Anti-obesity composition

Abstract

The present invention addresses the problem of providing a technology for anti-obesity. This problem is solved by an anti-obesity composition containing a peptide comprising Leu-Asp-Gln-Trp (SEQ ID NO: 1), an anti-obesity composition containing a peptide comprising Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2), or an anti-obesity composition containing a peptide comprising Leu-Asp-Gln-Trp (SEQ ID NO: 1) and a peptide comprising Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2).

Description

[Technical Field]

[0001] The present invention relates to an anti-obesity composition. [Background technology]

[0002] In recent years, there has been a significant increase in the number of people suffering from lifestyle-related diseases caused by increased body fat, such as hypertension, dyslipidemia (hyperlipidemia), and diabetes, making "extending healthy life expectancy" a global issue. Various anti-obesity agents are being investigated to alleviate these increasingly serious lifestyle-related diseases caused by increased body fat, but there is a demand for agents that not only have anti-obesity effects but are also particularly safe. For this reason, research into the use of food-derived materials is also progressing, and there are hopes for their application as functional ingredients.

[0003] Reported diseases, signs, symptoms, disorders, and the like that can be prevented or treated by anti-obesity treatment include, for example, impaired glucose tolerance (type II diabetes, impaired glucose tolerance, etc.), dyslipidemia, hypertension, hyperuricemia, gout, coronary artery disease (myocardial infarction, angina pectoris, etc.), cerebral infarction (cerebral thrombosis, transient ischemic attack (TIA), etc.), fatty liver (non-alcoholic fatty liver disease (NAFLD)), menstrual disorders, infertility, sleep apnea syndrome (SAS), obesity-hypoventilation syndrome, musculoskeletal diseases (osteoarthritis (sites: knee, hip, etc.), spondylosis osteoarthritis, osteoarthritis of the fingers, etc.), and obesity-related kidney disease (Non-Patent Document 1).

[0004] Under such circumstances, it has been reported that a tripeptide consisting of Asp-Gln-Trp activates the β-oxidation of fatty acids and suppresses fat production, thereby inhibiting fat accumulation (Non-Patent Document 2).

[0005] It has also been reported that a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1), which has Leu (L) at the N-terminus of the tripeptide, has antioxidant activity (Non-Patent Document 3) and angiotensin I-converting enzyme inhibitory activity (Non-Patent Document 4). However, it is not known that the peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1) has an anti-obesity effect.

[0006] It has also been reported that a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2) has dipeptidyl peptidase 4 inhibitory activity (Non-Patent Document 5). However, it is not known that the peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) has an anti-obesity effect. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Journal of the Japanese Society of Internal Medicine, Vol. 107, No. 2, Obesity Treatment Guidelines 2016 [Non-patent document 2] Journal of Functional Foods 85 (2021) 104618 [Non-patent document 3] International Dairy Journal 21 (2011) 214-221 [Non-patent document 4] LWT 156 (2022) 112984 [Non-patent document 5] Peptides 54 (2014) 39-48 Summary of the Invention [Problem to be solved by the invention]

[0008] The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a technique for anti-obesity. [Means for solving the problem]

[0009] As a result of intensive research conducted by the present inventors to solve the above problems, they discovered that a peptide consisting of a specific amino acid sequence has an anti-obesity effect, leading to the completion of the present invention.

[0010] The present invention provides an anti-obesity composition comprising a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1). The present invention can also provide an anti-obesity composition comprising the peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2). In any of the above compositions, the anti-obesity effect is preferably achieved by reducing body fat. In addition, in a preferred embodiment, the compositions contain the peptide in the form of a whey protein hydrolysate. The present invention can also provide an anti-obesity composition comprising a peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2). In addition, in a preferred embodiment, the compositions are all food and drink compositions. In addition, in a preferred embodiment, each of the compositions is a pharmaceutical composition. In addition, in a preferred embodiment, the compositions are all used for at least one purpose selected from the group consisting of reducing waist circumference, reducing body fat, reducing body weight, and reducing BMI. [Effects of the Invention]

[0011] According to the present invention, a technique for anti-obesity can be provided. The composition according to the present invention can be prepared in the form of a food or drink, a medicine, or a feed, and can be easily ingested by a subject. [Brief explanation of the drawings]

[0012] [Figure 1]1 is a graph showing test results of Test Example 1 according to one embodiment of the present invention. [Figure 2] 1 is a graph showing test results of Test Example 1 according to one embodiment of the present invention. [Figure 3] 1 is a graph showing test results of Test Example 2 according to one embodiment of the present invention. [Figure 4] 1 is a graph showing test results of Test Example 2 according to one embodiment of the present invention. [Figure 5] 1 is a graph showing test results of Test Example 3 according to one embodiment of the present invention. [Figure 6] 1 is a graph showing test results of Test Example 4 according to one embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention will be described in detail below. However, the present invention is not limited to the following embodiments and can be freely modified within the scope of the present invention.

[0014] In this specification, the peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1) may be referred to as the "peptide of the present invention" or "LDQW." The peptide consisting of Asp-Gln-Trp may be referred to as "DQW." Furthermore, in this specification, the "anti-obesity composition comprising a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1)" according to the present invention may be referred to as the "composition of the present invention." Additionally, in this specification, the terms "ingestion," "take," and "ingested" may be replaced with the terms "administration," "administered," and "administered," respectively. Furthermore, the composition of the present invention may be a mixture, and its components may be homogeneous or heterogeneous. Furthermore, the composition of the present invention may contain the peptide of the present invention alone, or may contain other ingredients as long as they exert an anti-obesity effect. In this specification, percentages are expressed by mass unless otherwise specified.

[0015] <Composition for anti-obesity containing a peptide consisting of LDQW> One aspect of the present invention is a composition for anti-obesity, which contains a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1). Here, Leu (L) represents a leucine residue, Asp (D) represents an aspartic acid residue, Gln (Q) represents a glutamine residue, and Trp (W) represents a tryptophan residue. It is preferable that all amino acids are L-type amino acids.

[0016] The peptide of the present invention may be its physiologically acceptable salt or solvate. That is, the composition of the present invention may contain the peptide of the present invention, or its physiologically acceptable salt or solvate, and may contain one or more of these. The "physiologically acceptable salt" is not particularly limited as long as it is physiologically acceptable. For example, alkali metal salts (such as sodium salts and potassium salts), alkaline earth metal salts (such as calcium salts and magnesium salts), etc. can be mentioned. The "physiologically acceptable solvate" is not particularly limited as long as it is physiologically acceptable. For example, solvates such as water and alcohol solvents (such as ethanol) can be mentioned. The solvate is preferably low-toxic and water-soluble. In addition, the solvate includes the solvate of the "physiologically acceptable salt".

[0017] The peptide of the present invention can be obtained, for example, (1) A method of decomposing a protein or peptide containing the peptide of the present invention in an amino acid sequence by an enzyme or the like, and separating and purifying it from the obtained decomposition product. (2) A method of synthesizing the peptide of the present invention by a peptide synthesis method, and then separating and purifying the peptide of the present invention from the obtained crude synthesis product (chemical synthesis method). (3) A method of extracting from a plant, animal or microorganism that produces or is modified to produce the peptide of the present invention, and separating and purifying it from the obtained extract (biosynthesis method). and the like.

