JP2009079045A - Composition stably containing peptide - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a composition which stably contains a peptide comprising a specific amino acid sequence. <P>SOLUTION: The present invention provides a composition comprising (A) one or more compounds selected from mucopolysaccharides, and (B) one or more compounds selected from the group consisting of a peptide represented by formula I: Leu-Glu-His-Ala, its derivatives and its salts, and the like. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a composition stably containing a peptide consisting of a specific amino acid sequence, and an invention related thereto.

A peptide comprising a specific amino acid sequence (Patent Document 1) or a product obtained by degrading soybean protein with a protease (preferably thermolysin) (Patent Document 2) is selected from the group consisting of collagen and hyaluronic acid A therapeutic or preventive agent for a disease caused by a decrease in the amount of collagen and / or hyaluronic acid; a food for improving or preventing a condition caused by a decrease in the amount of collagen and / or hyaluronic acid; Cosmetics for prevention and / or improvement of skin wrinkles or talmi due to exposure to ultraviolet rays, aging, etc .; cosmetics for prevention and / or improvement of skin elasticity or reduction of elasticity; etc. Is described. Furthermore, Patent Document 1 describes a method for treating soybean protein with a specific protease called thermolysin as a preferred method for preparing a peptide comprising the specific amino acid sequence.
However, Patent Documents 1 and 2 do not consider any stability of the specific peptide in a composition of a peptide having a useful specific amino acid sequence or a product obtained by degrading soybean protein with a protease. .

  By the way, proteins, peptides or amino acids are produced by Maillard reaction (browning reaction) in the presence of reducing sugars (for example, glucose, fructose, etc.) that can be used as additives for foods, pharmaceuticals, quasi drugs, cosmetics and the like. It is known that its content decreases when decomposed. The Maillard reaction is a series of reactions initiated by a reaction between a free amino group present in a protein, peptide or amino acid and a carbonyl group of a reducing sugar. As a method for suppressing the degradation of protein, peptide or amino acid, for example, a method of oxidizing and removing reducing sugar from food by adding an enzyme capable of oxidizing the reducing group of sugar (Patent Document 3); A method of adding high ester pectin to peptides in the environment (Patent Document 4) is known, but a method of suppressing the degradation of proteins, peptides or amino acids with mucopolysaccharides is not known.

For example, Patent Document 5 discloses a skin characterized by hydrolyzing a bean seed such as soybean or an aqueous extract of a processed product thereof with a proteolytic enzyme and using a fraction having a molecular weight of 10,000 or less as an active ingredient. An external preparation is described. More specifically, an essence (formulation example 6) containing soybean protein degradation product by hydrolase, hyaluronic acid which is a kind of mucopolysaccharide, and the like is described. Patent Document 6 discloses hydrolyzed proteins and the like; at least one organic powder or surface-treated organic powder; and a dermatologically acceptable carrier, wherein the carrier is in the form of an emulsion. A featured topical personal care composition is described. More specifically, topical creams (Table 5) containing sodium hyaluronate, a kind of mucopolysaccharide, hydrolyzed soy protein and the like are described.
However, in the external preparation for skin described in Patent Document 5 and the composition described in Patent Document 6, stability of the hydrolyzed soy protein and the peptide contained therein is not considered at all. In addition, there is no description or suggestion of a method for suppressing degradation of peptides contained therein or using mucopolysaccharides.

International Publication No. 2006/101187 Pamphlet International Publication No. 2007/049400 Pamphlet International Publication No. 02/39828 Pamphlet JP 2005-192557 A Japanese Patent Laid-Open No. 9-025225 Japanese translation of publication 2005-516048

The present inventors have intensively studied to blend a peptide having a specific amino acid sequence and a product obtained by degrading a protein containing the peptide with a protease into foods, pharmaceuticals, quasi drugs, cosmetics, and the like. As a result, it has been found that a peptide having a specific amino acid sequence and a product obtained by degrading a protein containing the peptide with a protease are degraded over time, and the content thereof is reduced. Degradation of a peptide consisting of a specific amino acid sequence can be used as an additive for foods, pharmaceuticals, quasi drugs, cosmetics, etc., such as sugar (eg, glucose, sucrose, fructose, etc.) or sugar alcohol (eg, sorbitol) , Erythritol, maltitol, etc.).
When a peptide consisting of a specific amino acid sequence showing various useful actions is blended in foods, pharmaceuticals, quasi drugs, cosmetics, etc., it is important to suppress the degradation. Therefore, an object of the present invention is to suppress degradation of a peptide consisting of a specific amino acid sequence and to suppress a decrease in the content thereof.

  As a result of intensive studies to suppress the degradation of a peptide consisting of a specific amino acid sequence and to suppress a decrease in the content thereof, the present inventors have used a specific amino acid sequence by combining one or more selected from mucopolysaccharides. The present inventors have found that it is possible to suppress a decrease in the content of the peptide consisting of

Accordingly, the present invention provides the following.
(1) (A) one or more selected from mucopolysaccharides;
(B) A composition containing one or more selected from the group consisting of a peptide represented by Formula I: Leu-Glu-His-Ala, derivatives thereof, and salts thereof.
(2) one or more selected from (A) mucopolysaccharides;
(B) A composition containing a protein protease hydrolyzate containing at least one selected from the group consisting of a peptide represented by the formula I: Leu-Glu-His-Ala, derivatives thereof and salts thereof .
(3) The composition according to (2) above, wherein the protein is soybean protein.
(4) The composition according to (2) or (3) above, wherein the protease is thermolysin.
(5) The composition according to any one of (1) to (4), further containing one or more selected from the group consisting of sugar and sugar alcohol.

  According to the present invention, since a decrease in the content of a peptide consisting of a specific amino acid sequence is suppressed, a stable composition containing the peptide can be provided.

  Hereinafter, the present invention will be described in detail. It should be understood that the terms used in the present specification are used in the meaning normally used in the art unless otherwise specified.

  The present invention comprises (A) one or more selected from mucopolysaccharides, and (B) a peptide represented by Formula I: Leu-Glu-His-Ala (SEQ ID NO: 1), a derivative thereof, and a salt thereof. It is a composition containing 1 or more types selected from a group.

  In another embodiment, the present invention comprises (A) one or more selected from mucopolysaccharides, and (B) a peptide represented by Formula I: Leu-Glu-His-Ala, a derivative thereof, and a salt thereof. It is a composition containing a protease hydrolyzate of a protein containing one or more selected from the group.

