Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/04—Linear peptides containing only normal peptide links
  • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
  • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
  • A23L33/17—Amino acids, peptides or proteins
  • A23L33/18—Peptides; Protein hydrolysates
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
  • C07K14/70571—Receptors; Cell surface antigens; Cell surface determinants for neuromediators, e.g. serotonin receptor, dopamine receptor
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P21/00—Preparation of peptides or proteins
  • C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Y—ENZYMES
  • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• the present invention relates to peptides and compositions containing the peptides as active ingredients.
• Patent Document 1 discloses peptides in which hydrophobic amino acids are adjacent to tyrosine, phenylalanine, tryptophan, or histidine as peptides having anxiolytic effects.
• Patent Document 2 describes (i) amino acid sequence LSSTQAQQSY, (ii) amino acid sequence LSSTQAQQSW, or (iii) amino acid sequence as peptides for improving or treating low motivation, depression or depressive mood disorders or symptoms based on them. Peptides consisting of any of the sequences LSSTQAQQSF are disclosed.
• the main purpose of the present technology is to provide a peptide having various actions, which can be easily ingested on a daily basis and has few side effects, and a composition containing the peptide as an active ingredient.
• a peptide consisting of the following amino acid sequence (a) or (b) is provided.
• the present technology also provides a composition containing the peptide as an active ingredient.
• the composition according to the present technology may be food or drink.
• a composition according to the present technology may be a pharmaceutical.
• the composition according to the present technology may be used to treat, prevent and/or improve depression, bipolar disorder, schizophrenia, anxiety, or sleep disorders.
• the peptide NLPPLT has excellent anti-anxiety effects, sleep-improving effects, anti-depressive effects, and improving effects on learning disorders and/or memory disorders
• various An active peptide and a composition containing the peptide as an active ingredient can be provided. Note that the effects described here are not necessarily limited, and may be any of the effects described in this specification.
• the peptide according to the present technology has an amino acid sequence (SEQ ID NO: 1) represented by Asn-Leu-Pro-Pro-Leu-Thr.
• Asn (N) indicates an L-asparagine residue
• Leu (L) an L-leucine residue
• Pro (P) an L-proline residue
• Thr (T) an L-threonine residue.
• the peptide according to the present technology has an amino acid string in which one or several amino acids are substituted, deleted, or added to the amino acid sequence shown in SEQ ID NO:1. In this case, the number of amino acids to be substituted, deleted or added is preferably 2 or less.
• the peptide according to the present technology has one or more actions selected from anxiolytic action, sleep improvement action, antidepressant action, and action for improving learning impairment and/or memory impairment.
• a single amino acid may be substituted, deleted, or added.
• the peptide according to the present technology may be a salt of peptide NLPPLT. Examples of the salts include alkali metals such as potassium and sodium; alkaline earth metals such as calcium and magnesium;
• Examples of methods for producing peptide NLPPLT include, but are not limited to, the following methods. Specifically, for example, a protein or peptide containing an amino acid sequence represented by Asn-Leu-Pro-Pro-Leu-Thr (SEQ ID NO: 1) is decomposed by hydrolysis or the like, and separated from the resulting decomposed product.
• Method of obtaining by purification Method of synthesizing peptide NLPPLT by a peptide chemical synthesis method and then separating and purifying peptide NLPPLT from the resulting synthetic product; Plants and animals that produce peptide NLPPLT and peptides containing the same and a method of extracting from microorganisms and separating and purifying from the obtained extract.
• Peptides according to the present technology can be produced, for example, by appropriately hydrolyzing proteins such as whey proteins with acids, alkalis, enzymes, or the like.
• a method for obtaining the peptide NLPPLT by hydrolyzing the raw material protein with a hydrolase is exemplified below.
• the protein is dissolved, dispersed or suspended in water before hydrolyzing the raw material protein with an enzyme.
• the raw material protein is not particularly limited as long as it is a protein containing the peptide according to the present technology in its structure and can produce the peptide according to the present technology when appropriately digested with a hydrolase.
• the protein include those derived from animals, plants, and microorganisms, and in the present technology, whey protein, which is available in large quantities, is preferred.
• the whey protein may be a commercially available product, or a whey protein concentrate, a whey protein isolate, or a whey protein isolate separated and purified from whey protein by a conventional method (e.g., ultrafiltration, ion exchange method, etc.).