[0018] First, the method (1) will be explained. The protein or peptide containing the peptide of the present invention in its amino acid sequence is not particularly limited, as long as it is hydrolyzed with one or a combination of enzymes, acids, and alkalis, and contains the peptide of the present invention in a hydrolyzate thereof, for example, whey protein. The enzymes can be used singly or in combination of multiple types, the acids can be used singly or in combination of multiple types, and the alkalis can be used singly or in combination of multiple types.

[0019] The whey protein is a protein contained in whey (e.g., whey powder, desalted whey powder, etc.) separated by a known method from milk (e.g., cow's milk, skim milk, etc., which may be commercially available products). Examples of the whey protein in the present invention include separated and purified whey protein concentrate (WPC), whey protein isolate (WPI), and mixtures of these in any ratio. The whey protein in the present invention is preferably derived from cow's milk. Furthermore, the whey protein in the present invention is preferably whey protein concentrate (WPC). When the whey protein is derived from cow's milk, it is generally known to contain 15 to 25% α-lactalbumin, 50 to 65% β-lactoglobulin, 5 to 10% bovine serum albumin, lactoferrin, etc. Such protein compositions are similar for whey protein concentrate (WPC) and whey protein isolate (WPI).

[0020] The enzyme is preferably a protease. Therefore, when the "protein or peptide comprising the peptide of the present invention in its amino acid sequence" is a whey protein and the enzyme is a protease, the method (1) involves hydrolyzing the whey protein comprising the peptide of the present invention in its amino acid sequence with the protease, and separating and purifying the peptide of the present invention from the resulting protein hydrolysate (i.e., whey protein hydrolysate). The whey protein hydrolysate is obtained by hydrolyzing the whey protein. Whey protein contains various components, and the whey protein hydrolysate includes those components that have been hydrolyzed.

[0021] Specific examples of the method for producing the whey protein hydrolysate include, but are not limited to, the following. First, whey protein, the raw material, is dispersed and dissolved in water or warm water to prepare a whey protein solution. While there are no limitations on the concentration of the whey protein solution, it is generally preferable to set the protein concentration within a range of about 2 to about 20%, preferably about 5 to about 15%, from the viewpoints of efficiency and operability. A concentration of 2% or higher improves production efficiency, while a concentration of 20% or lower is preferable because it can suppress the risk of a decrease in decomposition efficiency, the risk of burning during heat treatment, and the risk of an increase in viscosity during cooling.

[0022] The pH of the whey protein solution is preferably adjusted using an acid or alkali solution to fall within the optimal pH range for the proteolytic enzyme. The pH is preferably adjusted to between 6.5 and 10.0 before hydrolysis. Pre-hydrolysis treatment steps, such as heat treatment and ion exchange treatment, may be added before or after pH adjustment, or both.

[0023] A protease is added to a whey protein solution whose pH has been appropriately adjusted to carry out hydrolysis. One or more types of protease may be used in combination. The protease is not particularly limited, but is preferably a protease, more preferably an endoprotease, and may be used in combination with an exoprotease as needed. Examples of the endoprotease include commercially available products such as Bioprase (manufactured by Nagase Biochemical Industry Co., Ltd.), Protin SD-AY10 (manufactured by Amano Enzyme Inc.), Protin NY100 (manufactured by Amano Enzyme Inc.), Protease N Amano (manufactured by Amano Enzyme Inc.), Neutrase (manufactured by Novo Nordisk), Alcalase (manufactured by Novo Nordisk Inc.), trypsin (manufactured by Novo Nordisk Inc.), chymotrypsin (manufactured by Novo Nordisk Inc.), papain (manufactured by Amano Enzyme Inc.), and bromelain (manufactured by Amano Enzyme Inc.). Examples of the exoprotease include commercially available products such as Protease A Amano (manufactured by Amano Enzyme Co., Ltd.), Sumizyme LP50D (manufactured by Shin-Nihon Kagaku Kogyo Co., Ltd.), and Flavorzyme (manufactured by Novo Nordisk).

[0024] The whey protein solution to which the protease has been added is preferably hydrolyzed under predetermined treatment conditions, specifically by incubating the solution for a predetermined period of time at an optimum temperature for the enzyme used. The optimum temperature is preferably 40 to 70°C, more preferably 50 to 60°C. The treatment time is preferably 2 to 24 hours, more preferably 3 to 12 hours. Furthermore, the pH of the whey protein solution at this time is preferably adjusted to 4 or more and 10 or less, more preferably 5 or more and 9 or less, and even more preferably 6 or more and 8 or less. The enzymes can be added simultaneously or at appropriate intervals, and immobilized enzymes can also be used. The pH of the solution can also be adjusted appropriately during hydrolysis to maintain the optimum pH for the enzyme.

[0025] When the whey protein solution is hydrolyzed by batch processing, the hydrolyzed solution is heated by a conventional method to inactivate the enzyme. It is preferable to appropriately select the heating temperature and holding time conditions that allow sufficient inactivation, taking into consideration the thermal stability of the enzyme used. After the heat treatment, the solution is cooled by a conventional method, and can be directly or, if necessary, concentrated and dried to obtain a powder product.

[0026] When the "protein or peptide containing the peptide of the present invention in its amino acid sequence" is decomposed with an acid or alkali, the pH of the solution may be adjusted for treatment. The pH is preferably 5 or less or 9 or more, and more preferably 4 or less or 10 or more. The acid or alkali decomposition product can be obtained by leaving or stirring the mixture at room temperature for several minutes or more, preferably 5 minutes to 1 hour. Here, "room temperature" means about 4 to 40°C, preferably 10 to 30°C.

[0027] The hydrolyzed product obtained by hydrolysis with one or a combination of enzymes, acids, and alkalis as described above may be used in an unpurified state. Thus, the composition of the present invention may contain the peptide of the present invention as a whey protein hydrolysate.

[0028] On the other hand, the degradation products may be appropriately subjected to known separation and purification techniques, for example, molecular weight fractionation may be performed to obtain a fraction corresponding to the molecular weight of the peptide of the present invention. As a method for molecular weight fractionation, for example, ultrafiltration, gel filtration, etc. can be employed, which can increase the removal rate of peptides and free amino acids with unnecessary molecular weights. In the case of ultrafiltration, a desired ultrafiltration membrane can be used, and in the case of gel filtration, a gel filtration agent used in the desired size exclusion chromatography can be used. Furthermore, known separation and purification methods can be used for desalting, removing impurities, and increasing purity. Examples include various chromatographic methods such as ion exchange chromatography, adsorption chromatography, reversed-phase chromatography, and partition chromatography, as well as solvent precipitation, salting out, and partitioning between two liquid phases. The separated and purified peptide fractions can be subjected to peptide identification by mass spectrometry or the like to confirm whether or not they contain the peptide of the present invention.

[0029] The content of the peptide of the present invention in the degradation products is not particularly limited, but from the viewpoint of further exerting the effects of the present invention, the lower limit is preferably 0.001% by mass or more, 0.005% by mass or more, and more preferably 0.01% by mass or more. On the other hand, from the viewpoint of production efficiency of the degradation products, the upper limit is preferably 40% by mass or less, more preferably 20% by mass or less, even more preferably 10% by mass or less, and even more preferably 5% by mass or less. For example, the content is within the range of 0.001% to 40% by mass, 0.005% to 20% by mass, 0.01% to 10% by mass, or 0.01% to 5% by mass.

