Collagen production promoter

Abstract

The objective is to provide a collagen production promoter that can efficiently promote collagen production. [Solution] A collagen production promoter containing a 3-hydroxypyridinium derivative represented by the following general formula (1) as an active ingredient. TIFF2026076348000020.tif49159 In the formula, A1 independently represents a hydroxyl group, an alkoxy group having 1 to 10 carbon atoms, a peptide structure having 1 to 100 amino acids linked at the N-terminus, a hydrogen atom, an amino group, or a hydrocarbon group having 1 to 10 carbon atoms, while A2 independently represents a hydrogen atom, a hydrocarbon group having 1 to 10 carbon atoms, an acyl group, or a peptide structure having 1 to 100 amino acids linked at the C-terminus.

Description

[Technical Field]

[0001] The present invention relates to a collagen production promoter, and to cosmetic compositions, food and beverage compositions, cell culture compositions, wound healing pharmaceutical compositions, and bone formation promoting compositions containing the collagen production promoter. [Background technology]

[0002] Collagen is present in all multicellular animals and is the most abundant protein in vertebrates, existing extracellularly. It is known to act as a major component that resists strain forces in connective tissues such as bones, teeth, cartilage, tendons, ligaments, skin, and the fibrous structure of blood vessels, forming insoluble fibers that are extremely resistant to tension.

[0003] Furthermore, while it is known that collagen consists of three polypeptide chains, with each peptide chain being crosslinked by a compound having a 3-hydroxypyridinium structure, there have been no reports whatsoever of its degradation products, 3-hydroxypyridinium derivatives, exhibiting physiological effects via receptors.

[0004] Furthermore, collagen is an important component for skin elasticity and moisturizing function, and a decrease in collagen levels is known to cause a decline in skin firmness and elasticity, as well as the appearance of aging-like symptoms such as rough skin and wrinkles. For this reason, research has been conducted on food ingredients, cosmetic ingredients, and pharmaceutical compositions that increase the amount of collagen in the skin, with the aim of maintaining skin health or suppressing skin aging.

[0005] On the other hand, bone metabolism is normally maintained by a balance between the activity of osteoclasts, which break down old bone, and osteoblasts, which create new bone. It is known that calcium phosphate is deposited around type I collagen fibers secreted by osteoblasts to form hydroxyapatite, which is how bone is formed.

[0006] In recent years, with the aging population and the increase in underlying diseases such as diabetes, there has been a growing trend of patients with various bone diseases caused by decreased bone mass and bone quality, such as fractures and osteoporosis. As mentioned above, bone mass and bone quality are maintained in healthy individuals by a balance between bone resorption (bone breakdown) and bone formation. However, it is known that bone resorption becomes dominant due to factors such as aging and decreased hormone secretion due to menopause, leading to brittle bones. In response to this, treatments such as administering oral calcium preparations, vitamin D3 preparations, and parathyroid hormone preparations to suppress the activity of osteoclasts are commonly implemented.

[0007] Regarding collagen production promoters, for example, Patent Document 1 describes compounds represented by 1-acetoxy-1-(2,4-diacetoxyphenyl)-2-propene. Patent Document 2 describes an anti-aging topical skin preparation characterized by containing a synthetic peptide or derivative thereof composed of 3 to 8 amino acids and hydrolyzed collagen or a derivative thereof having a molecular weight in the range of 500 to 5000. Furthermore, pyridinoline, a representative compound of the present invention, is a known collagen-degrading component, as described in Non-Patent Document 1, and is used as a diagnostic agent for osteoporosis, as described in Patent Documents 3 and 4. [Prior art documents] [Patent Documents]

[0008] [Patent Document 1] Japanese Patent Publication No. 2009-79004 [Patent Document 2] Japanese Patent Publication No. 2011-042613 [Patent Document 3] Special Publication No. 05-502942 [Patent Document 4] Japanese Patent Application Publication No. 05-209870 [Non-patent literature]

[0009] [Non-Patent Document 1] Daisaburo Fujimoto, Takahiko Moriguchi, Torao Ishida, and Hiroshi Hayashi:'The structure of pyridinoline, a collagen crosslink.'Biochem. Biophysics. Res. Commun. 84, 52-57 (1978). [Overview of the project] [Problems that the invention aims to solve]

[0010] While commonly used treatments for bone diseases such as fractures and osteoporosis involve the administration of oral calcium preparations, vitamin D3 preparations, and parathyroid hormone preparations, these methods have side effects such as loss of appetite, general fatigue, and osteonecrosis of the jaw. Therefore, there has been a need for treatments that directly act on osteoblasts to increase collagen synthesis and promote bone formation. Furthermore, even with commercially available collagen peptides, large quantities are required to experience their effects, highlighting the need for ingredients that can efficiently promote collagen production.

[0011] The present invention aims to provide a novel collagen production promoter that can efficiently promote collagen production. Furthermore, it aims to provide cosmetic compositions, food and beverage compositions, cell culture compositions, wound healing pharmaceutical compositions, and bone formation promoting compositions containing the collagen production promoter. [Means for solving the problem]

[0012] As a result of diligent research to solve the above problems, the inventors of the present invention discovered that 3-hydroxypyridinium derivatives have the ability to promote collagen production, and thus completed the present invention. In other words, the present invention relates to the following collagen production promoters, cosmetic compositions, food and beverage compositions, cell culture compositions, wound healing pharmaceutical compositions, bone formation promoting compositions, and collagen degradation products.

[0013] The collagen production promoter of the present invention for solving the above problems is characterized by containing a 3-hydroxypyridinium derivative represented by the following general formula (1) as an active ingredient.

Chemical formula

Chemical formula

[0014] Furthermore, according to one embodiment of the collagen production promoter of the present invention, the 3-hydroxypyridinium derivative is characterized by containing 0.00001% by weight or more. This characteristic allows for the effective promotion of collagen production.

[0015] Furthermore, the collagen production promoter of the present invention is characterized by containing a 3-hydroxypyridinium derivative derived from a natural product. This characteristic allows for the acquisition of 3-hydroxypyridinium derivatives from natural products, eliminating the need for impurity removal such as catalysts, thus providing the advantage of easily obtaining pure materials.

[0016] Furthermore, according to one embodiment of the collagen production promoter of the present invention, it is characterized by being a cosmetic composition. This characteristic allows for the promotion of collagen production in skin that has lost firmness and elasticity due to a decrease in collagen content, thereby more effectively demonstrating the effects of the present invention, such as contributing to maintaining skin health or suppressing skin aging.

[0017] Furthermore, according to one embodiment of the collagen production promoter of the present invention, it is characterized by being a food and beverage composition. This characteristic allows for easy promotion of collagen production through oral administration, contributing to maintaining skin health, inhibiting skin aging, preventing osteoporosis, and further enhancing the effects of the present invention, such as contributing to extending the lifespan of pets and other animals.

[0018] Furthermore, according to one embodiment of the collagen production promoter of the present invention, it is characterized by being a cell culture composition. This characteristic allows for the simple promotion of collagen production in regenerative medicine and other applications, and further demonstrates the effects of the present invention, such as contributing to alternative testing to animal experiments.

[0019] Furthermore, according to one embodiment of the collagen production promoter of the present invention, it is characterized as a pharmaceutical composition for wound healing. This characteristic allows for the easy promotion of collagen production in regenerative medicine and other applications, thereby more effectively demonstrating the benefits of the present invention, such as contributing to skin regeneration in the treatment of lacerations and burns.

[0020] Furthermore, according to one embodiment of the collagen production promoter of the present invention, it is characterized by being a composition for promoting bone formation. This characteristic allows for a simple promotion of collagen production in the treatment of osteoporosis and other conditions, thereby more effectively demonstrating the benefits of the present invention, which contributes to the promotion of bone formation.

[0021] Furthermore, the present invention is a collagen hydrolysate characterized by containing 0.015% by weight or more of a 3-hydroxypyridinium substructure represented by the following general formula (3). [ka] This characteristic allows for the effective promotion of collagen production.

[0022] Furthermore, the present invention is a collagen hydrolysate characterized by containing 0.07% by weight or more of 3-hydroxypyridinoline represented by the following general formula (4). [ka] This characteristic allows for the effective promotion of collagen production. [Effects of the Invention]

[0023] According to the present invention, collagen production can be promoted simply, easily, and efficiently, and it can be used for maintaining skin health, suppressing skin aging, and treating conditions such as osteoporosis. [Brief explanation of the drawing]

[0024] [Figure 1] Purity testing of purified PYR by reverse-phase HPLC [Figure 2] Structural identification of PYR by mass spectral analysis [Figure 3] Results of RNA expression analysis of type I collagen type A in human tubular epithelial cells [Figure 4] TGFβ1 RNA expression level analysis results for type I collagen type A [Figure 5] Data demonstrating the osteoblast differentiation-promoting effect of CHHP [Figure 6] RT-PCR data on the effect of CHHP on promoting osteoblast collagen production. [Modes for carrying out the invention]

[0025] In describing the details of the present invention, specific examples will be given, but the invention is not limited to the following, and can be implemented with appropriate modifications, as long as it does not deviate from the spirit of the invention.

[0026] [Collagen production promoter] The collagen production promoter of the present invention is characterized by containing a 3-hydroxypyridinium derivative represented by the following general formula (1) as an active ingredient. [ka] (In formula (1), A 1 Each of these independently consists of a hydroxyl group, an alkoxy group with 1 to 10 carbon atoms, a peptide structure with 1 to 100 amino acids linked at the N-terminus, a hydrogen atom, and an amino group (-NR). 4 2) Or a hydrocarbon group having 1 to 10 carbon atoms, A 2 Each of these independently consists of a hydrogen atom, a hydrocarbon group with 1 to 10 carbon atoms, and an acyl group (-COR). 5 ), or a peptide structure with 1 to 100 amino acids linked at the C-terminus, where R is independently a hydrogen atom or a hydrocarbon group with 1 to 10 carbon atoms, 1 Each of these is independently a single bond, or a hydroxyl group, an amino group (-NR) 4 2) A divalent hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of and an oxa group (-O-), R 2 These are independently a hydroxyl group, a carboxyl group, and an amino group (-NR) 4 A hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of 2), R 3 These are a hydrogen atom, a structure represented by the following formula (2), or a hydroxyl group, a carboxyl group, and an amino group (-NR 4 A hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of 2), R 4 Each independently contains a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms, R 5(where m represents a hydrocarbon group with 1 to 10 carbon atoms, and m and n are independent integers from 0 to 4. However, m and n are values ​​such that m + n is an integer from 0 to 4.) [ka] (In formula (2), A 1 A in equation (1) 1 Independently, a hydroxyl group, an alkoxy group with 1 to 10 carbon atoms, a peptide structure with 1 to 100 amino acids linked at the N-terminus, a hydrogen atom, and an amino group (-NR) 4 2) Or a hydrocarbon group having 1 to 10 carbon atoms, A 2 A in equation (1) 2 Independently, hydrogen atoms, hydrocarbon groups with 1 to 10 carbon atoms, and acyl groups (-COR 5 ), or a peptide structure with 1 to 100 amino acids linked at the C-terminus, where R is independently a hydrogen atom or a hydrocarbon group with 1 to 10 carbon atoms in formula (1), 4 R in equation (1) 4 Each of them independently contains a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms, R 5 R in equation (1) 5 Independently, a hydrocarbon group with 1 to 10 carbon atoms, R 6 is a single bond, or a hydroxyl group and an amino group (-NR 4 (This represents a divalent hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of (2)). This collagen production promoter allows for the simple, easy, and efficient promotion of collagen production.

[0027] [Collagen production mechanism] Although the specific mechanism of collagen production in this invention has not been elucidated, it is thought that collagen production is promoted when the 3-hydroxypyridinium derivative of this invention is recognized by the Receptor for Advantage Glycation End Product (RAGE).

[0028] Advanced glycation end product receptors (RAGE) are proteins expressed in a wide range of cells, including smooth muscle cells, hepatocytes, nerve cells, vascular endothelial cells, and monocytes. As exemplified by Fischer's lock-and-key model, the ligand-binding site of a protein is thought to have a structure that is firmly complementary to the ligand. Therefore, if the ligand has a 3-hydroxypyridinium substructure, it is thought to be selectively recognized by the advanced glycation end product receptor (RAGE) without being affected by steric hindrance from substituents, etc.

[0029] [3-Hydroxypyridinium derivatives] The 3-hydroxypyridinium derivative used in the present invention is represented by the following formula (1). [ka] (In formula (1), A 1 Each of these independently consists of a hydroxyl group, an alkoxy group with 1 to 10 carbon atoms, a peptide structure with 1 to 100 amino acids linked at the N-terminus, a hydrogen atom, and an amino group (-NR). 4 2) Or a hydrocarbon group having 1 to 10 carbon atoms, A 2 Each of these independently consists of a hydrogen atom, a hydrocarbon group with 1 to 10 carbon atoms, and an acyl group (-COR). 5 ), or a peptide structure with 1 to 100 amino acids linked at the C-terminus, where R is independently a hydrogen atom or a hydrocarbon group with 1 to 10 carbon atoms, 1 Each of these is independently a single bond, or a hydroxyl group, an amino group (-NR) 4 2) A divalent hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of and an oxa group (-O-), R 2 These are independently a hydroxyl group, a carboxyl group, and an amino group (-NR) 4 A hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of 2), R 3 These are a hydrogen atom, a structure represented by the following formula (2), or a hydroxyl group, a carboxyl group, and an amino group (-NR 4A hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of 2), R 4 Each independently contains a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms, R 5 (where m represents a hydrocarbon group with 1 to 10 carbon atoms, and m and n are independent integers from 0 to 4. However, m and n are values ​​such that m + n is an integer from 0 to 4.) [ka] (In formula (2), A 1 A in equation (1) 1 Independently, a hydroxyl group, an alkoxy group with 1 to 10 carbon atoms, a peptide structure with 1 to 100 amino acids linked at the N-terminus, a hydrogen atom, and an amino group (-NR) 4 2) Or a hydrocarbon group having 1 to 10 carbon atoms, A 2 A in equation (1) 2 Independently, hydrogen atoms, hydrocarbon groups with 1 to 10 carbon atoms, and acyl groups (-COR 5 ), or a peptide structure with 1 to 100 amino acids linked at the C-terminus, where R is independently a hydrogen atom or a hydrocarbon group with 1 to 10 carbon atoms in formula (1), 4 R in equation (1) 4 Each of them independently contains a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms, R 5 R in equation (1) 5 Independently, a hydrocarbon group with 1 to 10 carbon atoms, R 6 is a single bond, or a hydroxyl group and an amino group (-NR 4 (This represents a divalent hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of (2)).

[0030] In the present invention, the 3-hydroxypyridinium derivative represented by general formula (1) may include not only the compound itself, but also its isomers, mixtures thereof, pharmaceutically acceptable salts, solvates, and prodrugs, as is common technical knowledge. Examples of prodrugs include those described in Japanese Patent Publication No. 2016-519659. The prodrug structure relating to the hydroxyl group is not particularly limited, but includes esters such as acyloxyalkyl (e.g., acyloxymethyl, acyloxyethyl) esters, alkoxycarbonyloxyalkyl esters, alkyl esters, aryl esters, phosphate esters, sulfonic acid esters, sulfate esters, and disulfides containing esters, ethers (e.g., alkyl ethers), amides, carbamates, hemisuccinates, dimethylaminoacetates, and phosphorylmethylcarbonyl.

