Composition for promoting energy metabolism

Abstract

The purpose of the present invention is to provide a composition for promoting energy metabolism. The present invention relates to a composition for promoting energy metabolism, the composition containing a peptide comprising the amino acid sequence of Ala-Glu-Tyr-Tyr (SEQ ID NO: 1) or a salt thereof.

Description

Composition for promoting energy metabolism

[0001] The present invention relates to a composition for promoting energy metabolism, etc.

[0002] Energy metabolism, including energy production, is one of the functions necessary to maintain life. Most energy production occurs within mitochondria, a type of intracellular organelle. Sugars, lipids, amino acids, and other nutrients ingested from food are metabolized through glycolysis, the pentose phosphate pathway, and the citric acid cycle and converted into NADH (nicotinamide adenine dinucleotide). Electrons from NADH are then transferred to mitochondrial respiratory chain proteins (electron transport chain), producing energy in the form of ATP (adenosine triphosphate).

[0003] Generally, energy metabolic function declines with age, leading to increased fatigue and decreased vitality. Energy metabolism also declines due to continued stress and irregular lifestyles. Because energy is constantly required in humans to maintain activity, there is a demand for substances that promote energy metabolism, are highly safe, and can be taken continuously over a long period of time.

[0004] Patent Document 1 discloses a composition for stimulating TRPV1, which contains a cyclic dipeptide or a salt thereof having amino acids as constituent units as an active ingredient. Patent Document 1 describes that use of the composition can achieve effects such as promoting energy consumption and promoting body heat production via stimulation of TRPV1 (Transient Receptor Potential Cation Channel, subfamily V, member 1).

[0005] International Publication No. 2017 / 014120

[0006] An object of the present invention is to provide a composition for promoting energy metabolism.

[0007] As a result of intensive research conducted in light of the above problems, the present inventors have found that a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof, has an energy metabolism-promoting effect.

[0008] That is, the present invention relates to the following compositions for promoting energy metabolism, etc., but is not limited thereto. [1] A composition for promoting energy metabolism, comprising a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof. [2] The composition according to [1] above, which is used for at least one selected from the group consisting of increasing energy metabolism, increasing metabolic rate, increasing resting metabolic rate, increasing basal metabolic rate, increasing heat production, increasing vitality, preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy. [3] The composition according to [1] or [2] above, further comprising Cyclo(Pro-Thr) or a salt thereof. [4] The composition according to any of [1] to [3] above, which is an oral composition. [5] The composition according to any of [1] to [4] above, which is a food or drink, or an oral pharmaceutical. [6] The composition according to any one of [1] to [5] above, which is labeled with at least one function selected from the group consisting of "increased metabolism," "increased metabolic rate," "increased resting metabolic rate," "increased basal metabolic rate," "increased energy level," "increased energy utilization," "increased heat production," "increased thermal response," "increased vitality," "prevention of fatigue," "reduction of fatigue," "prevention of fatigue," and "reduction of fatigue." [7] Use of a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1) or a salt thereof for promoting energy metabolism. [8] A method for promoting energy metabolism, which comprises administering a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1) or a salt thereof.

[0009] According to the present invention, a composition for promoting energy metabolism can be provided.

[0010] Figure 1A is a graph showing the basal respiration rate (oxygen consumption rate (OCR)) of C2C12 cells treated with a test substance for 4 hours, and Figure 1B is a graph showing the ATP-related respiration rate (OCR) of C2C12 cells treated with a test substance for 4 hours.

[0011] The composition for promoting energy metabolism of the present invention comprises a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof. The composition for promoting energy metabolism of the present invention will hereinafter also be referred to as the composition of the present invention. The composition of the present invention typically comprises, as an active ingredient, a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof. In this specification, the peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1) will sometimes be referred to as AEYY.

[0012] In the present invention, energy metabolism includes energy production and energy utilization, and is preferably energy production. Energy production refers to the production of energy necessary to maintain vital activities. Energy utilization refers to the utilization of energy in vital activities. In the present invention, the promotion of energy metabolism may be the promotion of energy production and / or the promotion of energy utilization, and is preferably the promotion of energy production. Promotion of energy metabolism includes, for example, increasing the amount of energy production and increasing the amount of energy utilization. In one aspect, the composition of the present invention can be preferably used as a composition for promoting energy production.