• whey protein and whey eg, whey powder, desalted whey powder, etc.
• the treatment method differs depending on the properties of the raw material protein. If the raw material protein is soluble, the raw material protein may be dispersed in water or warm water and dissolved. The protein may be homogenized by mixing and stirring.
• an alkaline agent or an acid agent may be added to the solution containing the protein to adjust the pH. This pH is preferably adjusted to or near the optimum pH for the hydrolase to be used.
• the alkali agent or acid agent is not particularly limited, and those acceptable for pharmaceuticals or food and drink may be used.
• alkaline agents include hydroxides such as sodium hydroxide and calcium hydroxide; carbonates such as potassium carbonate; and these may be alkali metal salts or alkaline earth metal salts.
• acid agents include inorganic acids such as hydrochloric acid and phosphoric acid; and organic acids such as citric acid, acetic acid and formic acid. In the present technology, one or more of these can be used as appropriate.
• it is preferable to sterilize the solution containing the protein by heating at 70 to 90° C. for about 15 seconds to 10 minutes from the viewpoint of preventing spoilage due to contamination with various bacteria.
• a predetermined amount of hydrolase is added to the solution containing the protein and reacted at a temperature of about 10 to 85° C. for 0.1 to 48 hours to obtain a hydrolyzate.
• the solution is maintained at an appropriate temperature, for example, 30 to 60° C., preferably 45 to 55° C., depending on the type of enzyme, to initiate protein hydrolysis. do.
• the reaction may be continued until the desired decomposition rate is reached while monitoring the decomposition rate of the enzymatic reaction.
• the hydrolase reaction is terminated, for example, by deactivating the enzyme in the hydrolyzate, and can be performed by a conventional heat deactivation treatment.
• the heating temperature and holding time of the heat deactivation treatment can be appropriately set in consideration of the thermal stability of the enzyme used, and the conditions that can be sufficiently deactivated can be set as appropriate.
• the temperature range is 80 to 130 ° C. for 30 minutes. Hold times of ⁇ 2 seconds can be performed.
• the hydrolase is not particularly limited, it is preferably an enzyme that can hydrolyze the raw material protein to generate the peptide according to the present technology. Moreover, you may use a hydrolase individually or in combination of 2 or more types. When using two or more enzymes, each enzymatic reaction may be carried out simultaneously or separately. Specifically, it is preferable to use endopeptidase, trypsin, and papain in combination as the enzymes that can be generated, and it is more preferable to use a mixture of these three types of enzymes.
• endopeptidase examples include those derived from microorganisms and animals, and specific examples include protease derived from Bacillus subtilus and protease derived from animal pancreas.
• Commercially available products can be used as the protease.
• commercially available proteases include Bacillus-derived proteases such as Protease N Amano (manufactured by Amano Enzymes), Bioplase SP-20 (manufactured by Nagase & Co.), and Neutrase (manufactured by Novozymes Japan); Animal pancreas-derived proteases such as OS (manufactured by Novozymes Japan) and the like can be mentioned.
• Examples of the trypsin include PTN6.0S (manufactured by Novozymes Japan) and the like, and examples of the papain include papain W-40 (manufactured by Amano Enzymes) and purified papain (manufactured by Mitsubishi Chemical). etc.
• the decomposition rate of the raw material protein is calculated by measuring the total nitrogen content of the sample by the Kjeldahl method, measuring the formol nitrogen content of the sample by the formol titration method, and calculating the decomposition rate from these measured values by the following formula (1 ).
• peptides according to the present technology it is preferable to isolate or purify the peptide according to the present technology from the hydrolyzate.
• Purification of peptides according to the present technology can be performed by the same techniques as those usually used for oligopeptide purification, such as ion-exchange chromatography, adsorption chromatography, reversed-phase chromatography, partition chromatography, and gel filtration chromatography.
• Various chromatographic methods such as solvent precipitation, salting out, and partitioning between two liquid phases can be appropriately combined.
• the fraction containing the target substance can be determined using the aforementioned various actions as indicators. Also, the active components of those fractions can be identified by mass spectrometry.
• Peptides according to the present technology can also be produced by chemical synthesis.
• Chemical synthesis of peptides according to the present technology can be performed by a liquid-phase method or a solid-phase method that is commonly used for synthesis of oligopeptides.