[0031] A in equations (1) and (2) 1 The phrase "alkoxy group having 1 to 10 carbon atoms" means that the hydrocarbon group in the alkoxy group may have a branched structure, a cyclic structure, or a carbon-carbon unsaturated bond (carbon-carbon double bond, carbon-carbon triple bond), and may be any of saturated hydrocarbon groups, unsaturated hydrocarbon groups, aromatic hydrocarbon groups, etc. The number of carbon atoms in the alkoxy group is usually 6 or more when the hydrocarbon group of the alkoxy group is an aromatic hydrocarbon group. On the other hand, when the hydrocarbon group of the alkoxy group is not an aromatic hydrocarbon group, it is preferably 8 or less, more preferably 6 or less, and may also be 3 or less. Specific examples of "alkoxy groups with 1 to 10 carbon atoms" include methoxy group, ethoxy group, n-propoxy group, i-propoxy group, n-butoxy group, i-butoxy group, s-butoxy group, t-butoxy group, n-pentyloxy group, 2,2-dimethylpropoxy group, cyclopentyloxy group, n-hexyloxy group, cyclohexyloxy group, n-heptyloxy group, 2-methylpentyloxy group, n-octyloxy group, 2-ethylhexyloxy group, n-nonyloxy group, n-decyloxy group, phenyloxy group, vinyloxy group, etc.

[0032] A in equations (1) and (2) 1 , A 2 , R, R 4 , R 5 The term "carbon hydrocarbon group having 1 to 10 carbon atoms" means that it may have a branched structure, a cyclic structure, and a carbon-carbon unsaturated bond (carbon-carbon double bond, carbon-carbon triple bond), and may be any of saturated hydrocarbon groups, unsaturated hydrocarbon groups, aromatic hydrocarbon groups, etc. The number of carbon atoms in the hydrocarbon group is usually 6 or more when the hydrocarbon group is an aromatic hydrocarbon group. On the other hand, when the hydrocarbon group is not an aromatic hydrocarbon group, it is preferably 8 or less, more preferably 6 or less, and may also be 3 or less. Specific examples of "carbon hydrocarbon groups having 1 to 10 carbon atoms" include methyl group, ethyl group, n-propyl group, i-propyl group, cyclopropyl group, n-butyl group, i-butyl group, s-butyl group, t-butyl group, 2,2-dimethyl-1-propyl group, cyclopentyl group, n-hexyl group, cyclohexyl group, n-heptyl group, 2-methylpentyl group, n-octyl group, 2-ethylhexyl group, n-nonyl group, n-decyl group, phenyl group, vinyl group, and the like.

[0033] R in equation (1) 1 of "hydroxyl group, amino group (-NR 4 2) A divalent hydrocarbon group having 1 to 10 carbon atoms which may have at least one functional group selected from the group consisting of an oxa group (-O-)" means a hydrocarbon group having two bonding sites. 4 2) The phrase "may have at least one functional group selected from the group consisting of and an oxa group (-O-)" means that the hydrogen atoms of the hydrocarbon group may be substituted with hydroxyl groups or amino groups, and that an oxa group may be introduced into the carbon skeleton of the hydrocarbon group. The hydrocarbon may preferably be 8 or less, more preferably 6 or less, and also 3 or less. Specific examples of "divalent hydrocarbon groups having 1 to 10 carbon atoms that may have at least one functional group selected from the group consisting of hydroxyl groups, amino groups, and oxa groups (-O-)" include methylene groups, oxymethylene groups, ethylene groups, dimethyl ether groups, propylene groups, butylene groups, hydroxymethylene groups, hydroxyethylene groups, 1-hydroxypropanylene groups, 2-hydroxypropanylene groups, hydroxybrene groups, aminomethylene groups, aminoethylene groups, 1-aminopropanyl groups, 2-aminopropanyl groups, aminobutylene groups, vinylene groups, and the like.

[0034] R in equation (2) 6 of "hydroxyl group and amino group (-NR 4 The phrases "may have at least one functional group selected from the group consisting of hydroxyl groups and amino groups" and "a divalent hydrocarbon group having 1 to 10 carbon atoms" in "a divalent hydrocarbon group having at least one functional group selected from the group consisting of 2)" are the same as those described above. Specific examples of "divalent hydrocarbon groups having 1 to 10 carbon atoms that may have at least one functional group selected from the group consisting of hydroxyl groups and amino groups" include methylene group, ethylene group, propylene group, butylene group, hydroxymethylene group, hydroxyethylene group, 1-hydroxypropanylene group, 2-hydroxypropanylene group, hydroxybrene group, aminomethylene group, aminoethylene group, 1-aminopropanyl group, 2-aminopropanyl group, aminobutylene group, vinylene group, etc.

[0035] A in equations (1) and (2) 1 The phrase "peptide structures with 1 to 100 amino acids linked at the N-terminus" means that it includes not only peptide structures composed of two or more amino acids, but also amino acid structures consisting of one amino acid. Furthermore, "linked at the N-terminus" means A 1 This means that the carbonyl group (>C=O) to which it is attached is bonded at the N-terminus of the peptide structure or at the amino group of the amino acid structure. Specific examples of an amino acid structure include essential amino acid structures such as glycine, alanine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, serine, threonine, asparagine, glutamine, tyrosine, cysteine, lysine, arginine, histidine, aspartic acid, and glutamic acid structures; modified amino acid structures such as hydroxylysine, allicin, hydroxyallicin, and hydroxyproline structures; and amino acid structures included in collagen. Particularly preferred are glycine, proline, hydroxyproline, lysine, hydroxylysine, allicin, and hydroxyallicin structures. When the "peptide structure with 1 to 100 amino acids linked at the N-terminus" is a peptide structure composed of 2 or more amino acids, the number of amino acids is preferably 50 or less, more preferably 20 or less, even more preferably 10 or less, and most preferably 3 or less. Examples of amino acids that make up a peptide structure composed of two or more amino acids include essential amino acids such as glycine, alanine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, serine, threonine, asparagine, glutamine, tyrosine, cysteine, lysine, arginine, histidine, aspartic acid, and glutamic acid, as well as denatured amino acids such as hydroxylysine, allicin, hydroxyallicin, and hydroxyproline, and amino acids included in the structure of collagen.

[0036] A in equations (1) and (2) 2 The phrase "peptide structures with 1 to 100 amino acids linked at the C-terminus" means that it includes not only peptide structures composed of two or more amino acids, but also amino acid structures consisting of one amino acid. Furthermore, "linked at the C-terminus" means A 2 This means that the amino group to which it is attached (>NR) is linked either at the C-terminus of the peptide structure or at the carbonyl group of the amino acid structure. Specific examples of an amino acid structure include essential amino acid structures such as glycine structure, alanine structure, valine structure, leucine structure, isoleucine structure, methionine structure, proline structure, phenylalanine structure, tryptophan structure, serine structure, threonine structure, asparagine structure, glutamine structure, tyrosine structure, cysteine structure, lysine structure, arginine structure, histidine structure, aspartic acid structure, glutamic acid structure, etc., modified amino acid structures such as hydroxylysine structure, alicine structure, hydroxyalicine structure, hydroxyproline structure, etc., and amino acid structures contained in the structure of collagen. Particularly preferred are glycine structure, proline structure, hydroxyproline structure, lysine, hydroxylysine structure, alicine structure, hydroxyalicine structure. When the "peptide structure having 1 to 100 amino acids bound at the N-terminus" is a peptide structure composed of two or more amino acids, the number of amino acids is preferably 50 or less, more preferably 20 or less, still more preferably 10 or less, and most preferably 3 or less. Examples of the types of amino acids in a peptide structure composed of two or more amino acids include, similarly, essential amino acids such as glycine, alanine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, serine, threonine, asparagine, glutamine, tyrosine, cysteine, lysine, arginine, histidine, aspartic acid, glutamic acid, etc., modified amino acids such as hydroxylysine, alicine, hydroxyalicine, hydroxyproline, etc., and amino acids contained in the structure of collagen.

[0037] R in formula (1) 2 and R 3 's "hydrocarbon group having 1 to 10 carbon atoms which may have at least one functional group selected from the group consisting of a hydroxyl group, a carboxyl group and an amino group (-NR 4 2)"'s "which may have at least one functional group selected from the group consisting of a hydroxyl group, a carboxyl group and an amino group (-NR 4 2)" and "hydrocarbon group" are synonymous with those described above. The number of carbon atoms in a hydrocarbon group is usually 6 or more when the hydrocarbon group is an aromatic hydrocarbon group. On the other hand, when the hydrocarbon group is not an aromatic hydrocarbon group, the number of carbon atoms is preferably 8 or less, more preferably 6 or less, and may also be 3 or less. "Hydroxyl group, carboxyl group and amino group (-NR 4 Specific examples of "carbon hydrocarbon groups having 1 to 10 carbon atoms that may have at least one functional group selected from the group consisting of (2)" include methyl group, ethyl group, n-propyl group, i-propyl group, cyclopropyl group, n-butyl group, i-butyl group, s-butyl group, t-butyl group, 2,2-dimethyl-1-propyl group, cyclopentyl group, n-hexyl group, cyclohexyl group, n-heptyl group, 2-methylpentyl group, n-octyl group, 2-ethylhexyl group, n-nonyl group, n-decyl group, phenyl group, vinyl group, 2-hydroxyethyl group, and 3-hydroxypropyl group. Examples include groups such as 2-carboxylethyl group, 4-carboxylbutyl group, 5-carboxypentyl group, 2-hydroxy-4-amino-5-carboxypentyl group, 2-amino-2-carboxyethyl group, and 3-amino-3-carboxypropyl group, with 2-hydroxyethyl group, 3-hydroxypropyl group, 2-carboxylethyl group, 4-carboxylbutyl group, 5-carboxypentyl group, 2-hydroxy-4-amino-5-carboxypentyl group, 2-amino-2-carboxyethyl group, and 3-amino-3-carboxypropyl group.

[0038] Furthermore, the 3-hydroxypyridinium derivative represented by formula (1) has a counteranion. Examples of counteranions include halogen ions such as chloride ions and bromide ions, acetate ions, and trifluoroacetate ions.

[0039] The molecular weight of the 3-hydroxypyridinium derivative represented by formula (1) is 10,000 or less, may be 5,000 or less, may be 1,000 or less, may be particularly 700 or less, may be 500 or less, and may be less than 500, 480 or less, or 450 or less.

[0040] Specific examples of 3-hydroxypyridinium derivatives represented by formula (1) include compounds having structures represented by the following formulas (1a) to (1e).

[0041] [ka] In the formula, each i is an independent integer between 1 and 10, preferably between 1 and 5. Furthermore, each j is an integer between 1 and 10, preferably between 1 and 5.

[0042] The collagen production promoter of the present invention may consist of only one compound corresponding to the 3-hydroxypyridinium derivative represented by formula (1), or it may contain two or more compounds corresponding to the 3-hydroxypyridinium derivative represented by formula (1). It is characterized by containing 0.00001% by weight or more of the 3-hydroxypyridinium derivative represented by formula (1) as an active ingredient (or, in the case of containing two or more compounds corresponding to the 3-hydroxypyridinium derivative represented by formula (1), as the layer content of the compounds), preferably 0.0001% by weight or more, more preferably 0.001% by weight or more, particularly preferably 0.01% by weight or more, and most preferably 0.1% by weight or more. Even more preferably 1% by weight or more, and more preferably 5% by weight or more. There is no particular upper limit to the content of the collagen production promoter, but it is preferably 50% by weight or less, more preferably 80% by weight or less, and most preferably 100% by weight. While the collagen production-promoting ability is exhibited by derivatives containing a 3-hydroxypyridinium substructure, increasing the content of the 3-hydroxypyridinium substructure increases the number of molecules recognized by RAGE, thereby more effectively exerting the collagen production effect.

[0043] The 3-hydroxypyridinium substructure is represented by the structure shown in formula (3) below. [ka]

[0044] The collagen production promoter of the present invention may contain other components, but it is preferable that the 3-hydroxypyridinium derivative represented by formula (1) is present in an amount of 0.05% by weight or more of all components contained in the collagen production promoter. More preferably, it is 0.01% by weight or more, particularly preferably 0.1% by weight or more, even more preferably 1% by weight or more, and particularly preferably 10% by weight or more.

[0045] Other ingredients included in collagen enhancers include amino acids, proteins, collagen-derived components that do not contain 3-hydroxypyridinium derivatives, and vitamin C.

[0046] The form of the collagen production promoter is not particularly limited and can be determined according to the dosage form, shape, and use of the formulation or product to which it is applied. For example, it can be prepared as an aqueous solution, glucose solution, buffer solution, biocompatible solution, liquid, emulsion, cream, powder, granules, pills, ointment, etc.

[0047] The target cells are not limited as long as the effects of the present invention are achieved, but it can act on human proximal tubular epithelial cells, osteoblasts, epidermal epithelial cells, and fibroblasts to significantly promote collagen production.

[0048] [Method for producing 3-hydroxypyridinium derivatives] The 3-hydroxypyridinium derivative in the present invention may be produced by organic synthesis or by purifying a natural product. For example, it can be obtained by purifying a natural product, such as by decomposing collagen.

[0049] (Method of production by organic synthesis) Methods for producing it by organic synthesis include, for example, a method of reacting glyceraldehyde with 6-aminohexanoic acid, and a method of reacting Nα-protected-O-protected-lysine with 5,6-epoxy-2-N-protected-O-protected-(2S)-2-aminohexanoic acid ester, as described in Japanese Patent Publication No. 10-195052. Furthermore, these derivatives can also be obtained by methods such as adding various substituents to commercially available 3-hydroxypyridine derivatives. For example, 3-hydroxypyridinium derivatives can be easily obtained by reacting 3-hydroxypyridine with 6-bromohexanoic acid.

[0050] (Method of production by collagen decomposition) Collagen can be broken down by treating it with acids, alkalis, enzymes, etc.

[0051] The collagen mentioned above is not particularly limited and includes, for example, collagen derived from mammals such as cows and pigs, collagen derived from fish such as sharks, and collagen derived from birds such as chickens and ostriches. Collagen is abundant in the bones, skin, and scales of the aforementioned mammals, fish, and birds, and can be obtained by subjecting these various materials to conventionally known treatments such as degreasing, demineralization, and extraction.

[0052] ·Hydrolysis conditions The conditions for the hydrolysis reaction are not particularly limited, but for example, a pH of 2.5 to 8.0 is preferred. More preferably, a pH of 2.5 to 5.5 is preferred. The decomposition temperature is not particularly limited, but for example, it is 10°C to 100°C, preferably 20°C to 50°C, and especially preferably 25°C to 30°C. The duration of the hydrolysis reaction is not particularly limited, but can be, for example, 1 minute to 200 hours, preferably 60 minutes to 100 hours, and particularly preferably 120 minutes to 24 hours.

