation exchange resin (preferably a strongly acidic cation exchange resin), an electrodialysis method, an ultrafiltration membrane method, a loose reverse osmosis membrane method, etc. It is preferable to desalinate and adjust the pH and calcium concentration as appropriate. For desalting, either a column method or a batch method may be employed. Further, the aqueous protein solution may be heat sterilized as appropriate before desalting.

Next, the protein aqueous solution is subjected to a hydrolysis treatment. Examples of the hydrolysis treatment include enzyme treatment, acid treatment, alkali treatment, heat treatment, etc., and two or more of these treatments may be combined as appropriate.
The proteolytic enzymes of the present disclosure include, for example, plant-derived, animal-derived, microbial-derived, and the like, and can be used singly or in combination of two or more thereof. As the protease, endoprotease is suitable.
Examples of the endoprotease include serine protease, metalloprotease, cysteine protease, and aspartic protease, and one or more of these can be selected and used. Among these, it is preferable to use serine protease and/or metalloprotease.
Furthermore, proteases are classified into alkaline proteases, neutral proteases, and acidic proteases. Among these, it is preferable to use neutral proteases.

A commercially available product can be used as the protease. Examples of the proteolytic enzyme include Bioprase (manufactured by Nagase ChemteX), Proleather (manufactured by Amano Enzymes), Protease S (manufactured by Amano Enzymes), PTN6.0S (manufactured by Novozymes), and Sabinase (manufactured by Novozymes). , GODO B. A. P (manufactured by Godo Shusei Co., Ltd.), Protease N (manufactured by Amano Enzyme Co., Ltd.), GODO B. N. P (manufactured by Godo Shuusei Co., Ltd.), Neutrase (manufactured by Novozymes Co., Ltd.), Alcalase (manufactured by Novozymes Co., Ltd.), Trypsin (manufactured by Novozymes Co., Ltd.), Chymotrypsin (manufactured by Novozymes Co., Ltd.), Subtilisin (manufactured by Novozymes Co., Ltd.), Papain (manufactured by Amano Enzyme Co., Ltd.) ), bromelain (manufactured by Amano Enzymes), etc., and one or more enzymes may be selected from these and used.
Among these, one or more neutral proteases selected from subtilisin (e.g., bioplase), trypsin (e.g., PTN6.0S), and bachillolysin (e.g., protease N) are preferred, and more preferred. is used in combination of these three types.

The amount of endoprotease used for the protein is not particularly limited and may be adjusted appropriately depending on the substrate concentration, enzyme titer, reaction temperature, reaction time, etc., but in general, the amount of endoprotease used per 1 g of protein in the protein is ,000 to 11,000 active units.
A desired peptide can be obtained by appropriately adjusting the conditions for hydrolysis by the protease.

Before the hydrolysis by the proteolytic enzyme, the pH of the raw protein solution can be adjusted to the optimum pH for the enzyme used using food-grade salts such as potassium carbonate and sodium hydroxide. The pH of the raw protein solution is preferably adjusted to 5-10, more preferably 7-10.

The reaction temperature of the proteolytic enzyme is desirably within the optimum temperature range of the enzyme used, preferably 30 to 60°C, more preferably 40 to 60°C.
The reaction retention time of the protease may be, for example, 0.5 to 24 hours, and preferably 1 to 24 hours, as long as the reaction is continued until a desired decomposition rate is reached while monitoring the decomposition rate of the enzymatic reaction. 15 hours, more preferably 3 to 10 hours. In particular, when the casein protein is used as a raw material, the decomposition rate is preferably 20 to 30%.

The decomposition rate of the raw protein is calculated by measuring the total nitrogen content of the sample using the Kjeldahl method (edited by the Japan Food Industry Association, "Food Analysis Methods", p. 102, Korin Co., Ltd., 1981), and by measuring the total nitrogen content of the sample. The amount of formol nitrogen in the sample was measured by the titration method (edited by Mitsuda et al., "Food Engineering Experiment Book", Vol. 1, p. 547, Yokendo, 1970), and the decomposition rate was calculated from these measured values using the following formula: do.
Decomposition rate (%) = (formol nitrogen amount/total nitrogen amount) x 100