[0013] Most energy production occurs within mitochondria, one of the intracellular organelles. In one embodiment, the energy metabolism is preferably mitochondrial energy metabolism, and more preferably mitochondrial energy production. For example, mitochondrial energy metabolism can be evaluated by continuously measuring the oxygen consumption rate (OCR) used by mitochondria during ATP synthesis in the cells being measured using an extracellular flux analyzer while adding an ATP synthase inhibitor (e.g., oligomycin, rotenone, etc.) and an uncoupler (e.g., carbonyl cyanide-4-trifluoromethoxyphenylhydrazone (FCCP)). For example, the mitochondrial energy metabolism rate can be evaluated by analyzing the basal respiration rate and ATP-related respiration rate, which are considered to be major indicators in the evaluation of mitochondrial function. For example, if the addition of a test substance increases basal respiration and / or ATP-related respiration in mitochondria compared to when the test substance is not added, it can be said that the test substance has the effect of promoting mitochondrial energy metabolism.

[0014] The peptide (AEYY) consisting of the amino acid sequence of SEQ ID NO: 1 is a linear tetrapeptide consisting of the amino acid sequence alanine-glutamic acid-tyrosine-tyrosine. In this specification, the amino acid sequence of a linear peptide is represented in accordance with the convention for notating peptides, with the N-terminus (amino terminus) at the left and the C-terminus (carboxyl terminus) at the right. When an amino acid can have optical isomers, the L-amino acid is shown unless otherwise specified.

[0015] In the present invention, the salt of AEYY is not particularly limited as long as it is a pharmacologically acceptable salt or a salt acceptable for food and drink, and may be either an acidic salt or a basic salt. Examples of acidic salts include inorganic acid salts such as hydrochloride, sulfate, nitrate, and phosphate; and organic acid salts such as acetate, citrate, maleate, malate, oxalate, lactate, succinate, fumarate, and propionate. Examples of basic salts include alkali metal salts such as sodium salt and potassium salt; and alkaline earth metal salts such as calcium salt and magnesium salt.

[0016] As shown in the examples below, AEYY or a salt thereof has the effect of promoting energy metabolism in mitochondria. AEYY or a salt thereof has the effect of promoting energy metabolism and can be used as an active ingredient for promoting energy metabolism. In one aspect, the composition of the present invention can be used to promote energy metabolism in mitochondria.

[0017] In the present invention, the origin and production method of AEYY or a salt thereof are not particularly limited. AEYY or a salt thereof can be produced according to known methods. AEYY can be produced, for example, by known peptide synthesis methods. When AEYY is obtained by chemical synthesis, it can be synthesized by either solid-phase or liquid-phase methods. The synthesized peptide can be purified by conventional purification methods such as reverse-phase high-performance liquid chromatography, chromatography using an ion exchange resin or a highly porous polymer resin, or affinity chromatography. AEYY can also be biosynthesized using a microorganism. When biosynthesized using a microorganism, an expression vector containing a gene encoding AEYY is prepared, and the expression vector is introduced into a host microorganism to obtain a transformant. The transformant is cultured, and AEYY can be purified from the culture. The host microorganism is not particularly limited, and for example, Escherichia coli, yeast, etc. can be used. Separation and purification from the culture can be performed by conventional purification methods. AEYY can also be produced by enzymatic, chemical, or thermal hydrolysis of a protein containing the amino acid sequence of the peptide. In one embodiment, AEYY may be incorporated into the composition using a protein hydrolysate containing AEYY, or may be incorporated into the composition using a concentrate, dry powder, or a more purified form of the protein hydrolysate. Salts of peptides such as AEYY can be easily prepared by those skilled in the art using any method known in the art.

[0018] Preferably, the composition of the present invention further comprises Cyclo(Pro-Thr) or a salt thereof. Cyclo(Pro-Thr) (cycloprolylthreonine) is a cyclic dipeptide having a structure in which proline (Pro) and threonine (Thr) are condensed together. As used herein, the term "cyclic dipeptide" refers to a compound characterized by having amino acids as its constituent units and having a diketopiperazine structure formed by dehydration condensation between the amino group of the N-terminal amino acid and the carboxyl group of the C-terminal amino acid. In this specification, the order in which the amino acids are listed does not matter as long as the amino acid composition of the cyclic dipeptide is the same; Cyclo(Pro-Thr) and Cyclo(Thr-Pro) (cyclothreonylproline) refer to the same cyclic dipeptide.

[0019] The salt of Cyclo(Pro-Thr) is not particularly limited as long as it is a pharmacologically acceptable salt or a salt acceptable for use in foods and beverages, and may be either an acidic salt or a basic salt. Examples of the acidic salt and basic salt include the acidic salts and basic salts exemplified above as the salts of AEYY.

[0020] Cyclo(Pro-Thr) or a salt thereof has the effect of promoting energy metabolism in mitochondria. Cyclo(Pro-Thr) or a salt thereof has the effect of promoting energy metabolism and can be used as an active ingredient for promoting energy metabolism. In one aspect, Cyclo(Pro-Thr) or a salt thereof can be preferably used for promoting energy metabolism in mitochondria.