In the present invention, one or more selected from mucopolysaccharides are used.
Mucopolysaccharide is a polysaccharide containing amino sugar and uronic acid in its basic skeleton, and is obtained from animal tissues. Specific examples of the mucopolysaccharide include hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparan sulfate, heparin, keratan sulfate I and II, and derivatives thereof, and salts thereof.

  As salts of hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparan sulfate, heparin, keratan sulfate I and II, and derivatives thereof, pharmaceutically and physiologically acceptable salts are preferable. Examples include alkali metal salts such as sodium and potassium; alkaline earth metal salts such as magnesium and calcium; ammonium salts; alkanolamine salts such as monoethanolamine; and preferably alkali metal salts such as sodium salts It is.

The origin of the mucopolysaccharide used in the present invention is not particularly limited, and those that can be usually used in the field of pharmaceuticals, quasi drugs, cosmetics, and foods can be used without particular limitation.
In the present invention, mucopolysaccharides may be used singly or in combination of two or more.
Moreover, when there are commercially available mucopolysaccharides, the commercially available products may be used as they are.

  Examples of derivatives such as hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparan sulfate, heparin, keratan sulfate I and II include ester derivatives in which the hydrogen atom of the carboxyl group is substituted with an alkyl group, alkenyl group, aryl group, etc .; An ester derivative in which a hydrogen atom is substituted with an acyl group, an ether derivative in which an alkyl group, an alkenyl group, an aryl group, or the like is substituted; an amide derivative in which the hydrogen atom of an amino group is substituted with an acyl group, an alkyl group, And amine derivatives substituted with an alkenyl group, an aryl group, and the like. As these derivatives, those having 1 to 6 substitutions per structural unit (disaccharide) are usually used, and those having 2 to 4 substitutions are preferable.

  Examples of ester derivatives include those in which a hydrogen atom of a carboxyl group is substituted with an alkyl group, an alkenyl group, an aryl group, or the like; a hydrogen atom of a hydroxyl group is substituted with an acyl group such as an alkanoyl group or an arylcarbonyl group; Can be mentioned.

As an alkyl group, a C1-C12 alkyl group is mentioned, Preferably it is a C1-C6 alkyl group. Specific examples include methyl group, ethyl group, propyl group, isopropyl group, butyl group, s-butyl group, t-butyl group, isobutyl group, pentyl group, hexyl group and the like.
Examples of the alkenyl group include alkenyl groups having 2 to 12 carbon atoms, preferably alkenyl groups having 2 to 6 carbon atoms. Specific examples include a vinyl group and an allyl group.
These alkyl group and alkenyl group may be substituted, and as the substituent, an alkoxy group having 1 to 6 carbon atoms (eg, methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, s-butoxy group) Group, t-butoxy group, isobutoxy group, pentyloxy group, hexyloxy group, etc.), halogen atom (eg, fluorine atom, chlorine atom, bromine atom, iodine atom), amino group, hydroxy group and the like.

  Examples of the aryl group include aryl groups having 6 to 12 carbon atoms. Specific examples include a phenyl group and a naphthyl group (1-naphthyl group, 2-naphthyl group). In addition, the aryl group may be substituted, and examples of the substituent include alkyl groups having 1 to 6 carbon atoms (eg, methyl group, ethyl group, propyl group, isopropyl group, butyl group, s-butyl group, t -Butyl group, isobutyl group, pentyl group, hexyl group, etc.), C1-C6 alkoxy group (eg, methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, s-butoxy group, t-butoxy group) Group, isobutoxy group, pentyloxy group, hexyloxy group, etc.), halogen atom (eg, fluorine atom, chlorine atom, bromine atom, iodine atom), amino group, hydroxy group and the like.

The alkanoyl group includes an alkanoyl group having 1 to 12 carbon atoms, and preferably an alkanoyl group having 1 to 7 carbon atoms. Specific examples include an acetyl group, a propionyl group, and a butyryl group. Further, the alkanoyl group may be substituted, and as the substituent, an alkoxy group having 1 to 6 carbon atoms (eg, methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, s-butoxy group, t-butoxy group, isobutoxy group, pentyloxy group, hexyloxy group and the like), halogen atom (eg, fluorine atom, chlorine atom, bromine atom, iodine atom), amino group, hydroxy group and the like.
Examples of the arylcarbonyl group include an arylcarbonyl group having 7 to 15 carbon atoms, and an arylcarbonyl group having 7 to 12 carbon atoms is preferable. Specific examples include a benzoyl group and a naphthylcarbonyl group (1-naphthylcarbonyl group, 2-naphthylcarbonyl group). In addition, the arylcarbonyl group may be substituted, and examples of the substituent include alkyl groups having 1 to 6 carbon atoms (eg, methyl group, ethyl group, propyl group, isopropyl group, butyl group, s-butyl group, t-butyl group, isobutyl group, pentyl group, hexyl group, etc.), C1-C6 alkoxy group (eg, methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, s-butoxy group, t- Butoxy group, isobutoxy group, pentyloxy group, hexyloxy group, etc.), halogen atom (eg, fluorine atom, chlorine atom, bromine atom, iodine atom), amino group, hydroxy group and the like.

Examples of ether derivatives include those in which a hydrogen atom of a hydroxyl group is substituted with an alkyl group, an alkenyl group, an aryl group, or the like.
The “alkyl group”, “alkenyl group” and “aryl group” exemplified as the substituent of the ether derivative are the “alkyl group”, “alkenyl group” and “aryl group” exemplified as the substituent of the ester derivative, respectively. The same thing is mentioned.

Examples of the amide derivative include those in which a hydrogen atom of an amino group is substituted with an acyl group such as an alkanoyl group or an arylcarbonyl group.
Examples of the “alkanoyl group” and “arylcarbonyl group” exemplified as the substituent of the amide derivative include those similar to the “alkanoyl group” and “arylcarbonyl group” exemplified as the substituent of the ester derivative.

Examples of the amine derivative include those in which a hydrogen atom of an amino group is substituted with an alkyl group, an alkenyl group, an aryl group, or the like.
Examples of the “alkyl group”, “alkenyl group” and “aryl group” exemplified as the substituent of the amine derivative are the “alkyl group”, “alkenyl group” and “aryl group” exemplified as the substituent of the ester derivative, respectively. The same thing is mentioned.