Hydrolysis by the proteolytic enzyme may be completed by heating the enzyme to inactivate it. The heating temperature and holding time of the heat inactivation treatment can be appropriately set to conditions that can sufficiently inactivate the enzyme, taking into consideration the thermal stability of the enzyme used. For example, when deactivating at 100°C or higher (preferably 110 to 130°C), it is preferable to perform the deactivation for 1 to 3 seconds, and when deactivating at a temperature lower than 100°C to 60°C or higher, it is preferably performed for 3 to 40 minutes. .
As the heat treatment method, either a batch method or a continuous method can be used. As the continuous method, methods such as a plate heat exchange method, an infusion method, and an injection method can be used.
Note that the above-mentioned heat deactivation treatment can also be used in combination as a sterilization treatment for the hydrolyzate, and a conventional heat treatment method or the like can be used.
After the hydrolysis is completed, the pH of the decomposition solution is preferably adjusted to 6 to 8, more preferably 7.0±0.5, and still more preferably 7.0±0.3, if necessary.

In addition, in the production of protein decomposition products according to the present invention, when a solution whose calcium concentration has not been adjusted is hydrolyzed, the resulting decomposed solution may be desalted as described above to adjust the calcium concentration. good. Next, the enzyme is deactivated by heating in a conventional manner. The reaction heating temperature and reaction holding time can be appropriately set to conditions that can sufficiently deactivate the enzyme, taking into consideration the thermal stability of the enzyme used. After inactivation by heating, it can be cooled by a conventional method and used as it is, or if necessary, it can be concentrated to obtain a concentrated liquid, and it is also possible to dry the concentrated liquid to obtain a powder product.

Furthermore, when the aqueous protein solution is hydrolyzed by acid treatment or alkali treatment, the pH of the aqueous protein solution may be adjusted. In the case of the treatment by pH adjustment, the pH of the aqueous protein solution is preferably pH 5 or less or pH 9 or more, more preferably pH 4 or less or pH 10 or more. By leaving or stirring the pH-treated aqueous solution at room temperature for several minutes or more, preferably from 5 minutes to 1 hour, an acid- or alkali-treated hydrolyzate can be obtained. Here, "room temperature" is about 4 to 40°C, preferably 10 to 30°C.
Further, the aqueous protein solution may be hydrolyzed by heat treatment. The pH of this aqueous protein solution may be unadjusted, or the pH may be adjusted (specifically, acidic (pH 5 or lower), neutral (pH 6 to 8), or alkaline (pH 8 or higher)). The heat treatment may be performed at a temperature of about 4 to 100° C. under the same conditions as the acid-alkali treatment described above.

Although a protein hydrolyzate such as casein can be obtained as described above, a commercially available casein hydrolyzate may also be used. It has been confirmed that commercially available casein hydrolysates usually contain MKP peptides.

Although the content of MKP peptide in the protein hydrolyzate is not particularly limited, the lower limit thereof is preferably 0.001% by mass or more, and 0.005% by mass from the viewpoint of better exerting the efficacy of the present invention. % or more, more preferably 0.01% by mass or more, and the upper limit thereof is preferably 1% by mass or less, more preferably 0.5% by mass, from the viewpoint of production efficiency of the decomposed product of the present invention. % or less, more preferably 0.4% by mass or less, even more preferably 0.3% by mass or less.

In addition, in the present invention, the target peptide content can be measured by the following method.
(a) The sample powder was diluted and dissolved in a 0.2% formic acid aqueous solution to a concentration of 1.0 mg/mL, crushed by ultrasonic waves for 10 minutes, and then filtered with a PVDF filter (manufactured by Millipore) with a diameter of 0.22 μm. A powder solution is prepared, and LC/MS analysis is performed under the following measurement conditions. On the other hand, several solutions of a chemically synthesized standard peptide (manufactured by Peptide Research Institute) of the peptide to be measured are prepared at different concentrations, and LC/MS analysis is performed under the following measurement conditions to create a calibration curve.
Among the peaks in the analysis of the powder solution, those whose molecular weight and retention time match those of the standard peptide are identified as having the same sequence as the standard peptide. By comparing the peak area of the standard peptide and the peak area of the sample powder, the content of the target peptide in the powder solution is determined.

(b) Target peptide content (mg/1 g of casein hydrolyzate)
Target peptide content (mg/1 g of casein hydrolyzate) = [measured value of target peptide in the obtained casein hydrolyzate (mg)]/[mass of the obtained casein hydrolyzate (g)]
[Measurement value (mg) of the target peptide in the obtained casein hydrolyzate] is the measurement value of the target peptide in the sample by the following "LC/MS".

(c) Equipment used for LC/MS Mass spectrometer: TSQ Quantum Discovery MAX (manufactured by Thermo Fisher Scientific).
High performance liquid chromatograph: Prominence (manufactured by Shimadzu Corporation), column: XBridge BEH300 C18 φ2.1 mm×250 mm, 3.5 μm (manufactured by Waters Corporation).