[0030] Next, the above-mentioned methods (2) and (3) will be explained. The peptides of the present invention can also be produced by chemical synthesis or biosynthesis. Chemical synthesis of peptides can be carried out by liquid-phase or solid-phase methods commonly used for peptide synthesis. The synthesized peptides can be deprotected as needed, and unreacted reagents and by-products can be removed to isolate the peptides of the present invention. Such peptide synthesis can be carried out using a commercially available peptide synthesizer. Peptide biosynthesis can be carried out by a conventional method, such as by introducing a peptide expression vector into a host organism and allowing it to be produced and secreted. Furthermore, the peptide of the present invention produced by chemical synthesis or biosynthesis may be appropriately subjected to known separation and purification techniques. For example, molecular weight fractionation may be performed as described above to obtain a fraction corresponding to the molecular weight of the peptide of the present invention.

[0031] The subject to be ingested with the composition of the present invention is a subject in need of prevention or improvement of obesity. The subject includes humans and non-human mammals. Examples of non-human mammals include cows, goats, sheep, pigs, monkeys, dogs, cats, rats, mice, hamsters, guinea pigs, etc. The following describes the case where the subject is a human.

[0032] Subjects who take the composition of the present invention are not limited to subjects who are obese at the time of taking the composition of the present invention, but also include subjects who are not obese at the time of taking the composition of the present invention.

[0033] In the present invention, "anti-obesity" refers to a concept that includes preventing obesity and improving obesity. In the present invention, "obesity" refers to a condition in which the Body Mass Index (BMI) (i.e., the value obtained by dividing body weight (kg) by the square of height (m)) is 25 kg / m 2 or above, or a BMI of 25 kg / m 2 Even if the body fat level is less than 100 mg / kg, the body may be in a state of excessive accumulation of body fat (visceral fat and / or subcutaneous fat). Specifically, a BMI of 25 kg / m 2 A BMI of 25 kg / m or more is usually considered obese. 2 Even if the waist circumference is less than 85 cm, there may be cases where excess body fat (visceral fat and / or subcutaneous fat) has accumulated. For example, a person may be judged to be obese if their waist circumference is 85 cm or more, or if their waist circumference is 90 cm or more, or if their visceral fat area is 100 cm or more. 2In the above cases, a person may be determined to be obese, and if the body fat percentage is 20% or higher for men, a person may be determined to be obese, and if the body fat percentage is 28% or higher for women, a person may be determined to be obese.

[0034] The peptides of the present invention have anti-obesity effects. That is, for example, when a subject ingests the peptides of the present invention, a greater anti-obesity effect is exhibited than when a placebo is ingested (or before ingesting the peptides of the present invention, or when not ingesting the peptides of the present invention). Note that, although the anti-obesity effect is not usually exhibited when a subject ingests a placebo (or before ingesting the peptides of the present invention, or when not ingesting the peptides of the present invention), it is acceptable for the effect to be exhibited as long as it is greater when the peptides of the present invention are ingested. From the above, the peptide of the present invention can be used as an active ingredient of an anti-obesity composition. In other words, the composition of the present invention can contain the peptide of the present invention as an active ingredient.

[0035] The present invention also includes embodiments in which a subject who ingests the composition of the present invention exhibits a stronger anti-obesity effect than a subject who ingests a composition that does not contain the peptide of the present invention (e.g., a placebo). That is, the anti-obesity effect of the present invention may be evaluated as being stronger than the anti-obesity effect exhibited in a subject who ingests a composition that does not contain the peptide of the present invention (e.g., a placebo). Although the anti-obesity effect is usually not exhibited when a subject ingests a composition that does not contain the peptide of the present invention (e.g., a placebo), it is acceptable for the anti-obesity effect to be exhibited as long as it is stronger when the subject ingests the composition of the present invention.

[0036] Anti-obesity in the present invention includes, for example, anti-obesity due to body fat reduction (e.g., inhibition of body fat accumulation, promotion of body fat breakdown, etc.). Therefore, compositions containing the peptides of the present invention can also be used for body fat reduction (e.g., inhibition of body fat accumulation, promotion of body fat breakdown, etc.). That is, the present invention can also provide compositions for body fat reduction (e.g., inhibition of body fat accumulation, promotion of body fat breakdown, etc.), which contain the peptides of the present invention. The body fat refers to visceral fat and / or subcutaneous fat. That is, examples of the body fat reduction include visceral fat reduction (e.g., suppression of visceral fat accumulation, promotion of visceral fat decomposition, etc.) and subcutaneous fat reduction (e.g., suppression of subcutaneous fat accumulation, promotion of subcutaneous fat decomposition, etc.). Since the anti-obesity effect achieved in the present invention results in reduced BMI, reduced waist circumference, and reduced body weight, compositions containing the peptides of the present invention can also be used for these purposes. That is, the present invention can also provide a composition for reducing BMI, which contains the peptides of the present invention. The present invention can also provide a composition for reducing waist circumference, which contains the peptides of the present invention. The present invention can also provide a composition for reducing body weight, which contains the peptides of the present invention.

[0037] The composition of the present invention has an anti-obesity effect and can be used for the prevention or treatment of diseases, symptoms, conditions, disorders, etc. that can be prevented or treated by anti-obesity. In the present invention, the term "prevention" refers to preventing or delaying the onset of a disease, symptom, condition, disorder, etc. in a subject, or reducing the risk of the onset of a disease, symptom, condition, disorder, etc. in a subject. The subject may have one or more diseases, symptoms, conditions, disorders, etc.

[0038] The uses of this embodiment may be therapeutic or non-therapeutic. "Non-therapeutic purposes" refers to actions that do not include medical procedures, i.e., actions that do not include treatments on the human body through therapy. Examples include health promotion and cosmetic procedures.

[0039] When the composition of the present invention is used for non-therapeutic purposes, the target may be healthy individuals. A healthy individual may mean an individual who is not obese at the time of ingesting the composition of the present invention, an individual who is not suffering from a disease, symptom, symptom, or disorder caused by obesity, an individual who is concerned about body fat, an individual who is slightly obese, an individual who is concerned about weight, an individual who is concerned about BMI, or the like. When the composition of the present invention is used for non-therapeutic purposes, it is possible to prevent obesity and prevent a disease, symptom, symptom, or disorder caused by obesity in healthy individuals. Examples of diseases, signs, symptoms, or disorders caused by obesity include obesity, impaired glucose tolerance (type II diabetes, impaired glucose tolerance, etc.), dyslipidemia, hypertension, hyperuricemia, gout, coronary artery disease (myocardial infarction, angina pectoris, etc.), cerebral infarction (cerebral thrombosis, transient ischemic attack (TIA), etc.), fatty liver (non-alcoholic fatty liver disease (NAFLD)), menstrual disorders, infertility, sleep apnea syndrome (SAS), obesity-hypoventilation syndrome, musculoskeletal diseases (osteoarthritis (sites: knee, hip, etc.), spondylosis osteoarthritis, osteoarthritis of the fingers, etc.), and obesity-related kidney disease.

[0040] When the composition of the present invention is used for therapeutic purposes, the subject can be a non-healthy individual. When the composition of the present invention is used for therapeutic purposes, it becomes possible to treat diseases, signs, symptoms, disorders, etc. caused by obesity in unhealthy individuals.

[0041] The content of the peptide of the present invention in the composition of the present invention is appropriately determined depending on the embodiment of the composition, but is preferably 0.000001% or more, more preferably 0.000005% or more, even more preferably 0.00001% or more, even more preferably 0.0001% or more, and is preferably 100% or less, more preferably 20% or less, and even more preferably 10% or less, in terms of the total amount. For example, the total amount is within the range of 0.000001% to 100%, 0.000005% to 20%, 0.00001% to 10%, 0.0001% to 10%, or 0.001% to 10%, etc.