[0021] In the present invention, the origin and production method of Cyclo(Pro-Thr) or a salt thereof are not particularly limited. Cyclo(Pro-Thr) can be produced according to known methods. Cyclo(Pro-Thr) may be derived from a natural product, chemically synthesized, produced by an enzymatic method or a microbial fermentation method, or synthesized by dehydrating and cyclizing a linear dipeptide. For example, a heat-treated peptide containing Cyclo(Pro-Thr) can be obtained by heating a protein hydrolysate such as collagen hydrolysate. Cyclo(Pro-Thr) may be incorporated into a composition using a protein hydrolysate containing Cyclo(Pro-Thr) or a heat-treated product thereof, or a concentrate, dried powder, or a highly purified product of the protein hydrolysate or the heat-treated product thereof may be incorporated into a composition. Commercially available Cyclo(Pro-Thr) or a salt thereof may also be used.

[0022] The composition of the present invention preferably contains AEYY or a salt thereof, and Cyclo(Pro-Thr) or a salt thereof as active ingredients. The composition of the present invention may contain AEYY or a salt thereof as the active ingredient for promoting energy metabolism, but preferably contains AEYY or a salt thereof, and Cyclo(Pro-Thr) or a salt thereof as the active ingredients for promoting energy metabolism. When AEYY or a salt thereof and Cyclo(Pro-Thr) or a salt thereof are used in combination, a higher energy metabolism promoting effect can be obtained.

[0023] In one aspect, the compositions of the present invention can be preferably used to increase energy metabolism, metabolic rate, resting metabolic rate, basal metabolic rate, heat production, vitality, etc. Metabolic rate (metabolic rate) refers to the rate at which an organism or individual uses energy. It represents the total energy consumption of various physiological processes in an organism, including those that maintain basic bodily functions at rest or during physical activity. Resting metabolic rate refers to energy utilization at rest (after food absorption). It represents the energy required for functions such as respiration, blood flow, and material transport in the human body under normal conditions. Basal metabolic rate is the minimum energy requirement required to maintain life. Thermogenesis refers to the process by which the body produces heat. This includes the production and release of heat energy as a result of metabolic and physiological processes. Vitality refers to a state of strong physical strength, vitality, or energy.

[0024] In one aspect, the composition of the present invention can be used for preventing or ameliorating a condition or disease associated with decreased energy metabolism. Such conditions or diseases include conditions or diseases accompanied by or caused by decreased energy metabolism. Conditions or diseases accompanied by or caused by decreased energy metabolism include, for example, fatigue (particularly physical fatigue), malaise, decreased vitality, weight gain, etc. As used herein, "preventing a condition or disease" includes preventing the onset of a condition or disease, delaying the onset of a condition or disease, reducing the incidence of a condition or disease, reducing the risk of developing a condition or disease, etc. "Ameliorating a condition or disease" includes recovering a subject from a condition or disease, alleviating the symptoms of a condition or disease, improving the symptoms of a condition or disease, delaying or preventing the progression of a condition or disease, etc.

[0025] In one aspect, the composition of the present invention can be preferably used for preventing or improving fatigue, preventing or improving lethargy, etc., and can be more preferably used for preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy.

[0026] In one aspect, the composition of the present invention can be preferably used for at least one selected from the group consisting of increasing energy metabolism, increasing metabolic rate, increasing resting metabolic rate, increasing basal metabolic rate, increasing heat production, increasing vitality, preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy.

[0027] The composition of the present invention can be used for either therapeutic (medical) or non-therapeutic (non-medical) purposes. Non-therapeutic is a concept that does not include medical procedures, i.e., human surgery, treatment, or diagnosis. The composition of the present invention can be provided in the form of an agent, for example, but is not limited to this form. The agent can also be provided as a composition as is, or as a composition containing the agent. In one aspect, the composition for promoting energy metabolism can also be referred to as an energy metabolism promoter.

[0028] The composition of the present invention may be administered orally or parenterally, but is preferably an oral composition. The composition of the present invention can be in the form of, for example, a food or drink, a pharmaceutical product, a quasi-drug, or a feed, with a food or drink or a pharmaceutical product being preferred, a food or drink or an oral pharmaceutical product being more preferred, and a food or drink being even more preferred. The composition of the present invention can also be added to a food or drink, a pharmaceutical product, a quasi-drug, a feed, or the like. The form of the composition of the present invention is not particularly limited, and may be any of a solid (e.g., powder, granules, tablet, etc.), a liquid, a paste, etc. In the composition of the present invention, components other than the above-mentioned peptide or a salt thereof can be appropriately selected depending on the form of the composition, etc.