  In the composition of the present invention, the average molecular weight of the mucopolysaccharide is not particularly limited, but is usually 1,000 to 4,000,000, preferably 1,000 to 1,200,000, more preferably 5,000 to 1,000,000, particularly preferably 5,000 to 400,000. It is.

Specific examples of the mucopolysaccharide used in the present invention include hyaluronic acid, sodium hyaluronate, potassium hyaluronate, magnesium hyaluronate, calcium hyaluronate, acetylated hyaluronic acid, sodium acetylated hyaluronate, and acetylated hyaluronic acid. Potassium, acetylated magnesium hyaluronate, acetylated calcium hyaluronate, chondroitin sulfate, sodium chondroitin sulfate and the like, and mixtures thereof can be used.
The mucopolysaccharide used in the present invention is preferably sodium hyaluronate, sodium chondroitin sulfate, etc. from the viewpoints of solubility and stability.

  The amount of mucopolysaccharides to be blended in the composition of the present invention is not particularly limited as long as the effect of the present invention can be obtained, but is usually 0.0005% by weight or more, preferably 0.001% by weight based on the entire composition. Or more, more preferably 0.01% by weight or more, further preferably 0.05% by weight or more, particularly preferably 0.1% by weight or more, and the upper limit of blending is preferably 5% by weight or less, more preferably 1% by weight. Hereinafter, it is more preferable that the content be 0.5% by weight or less, and particularly preferably 0.25% by weight or less.

In the present invention, one or more selected from the group consisting of a peptide represented by the formula I: Leu-Glu-His-Ala, a derivative thereof, and a salt thereof are used. Sometimes referred to as “specific peptides”).
Moreover, in this specification, a peptide sequence may be described by the single letter abbreviation of an amino acid, for example, the peptide represented by the said formula I may be called LEHA.

  In the present specification, the “peptide derivative” means, for example, acetylation, palmitoylation, myristylation, amidation, acrylation, dansylation, biotinylation, phosphorylation, succinylation, anilideation, benzyloxylation of peptides. Carbonylation, formylation, nitration, sulfonation, aldehyde formation, cyclization, glycosylation, monomethylation, dimethylation, trimethylation, guanidylation, amidineation, maleylation, trifluoroacetylation, carbamylation, trinitrophenyl Derivatized, nitrotroponylated or acetoacetylated derivatives. Among these, palmitoylation is preferable because it is expected to have high permeability to cells, and N-terminal acetylation, C-terminal amidation, and C-terminal methylation are exopeptidases that degrade peptides from the terminal. It is preferable because it is expected to be resistant to, and to be stable in the living body.

  In the present specification, the salt of the peptide or its derivative is preferably a pharmacologically or physiologically acceptable salt of the peptide or its derivative. For example, alkali metal salts such as sodium salts and potassium salts of peptides or derivatives thereof; alkaline earth metal salts such as calcium salts and magnesium salts; ammonium salts; salts with inorganic acids such as hydrochlorides, sulfates and nitrates; acetic acid Salt, citrate, maleate, malate, oxalate, lactate, succinate, fumarate, propionate, formate, benzoate, picrate, benzenesulfonate, etc. Salts with organic acids; preferred are ammonium salts, hydrochlorides, sulfates and acetates, and more preferred are ammonium salts and acetates.

  The peptides used in the present invention can be prepared by methods known in the art. For example, the peptide of the present invention may be synthesized by a chemical synthesis method (eg, a solid phase method (eg, Fmoc method), a liquid phase method, etc.), or may be produced by a method such as gene recombinant expression. . In addition, although the amino acid which comprises the peptide of this invention may be a L body or a D body, Preferably it is a L body.

  Furthermore, the peptide of the present invention can also be prepared by cutting out a peptide comprising the target amino acid sequence from the amino acid sequence of the protein containing the target amino acid sequence by a known means such as protease treatment. For example, examples of the protein containing the LEHA sequence include proteins as shown in Table 1 below.

  A person skilled in the art can appropriately select an appropriate protease in order to cut out a peptide comprising the target amino acid sequence from the amino acid sequence of the protein containing the target amino acid sequence in consideration of the sequence specificity of the protease.

  The protease used in the present invention is not particularly limited. For example, thermolysin, papain, bromelain, trypsin, chymotrypsin, pancreatin, subtilin, protease M “Amano” G (derived from Aspergillus oryzae: manufactured by Amano Enzyme Co., Ltd.) Etc. Only one type of protease may be used in the present invention, or two or more types may be used in combination. Among these, thermolysin is preferable from the viewpoint of obtaining higher collagen or hyaluronic acid production promoting effect.

  Thermolysin (EC3.4.24.27) is a heat-resistant protease produced by a heat-resistant bacterium called Bacillus thermoproteolyticus. Thermolysin is generally known to cleave peptide bonds on the amino group side of hydrophobic amino acid residues having large side chains (for example, isoleucine, leucine, valine, phenylalanine, methionine, alanine, etc.).

  As the protease, a commercially available product can be suitably used. For example, thermolysin is easily available from a manufacturer such as Daiwa Kasei Co., Ltd. In the present invention, a protease known in the art as a protease having peptide cleavage characteristics (such as cleavage sequence specificity) equivalent to thermolysin can be used as thermolysin.

  For example, in order to excise the LEHA sequence from the carrot-derived sequence, thermolysin (derived from Bacillus thermoproteolyticus) and chymotrypsin (derived from bovine pancreas) can be used in combination. Further, for example, in order to cut out the LEHA sequence from the potato-derived sequence, protease M “Amano” G (derived from Aspergillus oryzae: manufactured by Amano Enzyme Co., Ltd.) and the aforementioned thermolysin can be used. Further, for example, in order to cut out the LEHA sequence from the rice-derived sequence, the protease M “Amano” G and the thermolysin are used in combination. Further, for example, in order to cut out the LEHA sequence from the soybean-derived sequence, use of the thermolysin is exemplified. In addition, for example, in order to cut out the LEHA sequence from the kidney bean-derived sequence, the chymotrypsin and the thermolysin are used in combination. Moreover, for example, in order to cut out the LEHA sequence from the cassava-derived sequence, the chymotrypsin and the thermolysin are used in combination. Further, for example, in order to excise the LEHA sequence from the sequence derived from the river clover, it is possible to use trypsin (derived from porcine pancreas) and the thermolysin together. The combination of the protein containing the target amino acid sequence and the protease is not limited to the above combination, but a preferable combination includes a combination of soybean protein and thermolysin.