[0042] The intake amount of the composition of the present invention is determined appropriately depending on the form of the composition, the method of use, the subject, the subject's age, sex, and other conditions, but is not particularly limited as long as an anti-obesity effect is exhibited in the subject who ingests it. The total amount of the peptide of the present invention per day is preferably 0.001 mg or more, more preferably 0.005 mg or more, and even more preferably 0.01 mg or more, and is preferably 5 g or less, more preferably 1 g or less, and even more preferably 0.5 g or less. For example, the total amount per day is within the range of 0.001 mg to 5 g, 0.005 mg to 1 g, or 0.01 mg to 0.5 g.

[0043] The composition of the present invention can be taken once a day or in divided doses, or once every few days or weeks, but is preferably taken daily. For example, daily intake for 12 weeks or more is preferred for reducing BMI, and daily intake for 8 weeks or more is preferred for reducing waist circumference, and daily intake for 12 weeks or more is also preferred.

[0044] The composition of the present invention can be taken orally, but is not limited to this, and may also be taken nasally, or via a gastrostomy or enterostomy tube, for example, or may be administered to a subject via a nasogastric tube.

[0045] The composition of the present invention can be used, for example, as a food or drink composition, a feed composition, or a pharmaceutical composition. For example, it can be provided as an anti-obesity food and drink composition comprising the peptide of the present invention. Hereinafter, this may be referred to as the "food and drink composition of the present invention." It can also be provided as an anti-obesity feed composition comprising the peptide of the present invention. Hereinafter, this may be referred to as the "feed composition of the present invention." It can also be provided as an anti-obesity pharmaceutical composition comprising the peptide of the present invention. Hereinafter, this may be referred to as the "pharmaceutical composition of the present invention."

[0046] <Food and drink composition of the present invention> The food and drink composition of the present invention is not particularly limited as long as it contains the peptide of the present invention. The food and drink composition may be in any form, such as liquid, paste, gel-like solid, powder, etc., and may include, in addition to tablet confectionery and liquid food, wheat flour products such as bread, macaroni, spaghetti, noodles, cake mix, fried chicken flour, breadcrumbs, etc.; instant noodles, cup noodles, retort / prepared foods, canned foods, microwave foods, instant soup / stew, instant miso soup / cleaning liquid, canned soup, freeze-dried foods, other instant foods, etc.; canned agricultural products, canned fruit, etc. processed agricultural products such as jams and marmalades, pickles, boiled beans, dried agricultural goods, and cereals (processed grain products); processed seafood products such as canned seafood, fish ham and sausage, fish paste products, seafood delicacies, and tsukudani (simmered fish dishes); processed livestock products such as canned livestock paste, livestock ham and sausage; processed milk, milk drinks, yogurt, lactic acid bacteria drinks, cheese, ice cream, infant formula, cream, and other dairy products; fats and oils such as butter, margarines, and vegetable oils; soy sauce, miso, sausage Basic seasonings such as seasonings, processed tomato seasonings, mirin, vinegars, etc.; complex seasonings and foods such as cooking mixes, curry bases, sauces, dressings, noodle soups, spices, and other complex seasonings; frozen foods such as frozen ingredients, semi-cooked frozen foods, and cooked frozen foods; confectioneries such as caramel, candy, gummies, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice snacks, bean snacks, dessert sweets, jellies, and other sweets. Subcategories include carbonated drinks, natural fruit juices, fruit juice drinks, soft drinks with fruit juice, fruit pulp drinks, fruit drinks with fruit pieces, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, alcoholic drinks, and other beverages, as well as other commercially available foods such as baby food, furikake, and ochazuke nori seaweed; infant formula; enteral nutritional foods; special purpose foods, health functional foods (foods for specified health uses, foods with nutrient functions, foods with functional claims); and nutritional supplements. The food and drink composition of the present invention may also be a supplement, for example, a tablet supplement, which allows the intake of the peptide of the present invention without being affected by other foods in terms of the daily meal amount and calorie intake.

[0047] The food and beverage composition of the present invention can be produced by adding the peptide of the present invention to the raw materials of a normal food or beverage, and can be produced in the same manner as normal food and beverages except for the addition of the peptide of the present invention. The addition of the peptide of the present invention may be carried out at any stage in the production process of the food and beverage composition.

[0048] The food and drink composition of the present invention also includes raw materials for producing the food and drink composition, and food additives and the like that are added to the food and drink composition during or after the production process of the food and drink composition.

[0049] Furthermore, the food and beverage compositions of the present invention can contain known or future components with prebiotic effects or components that supplement prebiotic effects (hereinafter referred to as "prebiotics"), as long as they do not impair the effects of the present invention. For example, the food and beverage compositions of the present invention can be produced by blending the peptides of the present invention with prebiotics such as various proteins, such as whey protein, casein protein, soybean protein, or pea protein, or mixtures or hydrolysates thereof; amino acids, such as leucine, valine, isoleucine, or glutamine; vitamins, such as vitamin B6 or vitamin C; creatine; citric acid; fish oil; or oligosaccharides, such as isomaltooligosaccharides, xylooligosaccharides, soybean oligosaccharides, fructooligosaccharides, or lactulose.

[0050] The content of the peptide of the present invention in the food and beverage composition of the present invention is appropriately determined depending on the embodiment of the food and beverage composition, but is preferably 0.000001% or more, more preferably 0.000005% or more, even more preferably 0.00001% or more, even more preferably 0.0001% or more, and is preferably 100% or less, more preferably 20% or less, and even more preferably 10% or less, in terms of total amount. For example, the total amount is within the range of 0.000001% to 100%, 0.000005% to 20%, 0.00001% to 10%, 0.0001% to 10%, or 0.001% to 10%, etc.

[0051] The intake amount of the food and beverage composition of the present invention is appropriately determined depending on the form of the food and beverage composition, the method of use, the subject, the subject's age, sex, and other conditions, but is not particularly limited as long as an anti-obesity effect is exhibited in the subject who ingests it. The total amount of the peptide of the present invention per day is preferably 0.001 mg or more, more preferably 0.005 mg or more, and even more preferably 0.01 mg or more, and is preferably 5 g or less, more preferably 1 g or less, and even more preferably 0.5 g or less. For example, the total amount per day is within the range of 0.001 mg to 5 g, 0.005 mg to 1 g, or 0.01 mg to 0.5 g.

[0052] The food and drink composition of the present invention may be ingested alone or together with another food and drink composition or food or drink, or a pharmaceutical composition or medicine. For example, it may be ingested together with another anti-obesity food and drink composition or food or drink, or an anti-obesity pharmaceutical composition or medicine, etc.

[0053] The food and drink composition of the present invention can be sold as a food and drink composition or food and drink labeled with an application such as anti-obesity. Needless to say, any other wording may be used as long as it expresses an effect secondary to the above-mentioned application.