(d) LC/MS measurement conditions Mobile phase A: 0.2% by weight formic acid-aqueous solution Mobile phase B: 0.2% by weight formic acid-acetonitrile solution Time program: 2%B (0.0 min) - 25%B ( 5.0 minutes) - 65% B (5.1 minutes) - 65% B (10 minutes) - 85% B (10.1 minutes) - 85% B (13.0%) - 2% B (13. 1 minute) - STOP (30.0 minutes).
Sample injection amount: 10 μL, column temperature: 40°C, liquid flow rate: 200 μL/min
Analysis mode: SRM measurement.
Product Mass: m/z = 260.10 (Parent m/z = 375.21)

The obtained protein hydrolyzate can exhibit its efficacy even when used in an unpurified state. That is, the protein hydrolyzate may be ingested or administered in the form of food/drinks or pharmaceuticals, which will be described later.
Further, the obtained protein hydrolyzate may be subjected to appropriate known separation and purification. For example, the obtained protein hydrolyzate can be subjected to molecular weight fractionation to obtain a protein hydrolyzate containing a fraction corresponding to the molecular weight of the MKP peptide according to the present invention.
As the molecular weight fractionation, methods such as ultrafiltration and gel filtration can be employed, thereby increasing the removal rate of peptides and free amino acids with unnecessary molecular weights.
In the case of ultrafiltration, a desired ultrafiltration membrane may be used, and in the case of gel filtration, a desired gel filtration agent used for size exclusion chromatography may be used.
Furthermore, in order to desalt, remove impurities, and increase purity, various chromatography methods such as ion exchange chromatography, adsorption chromatography, reversed phase chromatography, and partition chromatography, as well as solvent Methods such as precipitation, salting out, partitioning between two liquid phases, etc. may also be used.

The peptides of the separated and purified peptide fraction can be identified by mass spectrometry for the purpose of confirming whether the MKP peptide is contained.
The MKP peptide thus obtained can be used as a peptide solution, or, if necessary, the solution can be concentrated by a known method and used as a concentrate. Alternatively, the concentrate can be dried by a known method and used as a powder.

(2) Method for obtaining by chemical synthesis MKP peptide can also be produced by chemical synthesis or biosynthesis.
Chemical synthesis of peptides can be performed by a liquid phase method or a solid phase method commonly used for oligopeptide synthesis. The synthesized peptide can be deprotected as necessary to remove unreacted reagents, by-products, etc., and isolate the MKP peptide. Such peptide synthesis can be performed using a commercially available peptide synthesizer.
Peptide biosynthesis can be carried out by a conventional method such as introducing a peptide expression vector into a host organism and causing production and secretion.

The compositions of the present invention can improve construction behavior.
Here, the act of composition refers to the ability to form a single coherent object by combining multiple constituent elements.
The improvement in compositional behavior can be confirmed, for example, by an improvement in the score in the compositional behavior item of ADAS-cog (Alzheimer's Disease Assessment Scale-cog).
Compositions of the present invention improve organization, thereby improving the ability to remember and recall directions and/or how to use an item.

Compositional apraxia, in which the ability to perform compositional acts is reduced, is known to be one of the characteristic symptoms of Alzheimer's disease (Non-Patent Document 4). From this, it is expected that when compositional behavior is improved by the composition of the present invention, brain dysfunctions such as Alzheimer's dementia, vascular dementia, vascular disorders, and cerebral amyloid angiopathy can be prevented. Here, prevention of brain dysfunction refers to preventing the occurrence of a disease or symptom, delaying its occurrence, and reducing the risk of its occurrence in applicable subjects who are not suffering from (onset of) brain dysfunction. Including lowering.

The composition of the present invention has been used as a food product and is highly safe. Furthermore, there is little concern about side effects even when administered continuously for a long period of time. Furthermore, it is highly safe when used in combination with other drugs.