[0029] In addition to the above-mentioned peptide or a salt thereof, the composition of the present invention may contain various diluents, acidulants, antioxidants, stabilizers, preservatives, flavorings, emulsifiers, colorants, seasonings, pH adjusters, nutritional enhancers, etc. that are permitted as additives to foods, beverages, pharmaceuticals, etc.

[0030] For example, when the composition of the present invention is used as a food or beverage, various foods and beverages can be prepared by blending AEYY or a salt thereof with ingredients that can be used in foods and beverages (e.g., food materials, food additives used as needed, etc.). The foods and beverages are not particularly limited, and examples thereof include general foods and beverages, health foods, functional foods, foods for specified health uses, health supplements, and foods for patients. The health foods, functional foods, foods for specified health uses, health supplements, and foods for patients can be used in various dosage forms such as liquids, fine granules, tablets, granules, powders, capsules, jellies, gels, chewable tablets, dry syrups, and liquid diets.

[0031] When the composition of the present invention is used as a pharmaceutical or quasi-drug, various dosage forms of pharmaceutical or quasi-drug products can be prepared by blending AEYY or a salt thereof with a pharmacologically acceptable carrier and, if necessary, additives. Such carriers, additives, etc. may be any pharmacologically acceptable carriers that can be used in pharmaceuticals or quasi-drugs, and may include, for example, one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, etc. The pharmaceutical or quasi-drug administration (ingestion) form may be oral or parenteral (transdermal, transmucosal, enteral, injection, etc.) administration. When the composition of the present invention is used as a pharmaceutical or quasi-drug, it is preferably an oral pharmaceutical or oral quasi-drug. Dosage forms for oral administration include, for example, liquids, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, jellies, gels, chewable tablets, etc. Dosage forms for parenteral administration include, for example, injections, drip infusions, ointments, lotions, patches, suppositories, nasal preparations, pulmonary preparations (inhalants), etc. The pharmaceutical product may be a pharmaceutical product for non-human animals.

[0032] When the composition of the present invention is used as a feed, AEYY or a salt thereof may be blended into the feed. The feed also includes feed additives. Examples of the feed include livestock feed for cows, pigs, chickens, sheep, horses, etc.; small animal feed for rabbits, rats, mice, etc.; and pet food for dogs, cats, small birds, etc.

[0033] When the composition of the present invention is used for, for example, a food or drink, a pharmaceutical product, a quasi-drug, a feed, etc., the method for producing it is not particularly limited, and it can be produced by a general method using AEYY or a salt thereof.

[0034] In one embodiment, the composition of the present invention is preferably a liquid composition, and more preferably a beverage. The beverage may be, for example, a functional beverage. The form of the beverage is not particularly limited, and may be a packaged beverage. The container for the packaged beverage is not particularly limited, and any container of any form or material may be used. For example, any commonly used container may be used, such as metal containers such as aluminum cans and steel cans; resin containers such as PET bottles; paper containers such as paper cartons; glass containers such as glass bottles; and wooden containers such as barrels. A packaged beverage is obtained by filling and sealing such a container with the beverage.

[0035] The content of AEYY or a salt thereof in the composition of the present invention is not particularly limited and can be set depending on the form, etc. In one embodiment, the content of AEYY or a salt thereof in the composition for promoting energy metabolism, in terms of AEYY, is preferably 0.0001 wt % or more, more preferably 0.0005 wt % or more, even more preferably 0.001 wt % or more, and particularly preferably 0.005 wt % or more, and is preferably 10 wt % or less, more preferably 5 wt % or less, even more preferably 0.5 wt % or less, and particularly preferably 0.1 wt % or less. In one embodiment, the content of AEYY or a salt thereof in the composition for promoting energy metabolism, in terms of AEYY, is preferably 0.0001 to 10 wt %, more preferably 0.0001 to 5 wt %, even more preferably 0.0005 to 0.5 wt %, particularly preferably 0.001 to 0.1 wt %, and most preferably 0.005 to 0.1 wt %. In one embodiment, when the composition for promoting energy metabolism is a liquid composition such as a beverage, the content of AEYY or a salt thereof in the composition, in terms of AEYY, is preferably 0.001 to 100,000 μg / mL, more preferably 0.001 to 50,000 μg / mL, even more preferably 0.005 to 5,000 μg / mL, even more preferably 0.01 to 5,000 μg / mL, particularly preferably 0.01 to 1,000 μg / mL, and most preferably 0.05 to 1,000 μg / mL. The term "quantity in terms of AEYY" or similar expressions refers to the amount of AEYY (a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1)), and in the case of a salt of AEYY, refers to the value obtained by multiplying the number of moles of the salt by the molecular weight of AEYY.