[0053] • Enzymatic decomposition Conditions for enzyme digestion by trypsin The enzyme digestion temperature by trypsin is not particularly limited, but it is usually carried out at, for example, 10°C to 100°C. Furthermore, a temperature of 30°C to 70°C is more preferable because collagen is broken down more effectively.

[0054] After hydrolysis, it is necessary to inactivate the enzymes, as is usually done. Specifically, although not limited to these methods, the enzymes can be inactivated by heating at 70-100°C for 1 second to 3 hours.

[0055] • Purification of decomposition products The degradation products obtained by hydrolysis can be purified through purification procedures such as column chromatography, cation exchange chromatography, solid-phase extraction, and reverse-phase HPLC.

[0056] Identification of purified products The obtained purified product is identified by fluorescence spectroscopy, NMR, GPC, and mass spectrometry, etc.

[0057] [Collagen breakdown product] The present invention relates to a collagen hydrolysate containing 0.015% by weight or more of the 3-hydroxypyridinium substructure represented by formula (3). Furthermore, the collagen hydrolysate of the present invention may contain 0.02% by weight or more of the 3-hydroxypyridinium substructure, further 0.03% by weight or more, further 0.05% by weight or more, further 0.1% by weight or more, further 0.2% by weight or more, and further 0.5% by weight or more. The collagen hydrolysate of the present invention contains a higher concentration of 3-hydroxypyridinium substructure compared to general collagen hydrolysates.

[0058] A collagen hydrolysate containing 0.015% by weight or more of the 3-hydroxypyridinium substructure of the present invention can be produced by decomposing collagen, and is obtained by separating and fractionating the 3-hydroxypyridinium derivative present as a cross-linking portion of collagen.

[0059] While the method of separation and fractionation is not limited, collagen degradation products containing 0.015% by weight or more of the 3-hydroxypyridinium substructure can be obtained by performing operations such as column chromatography, ion exchange chromatography, gel filtration chromatography, HPLC, electrophoresis, centrifugation, solid-liquid extraction, liquid-liquid separation, filtration, electrodialysis, GPC, distillation, recrystallization, reprecipitation, and distillation.

[0060] Furthermore, the present invention is a collagen hydrolysate containing 0.07% by weight or more of 3-hydroxypyridinoline represented by formula (4). The collagen hydrolysate of the present invention may also contain 0.08% by weight or more of 3-hydroxypyridinoline, further 0.1% by weight or more, further 0.3% by weight or more, further 0.5% by weight or more, further 1.0% by weight or more, and further 2.0% by weight or more. The collagen hydrolysate of the present invention contains a higher concentration of 3-hydroxypyridinoline compared to general collagen hydrolysates.

[0061] The collagen hydrolysate of the present invention containing 0.07% by weight or more of 3-hydroxypyridinoline can be produced by decomposing collagen, and is obtained by separating and fractionating the 3-hydroxypyridinoline present as a cross-linking portion of collagen.

[0062] While the method of preparative and fractionation is not limited, collagen hydrolysates containing 0.07% by weight or more of 3-hydroxypyridinoline can be obtained by performing operations such as column chromatography, ion exchange chromatography, gel filtration chromatography, HPLC, electrophoresis, centrifugation, solid-liquid extraction, liquid-liquid separation, filtration, electrodialysis, GPC, distillation, recrystallization, reprecipitation, and distillation.

[0063] [Analysis method for collagen breakdown products] A method for measuring the concentration of the 3-hydroxypyridinium substructure and 3-hydroxypyridinoline in collagen hydrolysates is, for example, to decompose or re-decompose collagen or collagen hydrolysates to the extent that the cross-linking component 3-hydroxypyridinoline can be isolated. The decomposition or re-decomposition method involves adding 6M hydrochloric acid to collagen or collagen hydrolysates and hydrolyzing them at 110°C under nitrogen purging for 24 hours, which can decompose them to the extent that 3-hydroxypyridinoline can be isolated. Furthermore, the concentration of 3-hydroxypyridinoline can be measured by measuring the area of ​​the fraction that has absorbance at the specific absorption wavelength of 3-hydroxypyridinoline, 295 nm. Furthermore, after determining the concentration of 3-hydroxypyridinoline, the concentration of the 3-hydroxypyridinium substructure in the collagen hydrolysate can be calculated by calculating the ratio of the molecular weight of the 3-hydroxypyridinium substructure to the molecular weight of 3-hydroxypyridinoline. For example, if 1 part by weight of 3-hydroxypyridinoline (PYR) is present in 100 parts by weight of collagen hydrolysate, then 0.212 parts by weight of the 3-hydroxypyridinium substructure is present. Molecular weight of the 3-hydroxypyridinium substructure (91 g / mol) Molecular weight of 3-hydroxypyridinoline (PYR) (429 g / mol)

[0064] [Common collagen breakdown products] In typical collagen hydrolysates, the content of 3-hydroxypyridinoline is less than 0.07% by weight. Furthermore, the content of the 3-hydroxypyridinium substructure is less than 0.0148% by weight. This is because it is known that each collagen molecule contains 0.16 3-hydroxypyridinoline molecules, and this can be calculated based on that information. For example, it can be calculated from the molecular weight of one collagen fiber (approximately 100,000), the molecular weight of 3-hydroxypyridinoline (229 g / mol), and the molecular weight of the 3-hydroxypyridinolium substructure (91 g / mol) (see Fujimoto et al J. Biochem. 83, 863-867 (1978)).

[0065] [Uses of collagen promoters] The applications of collagen promoters are not particularly limited and can be determined according to the purpose. For example, they can be used as cosmetic compositions or food compositions. Furthermore, they can be applied to living organisms to improve skin condition and promote wound healing. They can also be added to culture media and culture solutions to promote the formation of collagen-containing tissues and bone tissues such as artificial tissues and scaffold materials.

[0066] The collagen whose production can be promoted is not particularly limited and includes various types of collagen such as type I collagen and type II collagen, but it can be used in particular to promote the production of type I collagen, which is abundant in bones and skin.

[0067] Type I collagen Type I collagen consists of two α1 chains and one α2 chain, and as mentioned above, it is known to be abundant in bones and skin.

[0068] Furthermore, increasing the expression of the gene encoding type I collagen α1 chain (Col1a1) can promote collagen production, and it is known that promoting type I collagen production and maintaining the amount of type I collagen is effective in preventing and improving wrinkles and sagging. In addition, promoting type I collagen production is also effective in improving skin wound healing. Furthermore, it is known that 80% of the organic matter in bone is made up of type I collagen, and that calcium phosphate is deposited around type I collagen fibers secreted by osteoblasts to form hydroxyapatite, which then forms bone. Therefore, by promoting the production of type I collagen, bone regeneration can be achieved, making it possible to apply this to the treatment of osteoporosis and other conditions.

[0069] (Cosmetic composition) The cosmetic composition of the present invention is characterized by containing the above-mentioned collagen production promoter. The cosmetic composition of the present invention can promote collagen production in the epidermis and dermis of the skin, and can contribute to maintaining skin health or suppressing skin aging.

[0070] The cosmetic composition of the present invention can be prepared by preparing the above-mentioned collagen production promoter together with a suitable medium or carrier. The amount of collagen production promoter included is not limited, as it varies depending on the compound's structure, application site, the age and characteristics of the target population, and the form of the cosmetic composition. Therefore, it cannot be strictly defined. However, the amount of collagen production promoter included is not particularly limited; for example, it is preferably included in the composition at a concentration of 0.001% by weight or more, more preferably 0.01% by weight or more, particularly preferably 0.1% by weight or more, most preferably 1% by weight or more, and more preferably 3% by weight or more. The cosmetic composition of the present invention may contain other collagen-derived components (particularly peptides), but it is preferable that the compound represented by formula (1) and its derivatives account for 10% or more, preferably 20% or more, and most preferably 50% or more, of all collagen-derived components (particularly peptides) contained in the cosmetic composition of the present invention.

[0071] The cosmetic composition of the present invention may contain other ingredients, provided that they do not impair the effects of the present invention. Examples of other ingredients include whitening agents, anti-inflammatory agents, moisturizers, anti-aging agents, hair growth agents, sebum care / acne care agents, antibacterial / bacterial agents, astringents, and UV absorbers. Compounds that can improve collagen production, such as vitamin C, may also be included. Furthermore, the cosmetic composition of the present invention can be appropriately prepared in a suitable form, such as liquid, emulsion, cream, powder, granules, or microencapsulated, as desired. Furthermore, using the cosmetic composition of the present invention, various cosmetics and beauty products such as facial washes, soaps, cleansers, serums, foundations, all-in-one gels, sunscreens, body washes, antiperspirants, lotions, tonics, creams, emulsions, ointments, packs, masks, lipsticks, bath additives, and hair styling products can be manufactured in accordance with conventional methods.

[0072] The application area of ​​the cosmetic composition is not particularly limited as long as the effects of the present invention are achieved, and it can be used on various parts of the body, such as the face, body, and scalp. Furthermore, since the relationship between collagen production in scalp hair follicle stem cells and hair has become known, it can also be expected to be effective in preventing hair loss when applied to the scalp. The cosmetic composition of the present invention has the effect of significantly promoting collagen production in skin fibroblasts and can be used for maintaining skin health or inhibiting skin aging.

[0073] (Food and drink composition) The present invention is characterized by containing the above-mentioned collagen production promoter in the food and beverage composition. The food and beverage composition of the present invention can easily promote collagen production through oral administration and is effective in maintaining skin health, inhibiting skin aging, preventing osteoporosis, and preventing diseases in pets. In this invention, the term "food and beverage composition" refers to products that are ingested orally, such as food products, beverages, confectionery items, supplements, and pet food. These can contain the collagen production promoter of this invention, but their form is not particularly limited. They can be main dishes such as bread and noodles, side dishes such as cheese, sausages, ham, and processed seafood, beverages such as fruit juices, dairy drinks, and carbonated drinks, and confectionery items such as cakes, cookies, jellies, candies, puddings, and yogurt. Furthermore, the form of supplements is not particularly limited and can take the form of tablets, capsules, soft capsules, or nutritional drinks.

[0074] The amount of collagen production promoter in the food and beverage composition is not particularly limited, but is preferably 0.001% by weight or more, more preferably 0.01% by weight or more, particularly preferably 0.1% by weight or more, most preferably 1% by weight or more, and more preferably 3% by weight or more.

[0075] (Composition for cell culture) The cell culture composition of the present invention is characterized by containing the above-mentioned collagen production promoter. The cell culture composition of the present invention can easily generate collagen membranes and can contribute to regenerative medicine and alternatives to animal testing. The method for preparing the cell culture composition is not particularly limited, but for example, it can be produced by adding the collagen production promoter to a general culture medium. Alternatively, it can be produced by preparing the collagen production promoter as a component of the culture medium or culture solution.

[0076] The proportion of collagen production promoter in a cell culture composition varies depending on the type of cells, culture medium, and culture solution, and cannot be uniformly specified. However, it is preferable to contain 0.001% by weight or more in the composition, more preferably 0.01% by weight or more, particularly preferably 0.1% by weight or more, most preferably 1% by weight or more, and more preferably 3% by weight or more. The cell culture composition of the present invention may contain other collagen-derived components (particularly peptides), but it is preferable that the compound represented by formula (1) and its derivatives account for 10% by weight or more, preferably 20% by weight or more, and particularly preferably 50% by weight or more, of all collagen-derived components (particularly peptides) contained in the cell culture composition of the present invention.

[0077] The cell culture composition of the present invention may contain other components, as long as they do not impair the effects of the present invention. Examples of other components include inorganic salts, carbon sources, various vitamins, buffers, cell growth factors, amino acids, and serum. Compounds that can improve collagen production, such as DTPA, TPEN, or ACA, or compounds that enhance collagen production, such as vitamin C or its derivatives, may also be included.

[0078] The cell culture composition of the present invention has the effect of significantly promoting collagen production in cultured cells, and can be used, for example, for culturing artificial skin to promote the formation of artificial tissue. It can also be used for preparing scaffold materials for regenerative medicine to promote the formation of collagen-containing tissue.

[0079] (Wound healing pharmaceutical composition) The wound healing pharmaceutical composition of the present invention is characterized by containing the above-mentioned collagen production promoter. The wound healing pharmaceutical composition of the present invention is used to promote wound healing, such as skin regeneration in the treatment of lacerations and burns.

[0080] The wound healing pharmaceutical composition of the present invention can be prepared according to conventional methods. For example, it can be prepared by preparing the collagen production promoter together with a pharmaceutically acceptable carrier. The wound healing pharmaceutical composition of the present invention may contain pharmaceutically acceptable carriers, known additives, or other pharmaceutically active ingredients, as long as they achieve the effects of the present invention. Furthermore, compounds that enhance collagen production, such as vitamin C or its derivatives, may also be incorporated.

[0081] The proportion of the collagen production promoter in the wound healing pharmaceutical composition of the present invention varies depending on the application site, the age of the target, the form of the pharmaceutical composition, etc., and cannot be uniformly specified. However, for example, it is preferable to contain 0.001% by weight or more in the composition, more preferably 0.01% by weight or more, particularly preferably 0.1% by weight or more, most preferably 1% by weight or more, and more preferably 3% by weight or more. The wound healing pharmaceutical composition of the present invention may contain other collagen-derived components (particularly peptides), but it is preferable that the compound represented by formula (1) and its derivatives account for 10% by weight or more, preferably 20% by weight or more, and particularly preferably 50% by weight or more, of all collagen-derived components (particularly peptides) contained in the wound healing pharmaceutical composition of the present invention.

[0082] The wound-healing pharmaceutical composition of the present invention can be configured in a suitable form as desired. For example, it can be in the form of a dry powder, gel, cream, ointment, suspension, solution, or a biocompatible synthetic or natural solid matrix. Furthermore, it can be applied to a substrate such as a film and used by adhering it to the skin or other surfaces.

[0083] (Composition for promoting bone formation) The bone formation promoting composition of the present invention is characterized by containing the above-mentioned collagen production promoting agent. The bone formation promoting composition of the present invention can be used for the prevention and treatment of osteoporosis. The bone formation promoting composition of the present invention can be formulated for oral administration as a supplement, or it can be used by direct injection into the affected area. Examples of such formulations include tablets such as orally disintegrating tablets, chewable tablets, and dispersible tablets, as well as capsules, granules, powders, oral liquids, syrups, or oral jellies, aqueous solutions, glucose solutions, etc.

[0084] In the bone formation promoting composition of the present invention, pharmaceutically acceptable excipients, binders, lubricants, isotonic agents, stabilizers, preservatives, flavoring agents, solubilizers, or emulsifiers may be further added during formulation and drug formulation. The bone formation promoting composition of the present invention can be manufactured by conventionally known methods. For example, in the case of tablets, the formulation can be carried out in the following steps: weighing, primary mixing (uniformly mixing of active ingredients, excipients, binders, disintegrants, etc.), granulation, sieving, secondary mixing (mixing of lubricants, etc.), coating (if necessary), packaging, etc.