The subject to whom the composition of the present invention is administered (administered person) and the subject to which it is ingested (consumer) are not particularly limited as long as they are animals, but are usually humans. The subject may be an adult, a child, an infant, a newborn (including a low birth weight infant), etc., but the subject is usually an adult, and preferably an elderly person. Note that in the present invention, the term "elderly person" refers to a person who is 65 years old or older. Furthermore, if the target is a non-human animal, it may be an elderly animal corresponding to an elderly person.
Furthermore, the gender of the person taking and administering the composition of the present invention is not particularly limited.
Furthermore, the ingestor and recipient of the composition of the present invention are preferably those who do not have dementia, more preferably those who do not have brain dysfunction, and still more preferably elderly people who do not have dementia. Especially preferred are elderly people who do not have brain dysfunction. Dementia is usually progressive and cannot be completely cured, so it targets people who are in the stage of developing dementia. In addition, since mild cognitive impairment is a pre-dementia stage, it corresponds to a "state of brain dysfunction but not dementia" and can be a preferable ingestor and recipient of the present invention.
Here, not having dementia may mean not having been diagnosed with dementia by a doctor, or not having been diagnosed with dementia by a doctor, or by meeting the criteria for determining dementia using various neuropsychological tests. It may be that the above does not apply. If the ingestor (administrator) is a human, it should preferably be confirmed by being healthy (normal) on the Hasegawa Dementia Scale (HDS-R) or the Mini-Mental State Examination (MMSE). I can do it.

Another aspect of the invention is the use of MKP peptides in the manufacture of compositions for improving constitutive behavior.
Another aspect of the invention is the use of MKP peptides in improving constitutive behavior.
Another aspect of the invention is MKP peptides used for improving constitutive behavior.
Another aspect of the invention is a method of improving compositional behavior comprising administering an MKP peptide to an animal.

In this specification, "administering a peptide to an animal" may be synonymous with "ingesting a peptide to an animal." The intake may be voluntary (free intake) or forced (forced intake). Specifically, the administration step may be, for example, a step in which the peptide is blended into food, drink, or feed and supplied to the subject, thereby allowing the subject to freely ingest the peptide.

The timing of ingestion (administration) of the composition of the present invention is not particularly limited, and can be appropriately selected depending on the condition of the subject to be administered.

The amount of the composition of the present invention to be ingested (administered) is appropriately selected depending on the age, sex, condition, other conditions, etc. of the subject to be ingested (administered).
The intake (administration) amount of the composition of the present invention is preferably in the range of 0.1 mg/day to 1 g/day, and 0.1 mg/day to 0.1 g/day for adults, as the intake amount of the MKP peptide according to the present invention. A range of 5 mg is more preferred.
Incidentally, regardless of the amount and period of intake (administration), the composition of the present invention can be administered once a day or divided into multiple doses.

The period of intake (administration) of the composition of the present invention is not particularly limited, but the effect is likely to be obtained if it is preferably 12 weeks or more, more preferably 24 weeks or more. Furthermore, there is no particular upper limit to the intake (administration) period, and continuous, long-term intake (administration) is possible.

The ingestion (administration) route for the composition of the present invention may be either oral or parenteral, but oral is preferred. In addition, examples of parenteral ingestion (administration) include transdermal administration, intravenous injection, rectal administration, and inhalation.

When the composition of the present invention is used as a composition to be orally ingested, it is preferably in the form of a food or drink.

There are no particular restrictions on the form or properties of the food or drink as long as it does not impair the effects of the present invention and can be taken orally. It can be manufactured by a method.

Foods and beverages may be in the form of liquids, pastes, gel-like solids, powders, etc., such as tablets, liquid foods (nutritive foods for tube administration), bread, macaroni, spaghetti, noodles, cake mixes, etc. Flour products such as fried flour and bread crumbs; instant noodles, cup noodles, retort and cooked foods, cooked canned foods, microwave foods, instant soups and stews, instant miso soup and soups, canned soups, freeze-dried foods, and other instant foods, etc. Instant foods; Agricultural processed products such as canned agricultural products, canned fruits, jams and marmalade, pickles, boiled beans, dried agricultural products, cereals (processed grain products); Canned marine products, fish hams and sausages, seafood paste products, marine delicacies Processed seafood products such as boiled fish and fish; Processed livestock products such as canned livestock products, pastes, and meat hams and sausages; Processed milk, milk drinks, yogurts, lactic acid bacteria drinks, cheese, ice creams, powdered milk products, cream, Milk and dairy products such as other dairy products; Fats and oils such as butter, margarine, and vegetable oil; Basic seasonings such as soy sauce, miso, sauces, processed tomato seasonings, mirin, and vinegar; Cooking mixes, curry Complex seasonings and foods such as seasonings, sauces, dressings, noodle soups, spices, and other complex seasonings; Frozen foods such as raw frozen foods, semi-cooked frozen foods, and cooked frozen foods; Caramels, candy , chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice sweets, bean sweets, dessert sweets, jellies, and other sweets; Carbonated drinks, natural fruit juices, fruit juice drinks, and refreshing drinks with fruit juice. Beverages, pulp drinks, fruit drinks with fruit grains, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, alcoholic drinks, and other recreational drinks. , baby food, furikake, other commercially available foods such as ochazuke nori, infant formula, enteral nutritional food, and functional foods (foods for specified health uses, foods with nutritional function functions), and the like.