[0036] When the composition of the present invention contains Cyclo(Pro-Thr) or a salt thereof, the content thereof is not particularly limited and can be set depending on the form, etc. In one embodiment, the content of Cyclo(Pro-Thr) or a salt thereof in the composition for promoting energy metabolism, calculated as Cyclo(Pro-Thr), is, for example, preferably 0.0001 wt % or more, more preferably 0.0005 wt % or more, even more preferably 0.001 wt % or more, and particularly preferably 0.005 wt % or more, and is preferably 10 wt % or less, more preferably 5 wt % or less, even more preferably 0.5 wt % or less, and particularly preferably 0.1 wt % or less. In one aspect, the content of Cyclo(Pro-Thr) or a salt thereof in the composition for promoting energy metabolism, calculated as Cyclo(Pro-Thr), is, for example, preferably 0.0001 to 10 wt %, more preferably 0.0001 to 5 wt %, even more preferably 0.0005 to 0.5 wt %, particularly preferably 0.001 to 0.1 wt %, and most preferably 0.005 to 0.1 wt %. In one embodiment, when the composition for promoting energy metabolism is a liquid composition such as a beverage, the content of Cyclo(Pro-Thr) or a salt thereof in the composition, calculated as Cyclo(Pro-Thr), is preferably 0.001 to 100,000 μg / mL, more preferably 0.001 to 50,000 μg / mL, even more preferably 0.005 to 50,000 μg / mL, still more preferably 0.01 to 5,000 μg / mL, particularly preferably 0.01 to 1,000 μg / mL, and most preferably 0.05 to 1,000 μg / mL. The amount in terms of Cyclo(Pro-Thr) or similar expressions means the amount of Cyclo(Pro-Thr), and in the case of a salt of Cyclo(Pro-Thr), means the value obtained by multiplying the number of moles of the salt by the molecular weight of Cyclo(Pro-Thr).

[0037] The content of AEYY or a salt thereof, and the content of Cyclo(Pro-Thr) or a salt thereof in the composition of the present invention can be quantified by a known method, for example, liquid chromatography mass spectrometry (LC-MS / MS). The following measurement conditions can be used for LC-MS / MS measurement. As an apparatus, for example, an Agilent 1290 binary LC coupled with AB Sciex 5600+ TripleTOF system can be used. LC column: CORTECS T3 Column, 120 Å, 1.6 μm, 2.1 mm × 100 mm (Waters Corporation) Sample injection volume: 5 μL (LC chromatographic conditions) Column temperature: 40°C Mobile phase: Solution A: Water containing 0.1% formic acid Solution B: Acetonitrile containing 0.1% formic acid Gradient (solution B concentration): 0 to 1 min: Solution B 1% 1 to 8.5 min: Solution B 1% → 40% 8.5 to 13 min: Solution B 40% → 90% 13 to 15.5 min: Solution B 90% → 100% 15.5 to 17.8 min: Solution B 100% 17.8 to 18 min: Solution B 100% → 1% 18 to 20 min: Solution B 1%

[0038] When the composition of the present invention contains Cyclo(Pro-Thr) or a salt thereof, the weight ratio of AEYY or a salt thereof to Cyclo(Pro-Thr) or a salt thereof is not particularly limited. For example, the weight ratio of AEYY or a salt thereof (as calculated as AEYY):Cyclo(Pro-Thr) or a salt thereof (as calculated as Cyclo(Pro-Thr)) is preferably 1:500 to 500:1, and more preferably 1:50 to 50:1.

[0039] The dosage (also referred to as intake amount) of the composition of the present invention is not particularly limited, and may be an amount that can achieve the effect of promoting energy metabolism in a subject, and may be appropriately determined depending on the dosage form, administration method, subject body weight, etc.

[0040] When the composition of the present invention is ingested or administered to a human (adult) subject, the dose of AEYY or a salt thereof is preferably 0.01 mg or more, more preferably 0.1 mg or more, and preferably 10,000 mg or less, more preferably 1,000 mg or less, in terms of AEYY per day. In one embodiment, when the composition of the present invention is ingested or administered to a human (adult) subject, the dose of AEYY or a salt thereof is preferably 0.01 to 10,000 mg, more preferably 0.1 to 1,000 mg, in terms of AEYY per day. In the present invention, it is preferable to ingest or administer the above-mentioned amount of AEYY or a salt thereof once or more times a day, for example, once a day or in divided doses several times a day (e.g., 2 to 3 times a day). In one embodiment, it is preferable to ingest or administer the above-mentioned amount of AEYY or a salt thereof orally. In the case of a human (adult), it is preferable to ingest or administer the above-mentioned amount of AEYY or a salt thereof per 60 kg of body weight per day.