• the synthesized peptide is deprotected, if necessary, and unreacted reagents, by-products, etc. are removed to isolate the peptide according to the present technology.
• Synthesis of such peptides can be performed, for example, using a commercially available peptide synthesizer or the like.
• Obtaining the peptide of interest should be confirmed by using as an index any one or more actions selected from anxiolytic action, sleep improving action, antidepressant action, and improving action on learning and/or memory impairment. can be done.
• Peptide NLPPLT has an anti-anxiety effect as shown in Test Example 1 described later, and also has an effect via activation of serotonin 5-HT 1A receptor as shown in Test Example 2 described later, thus improving sleep. also has an effect. In addition, as shown in Test Example 3 below, it also has an antidepressant effect.
• the serotonin 5-HT 1A receptor is the most widely expressed of the various serotonin receptors and is involved in behaviors such as sleep, feeding, thermoregulation and anxiety. Therefore, it is conventionally known that activation of this serotonin 5-HT 1A receptor produces one or more actions selected from anxiolytic action, sleep improving action and antidepressant action. Therefore, the peptide according to the present technology can be used for anti-anxiety, sleep improvement, or anti-depression.
• the peptides according to the present technology can prevent, ameliorate, or treat various diseases and symptoms associated with serotonin 5-HT 1A receptors. Therefore, the peptide according to the present technology is used for methods of preventing, ameliorating and/or treating diseases and symptoms associated with serotonin 5-HT 1A receptors by ingesting or administering to animals including humans. can be used as an active ingredient.
• the term "improvement” refers to amelioration of a disease, symptom or condition; prevention or delay of worsening of a disease, symptom or condition; reversal, prevention or delay of progression of a disease or symptom; means preventing or delaying the onset of a disease or condition in an applicable subject, reducing the risk of a disease or symptom in an applicable subject, or maintaining health.
• Various diseases and symptoms associated with the serotonin 5-HT 1A receptor include, for example, depression, bipolar disorder, schizophrenia, anxiety neurosis, and sleep disorders.
• the peptide NLPPLT has an action mediated by AMPA receptor activation.
• AMPA receptors are a kind of glutamate receptors, are widely distributed in the central nervous system, and are known to be greatly involved in memory and learning. Therefore, it is conventionally known that activation of the AMPA receptor not only produces an antidepressant effect but also an ameliorating effect on learning impairment and/or memory impairment. Therefore, the peptide according to the present technology can be used not only for anti-anxiety, sleep improvement, or anti-depression, but also for improving learning disorders and/or memory disorders.
• non-therapeutic purpose is a concept that does not include medical practice, that is, treatment of the human body by treatment, and includes, for example, beauty practices and health promotion.
• compositions also provides a composition containing the aforementioned peptide according to the present technology as an active ingredient.
• the composition according to the present technology can be used as pharmaceuticals such as formulations, food and drink, feed, and the like.
• the peptide according to the present technology can be used for producing these various compositions.
• composition according to the present technology may be used in combination with any component, if necessary, in addition to the peptide according to the present technology.
• ingredients that are allowed in pharmaceuticals, food and drink, feeds, and the like can be used as appropriate.
• composition according to the present technology when used in pharmaceuticals, it can be prepared by adding the peptide according to the present technology to known pharmaceuticals, or new pharmaceuticals can be produced by mixing the peptides into raw materials for pharmaceuticals. can also In addition, when the peptide according to the present technology is used for pharmaceuticals, it may be used as it is, after being concentrated, or after being processed into a solid, liquid, granule, or powder form.
• the drug can be appropriately formulated into a desired dosage form according to an administration method such as oral administration or parenteral administration.
• the dosage form is not particularly limited, but in the case of oral administration, for example, solid preparations such as powders, granules, tablets, troches, and capsules; liquid preparations such as solutions, syrups, suspensions, emulsions, etc. can be
• parenteral administration for example, it can be formulated into suppositories, sprays, inhalants, ointments, patches, injections, and the like.
• formulation can be suitably implemented by a well-known method according to a dosage form.
• a pharmaceutical carrier When formulating, it may be formulated by adding a pharmaceutical carrier as appropriate.
• ingredients such as excipients, pH adjusters, colorants, and corrigents that are usually used for formulation can be used.