[0085] The bone formation-promoting composition of the present invention activates osteoblasts, and by local delivery to a patient's bone defect, it can repair bone or increase bone density. The drug carrier mixture used in this process is prepared by mixing it with, for example, calcium phosphate cement powder. Depending on the detailed embodiment, the bone composition of the present invention can be formed by combining the drug carrier mixture with a suitable bone matrix material. Alternatively, the bone repair composition may include a collagen sponge. The bone composition can then be applied to the treatment site, for example, by implanting or injecting it into the bone defect site. It can also be applied to implants.

[0086] The collagen production promoter content of the bone formation promoting composition of the present invention is not particularly limited, but is preferably 0.001% by weight or more, more preferably 0.01% by weight or more, particularly preferably 0.1% by weight or more, most preferably 1% by weight or more, and more preferably 3% by weight or more. The bone formation promoting composition of the present invention may contain other collagen-derived components (particularly peptides), but it is preferable that the compound represented by formula (1) and its derivatives account for 10% by weight or more, preferably 20% by weight or more, and particularly preferably 50% by weight or more, of all collagen-derived components (particularly peptides) contained in the bone formation promoting composition of the present invention.

[0087] The present invention will be described below with reference to examples, but the present invention is not limited to these examples. <Example 1> (1) Preparation of 3-hydroxypyridinoline (PYR) [ka] PYR was prepared according to the method described below. 1 g of bovine Achilles tendon collagen (Sigma) was mixed with 10 ml of 6 M hydrochloric acid (for automated amino acid analysis), and hydrolysis was carried out at 110°C for 24 hours under nitrogen displacement using a hydrolysis apparatus (Waters, Pico·Tag). The hydrolyzed product was filtered through a 0.2 μm pore size filter (ADVANTEC, DISMIC-25CS), and then the hydrochloric acid was removed and the product dried using a rotary evaporator (EYELA, N-1100, SB-1100, A1000S) to obtain collagen hydrolysate. The collagen hydrolysate was dissolved in 10 mM sodium acetate buffer, and then 10 M sodium hydroxide aqueous solution was added to adjust the pH to 4.0. This was subjected to cation exchange chromatography (FPC3500), and the target fraction was eluted with 10 mM sodium acetate buffer containing 0.5 M NaCl. The eluted fraction was concentrated using a rotary evaporator, dissolved in 0.1% HFBA, and subjected to solid-phase extraction using a Sep-PakC18·35cc Vac Cartridge (Waters, WAT043345). After eluting the non-adsorbed fraction and salts with 0.1% HFBA, the target fraction was eluted with 20% acetonitrile. The eluted fraction was concentrated using a rotary evaporator and dissolved in a small amount of 0.1% HFBA. This was filtered using a 0.45 μm pore size filter (ADVANTEC, DISMIC-03CP), then subjected to reverse-phase HPLC (Inertsil ODS-4, GL Science), and separated with 50% acetonitrile containing 0.1% HFBA. The fraction having absorbance at the specific absorption wavelength of PYR, 295 nm, was isolated and further purified by reverse-phase HPLC (Inertsil ODS-3, GL Science) followed by separation with 50% acetonitrile containing 0.1% HFBA to obtain PYR (general formula (4) above). The fluorescence spectra of the obtained compounds are shown in Figure 1. The mass spectra are shown in Figure 2. Specific fluorescence and mass spectral analysis confirmed that the obtained compound is PYR.

[0088] (2) Evaluation of collagen production promoting effect The collagen production-promoting effect of the compounds was evaluated using the following method. (2-1) Sample preparation Human proximal tubular epithelial cell line (HK-2, RIKEN Cell Bank) with a cell count of 1 × 10⁶ 6 Cells were seeded individually in a cell culture dish and cultured for 72 hours in DMEM medium containing 10% FBS. PYR was added to the cultured cells as the test compound and cultured for 24 hours. As a comparative example, samples were also prepared in a system without the addition of PYR.

[0089] Sample group Sample 1) Control (PYR-free) Sample 2) Addition of 10 μM PYR Sample 3) Addition of 50 μM PYR

[0090] (2-2) Measurement of collagen production The collagen production levels of the sample groups prepared in (2-1) above were analyzed by RT (reverse transcription)-PCR according to the following procedure. The culture medium was removed from the culture dish, washed twice with PBS solution for 5 minutes, and then 1 ml of TRIzol (ambion) was added to lyse the cells, which were then collected in a microcentrifuge tube (WATSON). 400 μl of chloroform was added and the mixture was stirred, followed by centrifugation to obtain the aqueous layer. 500 μl of isopropanol was added to the obtained aqueous layer fraction to precipitate the RNA. The supernatant was removed by centrifugation, and the obtained RNA was washed with 75% ethanol and then dissolved in water. 2 μg of RNA was used as a template, and (dT) 18 cDNA was synthesized by reverse transcription using primers and AMV Reverse Transcriptase (Takara Shuzo). Using the cDNA obtained from each sample group as a template, PCR was performed using primers specific to type I collagen a1 (Col1a1) and Hybridol DNA Polymerase. The amplified products after the reaction were subjected to agarose electrophoresis, and the band intensity was measured and evaluated. The expression intensity of GAPDH, which is constitutively expressed, was used as a control to correct the values.

[0091] Figure 3 shows the mRNA expression levels of type I collagen type A in human tubular epithelial cells obtained from PYR (Sample 2 and Sample 3). The intensity values ​​in the figure represent the percentage of the expression intensity of each sample relative to the control (set at 100). Table 1 shows the results of the staining intensity analysis. The intensity values ​​in the table represent the percentage of the staining intensity of each sample relative to the control (set at 100).

[0092] Expression level of type I collagen α1 (Col1a1) (average value) [Table 1]

[0093] Figure 3 and Table 1 show that PYR (Samples 2 and 3) significantly increased collagen production compared to the control (Sample 1), suggesting that it possesses collagen-enhancing activity.

[0094] (3) Evaluation of the effect of promoting TGFβ expression involved in collagen production The TGFβ expression-promoting effect of the compounds was evaluated using the following method. (3-1) Sample preparation Rat proximal tubular epithelial cell line (NRK52E, RIKEN Cell Bank) with a cell count of 1 × 10⁶ 6 Cells were seeded individually into cell culture dishes and cultured for 72 hours in DMEM medium containing 10% FBS. PYR was added to the cultured cells as the test compound and cultured for 24 hours. As a comparative example, samples were also prepared in a system without the addition of PYR.

[0095] Sample group Sample 4) Control (PYR-free) Sample 5) Addition of 10 μM PYR Sample 6) Addition of 50 μM PYR

[0096] (3-2) Measurement of collagen production The collagen production levels of the sample groups prepared in (3-1) above were analyzed by RT (reverse transcription)-PCR according to the following procedure. The culture medium was removed from the culture dish, washed twice with PBS solution for 5 minutes, and then 1 ml of TRIzol (ambion) was added to lyse the cells, which were then collected in a microcentrifuge tube (WATSON). 400 μl of chloroform was added and the mixture was stirred, followed by centrifugation to obtain the aqueous layer. 500 μl of isopropanol was added to the obtained aqueous layer fraction to precipitate the RNA. The supernatant was removed by centrifugation, and the obtained RNA was washed with 75% ethanol and then dissolved in water. 2 μg of RNA was used as a template, and (dT) 18 cDNA was synthesized by reverse transcription using primers and AMV Reverse Transcriptase (Takara Shuzo). Using the cDNA obtained from each sample group as a template, PCR was performed using TGFβ1-specific primers and Hybridol DNA Polymerase. The amplified products after the reaction were subjected to agarose electrophoresis, and the band intensity was measured and evaluated. The expression intensity of GAPDH, which is constitutively expressed, was used as a control to correct the values.

[0097] Figure 4 shows the results of the TGFβ1 mRNA expression level analysis of rat tubular epithelial cells obtained from PYR (Sample 5 and Sample 6). The intensity values ​​in the figure represent the percentage of the expression intensity of each sample, with the control set to 100. Table 1 shows the results of the staining intensity analysis. The intensity values ​​in the table represent the percentage of the staining intensity of each sample, with the control set to 100.

[0098] TGFβ1 expression level (average value) [Table 2]

[0099] Figure 4 and Table 2 show that PYR (Samples 5 and 6) significantly increased TGFβ1 expression compared to the control (Sample 4), indicating its collagen production-enhancing activity and thus its usefulness as a cosmetic and food composition. As described in non-patent literature (Jeon et al. J. Phys. Ther. Sci. 27, 1485-1490 2015), it is a well-known fact that increasing TGF-β expression and increasing type I collagen production are effective in wound healing. The collagen promoter of the present invention, which increases TGF-β expression and type I collagen expression, is useful as a pharmaceutical composition for wound healing.

[0100] <Example 2> (1) Preparation of 1-(5-carboxypentyl)-3-hydroxy-5-hydroxymethylpyridinium (CHHP) [ka]

[0101] 2M glyceraldehyde and 0.1M 6-aminohexanoic acid were reacted in 0.2M phosphate buffer (pH 7.4) at 50°C for 72 hours. The reaction solution was subjected to solid-phase extraction using a Sep-Pak C18 35cc Vac Cartridge (Waters, WAT043345). A non-adsorbent fraction eluted with water was obtained, concentrated using a rotary evaporator, dissolved in a small amount of water, and then trifluoroacetic acid (TFA) was added to adjust the pH to 2.0. This solution was again subjected to a Sep-Pak C18 35cc Vac Cartridge, and the fraction eluted with a 20% acetonitrile aqueous solution was concentrated. The concentrated solution was subjected to reverse-phase HPLC (Inertsil ODS-3, GL Science) and separated with 50% acetonitrile containing 0.05% TFA. By isolating the fraction with absorbance at the specific absorption wavelength of CHHP of 291 nm, purified CHHP (general formula (5) above) was obtained.

[0102] (2) Evaluation of the bone formation promoting effect (2-1) A mouse skull-derived cell line (MC3T3-E1, RIKEN BioResource Center) was used, with a cell count of 1 × 10⁶. 6 Cells were seeded into individual cells in a cell culture dish (NUNC) and cultured in αMEM medium containing 10% FBS, 25 μg / ml ascorbic acid, and 5 mM β-glycerophosphate to differentiate into osteoblast-like cells. CHHP was added as the test compound and cultured for 24 hours. The bone formation-promoting effect was evaluated based on its effect on promoting osteoblast differentiation or on promoting the production of bone matrix collagen. Osteoblast differentiation was evaluated by measuring the expression level of alkaline phosphatase using enzyme activity. MC3T3-E1 cells were cultured in differentiation medium for 14 days, followed by a further 7 days in differentiation medium supplemented with CHHP. After culture, the medium was removed, the dish was washed, and the cells were collected using a buffer containing a surfactant (0.1% NP-40) and a protease inhibitor (20 mM Tris, pH 7.5). The cells were then lysed by sonication to obtain a cell extract. The cell extract was added to a buffer containing an alkaline phosphatase substrate (pH 10.0) and reacted at 37°C for 1 hour. The product (p-nitrophenol) produced from the substrate after the reaction was evaluated by the increase in absorbance at 405 nm. Figure 4 shows data demonstrating the activity-enhancing effect of CHHP on alkaline phosphatase, i.e., its effect on promoting osteoblast differentiation (relative to the control without the additive).

[0103] (2-2) Furthermore, similar to human tubular epithelial cells, the effect of CHHP on promoting osteoblast collagen production was evaluated by RT-PCR, and the results are shown in Figure 5.

[0104] Sample group Sample 7) Control (CHHP-free) Sample 8) CHHP 1 μM added Sample 9) Addition of CHHP 10 μM

[0105] Expression level of type I collagen α1 (Col1a1) (average value) [Table 3]

[0106] Figures 5 and 6 and Table 3 show that CHHP (samples 8 and 9) significantly increased collagen production compared to the control (sample 7), suggesting that it also has an osteoblast differentiation-promoting effect.

[0107] As described in the non-patent literature (Lingqun Kong, Yoshiaki Deyama, Tomoya Kudo, Yoshitaka Yoshimura, Kuniaki Suzuki: "Effects of Estrogen on Various ATPases in Osteoblast-like MC3T3-E1 Cells," Hokkaido Dental Journal 33, 175-184 (2013) (http: / / hdl.handle.net / 2115 / 52456)), bone is formed through a process in which calcium phosphate and other substances accumulate in the collagen matrix produced by osteoblasts. Therefore, it is a well known fact that compounds that promote collagen production and differentiation of osteoblasts can increase bone mass and can serve as therapeutic and preventive drugs for osteoporosis. Furthermore, as stated in the same non-patent literature, the expression of alkaline phosphatase (ALP) increases in osteoblasts as they differentiate. From this, it is a well known fact that compounds that increase ALP activity have an osteoblast differentiation-promoting effect and can therefore serve as therapeutic and preventive drugs for osteoporosis. As described in the same non-patent document, mouse MC3T3-E1 cells have characteristics similar to human osteoblasts and are a commonly used cell line for functional analysis of osteoblasts. Therefore, it can be said that the promotion of collagen production and increased ALP expression in MC3T3-E1 cells can serve as indicators of bone formation promotion.

[0108] • Investigation of the interaction between the advanced glycation end product receptor (RAGE) and 3-hydroxypyridinium derivatives. (1) Analysis of RAGE ligand using the Biacore system Soluble RAGE was expressed as a recombinant protein by Escherichia coli and purified using affinity column chromatography. When the purified recombinant RAGE protein was immobilized on a sensor chip and pyridinoline and CHHP were added as analytes, a concentration-dependent binding response was detected. When kinetic analysis was performed, the dissociation constant K D was 179 μM (PYR) and 393 μM (CHHP). Furthermore, when analysis was also performed on a pyridinoline peptide having a peptide structure, a specific interaction was confirmed. From these results, it was demonstrated that 3-hydroxypyridinium derivatives specifically exhibit affinity for the receptor for advanced glycation end products (RAGE).

[0109] From these results, it was confirmed that the 3-hydroxypyridinium derivative of the present invention is specifically recognized by the receptor for advanced glycation end products (RAGE), and it was suggested that collagen production is promoted.

[0110] (Preparation of collagen degradation product containing 3-hydroxypyridinoline (PYR)) 10 L of 6 M hydrochloric acid was added to 1 kg of fish collagen, and hydrolysis was performed at 110°C for 24 hours. After the hydrolyzate was filtered through a filter with a pore size of 0.2 μm (manufactured by ADVANTEC), it was neutralized with a 10 M aqueous sodium hydroxide solution. Then, this was dried to obtain a collagen hydrolyzate. The collagen hydrolyzate was dissolved in 10 mM sodium acetate buffer, and then a 10 M aqueous sodium hydroxide solution was added to adjust the pH to 4.0. This was subjected to cation exchange chromatography, and the target fraction was eluted with 10 mM sodium acetate buffer containing 0.5 M NaCl. The eluted fraction was dried to obtain a PYR-containing collagen degradation product.

[0111] [Formulation Example] Next, examples of formulations in which the PYR-containing collagen hydrolysate of the present invention is applied to cleansing agents, skincare cosmetics, makeup cosmetics, haircare cosmetics, sunscreens / UV care cosmetics, foods, and beverages are shown. However, the technical scope of the present invention is not limited by these formulation examples.