  Soybean (Glycin max) is a leguminous annual plant with a height of about 60-70 cm. It is known that the seeds are often used as edible beans or processed into tofu, miso, soy sauce, etc.

  The soybean protein used in the present invention can be any protein derived from the soybean plant described above, but preferably can be any protein derived from the seed of the soybean plant.

  Therefore, in the present invention, the soybean plant itself, the seed of the soybean plant itself, the plant or the crushed or pulverized product of the seed may be used as the soybean protein, but preferably the protein from all the components in the soybean plant. What separated and refine | purified the component, More preferably, what isolate | separated and refine | purified the protein component from all the components in the seed of a soybean plant is used. Soy protein obtained by separation and purification as described above is substantially all kinds of proteins contained in soybean plants or soybean plant seeds as long as the protease hydrolyzate has the ability to promote collagen or hyaluronic acid production. May be included, and some types of proteins may be included.

  As the soy protein, commercially available products can also be suitably used. For example, Nisshin Cosmo Foods Co., Ltd., ADM Far East Co., Ltd., Showa Sangyo Co., Ltd., Fuji Oil Co., Ltd., Koyo Shokai Co., Ltd., etc. Are readily available from other manufacturers or suppliers.

  In the present specification, the seeds of soybean plants not only refer to the whole structure commonly referred to as soybean seeds but also include, for example, dehulled soybean seeds, defatted soybean seeds (powder), and snow flora (Okara) obtained from the whole soybean seeds. ) Etc.

  The reaction conditions used when the protein is hydrolyzed with a protease are not particularly limited, and can be appropriately selected by those skilled in the art according to common general technical knowledge. For example, when using a commercially available protease, reaction conditions can be selected according to the instructions for use. In general, reaction temperatures of 30-80 ° C, preferably 40-70 ° C, more preferably 50-60 ° C may be used. In general, a reaction time of 2 to 30 hours, preferably 3 to 24 hours, more preferably 10 to 20 hours, particularly preferably 12 to 18 hours may be used. The reaction pH is preferably near the optimum pH of the protease to be used. The means for stopping the reaction is not particularly limited, and known means can be used. Examples of such means include heat treatment such as heating at 85 ° C. for 15 minutes and heating at 100 ° C. for 5 minutes.

  After hydrolysis with protease, the target peptide can be obtained by purification by means known in the art. As such known means, for example, a strongly acidic ion exchange resin, octadecyl silica (ODS) resin, or the like can be used. For example, the peptide of interest can be purified by adsorbing an aqueous peptide solution after protease treatment to an ODS resin and eluting it with an organic solvent of any concentration (for example, acetonitrile). Alternatively, for example, an aqueous peptide solution after protease treatment is adsorbed onto a strongly acidic ion exchange resin, and an eluate having a salt concentration of about 0.18 M to 0.25 M (for example, sodium chloride, potassium chloride, etc.), more preferably about 0. The target peptide can be purified by elution with an eluent having a salt concentration of 20 M to 0.23 M.

  As described above, a peptide obtained by hydrolyzing a natural protein with a protease is more advantageous from the viewpoint of cost than a case where it is produced by a chemical synthesis method. Furthermore, it is considered that a peptide obtained by hydrolyzing a natural protein with a protease is safer for a living body. Therefore, the peptide thus obtained can be suitably used for pharmaceutical compositions, foods, sensitive skin cosmetics, feeds, and the like that require higher safety for application to living bodies.

  The derivative of the peptide of the present invention is prepared by a method known in the art according to a solid phase synthesis method by Fmoc method (LACarpino, GYHan, J. Am. Chem. Soc., 92, 5748 (1970)). obtain.

  Salts of the peptides of the present invention can also be made by those skilled in the art by any method known in the art.

  In the present invention, the specific peptides as described above may be used alone or in any combination of two or more.

  The blending amount of the specific peptides to be blended in the composition of the present invention is not particularly limited, but is usually 0.0001% by weight or more, preferably 0.00025% by weight or more, more preferably 0 based on the whole composition. .0005% by weight or more, more preferably 0.00075% by weight or more, particularly preferably 0.001% by weight or more, and the upper limit of blending is preferably 0.01% by weight or less, more preferably 0.0075% by weight or less, More preferably, it is 0.005 weight% or less, Most preferably, it is 0.0025 weight% or less.

  In addition, the weight ratio of the mucopolysaccharide and the specific peptide in the composition of the present invention is not particularly limited as long as the effect of the present invention can be achieved, but the specific peptide is usually 0 with respect to 1 part by weight of the mucopolysaccharide. .0002 to 20 parts by weight, preferably 0.002 to 5 parts by weight, more preferably 0.02 to 1 part by weight, and still more preferably 0.2 to 0.5 parts by weight.

In another embodiment of the present invention, the composition of the present invention comprises a protease hydrolyzate of a protein containing the specific peptides instead of the specific peptides [hereinafter referred to as “specific peptides”. It may be referred to as "protease degradation product containing".
Protease degradation products containing specific peptides are one of the above-mentioned methods for preparing specific peptides. Among the amino acid sequences of proteins containing the target amino acid sequence, a peptide comprising the target amino acid sequence is treated with protease, etc. It can be prepared by a method of cutting out by a known means. The protease degradation product thus prepared can be directly contained in the composition of the present invention without isolating specific peptides.
As a protease degradation product containing specific peptides, a thermolysin degradation product of soybean protein is preferable from the viewpoint of obtaining a higher collagen or hyaluronic acid production promoting effect.

  The protease degradation product containing the specific peptides obtained by the hydrolysis reaction as described above can be further processed by any method known to those skilled in the art, if necessary. For example, it is preferable to remove large solid particles in the hydrolyzate by a treatment such as filtration. Filtration conditions and the like are not particularly limited and may be appropriately selected by those skilled in the art according to common general technical knowledge. For example, when the filter paper is likely to be clogged, a filter aid or the like can be suitably used.

  Alternatively, the hydrolyzate can be pulverized by concentration under reduced pressure and then freeze-drying. The conditions and equipment used in the concentration under reduced pressure and lyophilization are not particularly limited and can be appropriately selected by those skilled in the art according to common general technical knowledge. The hydrolyzate thus powdered can be used as it is or after being dissolved in a solvent such as water.