• ingredients having known or future therapeutic, preventive and/or ameliorative effects on diseases and symptoms can also be used in combination.
• organic or inorganic carriers can be used as pharmaceutical carriers depending on the dosage form.
• carriers for solid preparations include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents and the like.
• Excipients include, for example, sugar derivatives such as lactose, sucrose, glucose, mannitol, sorbitol; starch derivatives such as corn starch, potato starch, ⁇ -starch, dextrin, carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, Hydroxypropylmethylcellulose, carboxymethylcellulose, cellulose derivatives such as carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light silicic anhydride, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate;
• binders examples include gelatin; polyvinylpyrrolidone; macrogol, etc., in addition to the above excipients.
• disintegrants include, in addition to the above excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and crosslinked polyvinylpyrrolidone.
• Lubricants include, for example, talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and gairou; ; 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 acid anhydride and silicic acid hydrate; be done.
• stabilizers 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.
• Flavoring agents include, for example, sweeteners, acidulants, flavoring agents, and the like.
• the carrier used includes a solvent such as water, a flavoring agent, and the like.
• the peptide according to the present technology can be added to a known food or drink, or a new food or drink can be prepared by mixing the peptide with the ingredients of the food or drink. You can also manufacture products. Moreover, when the peptide according to the present technology is used in food and drink, it may be used as it is, after being concentrated, or after being processed into a solid, liquid, granule, or powder form.
• Food and drink include tablets, liquid foods, feeds (including pets), regardless of whether they are liquid, paste, solid, or powder, as well as flour products, instant foods, processed agricultural products, and marine products.
• Examples include processed products, processed livestock products, milk/dairy products, oils and fats, basic seasonings, compound seasonings/foods, frozen foods, confectionery, beverages, and other commercially available foods.
• dairy products include fermented milk, milk drinks, lactic acid bacteria drinks, sweetened condensed milk, skim milk powder, sweetened milk powder, modified milk powder, cream, cheese, butter, and ice creams.
• Wheat flour products include, for example, bread, macaroni, spaghetti, noodles, cake mixes, fried chicken flour, and bread crumbs.
• instant foods include instant noodles, cup noodles, retort/cooked foods, cooked canned foods, microwave oven foods, instant soups/stews, instant miso soups/soup, canned soups, freeze-dried foods, and other instant foods. be done.
• processed agricultural products include canned agricultural products, canned fruits, jams and marmalades, pickles, boiled beans, dried agricultural products, and cereals (processed grain products).
• processed marine products include canned marine products, fish ham and sausages, fish paste products, marine delicacies, and tsukudani.
• processed livestock products include canned livestock products, pastes, livestock hams, sausages, and the like.
• Fats and oils include, for example, butter, margarines, and vegetable oils.
• Basic seasonings include, for example, soy sauce, miso, sauces, processed tomato seasonings, mirin, and vinegars. Examples include dressings, noodle soups, spices, and other compound seasonings.
• Frozen food includes, for example, material frozen food, half-cooked frozen food, cooked frozen food, and the like.
• Confectionery includes, for example, caramels, candies, chewing gums, chocolates, cookies, biscuits, cakes, pies, snacks, crackers, Japanese confections, rice confections, bean confections, dessert confections, and other confectionery.
• beverages include carbonated drinks, natural fruit juices, fruit juice drinks, soft drinks containing fruit juice, pulp drinks, fruit drinks containing fruit, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, and concentrated drinks.
• sports drinks, nutritional drinks, alcoholic beverages, and other beverages of choice include baby food, furikake, ochazuke seaweed, and the like.
• the food and drink defined by the present technology are provided and provided as food and drink labeled with insurance uses (for example, for anti-anxiety, sleep improvement, anti-depression, learning disability and/or memory impairment improvement, etc.). can be sold.
• insurance uses for example, for anti-anxiety, sleep improvement, anti-depression, learning disability and/or memory impairment improvement, etc.
• the act of "display” includes all acts for informing the consumer of the above-mentioned use, and if it is an expression that can remind or analogize the above-mentioned use, the purpose of the display, the display Regardless of the content, the object or medium to be displayed, all fall under the "display" act of this technology.
• the "indication" be expressed in such a way that the consumer can directly recognize the use.