[0112] (Prescription example 1) The soap was prepared according to the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Coconut fatty acids 20.00 Palm oil fatty acids 20.00 Glycerin 6.00 Sodium hydroxide (85%) 15.75 EDTA-2Na 0.20 Sodium citrate 0.50 water remainder

[0113] (Prescription example 2) A body wash (clear type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 10.00 Coconut fatty acids 5.00 Citric acid 1.00 Glycerin 3.00 K hydroxide 5.68 EDTA-4Na 0.20 Sodium gluconate 1.00 PEG-120 methyl glucose dioleate 2.00 water remainder

[0114] (Prescription example 3) A body wash (pearl type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 10.00 Sodium cocoyl methyl taurate, water 10.00 (NIKKOL CMT-30) Sodium laureth sulfate, water (27% aqueous solution) 22.20 PEG-150 distearate 3.00 (NIKKOL CDS-6000P) Sodium laureth-5 carboxylate, water 20.00 (NIKKOL AKYPO RLM 45NV) Lauryl betaine, water 10.00 (NIKKOL AM-301) BG 5.00 Glycol distearate 2.00 (NIKKOL Esthe Pearl 15V) Appropriate amount of citric acid EDTA-2Na 0.10 Preservative (appropriate amount) water remainder

[0115] (Prescription example 4) A body soap (a low-irritation body soap using amino acid surfactants) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 10.00 Cocoyl glutamate TEA, water (30% aqueous solution) 30.00 Sodium trideceth-4 carboxylate 5.00 (NIKKOL ECTD-3NEX) Sodium cocoamphoacetate, water () 10.00 (NIKKOL AM-101) PEG-50 Hydrogenated Castor Oil 0.50 (NIKKOL HCO-50) BG 5.00 EDTA-2Na 0.10 Preservative (appropriate amount) water remainder

[0116] (Prescription example 5) A body soap (a body soap containing both soap and anionic surfactant) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 10.00 Potassium coconut oil fatty acid and potassium myristate solution 40.00 Sodium lauroyl methylalanine, water 20.00 (NIKKOL Alaninate LN30) PEG-7 Glyceryl Cocoate 5.00 (NIKKOL TMGCO-7) Cocamide DEA 3.00 Glycerin 15.00 BG 5.00 EDTA-3Na 0.05 Preservative (appropriate amount) water remainder

[0117] (Prescription example 6) A facial cleanser was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Lauric acid 10.00 Palmitic acid 12.00 Stearic acid 10.00 Sodium Lauryl Methyl Taurate 5.00 (NIKKOL LMT) Glyceryl stearate (SE) 2.00 (NIKKOL MGS-BSEV) PEG-6, PEG32 10.00 Glycerin 10.00 Sorbitol (70% aqueous solution) 5.00 Olive fruit oil 1.00 (NIKKOL Olive Oil) Preservative (appropriate amount) K hydroxide 7.00 water remainder

[0118] (Prescription example 7) A facial cleansing foam (containing amino acid-based surfactants) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Citric acid 0.15 EDTA-2Na 0.05 BG 15.00 Glycerin 10.00 Cocamide DEA 3.00 Sodium cocoyl methyl taurate, water 4.00 (NIKKOL CMT-30) Sodium lauroyl methylalanine, water 8.00 (NIKKOL Alaninate LN-30) Cocamidopropyl betaine, water 8.00 (NIKKOL AM-3130N) Preservative (appropriate amount) Ethanol 3.00 water remainder

[0119] (Prescription example 8) A facial cleansing foam (containing phosphate surfactant) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Laureth-1 phosphate, water 7.50 (NIKKOL HOSTEN HLP-1) Arginine 4.40 DPG 4.00 Preservative (appropriate amount) Sodium cocoyl methyl taurate, water 5.00 (NIKKOL CMT-30) PEG-20 Glyceryl Triisostearate 4.00 (NIKKOL TGI-20) Pentylene glycol 2.00 Glyceryl caprylate 1.00 Glycerin 1.00 Chelating agent (appropriate amount) Citric acid 0.25 water remainder

[0120] (Prescription example 9) A cleansing oil (bicontinuous type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-10 Pentaisostearate 6.30 (NIKKOL Decaglyn 5-ISV) Polyglyceryl-6 laurate 11.70 (NIKKOL Hexaglyn 1-L) Polyglyceryl-10 Diisostearate 2.00 (NIKKOL Decaglyn 2-ISV) Methylheptyl laurate 58.30 (NIKKOL GS-MHL) Myristic acid 0.80 Mixed vegetable oil 3.00 (NIKKOL NATURAL OILS-1) Cyclopentasiloxane 10.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Ethanol 1.50 Preservative (appropriate amount) water remainder

[0121] (Prescription example 10) A cleansing cream (oil-in-water type, for both rinsing and wiping off) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: PEG-6 sorbitan stearate 2.00 (NIKKOL TS-106V) Polysorbate 60 0.80 (NIKKOL TS-10V) Glyceryl stearate 1.20 (NIKKOL MGS-BV2) Cetearyl alcohol 3.50 Squalane 45.00 (NIKKOL Sugar Squalane) Methylheptyl laurate 5.00 (NIKKOL GS-MHL) Dimethicone (350 mPa·s) 0.20 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 10.00 Glycerin 2.00 Carboxyvinyl polymer (2% aqueous solution) 5.00 (NTC-CARBOMER 380) Preservative (appropriate amount) Sodium stearylmethyltaurate 0.30 (NIKKOL SMT) K hydroxide 0.035 water remainder

[0122] (Prescription Example 11) A cleansing milk (oil-in-water type, for both rinsing and wiping) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Cyclopentasiloxane 5.00 Diphenylsiloxyphenyl trimethicone 5.00 Methylheptyl laurate 5.00 (NIKKOL GS-MHL) Squalane 3.00 (NIKKOL Sugar Squalane) PEG-7 Glyceryl Cocoate 10.00 (NIKKOL TMGCO-7) Tri(behenate / isostearate / eicosanedioate)glycerylcetearyl alcohol 0.50 Sorbitan isostearate 1.00 (NIKKOL SI-10RV) Sorbet-30 Tetraoleate 1.00 (NIKKOL GO-430 NV) Polysorbate 60 1.00 (NIKKOL TS-10V) Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Glycerin 10.00 BG 3.00 Carboxyvinyl polymer (2% aqueous solution) 10.00 (NTC-CARBOMER 381) (Acrylates / Alkyl Acrylate (C10-30) Crosspolymer (for high viscosity) (2% aqueous solution)) 5.00 EDTA-2Na 0.05 Preservative appropriate amount K Hydroxide 0.07 Water remainder

[0123] (Formulation Example 12) A cleansing gel (oil type) was prepared according to the following formulation. Component name Blending amount (mass%) Oil components: PPG-6 Decyltetradeceth-30 12.00 (NIKKOL PEN-4630) Octyldodeceth-25 4.00 Squalane 2.00 (NIKKOL Sugar Squalane) Cetyl Ethylhexanoate 6.40 (NIKKOL CIO) Triethylhexanoin 40.00 (NIKKOL Trifat S-308) Glycerin 12.00 Methyl Gluceth-10 3.00 (NIKKOL BMG-10) Aqueous components: PYR-containing Collagen Hydrolysate 1.00 Sorbitol (70% aqueous solution) 8.00 DPG 4.00 Water remainder

[0124] (Formulation Example 13) A cleansing gel (liquid crystal type) was prepared according to the following formulation. Component name Blending amount (mass%) PYR-containing Collagen Hydrolysate 1.00 PEG-40 Hydrogenated Castor Oil 8.00 (NIKKOL HCO-40) PEG-50 Hydrogenated Castor Oil 8.00 (NIKKOL HCO-50) Triethylhexanoin 60.00 (NIKKOL Trifat S-308) Glycerin 16.80 water remainder

[0125] (Prescription example 14) A cleansing gel (oil-in-water, rinse-off type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Isododecane 20.00 Cetyl ethylhexanoate 10.00 (NIKKOL CIO) Cyclopentasiloxane 10.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Compound emulsifier 4.50 (NIKKOL Nicomulus SE W) BG 5.00 Preservative (appropriate amount) K hydroxide 0.05 water remainder

[0126] (Prescription example 15) A cleansing gel (water-based type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: PEG-7 Glyceryl Cocoate 10.00 (NIKKOL TMGCO-7) PEG-20 Glyceryl Triisostearate 4.50 (NIKKOL TGI-20) PEG-20 sorbitan isostearate 3.50 (NIKKOL TI-10V) PEG-45 stearate 2.00 (NIKKOL MYS-45V) DPG 20.00 Preservative: appropriate amount Aqueous components: Collagen degradation product containing PYR 1.00 Methyl gluceth - 10 5.00 (NIKKOL BMG - 10) Carboxyvinyl polymer 0.40 (NTC - CARBOMER 380) EDTA - 2Na 0.10 Water: the balance

[0127] (Formulation Example 16) A cleansing lotion (wipe - off type) was prepared according to the following formulation. Component name Blending amount (mass%) Oil components: PEG - 7 glyceryl coconut oil fatty acid 4.00 (NIKKOL TMGCO - 7) PEG - 10 lauric acid 4.00 (NIKKOL MYL - 10) PEG - 50 hydrogenated castor oil 2.00 (NIKKOL HCO - 50) Preservative: appropriate amount Hydroxylated lecithin, glycerin 0.50 (NIKKOL Lecithinol SH50) Aqueous components: Collagen degradation product containing PYR 1.00 Cocoyl arginine ethyl PCA 0.10 Citric acid 0.005 Sodium citrate 0.03 EDTA - 2Na 0.03 Water: the balance

[0128] (Formulation Example 17) A makeup remover (two - layer type) was prepared according to the following formulation. Component name Blending amount (mass%) Oil components: Cyclopentasiloxane 20.00 ​Methylheptyl laurate 20.00 (NIKKOL GS-MHL) Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Ethanol 3.00 PEG-7 Glyceryl Cocoate 0.20 (NIKKOL TMGCO-7) PPG-4 Ceteth-10 0.10 (NIKKOL PBC-33) PEG-400 0.05 Sodium chloride 0.50 EDTA-3Na 0.02 Cocoyl arginine ethyl PCA 0.05 water remainder

[0129] (Prescription example 18) A vanishing cream (free of petroleum-based surfactants) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Stearic acid 15.00 Isopropyl palmitate 2.00 (NIKKOL IPP) Lanolin 1.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Sorbitol (70% aqueous solution) 8.00 K hydroxide 1.00 Preservative (appropriate amount) water remainder

[0130] (Prescription example 19) Vanishing cream was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polysorbate 60 2.00 (NIKKOL TS-10V) Sorbet-30 Tetraoleate 0.50 (NIKKOL GO-430NV) Glyceryl stearate, Glyceryl stearate (SE), PEG-10 stearate 0.50 (NIKKOL MGS-DEXV) Stearic acid 7.00 Cetanol 3.00 Cetyl palmitate 3.00 (NIKKOL N-SPV) Paraffin (135°F) 3.00 Jojoba seed oil 7.00 (NIKKOL Jojoba Oil S) Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 7.00 Preservative (appropriate amount) water remainder

[0131] (Prescription example 20) An emollient cream was prepared according to the following formula. Ingredient name Amount (mass%) Oily components: PEG-40 stearate 2.00 (NIKKOL MYS-40V) Glyceryl stearate, Glyceryl stearate (SE), PEG-10 stearate 5.00 (NIKKOL MGS-DEXV) Stearic acid 2.00 Cetanol 2.00 Squalane 12.00 (NIKKOL Sugar Squalane) Macadamia seed oil 4.00 (NIKKOL Macadamia Nut Oil) Dimethicone (300 mPa·s) 0.20 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 7.00 Preservative (appropriate amount) water remainder

[0132] (Prescription example 21) A moisturizing cream was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Functional lipid complex 3.00 (NIKKOL Nicolipid 81S) Cetanol 2.00 Caprylic / Capric Triglyceride 4.00 (NIKKOL Triester F-810) Low melting point wax 4.00 (NIKKOL Nicowax LM) Pyridoxine trihexyldecanoate 1.00 (NIKKOL VB6-IP) Ethylhexylglycerin, glyceryl caprate 0.40 (NIKKOL NicoGuard 88) Dimethicone (high polymerization) 1.00 Diphenylxyphenyl Trimethicone 2.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 5.00 Glycerin 5.00 Hydroxypropylmethylcellulose 0.20 Carbomer 0.30 (NTC-CARBOMER 380) Betaine 2.00 Arginine 0.30 water remainder

[0133] (Prescription example 22) A moisturizing cream (lecithin emulsified) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Hydrogenated lecithin 2.00 (NIKKOL Resinol S-10) Myristyl alcohol 2.00 Behenyl alcohol 3.00 (NIKKOL Behenyl Alcohol 80%) Cetyl palmitate 2.00 (NIKKOL N-SPV) Triethylhexanoin 10.00 (NIKKOL Trifat S-308) Squalane 8.00 (NIKKOL Sugar Squalane) Jojoba seed oil 1.00 (NIKKOL Jojoba Oil S) Dimethicone (350 mPa·s) 1.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 5.00 Glycerin 8.00 Xanthan gum 0.10 water remainder

[0134] (Prescription example 23) An anti-aging cream (liquid crystal type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Compound emulsifier 5.00 (NIKKOL Nicomulus LC) Cetearyl alcohol 3.00 Low melting point wax 0.20 (NIKKOL Nicowax LM) Squalane 5.00 (NIKKOL Sugar Squalane) Triethylhexanoin 5.00 (NIKKOL Trifat S-308) Pentaerythrityl tetraethylhexanoate 5.00 Cyclopentasiloxane 3.00 Dimethicone (350 mPa·s) 0.20 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Glycerin 10.00 Pentylene glycol 2.00 BG 8.00 Carbomer 0.20 (NTC-CARBOMER 380) Xanthan gum 0.10 Preservative (appropriate amount) K hydroxide 0.05 Sodium hyaluronate (1% aqueous solution) 1.00 Polysaccharide aqueous solution 5.00 (FUCOGEL 1.5P) water remainder

[0135] (Prescription example 24) A light cream (lecithin, high molecular weight emulsion) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Batyl alcohol 0.20 (NIKKOL Batyl Alcohol EX) Hydrogenated lecithin 0.30 (NIKKOL Resinol S-10) Glyceryl stearate 1.50 (NIKKOL MGS-BV2) Cetyl ethylhexanoate 1.00 (NIKKOL CIO) Diphenyldimethicone 3.00 Dimethicone (6 mPa·s) 4.00 Dimethicone (350 mPa·s) 1.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Compound emulsifier 5.00 (NIKKOL Nicomulus LH) Pentylene glycol 2.00 BG 4.00 Glycerin 5.00 Dipotassium glycyrrhizate 0.10 Preservative (appropriate amount) water remainder