[0054] The above-mentioned "indication" means all acts for informing consumers of the above-mentioned uses, and any indication that can recall or infer the above-mentioned uses falls under the "indication" of the present invention, regardless of the purpose of the indication, the content of the indication, the object or medium on which it is displayed, etc. However, it is preferable to display the use in an expression that allows consumers to directly recognize the above-mentioned uses. Specifically, examples include the act of describing the above-mentioned uses on the food and beverage composition or food and beverage product of the present invention or on the product packaging; the act of transferring, delivering, displaying for transfer or delivery, or importing the product or product packaging on which the above-mentioned uses are described; the act of describing the above-mentioned uses in advertisements, price lists, or transaction documents for the product and displaying or distributing them; or the act of describing the above-mentioned uses in information containing these contents and providing them by electromagnetic means (such as the Internet), and particularly displaying them on promotional materials at the point of sale such as packaging, containers, catalogs, pamphlets, POP, and other documents.

[0055] Furthermore, the labeling is preferably one approved by the government or the like (e.g., labeling approved under various government systems and made in a manner based on such approval). Examples include labeling as a health food, more specifically, a health food, a functional food, an enteral nutritional food, a food for special dietary uses, a food with nutrient functions, a quasi-drug, etc. Other examples include labeling approved by the Consumer Affairs Agency, such as a food for specified health uses, a food with nutrient functions, a food with functional claims, and labeling approved under similar systems. Examples of the latter include labeling as a food for specified health uses, a labeling as a conditional food for specified health uses, a labeling claiming to affect the structure or function of the body, a labeling claiming to reduce disease risk, and a labeling claiming functionality based on scientific evidence. More specifically, examples include labeling as a food for specified health uses (especially a labeling claiming health uses) as defined in the Cabinet Office Ordinance on the Permission for Labeling for Special Dietary Uses Provided in the Health Promotion Act (Cabinet Office Ordinance No. 57 of August 31, 2009), and similar labeling.

[0056] Examples of such indications include "For those concerned about body fat," "For those who are slightly obese," "For those concerned about weight (BMI)," "Reduces high BMI," "Helps improve high BMI," "Lose weight," "Reduces belly fat," "Lose weight and belly fat," "Reduces waist circumference," and "Makes fat consumption easier."

[0057] The food and beverage composition of the present invention can be used for any subject, including healthy and unhealthy individuals, but when it is made into a food or beverage labeled with a specific use or function, it is used for the above-mentioned non-therapeutic purposes.

[0058] <Feed composition of the present invention> Examples of the feed composition of the present invention include pet food, livestock feed, and fish feed. The feed composition of the present invention can be produced by mixing the peptide of the present invention with common feed or its raw materials, such as grains, cereals, bran, fish meal, bone meal, oils and fats, skim milk powder, whey, mineral feed, or yeast. The produced feed composition can be administered to common mammals, livestock, farmed fish, and pet animals.

[0059] The content of the peptide of the present invention in the feed composition of the present invention is appropriately determined depending on the embodiment of the feed composition, but is preferably 0.000001% or more, more preferably 0.000005% or more, even more preferably 0.00001% or more, even more preferably 0.0001% or more, and is preferably 100% or less, more preferably 20% or less, and even more preferably 10% or less, in total. For example, the total content is within the range of 0.000001% to 100%, 0.000005% to 20%, 0.00001% to 10%, 0.0001% to 10%, or 0.001% to 10%, etc.

[0060] The dosage of the feed composition of the present invention administered to a subject is determined appropriately depending on the form of the feed composition, the method of use, the subject, the subject's age, sex, and other conditions, but is not particularly limited as long as an anti-obesity effect is exhibited in the subject to which it is administered. The total amount of the peptide of the present invention per day is preferably 0.001 mg or more, more preferably 0.005 mg or more, and even more preferably 0.01 mg or more, and is preferably 5 g or less, more preferably 1 g or less, and even more preferably 0.5 g or less. For example, the total amount per day is within the range of 0.001 mg to 5 g, 0.005 mg to 1 g, or 0.01 mg to 0.5 g.

[0061] The feed composition of the present invention is administered to mammals other than humans and used for the above-mentioned therapeutic and non-therapeutic purposes.

[0062] The feed composition of the present invention may be administered alone or together with other feed compositions or feeds, or pharmaceutical compositions or medicines, for example, other anti-obesity feed compositions or feeds, or anti-obesity pharmaceutical compositions or medicines, etc.

[0063] <Pharmaceutical composition of the present invention> The pharmaceutical composition of the present invention is not particularly limited as long as it contains the peptide of the present invention. As the pharmaceutical composition of the present invention, the peptide of the present invention may be used as it is, or may be formulated by mixing it with a physiologically acceptable liquid or solid pharmaceutical carrier.

[0064] The dosage form of the pharmaceutical composition of the present invention is not particularly limited, and specific examples include tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, syrups, suppositories, injections, ointments, patches, eye drops, nasal drops, etc. In addition, additives commonly used as pharmaceutical carriers, such as excipients, binders, disintegrants, lubricants, stabilizers, flavorings, diluents, surfactants, or solvents for injections, can be used in the formulation.

[0065] As the pharmaceutical carrier, various organic or inorganic carriers can be used depending on the dosage form. Examples of carriers for solid preparations include excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents.

[0066] 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; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, carboxymethyl cellulose, and carboxymethyl cellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; and sulfate derivatives such as calcium sulfate.

[0067] Examples of binders include gelatin, polyvinylpyrrolidone, macrogol, and the like, in addition to the above-mentioned excipients.

[0068] Examples of disintegrants include the above-mentioned excipients as well as chemically modified starch or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and cross-linked polyvinylpyrrolidone.

[0069] Examples of lubricants include talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; veegum; waxes such as gaelt; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylates 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; and starch derivatives.

[0070] Examples of stabilizers include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; sorbic acid; and the like.

[0071] Examples of flavoring agents include sweeteners, acidulants, and fragrances. In the case of a liquid preparation for oral administration, examples of the carrier to be used include a solvent such as water, a flavoring agent, and the like.

[0072] The content of the peptide of the present invention in the pharmaceutical composition of the present invention is appropriately determined depending on the dosage form, dosage regimen, subject, age and sex of the subject, type and severity of the disease, symptom, condition or disorder, and other conditions, but is preferably 0.000001% or more, more preferably 0.000005% or more, even more preferably 0.00001% or more, even more preferably 0.0001% or more, and is preferably 100% or less, more preferably 20% or less, and even more preferably 10% or less, in terms of total amount. For example, the total amount is within the range of 0.000001% to 100%, 0.000005% to 20%, 0.00001% to 10%, 0.0001% to 10%, or 0.001% to 10%, etc.

[0073] The dosage of the pharmaceutical composition of the present invention administered to a subject is determined appropriately depending on the dosage form, dosage regimen, subject, age, and sex of the subject; the type and severity of the disease, symptom, condition, or disorder; and other factors, but is not particularly limited as long as an anti-obesity effect is exhibited in the subject to which it is administered. The total amount of the peptide of the present invention per day is preferably 0.001 mg or more, more preferably 0.005 mg or more, and even more preferably 0.01 mg or more, and is preferably 5 g or less, more preferably 1 g or less, and even more preferably 0.5 g or less. For example, the total amount per day is within the range of 0.001 mg to 5 g, 0.005 mg to 1 g, or 0.01 mg to 0.5 g.

[0074] The pharmaceutical composition of the present invention may be administered for the prevention of diseases, symptoms, conditions, disorders, etc. The administration form is preferably determined depending on the formulation, the subject, the subject's age and sex, the type and severity of the disease, symptoms, conditions, or disorders, and other conditions.

[0075] The pharmaceutical composition of the present invention is administered to unhealthy subjects and used for the above-mentioned therapeutic purposes.