  The protease degradation product containing the specific peptides used in the present invention may contain substantially all of a wide variety of peptides produced by hydrolyzing proteins with proteases, or Such a variety of peptides, with the presence or absence of specific peptides as an index, are obtained by further fractionation and purification by known methods (for example, peptides that hardly increase the ability to promote collagen and hyaluronic acid production). Excluding those etc.). However, for convenience, the soy protein is used as it is in a state containing substantially all of a wide variety of peptides obtained by hydrolysis with a protease.

  The average molecular weight of the protease degradation product containing the specific peptides used in the present invention is preferably 300 to 10,000. The average molecular weight is more preferably 400 to 5000, further preferably 500 to 3500, and even more preferably 550 to 3200, from the viewpoint of increasing the permeability to cells and obtaining a higher effect. . Accordingly, the average molecular weight can be, for example, 1000 to 2000. The average molecular weight of the hydrolyzate can be measured by any method known to those skilled in the art. For example, it can be easily measured by the gel permeation chromatography (GPC) method described in the Examples below.

  The amount of the specific peptide contained in the protease degradation product containing the specific peptide used in the present invention is not particularly limited, but is usually 0.01% by weight or more based on the total protease degradation product, preferably 0.025 wt% or more, more preferably 0.05 wt% or more, further preferably 0.075 wt% or more, particularly preferably 0.1 wt% or more, and the upper limit is preferably 1.0 wt% or less, More preferably, it is 0.75 weight% or less, More preferably, it is 0.5 weight% or less, Most preferably, it is 0.25 weight% or less.

  The blending amount of the protease degradation product containing the specific peptides to be blended in the composition of the present invention is not particularly limited, but is usually 0.1% by weight or more, preferably 0.25% by weight, based on the entire composition. Or more, more preferably 0.5% by weight or more, further preferably 0.75% by weight or more, particularly preferably 1.0% by weight or more, and the upper limit of blending is preferably 10% by weight or less, more preferably 7.5% by weight. It is good to set it as weight% or less, More preferably, it is 5 weight% or less, Most preferably, it is 2.5 weight% or less.

  In addition, the weight ratio of the mucopolysaccharide and the protease degradation product containing the specific peptide in the composition of the present invention is not particularly limited as long as the effect of the present invention can be achieved. However, the protein ratio is 1 part by weight of the mucopolysaccharide. The protease degradation product is usually in the range of 0.02 to 20000 parts by weight, preferably 0.2 to 2000 parts by weight, more preferably 2 to 200 parts by weight, and even more preferably 20 to 50 parts by weight.

  Since the degradation of the specific peptides is further accelerated in the presence of sugar or sugar alcohol, the protease degradation product containing the specific peptides or specific peptides is one or more selected from the group consisting of sugar and sugar alcohol [ Hereinafter, in the present specification, when these are collectively referred to as “saccharides”, it is particularly useful to further add mucopolysaccharide.

  As sugars and sugar alcohols, those that can be usually used in the field of pharmaceuticals, quasi drugs, cosmetics or foods can be used without any particular limitation.

  Examples of the sugar used in the present invention include glucose, fructose, maltose, glucose fructose liquid sugar, fructose glucose liquid sugar, high fructose liquid sugar, sucrose (sucrose) and the like.

  The sugar alcohol used in the present invention means a polyhydric alcohol in which the carbonyl group of the sugar is reduced, and specific examples include sorbitol, erythritol, maltitol, reduced palatinose, xylitol, lactitol, mannitol and the like.

In the present invention, saccharides may be used alone or in any combination of two or more.
Moreover, as for saccharides, when there exists a commercial item, you may use a commercial item as it is.

  The amount of the saccharide to be blended in the composition of the present invention is not particularly limited as long as the effect of the present invention can be achieved, but is usually 1% by weight or more, preferably 2.5% by weight or more, more preferably based on the whole composition. Is 5% by weight or more, more preferably 7.5% by weight or more, particularly preferably 10% by weight or more, and the upper limit of blending is preferably 30% by weight or less, more preferably 25% by weight or less, and further preferably 20% by weight. Hereinafter, the content is particularly preferably 15% by weight or less.

  The weight ratio of the mucopolysaccharide and the saccharide in the composition of the present invention is not particularly limited as long as the effect of the present invention can be achieved, but the mucopolysaccharide is usually 0.000017 to 5 parts by weight with respect to 1 part by weight of the saccharide. Preferably it is 0.0017 to 2.5 parts by weight, more preferably 0.017 to 1 part by weight, and still more preferably 0.17 to 0.5 parts by weight.

  The weight ratio of the specific peptide and saccharide in the composition of the present invention is not particularly limited as long as the effect of the present invention can be achieved, but the specific peptide is usually 0.0000033 to 0.01 wt. Parts, preferably 0.000033 to 0.009 parts by weight, more preferably 0.00033 to 0.008 parts by weight, and even more preferably 0.0033 to 0.0067 parts by weight.

  The weight ratio of the protease degradation product containing the specific peptide and the saccharide in the composition of the present invention is not particularly limited as long as the effect of the present invention can be achieved. However, the protease degradation containing the specific peptide with respect to 1 part by weight of the saccharide. The product is usually in the range of 0.0033 to 10 parts by weight, preferably 0.033 to 5 parts by weight, more preferably 0.33 to 2.5 parts by weight, and still more preferably 0.67 to 1 part by weight. It is good.

  In the composition of the present invention, in addition to the above-described mucopolysaccharides and specific proteases or protease degradation products containing the specific peptides, protease degradation containing mucopolysaccharides or specific peptides or specific peptides. Whitening ingredients, anti-inflammatory ingredients, antibacterial ingredients, cell activation ingredients, astringent ingredients, antioxidant ingredients, acne improvement, for the purpose of enhancing or supplementing the action of the product, or to add other useful actions to the composition of the present application Various components such as a component, a biological component synthesis promoting component such as hyaluronic acid, a blood circulation promoting component, a moisturizing component, and an anti-aging component can be used alone or in combination. Preferred are one or more of whitening components, anti-inflammatory components, antibacterial components, cell activation components, astringent components, antioxidant components, anti-aging components or moisturizing components. These components are not particularly limited as long as they can be used in the fields of pharmaceuticals, quasi-drugs, foods, and cosmetics, and arbitrary components can be appropriately selected and used.