[0136] (Prescription example 25) An anti-acne cream was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Ascorbyl tetrahexyldecanoate 10.00 (NIKKOL VC-IP) Hydrogenated lecithin 0.10 (NIKKOL Resinol S-10) Batyl alcohol 1.00 (NIKKOL Batyl Alcohol EX) Behenyl alcohol 2.00 (NIKKOL Behenyl Alcohol 65) Glyceryl stearate, PEG-60 glyceryl stearate 2.00 (NIKKOL MGS-150V) Cetyl palmitate 3.00 (NIKKOL N-SPV) Dimethicone (100 mPa·s) 0.20 Hydrogenated palm oil, palm kernel oil, palm oil 4.00 (NIKKOL Trifat PS-45H) Olefin oligomer 4.00 (NIKKOL Synselan 4SP) Tocopherol 0.10 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 5.00 Glycerin 3.00 Xanthan gum 0.04 Hydroxyethylcellulose 0.04 Citric acid 0.02 Sodium citrate 0.03 Preservative (appropriate amount) water remainder

[0137] (Prescription example 26) An eye cream was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Caprylic / Capric Triglyceride, Tocopheryl Retinoate 2.00 (NIKKOL Tocoretinoate-10) Hydrogenated lecithin 0.50 (NIKKOL Resinol S-10) Polyglyceryl-10 myristate 3.00 (NIKKOL Decaglyn 1-M) Oleylglyceryl 2.00 (NIKKOL Cerakill Alcohol) Cetyl palmitate 4.00 (NIKKOL N-SPV) Squalane 10.00 (NIKKOL Sugar Squalane) Triethylhexanoin 8.00 (NIKKOL Trifat S-308) Cetanol 6.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Glycerin 3.00 Xanthan gum 0.20 Preservative (appropriate amount) water remainder

[0138] (Prescription example 27) Massage cream was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Seteth-20 2.00 (NIKKOL BC-20) Glyceryl stearate 4.00 (NIKKOL MGS-BV2) Cetanol 2.00 Cetyl palmitate 2.00 (NIKKOL N-SPV) Vaseline 6.00 Squalane 30.00 (NIKKOL Sugar Squalane) Triethylhexanoin 5.00 (NIKKOL Trifat S-308) Meadowfoam oil 3.00 (NIKKOL Meadowfoam Oil) Dimethicone (300 mPa·s) 0.20 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Carbomer 0.20 (NTC-CARBOMER380) Glycerin 15.00 Preservative (appropriate amount) TEA 0.10 water remainder

[0139] (Prescription example 28) A massage cream (emulsified with beeswax and borax) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: PEG-20 Sorbitan Cocoate 3.00 (NIKKOL TL-10) Glyceryl stearate (SE) 2.70 (NIKKOL MGS-ASEV) Mineral oil (#70) 25.00 Ethylhexyl palmitate 22.00 (NIKKOL IOP) Beeswax 3.00 Stearic acid 1.30 Cetanol 1.00 Dimethicone (350 mPa·s) 0.10 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Carbomer 0.20 (NTC-CARBOMER381) BG 5.00 Preservative (appropriate amount) TEA 0.50 Sodium borate 0.50 water remainder

[0140] (Prescription example 29) A hand cream (water-based) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-2 isostearate 2.50 (NIKKOL DGMIS) Polyglyceryl polyricinoleate 1.50 (NIKKOL Hexaglyn PR-15) Squalane 4.00 (NIKKOL Sugar Squalane) Diphenylsiloxyphenoxytrimethicone 5.00 Glycerin 10.00 Tocopherol acetate 0.10 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Sodium alginate 0.10 BG 5.00 Mg sulfate 0.10 Preservative (appropriate amount) water remainder

[0141] (Prescription example 30) An emollient cream (amino acid gel emulsion, W / O type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-6 polyricinoleate 1.50 (NIKKOL Hexaglyn PR-15) Polyglyceryl-2 oleate 0.50 (NIKKOL DGMO-CV) Mineral oil (#70) 8.00 Triethylhexanoin 5.00 (NIKKOL Trifat S-308) Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Sodium glutamate 1.00 Glycerin 5.00 Sodium chloride 2.00 Preservative (appropriate amount) water remainder

[0142] (Prescription example 31) An emollient cream (organic modified clay mineral gel emulsion, W / Si type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: W / O compound emulsifier 4.00 (NIKKOL WO) Cyclopentasiloxane 7.00 Silicone-containing product 5.00 (NIKKOL SILBLEND-91) Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 5.00 Glycerin 25.00 Preservative (appropriate amount) Sodium hyaluronate (1% aqueous solution) 1.00 Polysaccharide aqueous solution (FUCOGEL1.5P) 1.00 Acetyl hydroxyproline (AHYP) 0.10 Sodium chloride 0.50 EDTA-2Na 0.10 water remainder

[0143] (Prescription example 32) The hand cream was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-10 myristate 1.20 (NIKKOL Decaglyn 1-M) Hydrogenated lecithin 0.50 (NIKKOL Resinol S-10) Glyceryl stearate, Glyceryl stearate (SE), PEG-10 stearate 1.00 (NIKKOL MGS-DEXV) Stearic acid 4.00 Cetanol 4.00 Cetyl palmitate 2.00 (NIKKOL N-SPV) Squalane 4.00 (NIKKOL Sugar Squalane) Hexyldecyl isostearate 2.00 (NIKKOL ICIS) Octyldodecyl myristate 4.00 (NIKKOL ODM-100) Dimethicone (300 mPa·s) 0.20 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 DPG 5.00 Preservative (appropriate amount) FOMBLIN (fluorine oil) emulsion base 3.00 (NIKKOL NET-HC-R) Silicone-based emulsion base 3.00 (NIKKOL NET-SG-60 A) water remainder

[0144] (Prescription example 33) A hand cream (natural hand cream) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-10 stearate 2.20 (NIKKOL Decaglyn 1-SV) Hydrogenated rapeseed alcohol 3.00 Macadamia seed oil 2.00 (NIKKOL Macadamia Nut Oil) Caprylic / Capric Triglyceride 5.00 (NIKKOL Triester F-810) Shea butter (Star Shea Butter Refined) 15.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Xanthan gum 0.10 Glycerin 10.00 BG 10.00 Preservative (appropriate amount) Ethanol 3.00 water remainder

[0145] (Prescription example 34) A moisturizing lotion (lecithin emulsified) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Hydrogenated lecithin 0.80 (NIKKOL Resinol S-10) Phytosterose 0.20 Stearyl alcohol 0.40 Behenyl alcohol 0.40 (NIKKOL Behenyl Alcohol 80%) Cetyl palmitate 0.80 (NIKKOL N-SPV) Triethylhexanoin 4.00 (NIKKOL Trifat S-308) Squalane 3.20 (NIKKOL Sugar Squalane) Jojoba seed oil 0.40 (NIKKOL Jojoba Oil S) Dimethicone (350 mPa·s) 0.40 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Xanthan gum 0.10 Glycerin 3.20 BG 5.00 Carbomer 0.06 (NTC-CARBOMER381) Preservative (appropriate amount) Arginine 0.07 water remainder

[0146] (Prescription example 35) A liquid crystal emulsion was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Compound emulsifier 4.00 (NIKKOL Nicomulus LC) Cetearyl alcohol 1.00 Olefin oligomer 4.00 (NIKKOL Synselan 4SP) Meadowfoam oil 1.00 (NIKKOL Meadowfoam Oil) Dimethicone (350 mPa·s) 0.50 Ethylhexylglycerin, glyceryl caprate 0.50 (NIKKOL NicoGuard 88) Water-based ingredients: PYR-containing collagen hydrolysate 1.00 BG 4.00 Glycerin 1.00 PEG-6, PEG-32 1.00 Lecithin, lysophosphatidic acid, lysolecithin 0.10 Carbomer 0.15 (NTC-CARBOMER381) Arginine 0.11 EDTA-2Na 0.10 Ethanol 5.00 Watermelon fruit extract, apple fruit extract, lentil fruit extract (Aqua-Speed) 3.00 water remainder

[0147] (Prescription example 36) A moisturizing lotion (made only from naturally derived ingredients) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-10 stearate 2.00 (NIKKOL Decaglyn 1-SV) Sorbitan sesquistearate 1.00 (NIKKOL SS-15V) Hydrogenated rapeseed alcohol 1.80 Shea butter (Star Shea Butter Refined) 2.00 Methylheptyl isostearate 3.00 (NIKKOL GS-MHIS) Methylheptyl laurate 3.00 (NIKKOL GS-MHL) Squalane 5.00 (NIKKOL Sugar Squalane) Mixed vegetable oil 1.00 (NIKKOL NATURAL OILS-1) Lavender oil 0.20 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Xanthan gum 0.10 BG 1.00 Glycerin 5.00 Preservative (appropriate amount) Polysaccharide aqueous solution (FUCOGEL1.5P) 1.00 water remainder

[0148] (Prescription example 37) A whitening lotion was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-10 Pentastearate 1.00 (NIKKOL Decaglyn 5-SV) PEG-40 stearate 1.00 (NIKKOL MYS-40V) Behenyl alcohol 1.70 (NIKKOL Behenyl Alcohol 65) Glyceryl stearate 1.00 (NIKKOL MGS-BV2) Triethylhexanoin 5.50 (NIKKOL Trifat S-308) Squalane 3.50 (NIKKOL Sugar Squalane) Jojoba seed oil 1.00 (NIKKOL Jojoba Oil S) Cyclopentasiloxane 5.00 Diphenyl dimethicone 3.50 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Xanthan gum 0.30 Sodium stearoyl methyl taurate 0.60 (NIKKOL SMT) DPG 2.00 BG 3.00 Preservative (appropriate amount) Magnesium ascorbyl phosphate 3.00 (NIKKOL VC-PMG) Polysaccharide aqueous solution (FUCOGEL1.5P) 0.50 Sodium hyaluronate (1% aqueous solution) 0.50 Sodium glutamate 0.50 EDTA-4Na 0.10 Sodium pyrosulfite 0.20 water remainder

[0149] (Prescription example 38) A lotion (α-gel lotion) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Cetyl phosphate 0.70 (NIKKOL Purefos α) Batyl alcohol 0.50 (NIKKOL Batyl Alcohol EX) Cetanol 1.50 Squalane 2.00 (NIKKOL Sugar Squalane) Dimethicone (6 mPa·s) 3.00 Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Arginine 0.50 Glycerin 10.00 BG 5.00 Betaine 0.50 Sodium hyaluronate (1% aqueous solution) 1.00 Carbomer 0.15 (NTC-CARBOMER381) Preservative (appropriate amount) Watermelon fruit extract, apple fruit extract, lentil fruit extract (Aqua-Speed) 1.00 water remainder

[0150] (Prescription example 39) The body milk was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Polyglyceryl-10 myristate 2.10 (NIKKOL Decaglyn1-M) Hydrogenated lecithin 0.60 (NIKKOL Resinol S-10) Stearic acid 0.50 Behenyl alcohol 1.60 (NIKKOL Behenyl Alcohol 65) Cetyl palmitate 0.60 (NIKKOL N-SPV) Hydrogenated polydecene 6.00 Cetyl ethylhexanoate 6.00 (NIKKOL CIO) Triethylhexanoin 6.00 (NIKKOL Trifat S-308) Water-based ingredients: PYR-containing collagen hydrolysate 1.00 Carbomer 0.10 (NTC-CARBOMER380) Glycerin 5.00 Preservative (appropriate amount) water remainder

[0151] (Prescription example 40) A softening lotion was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Glycerin 5.00 DPG 4.00 PEG-20 0.50 Xanthan gum 0.05 Citric acid 0.02 Sodium citrate 0.10 EDTA-2Na 0.05 BG 4.00 PEG-60 Hydrogenated Castor Oil 0.50 (NIKKOL HCO-60) Preservative (appropriate amount) Polysaccharide aqueous solution (FUCOGEL 1.5P) 2.00 Watermelon fruit extract, apple fruit extract, lentil fruit extract (Aqua-Speed) 3.00 water remainder

[0152] (Prescription example 41) A moisturizing lotion was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Glycerin 3.00 Sorbitol (70% aqueous solution) 2.00 DPG 4.00 Xanthan gum 0.05 Hydroxyethylcellulose 0.05 Acetyl hydroxyproline 5.00 Arginine 5.00 Sodium hyaluronate (1% aqueous solution) 1.00 Citric acid 0.05 Sodium citrate 0.01 PCA-Na (50% aqueous solution) 1.00 BG 5.00 Preservative (appropriate amount) water remainder

[0153] (Prescription example 42) An astringent lotion was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 PEG-60 Hydrogenated Castor Oil 1.00 (NIKKOL HCO-60) Ethanol 15.00 Preservative (appropriate amount) PEG-8 1.00 BG 5.00 Zinc paraphenolsulfonate 0.20 Citric acid 0.10 pH buffering agent (appropriate amount) Fragrance (appropriate amount) Dye appropriate amount water remainder

[0154] (Prescription example 43) A cleansing lotion was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 PEG-7 Glyceryl Cocoate 4.00 (NIKKOL TMGCO-7) PEG-10 Laurate 4.00 (NIKKOL MYL-10) PEG-50 Hydrogenated Castor Oil 2.00 (NIKKOL HCO-50) Preservative (appropriate amount) Hydrogenated lecithin, glycerin 0.50 (NIKKOL Resinol SH-50) Cocoyl arginine ethyl PCA 0.10 Citric acid 0.005 Sodium citrate 0.03 EDTA-2Na 0.03 water remainder

[0155] (Prescription example 44) Carmine lotion was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Dipotassium glycyrrhizate 0.10 PCA-Na 2.50 EDTA-2Na 0.10 BG 6.00 Preservative (appropriate amount) St. John's wort extract, burdock root extract, soapwort leaf extract, sage leaf extract, calendula flower extract, hop cone extract, lemon fruit extract, BG, water 4.00 (Viteren EGX-252 (BG)) Iron oxide 0.15 Zinc oxide 0.50 Kaolin 1.00 water remainder

[0156] (Prescription example 45) A lotion (nanoemulsion type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Hydroxyproline (L-hydroxyproline) 0.10 BG 1.00 DPG 1.00 PEG-32 0.30 PEG-150 0.20 Pentylene glycol 2.50 Citric acid 0.03 Sodium citrate 0.04 EDTA-2Na 0.10 Preservative (appropriate amount) Lipid-soluble vitamin C derivative-containing nanoemulsion base 5.00 (NIKKOL Solubble VCIP) water remainder

[0157] (Prescription example 46) A moisturizing gel (lecithin emulsified) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Hydrogenated lecithin 1.00 (NIKKOL Resinol S-10 PLUS) Glycerin 15.00 BG 6.00 PEG-32 2.00 Xanthan gum 0.08 (Acrylates / C10-30 Alkyl Acrylate) Crosspolymer (for high viscosity applications) 0.30 Hydroxyethylcellulose 0.08 Hexyl methylolpropionate 0.50 (NIKKOL NicoGuard 6) Diphenyl dimethicone 1.00 Cyclopentasiloxane 4.00 Arginine 0.30 Sodium hyaluronate (1% aqueous solution) 1.00 Polysaccharide aqueous solution (FUCOGEL1.5P) 3.00 Perilla leaf extract, ethanol 0.30 (Perilla extract NA FREE) water remainder