Moreover, it can also be used as feed as one aspect of food and drink products. Examples of the feed include pet food, livestock feed, and fish feed.
The form of the feed is not particularly limited, and in addition to the MKP peptide, for example, grains such as corn, wheat, barley, rye, and milo; vegetable oil meal such as soybean oil meal, rapeseed oil meal, coconut oil meal, and linseed oil meal; bran; Bran such as barley bran, rice bran, and defatted rice bran; Manufactured lees such as corn gluten meal and corn jam meal; Animal feed such as fish meal, skim milk powder, whey, yellow grease, and tallow; Torula yeast, brewer's yeast, etc. It may contain yeasts; mineral feeds such as tricalcium phosphate and calcium carbonate; oils and fats; simple amino acids; sugars, etc.

When the composition of the present invention is in the form of a food or drink (including feed), it may be provided and sold as a food or drink labeled with uses for improving constituent behavior, preventing decline in brain function, preventing decline in memory, etc. is possible. Furthermore, the MKP peptide according to the present specification can be used for producing these foods and drinks.

Such acts of "display" include all acts to inform the consumer of the above-mentioned use, and if the expression is such that it may remind or infer the above-mentioned use, the purpose of the display, the content of the display, Regardless of the object or medium to be displayed, all of them fall under the act of "display" in the present invention.
Moreover, it is preferable that the "display" be performed using expressions that allow the consumer to directly recognize the above-mentioned use. Specifically, the act of transferring, handing over, exhibiting for transfer or delivery, or importing food and drink products or products with the above-mentioned usage written on their packaging, advertisements related to products, price lists, or transaction documents. Examples include actions such as displaying or distributing information with the above-mentioned uses written thereon, or writing the above-mentioned uses on information containing these items and providing it by electromagnetic (Internet, etc.) method.

On the other hand, the display content is preferably a display approved by the government (for example, a display that has been approved based on various systems established by the government and is performed in a manner based on such approval). Further, it is preferable to attach such display contents to packaging, containers, catalogs, pamphlets, promotional materials at sales sites such as POP, and other documents.

In addition, "labeling" includes health foods, functional foods, enteral nutritional foods, special purpose foods, foods with health claims, foods for specified health uses, foods with nutritional function claims, foods with functional claims, quasi-drugs, etc. Display can also be mentioned. Among these, in particular, labeling approved by the Consumer Affairs Agency, for example, labeling approved under the system related to food for specified health uses, food with nutritional function claims, or food with functional claims, or a system similar to these, etc. can be mentioned. Specifically, labeling as a food for specified health uses, labeling as a food for specified health uses with conditions, labeling that it affects the structure or function of the body, labeling to reduce disease risk, and labeling as a food for specified health uses based on scientific evidence. More specifically, the Cabinet Office Ordinance on Permission for Special Purpose Labeling as stipulated in the Health Promotion Act (Cabinet Office Ordinance No. 57 of August 31, 2009) Typical examples include labeling as food for specified health uses (in particular, labeling for health uses) and similar labeling as stipulated in the Act.
Such displays include, for example, ``Supports the ability to memorize and recall directions and how to use objects'', ``Supports the ability to memorize and recall directions and how to use objects'', and ``Supports the ability to memorize and recall directions and how to use objects'', and ``Supports the ability to memorize and recall directions and how to use objects'', and ``Supports the ability to memorize and recall directions and how to use objects'', and ``Maintain the ability to memorize and recall how to use directions and objects'', ``Maintain the ability to memorize and recall how to use directions and objects'', ``Improve compositional behavior (ability to memorize and recall directions and how to use objects)'', `` Memorize directions, how to use objects, and improve your ability to recall.''"To improve and maintain the ability to memorize and recall directions and how to use objects,""For preventing cognitive decline,""Supporting cognitive function,""Preventing cognitive decline." An example of this is displaying a message such as "For those who wish to do so."