  In addition to the above components, the composition of the present invention may be appropriately mixed with components usually used in the field of pharmaceuticals, quasi drugs, cosmetics or foods, depending on the use or dosage form of the composition. Ingredients that can be blended are not particularly limited. For example, amino acids, alcohols (excluding sugar alcohols), polyhydric alcohols (excluding sugar alcohols), high molecular compounds such as gums and polysaccharides (mucopolysaccharides) ), Surfactants, solubilizing components, fats and oils, transdermal absorption promoting components, antiseptic / antibacterial / bactericidal agents, pH adjusting agents, chelating agents, antioxidants, enzyme components, binders, disintegrating agents, lubricants In addition to agents, fluidizing agents and cooling agents, minerals, cell activators, nutrient tonics, excipients, thickeners, stabilizers, preservatives, isotonic agents, dispersants, adsorbents, disintegration Adjuvants, wetting agents or humidity control agents, moisture-proofing agents, coloring agents, flavoring agents or fragrances, fragrances, reducing agents, solubilizers, solubilizing agents, foaming agents, thickening agents or thickening agents, solvents, Base, emulsifier, plasticizer, buffer, brightener, sweetener, acidulant, dietary fiber, fruit juice powder, Machine acids, seasonings, flavor components, swelling agent, bleaching agent, and the like coloring agent.

The composition of the present invention can take a dosage form usually used for pharmaceuticals, quasi drugs, cosmetics or foods, and is usually a solid, semi-solid or liquid. Specifically, tablets (including orally disintegrating tablets, chewable tablets, effervescent tablets, troches, jelly-like drops, coated tablets, chewable tablets, etc.), pills, granules, fine granules, powders, Hard capsules, soft capsules, dry syrups, liquids (including drinks, suspensions, syrups), gels, liposomes, extracts, tinctures, lemonades, ointments, jellys, etc. Can take form. In addition, pastes, mousses, gels, liquids, emulsions, creams, sheets (substrate support), aerosols can be added by blending other solvents and commonly used bases as necessary. It can be prepared in various desired forms such as spray.
These dosage forms can be produced by conventional methods in the art.

The composition of the present invention can be a composition for internal use or a composition for external use.
In addition to pharmaceuticals for internal use (including quasi-drugs), the composition for internal use includes confectionery, beverages, health foods, nutritional supplements (including balanced nutritional foods, supplements, etc.), functional nutritional foods, and special health use Includes food such as food. In food, specifically, milk, milk drink, lactic acid bacteria drink, soft drink with fruit juice, carbonated drink, vegetable juice drink, tea drink, ionic drink, sports drink, functional drink, vitamin supplement drink, nutrition supplement balance drink, Beverages such as jelly beverages, alcoholic beverages, soups; puddings such as custard pudding, milk pudding and pudding with fruit juice; gel foods such as jellys, dressings and creams; gums such as chewing gum and bubble gum (plates) Gum, sugar-coated grain gum, etc.), chocolates (solid chocolate, coated chocolate, chocolate with flavors such as strawberry chocolate, blueberry chocolate, melon chocolate), soft candy (caramel, nougat, gummy candy, marshmallow, etc.) ), Caramels such as toffee, biscuits (Hard biscuits, soft biscuits, soft cookies, crackers, semi-biscuit biscuits, etc.), cakes and other confectionery; ice cream, ice milk, lacto ice, sorbet, frozen confectionery such as ice confectionery; breads; noodles; liquid (water, milk , Juice, etc.) can be easily formed into beverages, such as powdered beverages, pastes, tablets and the like, foods for re-formation; supplements having the form of powders, capsules, tablets and the like.

  Moreover, as an external composition, cosmetics are contained in addition to external medicine (including quasi drugs). In cosmetics, specifically, makeup cosmetics such as foundation, lipstick, mascara, eye shadow, eyeliner, eyebrows and beauty nails; milky lotion, cream (including massage cream, pack cream, etc.), lotion, cosmetic liquid, Basic cosmetics such as oils and packs; cleansing agents such as facial cleansers, cleansing and body cleaning agents, bathing agents, and the like.

  The composition of the present invention comprises a therapeutic or preventive agent for a disease caused by a decrease in the amount of collagen and / or hyaluronic acid; a food for improving or preventing a condition caused by a decrease in the amount of collagen and / or hyaluronic acid; Cosmetics for prevention and / or improvement of skin wrinkles or talmi due to exposure, aging, etc .; cosmetics for prevention and / or improvement of skin elasticity or reduction of elasticity; etc. it can.

  EXAMPLES The present invention will be described in detail below based on examples, but the present invention is not limited to these examples.

Example 1
Preparation of LEHA peptide The peptide was synthesized by a solid phase synthesis method by the Fmoc method using an automatic peptide synthesizer (manufactured by Shimadzu Corporation: PSSM8). Subsequently, LEHA peptide was obtained by purifying by removing unreacted substances by preparative HPLC.
The purified product thus obtained was subjected to reverse phase high performance liquid chromatography for analysis [column: μBondasphere 5 μC18-100Å (inner diameter: 3.9 mm, length: 150 mm), manufactured by Waters; mobile phase: solvent A (0.1% Tri Fluoroacetic acid) and solvent B (0.1% trifluoroacetic acid, 90% acetonitrile) gradient (0 min (solvent B = 12%) to 20 min (solvent B = 17%)); flow rate: 1 ml / min; Detection method: Absorbance at a wavelength of 220 nm] showed a single sharp peak at 12.8 minutes and a purity of 99%.

Example 2
Preparation of Thermolysin Degradation Product of Soy Protein 50 g of defatted soybean powder (trade name Profam 974, manufactured by ADM Far East Co., Ltd.) was dispersed in 2 L of distilled water and adjusted to pH 8.5 with 0.1 N NaOH. 500 mg of thermolysin (product name “Samoase PC10F”, manufactured by Daiwa Kasei Co., Ltd.) was added thereto, and decomposition was performed at 60 ° C. for 15 hours. After the reaction, thermolysin was inactivated by boiling at 100 ° C. for 10 minutes. After allowing to cool, 25 g of a filter aid (Radiolite 500, Showa Chemical Industry Co., Ltd.) was added and stirred, followed by filtration.

  The filtrate obtained as described above was passed through a column packed with a strongly acidic ion exchange resin (trade name “Dowex 50W × 2, H + form, 50-100 mesh”, manufactured by Dow Chemical Company), and then 5 times the column. Wash with a volume of deionized water to remove non-peptide components. A 2M ammonia solution was passed through to elute the column adsorbing component, and the peptide fraction was collected. Ammonia was removed using an evaporator and further concentrated to dryness. Water was added thereto to dissolve the dried solid, and then centrifuged (10,000 rpm, 30 minutes) to remove insoluble matters. As a result of freeze-drying the supernatant, about 26 g of soybean protein thermolysin hydrolyzate was finally obtained as a thermolysin hydrolyzate of soybean protein.