[0158] (Prescription example 47) An all-in-one gel was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Polyglyceryl-6 polyricinoleate 0.50 (NIKKOL Hexaglyn PR-15) Dimethicone (10 mPa·s) 1.40 Dimethicone (350 mPa·s) 0.60 Cyclopentasiloxane 0.20 Carbomer 0.20 (NTC-CARBOMER380) (Acrylates / C10-30 Alkyl Acrylate) Crosspolymer (for high viscosity applications) 0.10 Hydroxypropylmethylcellulose 0.20 Glycerin 15.00 BG 6.00 Sorbitol (70% aqueous solution) 3.00 Ethanol 3.00 Hexyl dimethylolpropionate 0.70 (NIKKOL NicoGuard 6) K hydroxide 0.10 Nanoemulsion base containing vitamins, etc. 1.00 (NIKKOL Nicosome Vitamins ABCE) Nanoemulsion base containing Pseudozyma tsukubaensis / (olive oil / glycerin / soy protein) ferment, etc. 5.00 (NIKKOL Nicosome SML)

[0159] (Prescription example 48) An anti-aging serum was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 BG 2.00 DPG 3.00 Pentylene glycol 4.00 Glycerin 7.00 Xanthan gum 0.02 (Acrylates / C10-30 Alkyl Acrylate) Crosspolymer (for high viscosity applications) 0.10 (Acrylates / C10-30 Alkyl Acrylate) Crosspolymer (for low viscosity applications) 0.20 Preservative (appropriate amount) Arginine 0.10 Ascorbyl tetrahexyldecanoate 1.00 (NIKKOL VC-IP) Diphenyldimethicone 3.00 Cyclopentasiloxane 7.00 Tocopherol 0.10 EDTA-2Na 0.10 water remainder

[0160] (Prescription example 49) An anti-aging serum (containing liposomes) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Carbomer 0.54 (NTC-CARBOMER380) BG 4.00 Diglycerin 4.00 PEG-20 1.00 Maltitol (75% aqueous solution) 1.00 K hydroxide 0.18 Hydroxyproline (L-hydroxyproline) 0.50 Silicone-containing emulsion base 1.00 (NIKKOL NET-813-1) Liposome containing polyhydric alcohol 1.00 (NIKKOL Aquasome BH) Polysaccharide aqueous solution (FUCOGEL1.5P) 3.00 Sodium hyaluronate (1% aqueous solution) 3.00 Ethanol 3.00 Preservative (appropriate amount) water remainder

[0161] (Prescription example 50) A whitening serum (clear type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Xanthan gum 0.40 Hydroxyethylcellulose 0.40 BG 3.00 Glycerin 3.00 Hyaluronic acid (1% aqueous solution) 5.00 Preservative (appropriate amount) Magnesium ascorbyl phosphate 3.00 (NIKKOL VC-PMG) Sodium citrate 0.50 EDTA-4Na 0.10 water remainder

[0162] (Prescription example 51) A moisturizing serum (translucent type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Hydrogenated lecithin 0.60 (NIKKOL Resinol S-10) BG 6.00 PEG-32 1.00 Glycerin 15.00 Hydroxyethylcellulose 0.08 (Acrylates / C10-30 Alkyl Acrylate) Crosspolymer (for high viscosity applications) 0.20 Preservative (appropriate amount) Arginine 0.20 EDTA-2Na 0.10 Pyridoxine trihexyldecanoate 1.00 (NIKKOL VB6-IP) Cyclopentasiloxane 5.00 Batyl alcohol 0.10 (NIKKOL Batyl Alcohol EX) Tocopherol 0.10 water remainder

[0163] (Prescription example 52) A peel-off pack (gel type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Polyvinyl alcohol 15.00 Carboxymethylcellulose 5.00 BG 5.00 Appropriate amount of cushioning material Ethanol 12.00 Ores-20 0.50 (NIKKOL BO-20V) Fragrance (appropriate amount) Preservative (appropriate amount) water remainder

[0164] (Prescription example 53) The clay pack was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 25.00 Bentonite 12.00 Kaolin 13.00 Titanium dioxide 2.00 Ceteth-20 1.00 (NIKKOL BC-20V) BG 10.00 Glycerin 10.00 Preservative (appropriate amount) Polyvinyl alcohol 1.00 water remainder

[0165] (Prescription example 54) The cream pack was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 25.00 Squalane 10.00 (NIKKOL Sugar Squalane) Caprylic / Capric Triglyceride 5.00 (NIKKOL Triester F-810) Ascorbyl tetrahexyldecanoate 2.00 (NIKKOL VC-IP) Vaseline 3.00 Stearyl alcohol 3.00 PEG-55 stearate 0.80 (NIKKOL MYS-55V) Glyceryl stearate 2.20 (NIKKOL MGS-BV2) Cetyl palmitate 2.00 (NIKKOL N-SPV) Dimethicone (350 mPa·s) 0.50 Tocopherol 0.10 Hydroxyethylcellulose 0.10 BG 5.00 Glycerin 7.00 Preservative (appropriate amount) Titanium dioxide, aluminum hydroxide 2.00 Sodium stearoyl methyl taurate 0.05 (NIKKOL SMT) Citric acid 0.01 EDTA-2Na 0.05 Ethylgothioneine, Water (THIOTAINE) 1.00 water remainder

[0166] (Prescription example 55) The makeup base was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Compound emulsifier 4.00 (NIKKOL Nicomulus LC) Cetearyl alcohol 1.00 Squalane 5.00 (NIKKOL Sugar Squalane) Phenylentrimethicone 5.00 Polypropylsilsesquioxane (alkyl acrylate / dimethicone) copolymer, cyclopentasiloxane 1.00 Carbomer 0.15 (NTC-CARBOMER381) Hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer 0.30 EDTA-2Na 0.10 Preservative (appropriate amount) Arginine 0.05 BG 4.00 Glycerin 1.00 Pigment-grade titanium dioxide (appropriate amount) Red iron oxide (appropriate amount) (HDI / Trimethylol Hexyllactone) Crosspolymer, Silica, Polymethyl Methacrylate, Mica, Titanium Dioxide 3.00 Watermelon fruit extract, apple fruit extract, lentil fruit extract (Aqua-Speed) 3.00 Ethanol 5.00 water remainder

[0167] (Prescription example 56) An oil-based foundation was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Talc remaining Kaolin 15.00 Pigment-grade titanium dioxide 15.00 Red iron oxide (appropriate amount) Yellow iron oxide (appropriate amount) Black iron oxide (appropriate amount) Solid paraffin 3.00 Microcrystalline wax 6.00 Beeswax 2.00 Vaseline 12.00 Lanolin acetate 1.00 Squalane 6.00 (NIKKOL Sugar Squalane) Isopropyl palmitate 18.00 (NIKKOL IPP) Preservative (appropriate amount)

[0168] (Prescription example 57) Foundation (O / W type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Compound emulsifier 2.50 (NIKKOL Nicomulus 41) Glyceryl stearate 1.00 (NIKKOL MGS-BV2) Batyl alcohol 0.30 (NIKKOL Batyl Alcohol 100%) Cetearyl alcohol 1.00 Diethylaminohydroxybenzoyl hexyl benzoate 2.00 Ethylhexyl methoxycinnamate 10.00 Isononyl isononanoate 2.00 Cyclopentasiloxane 7.00 Dimethicone (10 mPa·s) 3.00 Pigment-grade titanium dioxide 6.00 Red iron oxide (appropriate amount) Yellow iron oxide (appropriate amount) Black iron oxide (appropriate amount) Glycerin 5.00 Dipropylene glycol 5.00 Silicate (Al / Mg) 0.50 Sodium stearoyl methyl taurate 0.50 (NIKKOL SMT) EDTA-2Na 0.05 Preservative (appropriate amount) Xanthan gum 0.30 water remainder

[0169] (Prescription example 58) Foundation (W / O type) was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Compound emulsifier 5.00 (NIKKOL NIKKOL WO) Silicone-containing product 2.50 (NIKKOL SILBLEND-91) Cyclopentasiloxane, (Dimethicone / Vinyl Dimethicone) Crosspolymer 2.00 Cyclopentasiloxane 20.00 Dimethicone (6 mPa·s) 2.50 Diphenylsiloxyphenyl trimethicone 2.50 (Alkyl acrylate / dimethicone) copolymer, cyclopentasiloxane 1.00 Pentaerythrityl tetraethylhexanoate 2.00 Diisostearyl malate 1.00 (NIKKOL DISM) (HDI / Trimethylol Hexyllactone) Crosspolymer, Silica, 3.00 Silicone-treated pigment-grade titanium dioxide 7.00 Silicone-treated red iron oxide (appropriate amount) Appropriate amount of silicone-treated yellow iron oxide Appropriate amount of silicone-treated black iron oxide Glycerin 8.00 BG 3.00 Sodium chloride 0.50 EDTA-2Na 0.05 Ethanol 3.00 Preservative (appropriate amount) water remainder

[0170] (Prescription example 59) A dual-purpose foundation was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Silicone-treated talc residue Silicone-treated mica 40.00 Silicone-treated titanium dioxide microparticles 5.00 Silicone-treated pigment-grade titanium dioxide 15.00 Silicone-treated red iron oxide (appropriate amount) Appropriate amount of silicone-treated yellow iron oxide Appropriate amount of silicone-treated black iron oxide Zinc stearate 0.10 Nylon powder 0.20 Squalane 4.00 (NIKKOL Sugar Squalane) Solid paraffin 0.50 Dimethicone (10 mPa·s) 4.00 Triethylhexanoin 5.00 (NIKKOL Trifat S-308) Ethylhexyl Methoxycinnamate 1.00 Preservative (appropriate amount)

[0171] (Prescription example 60) The powder foundation was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Sericite remaining Talc 5.00 Mica 10.00 Fine particle titanium dioxide 5.00 Pigment-grade titanium dioxide 10.00 Red iron oxide (appropriate amount) Yellow iron oxide (appropriate amount) Black iron oxide (appropriate amount) Squalane 1.50 (NIKKOL Sugar Squalane) Triethylhexanoin 5.00 (NIKKOL Trifat S-308) Ethylhexylglycerin, glyceryl caprate 0.40 (NIKKOL NicoGuard 88)

[0172] (Prescription example 61) The lipstick was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Appropriate amount of coloring agent Diisostearyl malate (remainder) Polyethylene 5.00 Ceresin 6.00 Microcrystalline wax 3.00 Triethylhexanoin 20.00 (NIKKOL Trifat S-308) Cetyl ethylhexanoate 7.00 (NIKKOL CIO) Hydrogenated polyisobutene 2.00 Antioxidant (appropriate amount)

[0173] (Prescription example 62) A clear lip gloss was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Dextrin palmitate 4.50 Mineral oil 25.00 Diisostearyl malate (remainder) (NIKKOL DISM) Polyglyceryl-2 Triisostearate 5.50 (NIKKOL DGTIS) Antioxidant (appropriate amount) Hydrogenated polyisobutene 40.00

[0174] (Prescription example 63) The nail enamel was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Nitrocellulose 14.00 (Phthalic anhydride / trimellitic anhydride / glycols) copolymer 16.00 Acetyl tributyl citrate 5.00 Ethyl acetate 20.00 Butyl acetate residue Ethanol (appropriate amount) Isopropanol (appropriate amount) Pigment (appropriate amount) Stearalkonium hectorite (appropriate amount)

[0175] (Prescription example 64) The shampoo was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Cocadopropyl betaine 10.00 (NIKKOL AM-3130 N) Sodium laureth sulfate, water 20.00 TEA laureth sulfate, water 10.00 Cocamide DEA 3.00 Glycol distearate 2.00 (NIKKOL Esthe Pearl 10V) Polyquaternium-10, sodium chloride (1.5% aqueous solution) 10.00 BG 3.00 Citric acid (10% aqueous solution) 1.50 EDTA-2Na 0.10 Preservative (appropriate amount) water remainder

[0176] (Prescription example 65) A mild shampoo was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Sodium cocoyl methyl taurate, water 25.00 (NIKKOL CMT-30) Sodium lauroyl methylalanine, water 15.00 (NIKKOL Alaninate LN-30) Lauryl betaine, water 20.00 (NIKKOL AM-301) Lauramide DEA 5.00 PEG-60 Hydrogenated Castor Oil 1.00 (NIKKOL HCO-60) Glycol distearate 2.00 (NIKKOL Esthe Pearl 15V) Polyquaternium-10, sodium chloride (1.5% aqueous solution) 10.00 BG 3.00 Citric acid (10% aqueous solution) 0.40 EDTA-2Na 0.10 Preservative (appropriate amount) water remainder

[0177] (Prescription example 66) A silicone-free shampoo was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 BG 4.00 Sorbitol, water 2.00 Polyquaternium-10 0.50 Citric acid (10% aqueous solution) 0.65 EDTA-2Na 0.10 Preservative (appropriate amount) Sodium cocoyl methyl taurate, water 20.00 (NIKKOL CMT-30) Sodium lauroyl methylalanine, water 15.00 (NIKKOL Alaninate LN-30) Cocamidopropyl betaine, water 15.00 (NIKKOL AM-3130N) Cocamide DEA 2.00 Laureth-2 1.00 (NIKKOL BL-2) Decyl glucoside, water 10.00 PEG-7 Glyceryl Cocoate 1.00 (NIKKOL TMGCO-7) Stearamidopropyldimethylamine 0.50 (NIKKOL Amidoamine MPS) Perilla leaf extract, ethanol, water 0.30 (Perilla extract NA FREE) Plant extract mixture 0.10 (Viteren EGX-232 (BG)) water remainder

[0178] (Prescription example 67) Dry shampoo was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Functional blended raw materials 2.00 (NIKKOL APP-CLEV) Glycerin fatty acid ester mixture 0.50 (NIKKOL NicoGuard DL) Ethanol 30.00 water remainder

[0179] (Prescription example 68) A scalp oil shampoo was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 (Caprylyl / Capryl) Glucoside, Water 10.00 (GreenAPG PC 0810) BG 4.00 Sorbitol, water 2.00 Guar hydroxypropyltrimonium chloride 1.00 Preservative (appropriate amount) Citric acid 0.65 EDTA-2Na 0.10 Sodium cocoyl methyl taurate, water 20.00 (NIKKOL CMT-30) Sodium lauroyl methylalanine, water 15.00 (NIKKOL Alaninate LN-30) Cocamidopropyl betaine, water 15.00 (NIKKOL AM-3130N) PG laurate 2.00 Polyglyceryl-10 Laurate 3.00 (NIKKOL Decaglyn 1-L) Squalane 0.30 (NIKKOL Sugar Squalane) Methylheptyl laurate 0.30 (NIKKOL GS-MHL) Argania spinosa kernel oil (argan oil) 0.30 Plant and seaweed extract mixture 1.00 (Viteren EGX-771BG)

[0180] (Prescription example 69) The conditioner was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Behentrimonium chloride, ethanol 2.40 (NIKKOL CA-2580) Distearyldimonium chloride, isopropanol, water 0.80 (NIKKOL CA-3475V) Cetanol 5.00 Oleyl alcohol 1.00 Glyceryl stearate 1.00 (NIKKOL MGS-BV2) Low melting point wax 1.00 (NIKKOL Nicowax LM) Olive fruit oil 2.00 (NIKKOL Olive Oil) Vaseline 8.00 Dimethicone (100 mPa·s) 1.00 Dimethicone (1000 mPa·s) 0.50 Tocopherol 0.05 Glycerin 6.00 Preservative (appropriate amount) water remainder