[0076] The pharmaceutical composition of the present invention may be administered alone or together with another pharmaceutical composition or drug, or a food and drink composition or food and drink, or a feed composition or feed. For example, it may be administered together with another anti-obesity pharmaceutical composition or drug, or an anti-obesity food and drink composition or food and drink, or an anti-obesity feed composition or feed, etc.

[0077] The present invention also includes the following aspects. <1-1> Use of a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1) for producing an anti-obesity composition. <1-2> A peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1) or a composition containing said peptide for use in anti-obesity. <1-3> A peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) or a composition containing the peptide for use in the prevention or treatment of diseases, signs, symptoms, disorders, etc. that can be prevented or treated by anti-obesity. <1-4> A method for preventing or ameliorating obesity, comprising the step of administering a prophylactically effective amount or an ameliorating effective amount of a peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) or a composition containing said peptide to a subject in need of prevention or amelioration. <1-5> A method for preventing or treating a disease, sign, symptom, disorder, etc. that can be prevented or treated by anti-obesity therapy, comprising the step of administering a prophylactically effective amount or a therapeutically effective amount of a peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) or a composition containing said peptide to a subject in need of prevention or treatment. Use of a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1) or a composition containing the peptide for anti-obesity. A peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1) or a composition containing the peptide for use in anti-obesity.

[0078] <An anti-obesity composition containing a peptide consisting of LKPTPEGDLEIL> Another aspect of the present invention is an anti-obesity composition containing a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2). Here, Leu (L) represents a leucine residue, Lys (K) represents a lysine residue, Pro (P) represents a proline residue, Thr (T) represents a threonine residue, Glu (E) represents a glutamic acid residue, Gly (G) represents a glycine residue, Asp (D) represents an aspartic acid residue, and Ile (I) represents an isoleucine residue. In addition, in this specification, the peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) may be described as "LKPTPEGDLEIL". This aspect incorporates the description of the aspect related to the "anti-obesity composition containing a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1)" already described.

[0079] Moreover, this aspect includes the following aspects. Use of a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) for manufacturing an anti-obesity composition. A peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) or a composition containing the peptide for use in anti-obesity. Use of a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) or a composition comprising the peptide for the prevention or treatment of diseases, syndromes, symptoms, disorders, etc. that can be prevented or treated by anti-obesity. <2-4>A method for preventing or improving obesity, comprising administering a prophylactically effective amount or an ameliorating effective amount of a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) or a composition comprising the peptide to a subject in need of prevention or amelioration. <2-5>A method for preventing or treating diseases, syndromes, symptoms, disorders, etc. that can be prevented or treated by anti-obesity, comprising administering a prophylactically effective amount or a therapeutically effective amount of a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) or a composition comprising the peptide to a subject in need of prevention or treatment. <2-6>Use of a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) or a composition comprising the peptide for anti-obesity. <2-7>A peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) or a composition comprising the peptide for use in anti-obesity.

[0080] Another aspect of the present invention is a composition for anti-obesity, comprising a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2). This embodiment invokes the explanation of the embodiment relating to "an anti-obesity composition comprising a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1)" and the explanation of the embodiment relating to "an anti-obesity composition comprising a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2)" already explained.

[0081] The preferred contents of the two peptides in the composition according to this embodiment are determined appropriately depending on the embodiment of the composition. However, the preferred contents of the two peptides in total are the same as those described in the embodiment of "An anti-obesity composition comprising a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1)." Furthermore, the preferred intake amount (or dosage amount) of the two peptides in the composition according to this embodiment is determined as appropriate depending on the form of the composition, the method of use, the subject, the subject's age, sex, and other conditions, and if it is a pharmaceutical composition, it is determined as appropriate depending on the dosage form, the method of use, the subject, the subject's age, sex, the type of disease, symptom, condition, or disorder, the severity thereof, and other conditions, but in any case, there are no particular limitations as long as an anti-obesity effect is exerted in the subject who takes (or is administered) it, and the preferred intake amount (or dosage amount) described in the embodiment according to "an anti-obesity composition comprising a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1)" is used for the total amount of the two peptides.

[0082] This aspect also includes the following aspects. <3-1> Use of a peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2) for producing an anti-obesity composition. <3-2> A peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2), or a composition containing these peptides, for use in anti-obesity. <3-3> A peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2), or a composition containing these peptides, for use in the prevention or treatment of a disease, sign, symptom, or disorder that can be prevented or treated by anti-obesity. <3-4> A method for preventing or ameliorating obesity, comprising the step of administering to a subject in need of prevention or amelioration a prophylactically effective amount or an ameliorating effective amount of a peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2), or a composition containing these peptides. <3-5> A method for preventing or treating a disease, sign, symptom, disorder, etc. that can be prevented or treated by anti-obesity therapy, comprising the step of administering to a subject in need thereof a prophylactically effective amount or a therapeutically effective amount of a peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2), or a composition containing these peptides. <3-6> Use of a peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2), or a composition containing these peptides, for anti-obesity. <3-7> A peptide consisting of Leu-Asp-Gln-Trp (sequence number 1) and a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (sequence number 2), used for anti-obesity, or a composition containing these peptides. [Example]

[0083] The present invention will be specifically described below using examples, but the present invention is not limited to these examples.

[0084] <Production Example 1 of Whey Protein Hydrolysate Containing LDQW and LKPTPEGDLEIL> Commercially available WPC (whey protein concentrate, Milei 80 (manufactured by Milei)) was dissolved in water at a concentration of 10%, and sodium hydroxide was added to adjust the pH of the solution to 7, thereby preparing an aqueous whey protein solution. To this aqueous whey protein solution, 5,000 units of protease N Amano (manufactured by Amano Enzyme Inc.) was added per 1 g of protein, and decomposition was carried out at 50 °C for 7 hours. Subsequently, heating was carried out at 90 °C for 10 minutes to inactivate the enzyme, and then powdered by a spray dryer to obtain a dried product of whey protein hydrolysate.

[0085] <Production Example 2 of Whey Protein Hydrolysate Containing LDQW and LKPTPEGDLEIL> Commercially available whey powder (NZ-GLP (manufactured by Fonterra)) was dissolved in water at a concentration of 20%, and sodium hydroxide was added to adjust the pH of the solution to 6, thereby preparing an aqueous whey protein solution. To this aqueous whey protein solution, 7,000 units of protin NY100 (manufactured by Amano Enzyme Inc.) and 1,000 units of alcalase (manufactured by Novo Nordisk) were added per 1 g of protein, and decomposition was carried out at 55 °C for 5 hours. Subsequently, heating was carried out at 90 °C for 10 minutes to inactivate the enzyme, clarified by a microfiltration membrane, and then powdered by a spray dryer to obtain a dried product of whey protein hydrolysate.

[0086] <Chemical Synthesis Example 1 of LDQW> Peptide LDQW was chemically synthesized by the Fmoc solid-phase synthesis method. After deprotection, it was purified by reverse-phase HPLC and then lyophilized.

[0087] <Chemical Synthesis Example 1 of LKPTPEGDLEIL> Peptide peptide LKPTPEGDLEIL was chemically synthesized by the Fmoc solid-phase synthesis method. After deprotection, it was purified by reverse-phase HPLC and then lyophilized.