  The average molecular weight of the decomposition product thus obtained was measured by the GPC method. 100 mg of a thermolysin degradation product of soybean protein after lyophilization was dissolved in 2.0 ml of 0.1 M sodium phosphate buffer (pH 7.0) to prepare a test solution. A column (φ2.6 × 100 cm) packed with Sephadex G25 (Medium type, manufactured by Amersham Biosciences) was equilibrated with the same 0.1 M sodium phosphate buffer (pH 7.0). The column was loaded with 2.0 ml of the test solution and eluted at a flow rate of 1.0 ml / min. Insulin (bovine pancreas origin, Sigma, molecular weight 5733), Insulin A chain (bovine pancreas origin, Sigma, molecular weight 2532), and Bradykinin (Sigma, molecular weight 1050) are used as peptide preparations with known molecular weights. It was. Peptides were detected at 214 nm, and the molecular weight distribution and average molecular weight were estimated from the elution time. As a result, the average molecular weight of the thermolysin degradation product of soybean protein obtained by the above method was estimated to be about 1500.

The LEHA peptide content of the thermolysin degradation product of soybean protein obtained by the above method was measured by a liquid chromatography method (measurement conditions are as described in Example 1 above).
As a result, it was estimated that the content of LEHA peptide in about 26 g of the thermolysin degradation product of soybean protein obtained by the above method was about 30 mg.

Example 3
Evaluation of the stability of LEHA peptide
1. Preparation of test solution According to the formulation described in Table 2, the thermolysin degradation product (average molecular weight 1500), hyaluronic acid (trade name: hyaluronic sun HA-F, manufactured by Kewpie Co., Ltd.) or chondroitin obtained in Example 2 Sodium sulfate (manufactured by Maruha Co., Ltd.) and saccharides were dissolved in purified water and the total amount was adjusted, and then the pH was adjusted with citric acid to prepare each test solution.

2. For LEHA peptide stability evaluation test examples and comparative examples 6-7, the residual ratio of the LEHA peptide after heating at 121 ° C. for 20 minutes is shown as 125 ° C. for test examples / comparative examples 1-4 and 8-10. The residual rate of LEHA peptide after heating for hours was measured, and in Test Example and Comparative Example 5, the residual rate of LEHA peptide after storage at 60 ° C. for 20 days was measured.
Specifically, the content of the LEHA peptide in each test solution was quantified by liquid chromatography (measurement conditions are as described in Example 1 above). Table 3 shows the residual ratio (%) of the LEHA peptide after the heat treatment with respect to immediately after the preparation. Moreover, the decomposition inhibition rate (%) was calculated by the following formula.
Decomposition suppression rate (%) = (1− (100−A) / (100−A ′)) × 100
A: Residual rate of test example A ': Residual rate of comparative example

  From the above results, it can be seen that the degradation of the LEHA peptide is remarkably suppressed by incorporating mucopolysaccharides such as hyaluronic acid and sodium chondroitin sulfate.

  Formulation examples are given below, but the present invention is not limited to these examples.

Formulation Example 1: Emulsion [Ingredient] [Ratio]
Hyaluronic acid 0.1
Thermolysin degradation product of soybean protein (average molecular weight 1500) 0.5
Squalane 2.0
Liquid paraffin 5.0
Cetanol 0.5
Glyceryl monostearate 2.0
POE (25) cetyl ether 2.0
Triethanolamine 0.8
Glycerin 4.0
1,3-butylene glycol 6.0
Preservative (Methylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water
100.0% by weight

Formulation Example 2: Cream [Ingredient] [Ratio]
Acetylated sodium hyaluronate 0.3
Soybean protein thermolysin degradation product (average molecular weight 1500) 1.0
Vaseline 1.0
Squalane 5.0
Liquid paraffin 10.0
Stearic acid 1.5
Stearyl alcohol 2.0
Glyceryl monostearate 2.0
POE (20) cetyl ether 3.0
Triethanolamine 1.0
Glycerin 6.0
1,3-butylene glycol 8.0
Preservative (Ethylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water
100.0% by weight

Formulation Example 3: Lotion [Ingredient] [Ratio]
Acetylated hyaluronic acid 0.05
Thermolysin degradation product of soybean protein (average molecular weight 1500) 0.5
POE (20) sorbitan monoisostearate 0.3
Succinic acid 0.2
Sodium succinate 0.5
Edetate trisodium 0.05
1,3-butylene glycol 6.0
Preservative (Methylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water
100.0% by weight

Formulation Example 4: Soft capsule [Ingredient] [Ratio]
Sodium hyaluronate 1.0
Thermolysin degradation product of soybean protein (average molecular weight 1500) 7.5
Melon extract 5.0
Ceramide-containing material 1.0
Haematococcus alga pigment extract (containing astaxanthin) 1.0
Gelatin 30.0
Glycerin 15.0
Safflower oil
100.0% by weight

Formulation Example 5: Beverages [Ingredients] [Ratio]
Sodium hyaluronate 0.01
Soybean protein thermolysin degradation product (average molecular weight 1500) 2.0
Royal Jelly Extract 1.0
Lemon juice 1.0
Vitamin B2 0.005
Vitamin B6 0.05
Vitamin C 0.5
Erythritol 5.0
Reduced maltose starch syrup 5.0
Stabilizer (pectin) 0.2
Sweetener (acesulfame potassium, sucralose) 0.008
Calcium lactate 0.05
Magnesium chloride 0.015
Potassium chloride 0.015
Perfume 0.025
Preservatives (sodium benzoate, butylparaben) appropriate amount acidulant appropriate amount
(PH adjusted to 3.5)
Purified water remaining
100.0% by weight

Formulation Example 6: Beverages [Ingredients] [Ratio]
Soybean protein thermolysin degradation product (average molecular weight 1500) 2.0
Shark cartilage extract (contains 40% chondroitin sulfate) 0.8
Methylsulfonylmethane 2.0
Fish collagen 0.1
Glucosamine 1.0
Honey 6.0
Maltitol 4.0
Calcium gluconate 0.3
Sweetener (acesulfame potassium, sucralose) 0.02
Preservatives (sodium benzoate, butylparaben) 0.05
Antioxidant (Berberry extract, enzyme-treated rutin) 0.04
Vitamin B2 0.005
Perfume 0.025
Sour seasoning
(Adjust pH to 3.6)
Purified water remaining
100.0% by weight