[0041] In one aspect, when the composition of the present invention is ingested or administered to a human (adult) subject, the dosage of Cyclo(Pro-Thr) or a salt thereof is preferably 0.01 to 10,000 mg, more preferably 0.1 to 1,000 mg, per day, calculated as Cyclo(Pro-Thr). In one aspect, the above-mentioned amount of Cyclo(Pro-Thr) or a salt thereof is preferably orally ingested or administered. In the case of a human (adult), the above-mentioned dosage is preferably an amount per 60 kg of body weight per day. In one aspect, the composition of the present invention may be an oral composition for ingesting or administering the above-mentioned amount of AEYY or a salt thereof, or AEYY or a salt thereof and Cyclo(Pro-Thr) or a salt thereof, per 60 kg of body weight per day to a human.

[0042] The composition of the present invention is preferably one that is ingested or administered continuously. It is expected that greater effects can be obtained by continuously ingesting or administering AEYY or a salt thereof. In one embodiment, the composition of the present invention is ingested or administered continuously for preferably one week or more, more preferably four weeks or more, and even more preferably eight weeks or more. AEYY, Cyclo(Pro-Thr) and salts thereof can be ingested as foods and beverages, and from the viewpoint of safety, it is believed that there are few problems with long-term ingestion, for example.

[0043] The subject (which may also be referred to as the administration subject) to whom the composition of the present invention is administered or ingested is not particularly limited. It is preferably a human or non-human mammal, more preferably a human. In one embodiment, the administration subject of the present invention includes a subject who needs or desires promotion of energy metabolism. In one embodiment, the administration subject of the present invention is preferably a subject who needs or desires at least one selected from the group consisting of increased energy metabolism, increased metabolic rate, increased resting metabolic rate, increased basal metabolic rate, increased heat production, increased vitality, prevention of fatigue, reduction of fatigue, prevention of lethargy, and reduction of lethargy. Furthermore, the administration subject of the present invention includes a subject who needs or desires prevention or amelioration of a disease or condition associated with decreased energy metabolism. In one embodiment, the administration subject of the composition of the present invention may be a healthy individual. For example, the composition may be used in healthy individuals for the purpose of promoting energy metabolism or preventing a disease or condition associated with decreased energy metabolism.

[0044] The composition of the present invention may be labeled with a function exhibited by promoting energy metabolism. Such a label is also referred to as a functional label. The label is not particularly limited. Examples of such labels include "increased metabolism," "increased metabolic rate," "increased resting metabolic rate," "increased basal metabolic rate," "increased energy level," "increased energy utilization," "increased heat production," "increased thermoresponsiveness," "increased vitality," "prevention of fatigue," "reduced fatigue," "prevention of fatigue," "reduced fatigue," and other labels or functional labels equivalent thereto. In one embodiment, the composition of the present invention is preferably labeled with at least one function selected from the group consisting of "increased metabolism," "increased metabolic rate," "increased resting metabolic rate," "increased basal metabolic rate," "increased energy level," "increased energy utilization," "increased heat production," "increased thermoresponsiveness," "increased vitality," "prevention of fatigue," "reduced fatigue," "prevention of fatigue," and "reduced fatigue." In one embodiment of the present invention, the composition of the present invention is preferably a food or beverage labeled with one or more of the above labels. The label may also indicate that the composition is used to achieve the above function. The label may be attached to the composition itself, or to the container or packaging of the composition.

[0045] The present invention also encompasses the following methods and uses: A method for promoting energy metabolism by administering a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof. A method for at least one selected from the group consisting of increasing energy metabolism, increasing metabolic rate, increasing resting metabolic rate, increasing basal metabolic rate, increasing heat production, increasing vitality, preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy, by administering a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof. Use of a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof, for promoting energy metabolism. Use of a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1) or a salt thereof for at least one selected from the group consisting of increasing energy metabolism, increasing metabolic rate, increasing resting metabolic rate, increasing basal metabolic rate, increasing heat production, increasing vitality, preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy. The method may be a therapeutic method or a non-therapeutic method. The use may be a therapeutic use or a non-therapeutic use. AEYY or a salt thereof can be used to promote energy metabolism, and can be preferably used for at least one selected from the group consisting of increasing energy metabolism, increasing metabolic rate, increasing resting metabolic rate, increasing basal metabolic rate, increasing heat production, increasing vitality, preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy.

[0046] In the above method, it is preferable to further administer Cyclo(Pro-Thr) or a salt thereof. In the above use, it is preferable to further use Cyclo(Pro-Thr) or a salt thereof. By administering or using AEYY or a salt thereof and Cyclo(Pro-Thr) or a salt thereof, a higher energy metabolism-promoting effect can be obtained.