[0181] (Prescription example 70) A non-silicone treatment was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Stearamidopropyldimethylamine 3.25 (NIKKOL Amidoamine MPS) Glyceryl stearate 0.50 (NIKKOL MGS-BV2) Stearyl alcohol 7.00 Sandalwood extract, Phellodendron amurense bark extract, Barley extract 1.00 (BOIS II) Methylheptyl laurate 2.00 (NIKKOL GS-MHL) Phytostearyl isostearate 3.00 Octyldodecanol 3.00 Vaseline 4.00 PG 5.00 Glutamic acid 1.05 Polyquaternium-47, Water 2.40 Preservative (appropriate amount) Perilla leaf extract, ethanol, water 0.30 (Perilla extract NA FREE) Plant extract mixture 0.10 (Viteren EGX-232 (BG)) Polysaccharide aqueous solution (FUCOGEL1.5P) 3.00 water remainder

[0182] (Prescription example 71) Hair wax was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Compound emulsifier 7.00 (NIKKOL Nicomulus LC) Cetearyl alcohol 5.00 Behenyl alcohol 2.00 (NIKKOL Behenyl Alcohol 65) Low melting point wax 1.00 (NIKKOL Nicowax LM) Beeswax 10.00 Squalane 1.00 (NIKKOL Sugar Squalane) Olefin oligomer 5.00 (NIKKOL Synselan 4SP) Triethylhexanoin 5.00 (NIKKOL Trifat S-308) Vaseline 5.00 Cyclopentasiloxane 2.00 Dimethicone (6 mPa·s) 2.00 Preservative (appropriate amount) BG 5.00 Glycerin 3.00 EDTA-2Na 0.10 PEG-115M 0.10 Carbomer 10.00 (NTC-CARBOMER 380) Arginine 0.20 (VP / VA) Copolymer, Ethanol Polyquanium-11, Water 1.00 water remainder

[0183] (Prescription example 72) A hair gel cream was prepared using the following formula. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 1.00 Compound emulsifier 4.50 (NIKKOL Nicomulus SE W) BG 8.00 Preservative (appropriate amount) K hydroxide 0.05 EDTA-2Na 0.03 Glycerin 5.00 Squalane 2.50 (NIKKOL Sugar Squalane) Cyclopentasiloxane 8.00 Dimethicone (10 mPa·s) 1.50 Amodimethicone 1.00 Cyclopentasiloxane, Dimethiconol 1.00 PEG / PPG-19 / 19 Dimethicone, Cyclopentasiloxane 1.00 Diethylaminohydroxybenzoyl hexyl benzoate, Ethylhexyl methoxycinnamate 1.50 Tocopherol 0.10 water remainder

[0184] (Prescription example 73) Sunscreen milk (non-chemical, W / O type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Fine particle titanium dioxide dispersion 30.00 (CM3K40VMJ) Fine particle zinc oxide dispersion 35.00 (CM3K50XZ4J) Isododecane 11.50 Trimethylsiloxysilicate, Cyclopentasiloxane 3.00 (HDI / Trimethylol Hexyllactone) Crosspolymer, Silica 3.00 Ethylhexyl palmitate 0.50 (NIKKOL IOP) (Vinyl dimethicone / methicone silsesquioxane) crosspolymer 2.00 PEG-9 Polydimethylsiloxyethyl Dimethicone 1.00 Water-based ingredients: PYR-containing collagen hydrolysate 5.00 Glycerin 5.00 Preservative (appropriate amount) Sodium citrate 0.20 EDTA-3Na 0.05 water remainder

[0185] (Prescription example 74) A sunscreen cream (non-chemical, W / O type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Fine particle titanium dioxide dispersion 35.00 (CM3K40T4J) Fine particle zinc oxide dispersion 34.00 (CM3KG60XZ4) Cyclopentasiloxane 9.55 Trimethylsiloxysilicate, Cyclopentasiloxane 3.00 (HDI / Trimethylol Hexyllactone) Crosspolymer, Silica 3.00 Ethylhexyl palmitate 0.50 (NIKKOL IOP) (Vinyl dimethicone / methicone silsesquioxane) crosspolymer 2.00 PEG-9 Polydimethylsiloxyethyl Dimethicone 1.00 Water-based ingredients: PYR-containing collagen hydrolysate 5.00 BG 3.00 Preservative (appropriate amount) Sodium citrate 0.20 EDTA-3Na 0.05 water remainder

[0186] (Prescription example 75) A sunscreen cream (oil-water type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Fine particle titanium dioxide dispersion 15.00 (CMIP40VMJ) Fine particle zinc oxide dispersion 15.00 (CMIP50XZ4J) Sorbitan stearate 2.00 (NIKKOL SS-10V) Polyglyceryl-10 myristate 0.50 (NIKKOL Decaglyn1-M) Tocopherol 0.10 Ethylhexyl methoxycinnamate 10.00 Dimethylaminohydroxybenzoyl hexyl benzoate 3.00 Water-based ingredients: PYR-containing collagen hydrolysate 5.00 BG 7.00 Sodium stearoyl methyl taurate 0.50 (NIKKOL SMT) (HDI / Trimethylol Hexyllactone) Crosspolymer, Silica 2.00 Xanthan gum (2% aqueous solution) 5.00 Sodium hyaluronate (1% aqueous solution) 5.00 Polysaccharide aqueous solution (FUCOGEL1.5P) 1.00 Preservative (appropriate amount) Compound emulsifier 3.00 (NIKKOL Nicomulus LH) Ethanol 5.00 water remainder

[0187] (Prescription example 76) A sunscreen gel cream (oil-water type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Fine particle titanium dioxide dispersion 22.50 (MCP55ZSJ) Diethylamino hydroxybenzoyl hexyl benzoate 2.80 Octocrylene 1.00 Sorbitan oleate 0.25 (NIKKOL SO-10V) Polyglyceryl-10 Diisostearate 0.75 (NIKKOL Decaglyn2-ISV) (Sodium acrylate / sodium acryloyldimethyl taurate) copolymer, isohexadecane, polysorbate 80, sorbitan oleate, water 2.00 Mixed vegetable oil 2.50 (NIKKOL NATURAL OILS-1) Trimethylsiloxysilicate, Dimethicone 3.00 Water-based ingredients: PYR-containing collagen hydrolysate 5.00 Glycerin 1.00 BG 1.00 Methyl methacrylate crosspolymer 2.00 Xanthan gum (2% aqueous solution) 5.00 Sodium hyaluronate (1% aqueous solution) 5.00 Ethanol 5.00 water remainder

[0188] (Prescription example 77) A BB cream (non-chemical, W / O type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Fine particle titanium dioxide dispersion 10.00 (CM3K40T4J) Fine particle zinc oxide dispersion 25.00 (CM3K50XZ4J) Cyclopentasiloxane 12.50 PEG-9 Polydimethylsiloxyethyl Dimethicone 1.00 Cyclopentasiloxane, (Dimethicone / Vinyl Dimethicone) Crosspolymer 2.00 (Vinyl dimethicone / methicone silsesquioxane) crosspolymer 3.00 Trimethylsiloxysilicate, Cyclopentasiloxane 3.00 (HDI / Trimethylol Hexyllactone) Crosspolymer, Silica 4.00 Silicone dispersion of iron oxide 15.00 (NIKKOL Fresh Color Base) Water-based ingredients: PYR-containing collagen hydrolysate 8.00 Glycerin 5.00 BG 2.00 Preservative (appropriate amount) EDTA-2Na 0.05 L-hydroxyproline 0.10 Sodium hyaluronate (1% aqueous solution) 1.00 Sodium chloride 1.00 water remainder

[0189] (Prescription example 78) CC cream (non-chemical, O / W type) was prepared using the following formula. Ingredient name Amount (mass%) Oily components: Fine particle titanium dioxide dispersion 27.50 (IOPP40VMJ) Sorbitan stearate 2.00 (NIKKOL SS-10V) Triethylhexanoin 2.00 (NIKKOL Trifat S-308) Cetearyl alcohol 0.50 PEG-60 Hydrogenated Castor Oil 0.50 (NIKKOL HCO-60) Polysorbate 80 0.20 (NIKKOL TO-10V) Tocopherol 0.20 Water-based ingredients: PYR-containing collagen hydrolysate 8.00 BG 4.00 Pentylene glycol 3.00 Methyl methacrylate cristpolymer 2.00 Glycerin 1.00 Sodium stearoyl methyl taurate 0.50 (NIKKOL SMT) Stearoxyhydroxypropylmethylcellulose 0.10 Xanthan gum (2% aqueous solution) 5.00 Sodium hyaluronate (1% aqueous solution) 2.00 Preservative (appropriate amount) Dipotassium glycyrrhizate 0.50 Citric acid 0.20 Sodium citrate 0.03 Silicone dispersion of iron oxide 10.00 (NIKKOL Fresh Color Base AQUA) water remainder

[0190] (Prescription example 79) Tablets Using a known fluid bed granulator, the above-mentioned PYR-containing collagen hydrolysate was granulated and coated with a 1% by mass starch aqueous solution to obtain PYR-containing collagen hydrolysate granules. The obtained PYR-containing collagen hydrolysate granules were mixed with other powders according to the following formulation, and then tablets (10 mmφ, 300 mg) were prepared using a tablet press. Ingredient name Amount (mass%) PYR-containing collagen hydrolysate granules 50.0 Lactose 10.0 Starch (for granulation) 0.5 Starch (for powder mixing) 12.0 Maltitol 10.0 Microcrystalline cellulose 15.0 Silicon dioxide 0.5 Magnesium stearate 2.0 By granulating and coating PYR-containing collagen hydrolysate with starch, tableting problems such as sticking were suppressed.

[0191] (Prescription example 80) Granule stick The above PYR-containing collagen hydrolysate was mixed according to the following formulation and then granulated using a known fluid bed granulator. A 1% by mass aqueous solution of dextrin was used for granulation. The resulting granules were filled into sticks to obtain granule sticks (1.2 g / packet). Ingredient name Amount (mass%) PYR-containing collagen hydrolysate 90.0 Dextrin (for granulation) 1.0 Vitamin B1 0.09 Vitamin B2 0.1 Vitamin B6 0.13 Vitamin B12 0.0002 Niacin 1.4 Pantothenic acid 0.42 Folic acid 0.017 Fat-coated vitamin C (VC 80%) 4.0 Pineapple powder flavoring 0.1 Sucrose 2.7328 Stevia 0.01 By using oil-coated vitamin C (manufactured by NOF Corporation), the acidity was suppressed, resulting in granules that were easy to eat. Furthermore, contact between vitamin C and PYR-containing collagen hydrolysates was prevented, thus preventing browning during long-term storage. Additionally, the inclusion of vitamin B2 results in a yellow coloration, which reduces the difference in discoloration during long-term storage.

[0192] (Prescription example 81) Beverages Ingredient name Amount PYR-containing collagen hydrolysate 5000mg Hyaluronic acid 10mg Placenta 10mg Ceramide 200 μg Coenzyme Q10 1mg Vitamin C 100mg Sweeteners (sucralose, acesulfame K) appropriate amount Acidulant (citric acid) appropriate amount 50mL water [Industrial applicability]

[0193] The collagen production promoter of the present invention can easily and efficiently promote collagen production and is useful for maintaining skin health, inhibiting skin aging, and treating conditions such as osteoporosis.

Claims

1. A collagen production promoter containing a 3-hydroxypyridinium derivative represented by the following general formula (1) as an active ingredient. 【Chemistry 1】 (In formula (1), A ４ is independently a hydroxyl group, an alkoxy group having 1 to 10 carbon atoms, a peptide structure having 1 to 100 amino acids bonded at the N-terminus, a hydrogen atom, an amino group (—NR ４ ２ ), or a hydrocarbon group having 1 to 10 carbon atoms. A ２ is independently a hydrogen atom, a hydrocarbon group having 1 to 10 carbon atoms, an acyl group (—COR ５ ), or a peptide structure having 1 to 100 amino acids bonded at the C-terminus. R is independently a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms. R １ is independently a single bond or a divalent hydrocarbon group having 1 to 10 carbon atoms which may have at least one functional group selected from the group consisting of a hydroxyl group, an amino group (—NR ４ ２ ), and an oxa group (—O—). R ２ is independently a hydrocarbon group having 1 to 10 carbon atoms which may have at least one functional group selected from the group consisting of a hydroxyl group, a carboxyl group, and an amino group (—NR ４ ２ ). R ３ is a hydrogen atom, a structure represented by the following formula (2), or a hydrocarbon group having 1 to 10 carbon atoms which may have at least one functional group selected from the group consisting of a hydroxyl group, a carboxyl group, and an amino group (—NR ４ ２ ). R ４ is independently a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms. R ５ is a hydrocarbon group having 1 to 10 carbon atoms. m and n independently represent integers of 0 to 4. However, m and n are numerical values such that m + n is an integer of 0 to 4.) 【Chemistry 2】 (In formula (2), A １ A in equation (1) １ Independently, a hydroxyl group, an alkoxy group having 1 to 10 carbon atoms, a peptide structure having 1 to 100 amino acids bonded at the N-terminus, a hydrogen atom, and an amino group (-NR ４ ２ ), or a hydrocarbon group having 1 to 10 carbon atoms, A ２ A in equation (1) ２ Independently, hydrogen atoms, hydrocarbon groups with 1 to 10 carbon atoms, and acyl groups (-COR) ５ ), or a peptide structure having 1 to 100 amino acids linked at the C-terminus, R is independently a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms in formula (1), R ４ R in equation (1) ４ Each of them independently contains a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms, R ５ R in equation (1) ５ A hydrocarbon group with 1 to 10 carbon atoms is independently attached to R ６ is a single bond, or a hydroxyl group and an amino group (-NR ４ ２ This represents a divalent hydrocarbon group having 1 to 10 carbon atoms, which may have at least one functional group selected from the group consisting of the following:

2. The collagen production promoter according to claim 1, characterized in that it contains 0.00001% by weight or more of the 3-hydroxypyridinium derivative as an active ingredient.

3. The collagen production promoter according to claim 1 or 2, characterized in that the 3-hydroxypyridinium derivative is derived from a natural product.

4. A cosmetic composition comprising a collagen production promoter according to any one of claims 1 to 3.

5. A food or beverage composition comprising a collagen production promoter according to any one of claims 1 to 3.

6. A cell culture composition comprising a collagen production promoter according to any one of claims 1 to 3.

7. A wound healing pharmaceutical composition comprising a collagen production promoter according to any one of claims 1 to 3.

8. A composition for promoting bone formation, comprising a collagen production promoter according to any one of claims 1 to 3.

9. A collagen hydrolysate characterized by containing 0.015% by weight or more of a 3-hydroxypyridinium substructure represented by the following general formula (3). 【Transformation 3】

10. A collagen hydrolysate characterized by containing 0.07% by weight or more of 3-hydroxypyridinoline represented by the following general formula (4). 【Chemistry 4】

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