[0088] <Confirmation of LDQW> The synthetic peptide obtained in the above-mentioned "Chemical Synthesis Example 1 of LDQW" was subjected to liquid chromatography (HPLC, manufactured by Shimadzu Corporation) and liquid chromatography-mass spectrometry (LC-MS, manufactured by Thermo Fisher Scientific). The purity (95% or higher) and molecular weight (560) of LDQW were confirmed. The analysis conditions of LC-MS are shown below. Also, the retention time (RT) of LDQW and the mass-to-charge ratio (m / z) observed by Full MS are shown in Table 1. In order to analyze LDQW in the whey protein hydrolyzate prepared in the above-mentioned "Production Example 1 of Whey Protein Hydrolyzate Containing LDQW and LKPTPEGDLEIL", qualitative and quantitative analyses of LDQW were performed using the LDQW prepared in the above-mentioned "Chemical Synthesis Example 1 of LDQW" as a control. As a result of analysis by LC-MS in the same manner as the synthetic peptide, it was revealed that the whey protein hydrolyzate contained LDQW at a concentration of 0.23 mass percent. [[ID=)3]]

[0089] <LC-MS Analysis Conditions> (Equipment) LC: Vanquish UHPLC System (manufactured by Thermo Fisher Scientific) MS: Q Excative Focus (manufactured by Thermo Fisher Scientific)

[0090] (LC) Column: ACQUITY UPLC BEH C18, 1.7 μm, 2.1 x 150 mm (manufactured by Waters) Column temperature: 40°C Buffer: A; 0.1% aqueous formic acid solution, B; acetonitrile containing 0.1% formic acid Elution program: 0 - 30 min, 98 - 70% A; 30 - 35 min, 70 - 10% A; 35 - 36 min, 10 - 98% A; 36 - 45 min, 98% A Flow rate: 0.2 mL / min Injection: 10 μL

[0091] (MS) Polarity: Positive Scan range: 120 to 1800 m / z MS resolution: 70,000 MS / MS resolution: 17,500 Sheath gas flow rate: 45 units Aux gas flow rate: 10 units Spray voltage: 3.5 kV Capillary temperature: 275℃ Aux gas heater temperature: 350℃

[0092]

Table 1

[0093] <Confirmation of LKPTPEGDLEIL> The synthetic peptide obtained in the above "Chemical Synthesis Example 1 of LKPTPEGDLEIL" was subjected to a liquid chromatograph (HPLC, manufactured by Shimadzu Corporation) and a liquid chromatography mass spectrometer (LC-MS, manufactured by Thermo Fisher Scientific) according to the above "LC-MS Analysis Conditions". The purity (95% or more) and molecular weight (1,324) of LKPTPEGDLEIL were confirmed. In addition, the retention time (RT) of LKPTPEGDLEIL and the mass-to-charge ratio (m / z) observed in Full MS are shown in Table 2. To analyze LKPTPEGDLEIL in the whey protein hydrolysate prepared in "Production Example 1 of Whey Protein Hydrolysate Containing LDQW and LKPTPEGDLEIL," qualitative and quantitative analyses of LKPTPEGDLEIL were performed using the LKPTPEGDLEIL prepared in "Chemical Synthesis Example 1 of LKPTPEGDLEIL" as a control. Analysis by LC-MS, similar to that for synthetic peptides, revealed that the whey protein hydrolysate contained LKPTPEGDLEIL at a concentration of 0.39 mass percent.

[0094] [Table 2]

[0095] <Test Example 1> The anti-obesity effect was evaluated using the presence or absence of neutral fat accumulation in mouse-derived 3T3-L1 cells (JCRB9014). Because 3T3-L1 cells are cells that differentiate into adipocytes through differentiation induction, 3T3-L1 cells before differentiation are referred to here as "3T3-L1 preadipocytes." This also applies to the test examples described below. 3T3-L1 preadipocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) high-glucose medium supplemented with 10.0% (v / v) Fetal Bovine Serum (FBS), 1% (v / v) penicillin / streptomycin, and 2 mmol / L L-Glutamine at 37°C in the presence of 5% CO2. 1 × 10 3T3-L1 preadipocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) high-glucose medium supplemented with 10.0% (v / v) Fetal Bovine Serum (FBS), 1% (v / v) penicillin / streptomycin, and 2 mmol / L L-Glutamine. 4 The cells were seeded onto a 24-well microplate at a cell number of 1000 cells / well, and after 72 hours of culture, the medium was replaced with a differentiation-inducing medium containing 0.5 mM isobutylmethylxanthine, 1 μg / mL insulin, and 0.25 μM dexamethasone. After 48 hours of culture, the medium in the untreated group (NC group) was replaced with "medium containing 1 μg / mL insulin," and in the test substance (whey protein hydrolysate containing LDQW and LKPTPEGDLEIL produced in the above "Production Example 1 of whey protein hydrolysate containing LDQW and LKPTPEGDLEIL," or synthetic LDQW prepared in the above "Chemical synthesis example 1 of LDQW")-added group, the medium was replaced with "culture medium containing 1 μg / mL insulin + a predetermined concentration of the test substance," and both were cultured for another 48 hours. The medium was then changed every two days. After 7 days from the start of differentiation induction, the culture medium was removed, the cells were washed twice with phosphate buffered saline (PBS), and fixed overnight at room temperature with 10% neutral buffered formalin. After washing the cells with ultrapure water, Oil Red O solution from a lipid assay kit (Cosmo Bio) was added and the cells were left at room temperature for 30 minutes to stain intracellular triglycerides. The dye was then extracted with the kit's extract and measured at an absorbance wavelength of 540 nm using a plate reader. Each group was tested in triplicate (n=3). The results are shown in Figure 1 (whey protein hydrolysates containing LDQW and LKPTPEGDLEIL) and Figure 2 (synthetic LDQW). Whey protein hydrolysates containing LDQW and LKPTPEGDLEIL (final concentrations of 1.0 mg / mL and 2.0 mg / mL) and synthetic LDQW (final concentrations of 3.0 μM and 30 μM) each exhibited an inhibitory effect on neutral fat accumulation. The neutral fat levels were expressed as mean values ​​± standard error relative to the NC group. Statistically significant differences were detected in comparison with the NC group, and * indicates a statistically significant difference of p<0.05. This test was also performed in Test Example 2, described below.

[0096] <Test Example 2> In vivo, triglycerides are broken down into glycerol and free fatty acids by lipolytic enzymes. The anti-obesity effects were evaluated using the amounts of glycerol and free fatty acids in the culture medium of mouse-derived 3T3-L1 cells (JCRB9014). 3T3-L1 preadipocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) high-glucose medium supplemented with 10.0% (v / v) Fetal Bovine Serum (FBS), 1% (v / v) penicillin / streptomycin, and 2 mmol / L L-Glutamine at 37°C in the presence of 5% CO2. 1 × 10 3T3-L1 preadipocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) high-glucose medium supplemented with 10.0% (v / v) Fetal Bovine Serum (FBS), 1% (v / v) penicillin / streptomycin, and 2 mmol / L L-Glutamine. 4 The cells were seeded onto a 24-well microplate at a cell number of 1000 cells / well, and after 72 hours of culture, the medium was replaced with a differentiation-inducing medium containing 0.5 mM isobutylmethylxanthine, 1 μg / mL insulin, and 0.25 μM dexamethasone. After 48 hours of culture, the medium in the untreated group (NC group) was replaced with "medium containing 1 μg / mL insulin," and in the group to which the test substance (synthetic LDQW prepared in "Example 1 of Chemical Synthesis of LDQW" above) had been added, the medium was replaced with "culture medium containing 1 μg / mL insulin + a predetermined concentration of the test substance," and both groups were cultured for another 48 hours. The medium was then changed every two days, and collected 7 days after the start of differentiation induction. The amount of glycerol in the collected medium was measured using a glycerol measurement kit (Cayman Chemical Co.), and the amount of free fatty acids in the collected medium was measured using a NEFA C-Test Wako Kit (Fujifilm Wako Co.). Each group was tested in triplicate (n = 3). The results are shown in Figure 3 (glycerol content) and Figure 4 (free fatty acid content). Synthetic LDQW (final concentrations 3.0 μM and 30 μM) increased the amount of glycerol and free fatty acids in the medium, demonstrating its effect in promoting triglyceride breakdown. The data obtained are shown as mean values ​​± standard error.