[0047] The promotion of energy metabolism and preferred embodiments thereof are the same as those of the composition of the present invention described above. In one embodiment, AEYY or a salt thereof can be preferably used for promoting energy metabolism in mitochondria.

[0048] In one embodiment, AEYY or a salt thereof can be used for preventing or improving conditions or diseases associated with decreased energy metabolism. A method for preventing or improving conditions or diseases associated with decreased energy metabolism, which comprises administering AEYY or a salt thereof, is also encompassed by the present invention. AEYY or a salt thereof can be preferably used for, for example, preventing or improving fatigue, preventing or improving lethargy, increasing vitality, etc., and can more preferably be used for preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy.

[0049] In the above-mentioned methods and uses, it is preferable to have a subject ingest or administer AEYY or a salt thereof (preferably AEYY or a salt thereof, and Cyclo(Pro-Thr) or a salt thereof) once or more times a day, for example, once to several times (e.g., 2 to 3 times) a day. The subject is preferably a human or a non-human mammal, more preferably a human. The above-mentioned uses are preferably used in humans or non-human mammals, more preferably humans.

[0050] In the above-mentioned methods and uses, AEYY or a salt thereof may be used in an amount sufficient to achieve an energy metabolism-promoting effect (which may also be referred to as an effective amount). The preferred dosage, administration method, and administration target of AEYY or a salt thereof are the same as those for the composition of the present invention described above. In the above-mentioned methods and uses, AEYY or a salt thereof and Cyclo(Pro-Thr) or a salt thereof are preferably used. The preferred dosage of Cyclo(Pro-Thr) or a salt thereof is the same as that for the composition of the present invention described above. AEYY or a salt thereof and Cyclo(Pro-Thr) or a salt thereof may be ingested or administered directly, or may be ingested or administered as a composition containing them. Preferably, a composition containing AEYY or a salt thereof and Cyclo(Pro-Thr) or a salt thereof is ingested or administered. In one aspect, it is preferable to ingest or administer the above-mentioned composition for promoting energy metabolism of the present invention.

[0051] AEYY or a salt thereof can be used for the production of foods and beverages, pharmaceuticals, quasi-drugs, feed, etc. used for promoting energy metabolism. In one aspect, the present invention also encompasses the use of AEYY or a salt thereof in the production of the above-mentioned composition for promoting energy metabolism. The present invention also encompasses the use of AEYY or a salt thereof in the production of a composition used for at least one selected from the group consisting of increasing energy metabolism, increasing metabolic rate, increasing resting metabolic rate, increasing basal metabolic rate, increasing thermogenesis, increasing vitality, preventing fatigue, reducing fatigue, preventing lethargy, and reducing lethargy. The present invention also encompasses the use of AEYY or a salt thereof, and Cyclo(Pro-Thr) or a salt thereof in the production of a composition for promoting energy metabolism.

[0052] In this specification, a numerical range expressed by a lower limit and an upper limit, i.e., "lower limit to upper limit," includes the lower limit and the upper limit. For example, a range expressed by "1 to 2" means 1 or more and 2 or less, including 1 and 2. In this specification, the upper limit and the lower limit may be any combination of ranges.

[0053] The present invention will be described in more detail below with reference to examples, but the scope of the present invention is not limited to these examples.

[0054] Example 1 The effect of test substances on promoting energy metabolism was evaluated using undifferentiated C2C12 cells. A mitochondrial stress assay was used to evaluate the effect on promoting energy metabolism through mitochondrial function and cellular energy utilization.

[0055] The following peptides and taurine were used as test substances: AEYY: a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1) Cyclo(Pro-Thr) (cycloprolyl threonine) Taurine (Sigma-Aldrich) AEYY and Cyclo(Pro-Thr) were synthesized by Genscript.

[0056] (Methods) Cell Culture and Treatment C2C12 cells (ATCC CRL-1772) were cultured in growth medium (Dulbecco's Modified Eagle Medium (DMEM) containing 10% (vol / vol) fetal bovine serum (Gibco, Thermo Fisher Scientific) and supplemented with 1% penicillin-streptomycin (Gibco, Thermo Fisher Scientific)) under a humidified atmosphere (5% CO ２ C2C12 cells were cultured at 2 × 10 per well in an XF24 polystyrene V7 microplate. ４ The cells were seeded and treated with the test substance for 4 hours. Specifically, for the test substance treatment, the cells were cultured for 4 hours in a growth medium containing the test substance (AEYY, Cyclo(Pro-Thr) or taurine) at a concentration of 16 mg / mL or 64 mg / mL. Cells not treated with the test substance (untreated cells) served as a control. The control group of cells was cultured for 4 hours in a growth medium without the test substance. After the culture, the growth medium in the cell culture microplate was replaced with warmed Seahorse XF DMEM assay medium. Before the assay, the cell culture microplate was placed in a CO ２ The cells were placed in an incubator at 37°C for 1 hour without feeding.