Formulation Example 7: Beverages [Ingredients] [Ratio]
Sodium hyaluronate 0.01
LEHA peptide 0.002
Haematococcus alga pigment extract (containing astaxanthin) 0.15
Grapefruit juice 2.0
Vitamin C 0.5
Erythritol 8.0
Water soluble dietary fiber 5.0
Stabilizer (soybean polysaccharide) 0.2
Sweetener (sucralose) 0.010
Perfume 0.020
Sour seasoning
Adjust pH to 3.0)
Purified water remaining
100.0% by weight

Formulation Example 8: Lotion [Ingredient] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 0.5
Sodium hyaluronate 0.2
1,3-butylene glycol 30.0
D-sorbitol 5.0
(Glycerylamidoethyl methacrylate /
Stearyl methacrylate) copolymer 0.5
Polyoxyethylene hydrogenated castor oil 50 0.4
Preservative (Methylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water remaining
100.0% by weight

Formulation Example 9: Lotion [Ingredient] [Ratio]
Soybean protein thermolysin degradation product (average molecular weight 1500) 1.0
Acetylated sodium hyaluronate 0.5
Whey / BG mixture 2.0
Polyethylene glycol 1500 0.4
Seaweed extract 0.05
2-Methacryloyloxyethyl phosphorylcholine
Butyl methacrylate copolymer (BG) 0.5
Preservative (Methylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water remaining
100.0% by weight

Formulation Example 10: Emulsion [Ingredient] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 1.5
Sodium hyaluronate 0.5
Concentrated glycerin 10.0
Sodium deoxyribonucleic acid 0.5
Liquid paraffin 15.0
Squalane 10.0
Polysorbate 60 4.0
Glycerol monostearate 1.0
Preservative (Methylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water remaining
100.0% by weight

Formulation Example 11: Cream [Ingredient] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 0.4
Hyaluronic acid 0.2
Medium chain fatty acid triglycerides 6.0
Behenyl alcohol 2.5
Aloe extract 0.005
Oat extract 0.01
Glycerol monostearate 1.2
Preservative (Ethylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water remaining
100.0% by weight

Formulation Example 12: Cream [Ingredient] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 0.8
LEHA peptide 0.002
Acetylated sodium hyaluronate 0.5
Concentrated glycerin 8.0
Squalane 6.0
Tori (Capryl / Caprin / Mistillin / Stearic acid)
Glycerides 0.7
Glucosamine hydrochloride 0.1
Kinetin 0.02
Polyoxyethylene hydrogenated castor oil 60 0.2
(Hydroxyethyl acrylate /
Acryloyldimethyltaurine Na) copolymer 1.5
Preservative (Methylparaben, Propylparaben) Appropriate amount Perfume Appropriate amount
Purified water remaining
100.0% by weight

Formulation Example 13: Soft capsule [Ingredient] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 5.0
Acetylated sodium hyaluronate 2.0
Elastin 1.0
Lycopene 2.0
Evening primrose oil 4.0
Gelatin 35.0
Glycerin 10.0
Titanium dioxide 0.5
Safflower oil
100.0% by weight

Formulation Example 14: Beverages [Ingredients] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 2.5
Hyaluronic acid 0.75
Royal Jelly 0.5
Vitamin C 0.5
Vitamin B1 0.01
Vitamin B6 0.05
Rose petal extract 0.25
Sweetener (sucralose) 0.05
Citric acid 0.5
Trisodium citrate 0.02
Perfume
Purified water remaining
100.0% by weight

Formulation Example 15: Tablet [Ingredient] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 5.0
Sodium hyaluronate 5.0
Glutathione-containing yeast 7.0
Cysteine peptide 3.0
Vitamin B2 0.5
Vitamin B6 2.0
Calcium stearate 1.0
Crystalline cellulose
100.0% by weight

Formulation Example 16: Coated tablets [Components] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 0.1
Sodium hyaluronate 0.2
Vitamin E 1.5
Lipoic acid 1.0
Coenzyme Q10 1.2
Sucrose fatty acid ester 3.0
Crystalline cellulose 35.0
Shellac 3.0
Lactose remaining
100.0% by weight

Formulation Example 17: Chewable tablets [ingredients] [ratio]
Soybean protein thermolysin degradation product (average molecular weight 1500) 1.0
Hyaluronic acid 0.1
Collagen peptide 5.0
Elastin 3.0
Sweetener 0.1
Perfume Trace amount Magnesium stearate 0.5
Xylitol 30.0
Mannitol remaining
100.0% by weight

Formulation Example 18: Granules [Ingredients] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 3.0
Acetylated sodium hyaluronate 0.5
Calcium pantothenate 5.0
Rutin 0.3
Vitamin C 0.1
Sweetener 0.1
Citric acid 0.3
Fragrance Trace amount Lactose 35.0
Crystalline cellulose
100.0% by weight

Formulation Example 18: Carbonated beverage [Ingredient] [Ratio]
Thermolysin degradation product of soybean protein (average molecular weight 1500) 5.0
Sodium hyaluronate 1.0
Swallow's nest extract 0.2
Alginic acid 0.5
L-leucine 0.15
L-Valine 0.10
L-isoleucine 0.12
Fructose glucose liquid sugar 9.0
Erythritol 5.0
Malic acid 0.3
Citric acid 0.3
Perfume
Purified water remaining
100.0% by weight

  SEQ ID NO: 1 is a peptide that can be used in the present invention.

Claims (5)

(A) one or more selected from mucopolysaccharides;
(B) A composition containing one or more selected from the group consisting of a peptide represented by Formula I: Leu-Glu-His-Ala, derivatives thereof, and salts thereof. (A) one or more selected from mucopolysaccharides;
(B) A composition containing a protein protease hydrolyzate containing at least one selected from the group consisting of a peptide represented by the formula I: Leu-Glu-His-Ala, derivatives thereof and salts thereof .   The composition of claim 2, wherein the protein is soy protein.   The composition according to claim 2 or 3, wherein the protease is thermolysin.   Furthermore, the composition as described in any one of Claims 1-4 containing 1 or more types selected from the group which consists of saccharide | sugar and sugar alcohol.