The composition of the invention can also be in the form of a pharmaceutical product.
The route of administration of the drug may be either oral or parenteral, but oral is preferred. In addition, examples of parenteral ingestion (administration) include transdermal administration, intravenous injection, rectal administration, and inhalation.
The pharmaceutical form can be appropriately formulated into a desired dosage form depending on the administration method. For example, in the case of oral administration, it can be formulated into solid preparations such as powders, granules, tablets, and capsules; liquid preparations such as solutions, syrups, suspensions, and emulsions. In the case of parenteral administration, it can be formulated into suppositories, ointments, injections, etc.
When formulating, in addition to the MKP peptide, components such as excipients, pH adjusters, coloring agents, and flavoring agents that are commonly used in formulating can be used. In addition, it is also possible to use other medicinal ingredients such as other medicinal ingredients and ingredients that are known or will be discovered in the future and have a constituent behavior improving effect.
In addition, formulation can be carried out by appropriately known methods depending on the dosage form. During formulation, carriers commonly used in formulation may be added as appropriate. Such carriers include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, and the like.

Examples of excipients include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, Cellulose derivatives such as hydroxypropyl methylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light silicic anhydride, synthetic aluminum silicate, magnesium aluminate metasilicate; phosphate derivatives such as calcium phosphate; carbonic acid Examples include carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate.

Examples of the binder include gelatin, polyvinylpyrrolidone, macrogol, and the like, in addition to the above-mentioned excipients.

Examples of the disintegrant include, in addition to the above excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone.

Examples of lubricants include talc; stearic acid; stearic acid metal salts such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and gay wax; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid. ; carboxylic acid sodium salts such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as silicic anhydride and silicic acid hydrate; starch derivatives, etc. It will be done.

Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

Examples of flavoring agents include sweeteners, acidulants, fragrances, and the like.
In addition, examples of carriers used in the case of liquid preparations for oral administration include solvents such as water.

The timing of ingesting the pharmaceutical of the present invention is not particularly limited, such as before meals, after meals, between meals, and before going to bed.

The present invention will be explained in more detail below using Examples, but the present invention is not limited to these Examples.

A randomized, double-blind, placebo-controlled, parallel-group comparison study was conducted in the following manner.
(1) Subjects A total of 268 adult men and women without dementia who were determined to be eligible to participate through screening were asked to participate as subjects after obtaining their consent. Table 1 shows the background of the subjects. Further, specific selection criteria and exclusion criteria are as follows.
Inclusion Criteria: Age 40 years or older; Hasegawa Simple Intelligence Scale (HDS-R) score 21 or higher Exclusion Criteria: Persons diagnosed with or suspected of having dementia; Persons suffering from schizophrenia, depression, etc. Persons suspected of having a mental disorder (judged using the shortened version of the Depression Scale for the Elderly (GDS-15-J), excluding those with a score of 6 or higher)
・Those who have a serious disease in the brain, liver, kidneys, heart, lungs, digestive system, blood, endocrine metabolic system, etc., or a history of such a serious disease ・Those who are pregnant, breastfeeding, or pregnant within the next six months・Persons with severe drug or food allergies, or a serious history of these ・Persons who have been judged ineligible as a subject by the study director

(2) Test method This study targeted middle-aged and elderly men and women without dementia, and compared ADAS-cog scores when they took 1 g/day of MKP peptide daily for 24 weeks, compared to taking a placebo (1 g/day of dextrin). We verified whether it would improve.
ADAS-cog measures ``word recall,'' ``oral language ability,'' ``aural comprehension of language,'' ``difficulty in voicing,'' ``following verbal commands,'' ``hand and object naming,'' ``compositional acts,'' and ``ideomotor It consists of 11 items: ``, ``orientation,'' ``word recognition,'' and ``ability to recall test teaching.'' The test assigns a total of 70 points to these items, and the lower the score, the more normal the cognitive function is judged to be. For statistical analysis, all the data obtained were analyzed for subjects aged 65 years or older. Analysis of covariance was performed using the pre-ingestion value as a covariate.

(3) Results At the 24th week of intake in subjects aged 65 years or older, the composition behavior of the MKP peptide group was significantly improved compared to the placebo group (Table 2).
Furthermore, no adverse events resulting from ingestion of the test food were observed, confirming the safety of the MKP peptide.

Claims (4)

Contains a peptide consisting of Met-Lys-Pro or a peptide comprising the above amino acid sequence ,
A composition for improving constituent behavior that is ingested or administered to a subject without dementia . 2. A composition according to claim 1 , which is used to improve the ability to remember and recall directions and/or how to use an article. The composition according to claim 1 or 2 , which is a food or drink. The composition according to claim 1 or 2 , which is a pharmaceutical.