[0057] Mitostress Assay: For the mitostress assay, oxygen consumption rate (OCR) was measured using a Seahorse XFe24 Analyser (Agilent Technologies) and XF Cell Mito Stress Test Reagents (Agilent Technologies). Baseline OCR (basal respiration) was measured four times for 3 min each. To quantify nonphosphorylatable respiration, 0.8 μM oligomycin was then added to each well, and OCR was measured three times for 3 min each. Next, to quantify maximal respiration, 5 μM carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) was added to each well, and OCR was measured four times for 3 min each. In the final step, to quantify non-mitochondrial respiration, 0.5 μM rotenone / antimycin A was added to each well and OCR was measured in triplicate for 3 min each. All experimental conditions were performed in quadruplicate.

[0058] Statistical Analysis All data are presented as mean ± standard error of the mean (SEM). Comparisons between different groups were analyzed by one-way analysis of variance (ANOVA) followed by the Tukey-Kramer test. A P value < 0.05 was considered statistically significant.

[0059] (Results) As described above, cells were treated with AEYY, Cyclo(Pro-Thr), and taurine to evaluate their effects on promoting energy metabolism. Taurine is known to have metabolic improving effects. Figures 1A and 1B show the effects of the test substances on mitochondrial energy metabolism in C2C12 cells (*: P<0.05 vs. control group). Figure 1A is a graph showing the basal respiration rate (oxygen consumption rate (OCR)) of C2C12 cells treated with the test substance for 4 hours. Figure 1B is a graph showing the ATP-related respiration rate (OCR) of C2C12 cells treated with the test substance for 4 hours. The vertical axis shows the percentage of OCR of the control group. Data are presented as mean ± SEM (N=9 or 12 for the AEYY-treated group, Cyclo(Pro-Thr), and taurine-treated groups, and N=18 for the control group).

[0060] Compared with the untreated control group, significant increases in basal respiration were observed in the taurine-treated group (64 mg / mL), AEYY-treated groups (16 mg / mL and 64 mg / mL), and Cyclo(Pro-Thr)-treated groups (16 mg / mL and 64 mg / mL) (Figure 1A). Furthermore, significant increases in ATP-related respiration were observed in the taurine-treated group (64 mg / mL), AEYY-treated groups (16 mg / mL and 64 mg / mL), and Cyclo(Pro-Thr)-treated groups (16 mg / mL and 64 mg / mL) compared with the untreated control (Figure 1B). AEYY and Cyclo(Pro-Thr) promoted cellular energy metabolism. These results indicate that AEYY and Cyclo(Pro-Thr) have the effect of increasing mitochondrial energy production. The energy metabolism promoting effects of AEYY and Cyclo(Pro-Thr) were higher than or equivalent to those of taurine.

[0061] AEYY and Cyclo(Pro-Thr) increased the oxygen consumption rate, an indicator of energy metabolism. These compounds promote energy metabolism. These peptides may contribute to increased energy production, which may in turn contribute to a feeling of energy.

Claims

1. A composition for promoting energy metabolism, comprising a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof.

2. The composition according to claim 1, which is used for at least one selected from the group consisting of increasing energy metabolism, increasing metabolic rate, increasing resting metabolic rate, increasing basal metabolic rate, increasing heat production, increasing vitality, preventing fatigue, reducing fatigue, preventing lethargy and reducing lethargy.

3. The composition according to claim 1 or 2, further comprising Cyclo(Pro-Thr) or a salt thereof.

4. The composition according to claim 1 or 2, which is an oral composition.

5. The composition according to claim 1 or 2, which is a food, beverage or oral medicine.

6. The composition of claim 1 or 2, wherein the composition is labeled with at least one function selected from the group consisting of "increasing metabolism," "increasing metabolic rate," "increasing resting metabolic rate," "increasing basal metabolic rate," "increasing energy levels," "increasing energy utilization," "increasing heat production," "increasing thermal response," "increasing vitality," "preventing fatigue," "reducing fatigue," "preventing lethargy," and "reducing lethargy." 7. Use of a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof, for promoting energy metabolism.

8. A method for promoting energy metabolism, which comprises administering a peptide consisting of the amino acid sequence Ala-Glu-Tyr-Tyr (SEQ ID NO: 1), or a salt thereof.

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