• the act of transferring, handing over, displaying for the purpose of transfer or delivery, importing products related to food and beverages or product packaging that describes the above-mentioned use, advertisements related to products, price lists or transaction documents Examples include the act of displaying or distributing information with the use described therein, or providing information containing such information with the use described therein by electromagnetic means (for example, the Internet, etc.).
• the content of the display is a display approved by the government (for example, a display that is approved based on various systems established by the government and performed in a manner based on such approval).
• a display that is approved based on various systems established by the government and performed in a manner based on such approval.
• labeling includes health food, functional food, food for the sick, enteral nutritional food, food for special dietary uses, food with health claims, food for specified health use, food with function claims, food with nutrient function claims, and medical products. Display as an off-the-shelf item is also possible.
• labeling approved by the Consumer Affairs Agency for example, labeling approved by the Foods for Specified Health Uses system, the Foods with Function Claims system, and similar systems can be mentioned. More specifically, labeling as food for specified health use, labeling as food for specified health use with certain conditions, labeling as food with function claims, labeling to the effect that it affects the structure and function of the body, labeling to reduce disease risk, etc. can be mentioned.
• a typical example is labeling as a food for specified health use (especially, labeling of health use) specified in the Ordinance for Enforcement of the Health Promotion Law (Ministry of Health, Labor and Welfare Ordinance No. 86 of April 30, 2003). ), labeling as food with function claims stipulated in the Food Labeling Law (Law No. 70 of 2013) and similar labeling.
• the words used to display the above-mentioned are only words such as "for anti-anxiety”, “for improving sleep”, “for anti-depression”, “for improving learning disorders and/or memory disorders”. It is not limited, and even if it is a word other than that, it is included in the scope of the present technology as long as it is a word that expresses an anti-anxiety effect, a sleep improvement effect, an antidepressant effect, a learning disability and / or a memory impairment improvement effect, etc. It goes without saying that Examples of such wording include indications based on various uses that make consumers aware of anti-anxiety effects, sleep-improving effects, anti-depressive effects, learning and/or memory-impairing effects, and the like. is.
• composition according to the present technology when used in feed, it can be prepared by adding the peptide according to the present technology to a known feed, or a new feed can be produced by mixing the peptide into feed ingredients. can also In addition, when the peptide according to the present technology is used in feed, it may be used as it is, after being concentrated, or after being processed into a solid, liquid, granular or powdered form.
• Raw materials for the feed include, for example, cereals such as corn, wheat, barley, and rye; brans such as bran, wheat bran, rice bran, and defatted rice bran; manufacturing lees such as corn gluten meal and corn jam meal; Animal feeds such as whey, fish meal, and bone meal; yeasts such as brewer's yeast; mineral feeds such as calcium phosphate and calcium carbonate; oils and fats; amino acids; Examples of the form of the feed include pet feed (for example, pet food), livestock feed, fish farm feed, and the like.
• the content or amount of the peptide according to the present technology in the composition according to the present technology is not particularly limited. is preferred.
• the administration or ingestion interval and the administration or ingestion period of the composition according to the present technology are not particularly limited.
• the peptide according to the present technology is believed to be capable of preventing, improving, or treating various diseases and symptoms associated with the serotonin 5-HT 1A receptor.
• the compositions can be used, for example, to treat, prevent and/or ameliorate depression, bipolar disorder, schizophrenia, anxiety, or sleep disorders.
• the present technology can also employ the following configuration.
• [1] Use of the following peptide (a) or (b) in an anti-anxiety composition.
• anti-anxiety composition Use of the peptide of (a) or (b) above for the production of
• [3] Use of the above peptide (a) or (b) for treatment, prevention and/or improvement of depression, bipolar disorder, schizophrenia, anxiety neurosis, or sleep disorder.
• a method for treating, preventing and/or improving depression, bipolar disorder, schizophrenia, anxiety neurosis, or sleep disorders comprising the peptide of (a) or (b) above as an active ingredient.
• Use of the above peptide (a) or (b) in a composition for improving learning disorders and/or memory disorders comprising the peptide of (a) or (b) above as an active ingredient.