[0097] <Test Example 3> The anti-obesity effect was evaluated using the presence or absence of neutral fat accumulation in mouse-derived 3T3-L1 cells (JCRB9014) as an indicator. 3T3-L1 preadipocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) high-glucose medium supplemented with 10.0% (v / v) Fetal Bovine Serum (FBS), 1% (v / v) penicillin / streptomycin, and 2 mmol / L L-Glutamine at 37°C in the presence of 5% CO2. 1 × 10 3T3-L1 preadipocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) high-glucose medium supplemented with 10.0% (v / v) Fetal Bovine Serum (FBS), 1% (v / v) penicillin / streptomycin, and 2 mmol / L L-Glutamine. 4 The cells were seeded onto a 24-well microplate at a cell number of 1000 cells / well, and after 72 hours of culture, the medium was replaced with a differentiation-inducing medium containing 0.5 mM isobutylmethylxanthine, 1 μg / mL insulin, and 0.25 μM dexamethasone. After 48 hours of culture, the medium in the untreated group (NC group) was replaced with "medium containing 1 μg / mL insulin," or in the group to which the test substance (synthetic DQW or synthetic LDQW prepared in "Example 1 of Chemical Synthesis of LDQW" was added), the medium was replaced with "culture medium containing 1 μg / mL insulin + a predetermined concentration of the test substance," and both groups were cultured for another 48 hours. The medium was then changed every two days. After 7 days from the start of differentiation induction, the culture medium was removed, the cells were washed twice with phosphate buffered saline (PBS), and fixed overnight at room temperature with 10% neutral buffered formalin. After washing the cells with ultrapure water, Oil Red O solution from a lipid assay kit (Cosmo Bio) was added and the cells were left at room temperature for 30 minutes to stain intracellular triglycerides. The dye was then extracted with the kit's extract and measured at an absorbance wavelength of 540 nm using a plate reader. Five samples were used for each group (n=5). The results are shown in Figure 5. It was revealed that synthetic LDQW exhibited a neutral fat accumulation inhibitory effect at lower concentrations than synthetic DQW. Specifically, it was revealed that synthetic LDQW exhibited the same neutral fat accumulation inhibitory effect as synthetic DQW at a concentration 1 / 10 lower than that of synthetic DQW. The neutral fat content was expressed as the mean ± standard error relative to the untreated group (NC group). Statistically significant differences (p<0.05) were determined by multiple comparison tests using the Turkey HSD method. In Figure 5, non-overlapping letters indicate significant differences between groups.

[0098] <Test Example 4> Using the synthetic LKPTPEGDLEIL prepared in the above "Chemical Synthesis Example 1 of LKPTPEGDLEIL" as the test substance, the anti-obesity effect was evaluated using the presence or absence of triglyceride accumulation in mouse-derived 3T3-L1 cells (JCRB9014) as an indicator in the same procedure as in Test Example 1. The results are shown in Figure 6. Synthetic LKPTPEGDLEIL (final concentrations of 4.3 μM and 43 μM) exhibited an inhibitory effect on neutral fat accumulation. The neutral fat content was expressed as the mean ± standard error relative to the NC group. Statistical significance was detected in comparison with the NC group, and *** indicates a statistically significant difference of p<0.001.

[0099] <Test Example 5> In a randomized, double-blind, parallel-group comparative study in humans, the anti-obesity effect of whey protein hydrolysate (WPH) containing LDQW at a concentration of 0.23 mass percent and LKPTPEGDLEIL at a concentration of 0.39 mass percent, which was produced in the above-mentioned "Production Example 1 of whey protein hydrolysate containing LDQW and LKPTPEGDLEIL," was examined using BMI as an indicator. The subjects were 71 men with an average age of approximately 52 years old, and were divided into a test group of 36 who took WPH and a control group of 35 who took a placebo. The test group was instructed to take 5g of WPH per day for 12 consecutive weeks, while the control group was instructed to take 5g of dextrin (manufactured by Matsutani Chemical Industry Co., Ltd.) per day instead of WPH for 12 consecutive weeks. The subjects' BMI was measured before (week 0) and after the final intake (week 12) of each sample. The BMI of the subjects in each group was as shown in Table 3. After 12 weeks of intake, the BMI of the test group was significantly reduced compared to the control group. The unit is kg / m 2 is. These results demonstrate that consuming whey protein hydrolysate for 12 weeks reduces BMI.

[0100] [Table 3]

[0101] <Test Example 6> In a randomized, double-blind, parallel-group comparative study in humans, the anti-obesity effect of whey protein hydrolysate (WPH) containing LDQW at a concentration of 0.23 mass percent and LKPTPEGDLEIL at a concentration of 0.39 mass percent, which was produced in the above-mentioned "Production Example 1 of whey protein hydrolysate containing LDQW and LKPTPEGDLEIL," was investigated using waist circumference as an indicator. The subjects were 177 healthy adult men and women with an average age of approximately 46 and a BMI of 25 to 30. They were divided into a test group of 90 who took WPH and a control group of 87 who took a placebo. The test group was instructed to ingest the test food, consisting of 1.3 g of powder containing 1 g of WPH dissolved in 100 mL of drinking water, once daily for 12 weeks. The control group was instructed to ingest 1.3 g of powder containing 1 g of dextrin (Matsutani Chemical Industry Co., Ltd.) dissolved in 100 mL of drinking water, once daily for 12 weeks. The subjects' waist circumferences were measured before (week 0), after 4 weeks, after 8 weeks, and after 12 weeks of ingestion. The waist circumferences of the subjects in each group are shown in Table 4. After 8 and 12 weeks of intake, the waist circumferences of the test group were significantly reduced compared to the control group. The unit is cm. These results demonstrate that ingesting the whey protein hydrolysate for at least 8 weeks significantly reduces waist circumference.

[0102] [Table 4]

Claims

1. An anti-obesity composition comprising a peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1).

2. An anti-obesity composition comprising a peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2).

3. The composition according to claim 1, wherein the anti-obesity effect is achieved by reducing body fat.

4. 3. The composition according to claim 1, wherein the peptide is a whey protein hydrolysate.

5. A peptide consisting of Leu-Asp-Gln-Trp (SEQ ID NO: 1), and A peptide consisting of Leu-Lys-Pro-Thr-Pro-Glu-Gly-Asp-Leu-Glu-Ile-Leu (SEQ ID NO: 2) An anti-obesity composition comprising:

6. The composition according to claim 1 or 2, which is a food or drink composition.

7. 3. The composition of claim 1 or 2, which is a pharmaceutical composition.

8. The composition according to claim 1 or 2, wherein the composition is used for at least one selected from the group consisting of reducing waist circumference, reducing body fat, reducing body weight, and reducing BMI.

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