• Production Example 1 Production of Peptide NLPPLT by Enzymatic Degradation of Whey Protein 1 kg of a commercially available whey protein concentrate was dissolved in 9 kg of purified water and sterilized at 75° C. for 15 seconds in a plate-type sterilizer. After that, sodium hydroxide was added to adjust the pH of the solution to 9.0, and then a protease was added for hydrolysis to obtain a solution containing NLPPLT. Thereafter, the enzyme was deactivated, concentrated by a conventional method, and dried to obtain about 1 kg of powdered whey protein hydrolyzate (hereinafter referred to as "WPH”) containing the peptide NLPPLT.
• WPH powdered whey protein hydrolyzate
• Test Example 1 Elevated Plus Maze Test An elevated plus maze consisting of a cross where two flat plates (arms) of 25 cm ⁇ 5 cm intersect was set at a height of 50 cm from the floor.
• One of the arms is a closed arm with a 15 cm high transparent enclosure, allowing mice to walk safely without fear of falling.
• the other arm is an open arm without such an enclosure, and since it has an open perimeter, mice that are afraid of heights usually do not like the open arm.
• anxiolytic treatment of mice tends to increase the time spent in open arms. Therefore, in the confirmation test of anxiolytic effects, the longer the mouse stays in the open arm or the more times the mouse enters the open arm, the higher the anxiolytic activity.
• Test Results Table 1 below shows the test results.
• the administration of WPH increased the time spent on the open arms and the number of times the animals entered the open arms, confirming that WPH has anxiolytic activity.
• peptide NLPPLT Since the administration of peptide NLPPLT increased the residence time on the open arm compared to the control, it was confirmed that peptide NLPPLT has anxiolytic activity.
• the dwell time on the open arms was reduced compared to administration of the peptide alone, at a rate comparable to the control. Became. Therefore, it was suggested that the action of the peptide NLPPLT was based on the action mediated by the activation of the serotonin 5-HT 1A receptor.
• the peptide NLPPLT since the peptide NLPPLT has an action mediated by activation of the serotonin 5-HT 1A receptor, it is not only an anxiolytic action but also one of the actions based on the activation of the serotonin 5-HT 1A receptor. It was also suggested that it is thought to have a sleep-improving effect.
• Test Example 2 Open field test An open field test was performed as a behavioral test used for screening anxiolytics. It utilizes the fact that administration of anxiolytics reduces the proportion of time that mice stay in the center of the field (hereinafter referred to as "staying time"). Behavior was observed for 5 minutes 30 minutes after administration.
• a circular field consisting of a plurality of concentric circles with the same center and different radii was used. The circle closest to the center of the field is taken as the center of the field, and the time for the mouse placed in the center of the field to stay in the center of the field is measured.
• the ratio (%) of staying time was calculated as an index of anxiolytic activity.
• Test Results Table 4 below shows the test results.
• peptide NLPPLT Since the administration of peptide NLPPLT increased the residence time compared to the control, it was confirmed that peptide NLPPLT has anxiolytic activity.
• Tail Suspension Test A tail suspension test used for screening antidepressants was performed. It is thought that when a mouse is suspended by its tail and placed under conditions in which it cannot escape, it will initially try to escape and move around, but over time it will despair and become immobile. On the other hand, mice treated with antidepressants show increased escape time and decreased immobility time. That is, when the immobility time is reduced, it is evaluated as having an antidepressant effect. Behavior was observed for 6 minutes 30 minutes after administration.
• Test Results Table 5 below shows the test results.
• WPH has antidepressant properties, since the administration of WPH reduced the duration of immobile behavior compared to controls.
• peptide NLPPLT Since the administration of peptide NLPPLT reduced the time of immobility manifestation compared to controls, it was confirmed that peptide NLPPLT has antidepressant properties.
• the action of peptide NLPPLT is based on the action mediated by AMPA receptor activation.
• AMPA receptors are known to be involved in antidepressant effects, which is consistent with the present peptide exhibiting antidepressant activity.
• this peptide exhibits memory impairment improving action and learning impairment improving action.
• the peptide according to the present technology can be used for anti-anxiety, sleep improvement, anti-depression, and improvement of learning and/or memory impairment.
• a composition containing the peptide as an active ingredient is useful for reducing anxiety symptoms, reducing mental stress, treating, preventing and/or improving depression, bipolar disorder, schizophrenia, anxiety neurosis, or sleep disorders. It is possible to use it in a wide range of fields such as pharmaceuticals or food and drink for the purpose.

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