HK1168036B - Collagen production accelerating composition - Google Patents

Description

Collagen production promoting composition

Technical Field

The present invention relates to a collagen production promoting composition comprising 1 or 2 or more compounds selected from the group consisting of D-aspartic acid and D-alanine, and derivatives and/or salts thereof, and a method for inhibiting and/or improving skin conditions comprising the step of administering the compounds.

Background

Type I collagen is one of the major proteins of the skin, and is 3-peptide chain proteins consisting of 2 α 1(I) peptide chains and 1 α 2(I) peptide chain. Type I collagen is produced by fibroblasts in the dermal layer of the skin, forming an extracellular matrix embedded in the fibroblasts. Both of normal (endocrinsic) aging and photoaging are accompanied by a decrease in collagen production and an increase in collagen degrading enzyme activity (non-patent documents 1 to 3). Here, it has been recognized that there is a correlation between promotion of collagen production and inhibition and/or improvement of skin conditions, such as wrinkle formation, due to general aging and photoaging. In fact, retinoids, which are derivatives of vitamin a, are known to have pharmacological effects on the skin of the face and upper arms, which are subjected to general aging and photoaging (non-patent documents 4 to 5). However, retinoids have side effects such as photosensitization and inflammatory reaction known as retinoid reaction (non-patent document 6). In addition, retinoids generally have strong photoreactivity, require light shielding for stable storage, and are easily decomposed by light transmitted into the body after administration to the skin under normal living conditions (non-patent document 3).

Documents of the prior art

Non-patent document

Non-patent document 1: takeda, k, et al, j.cell.physiol., 153: 450(1992)

Non-patent document 2: varani, j, et al, am.j.pathol, 158: 931(2001)

Non-patent document 3: varani, j, et al, am.j.pathol., 168: 1861(2006)

Non-patent document 4: kligman, a.m., et al, j.am.acad.dermaltol, 15: 836(1986)

Non-patent document 5: kligman, a.m., et al, j.am.acad.dermaltol, 29: 25(1993)

Non-patent document 6: mukherjee, s. et al, clin. interv. aging1: 327(2006)

Disclosure of Invention

Problems to be solved by the invention

Here, there is a need for the development of a novel composition having a collagen production promoting effect, which is free from side effects of retinoids and has high photostability.

Means for solving the problems

The present invention provides a collagen formation-promoting composition containing 1 or 2 or more compounds selected from the group consisting of D-aspartic acid and D-alanine, and derivatives and/or salts thereof.

The collagen production-promoting composition of the present invention is used for inhibiting and/or improving the skin condition according to circumstances.

In the collagen production-promoting composition of the present invention, the skin condition includes, but is not limited to, photoaging and/or wrinkles.

The collagen production-promoting composition of the present invention is used as a skin external preparation according to circumstances.

The collagen production-promoting composition of the present invention is used as a food according to circumstances.

The collagen production promoting composition of the present invention is, in some cases, a type I collagen production promoting composition.

The present invention provides a method for inhibiting and/or improving skin conditions, comprising the step of administering a collagen production-promoting composition comprising 1 or more than 2 compounds selected from the group consisting of D-aspartic acid and D-alanine, and derivatives and/or salts thereof.

Skin conditions inhibited and/or improved by the methods of the present invention include, but are not limited to, photoaging and/or wrinkles.

In the method of the present invention, the collagen production-promoting composition may be an external preparation for skin.

In the method of the present invention, the collagen production-promoting composition may be a food composition.

In the method of the present invention, the collagen production promoting composition is, in some cases, a type I collagen production promoting composition.

In the present specification, "salts" of D-aspartic acid and D-alanine refer to: any salt including metal salts, amine salts and the like, without impairing the collagen production promoting effect of D-aspartic acid and D-alanine. The aforementioned metal salt may include alkali metal salts, alkaline earth metal salts, and the like. The aforementioned amine salt may include triethylamine salt, benzylamine salt and the like.

In the present specification, "derivatives" of D-aspartic acid and D-alanine refer to: a compound in which D-aspartic acid and D-alanine molecules are covalently bonded to an arbitrary atomic group at an amino group, a carboxyl group, or a side chain, without impairing the collagen production promoting effect of D-aspartic acid and D-alanine. The optional group includes, but is not limited to, a protecting group such as N-phenylacetyl, 4' -Dimethoxytrityl (DMT), a biopolymer such as a protein, a peptide, a sugar, a lipid, a nucleic acid, etc., a synthetic polymer such as polystyrene, polyethylene, a polyvinyl compound, a polyester, etc., and a functional group such as an ester group. The ester group may include an aliphatic ester such as a methyl ester, an ethyl ester, or an aromatic ester.

Among amino acids, there are the L-form and the D-form which are optical isomers, natural proteins are proteins in which L-amino acids are bonded by peptide bonds, and only L-amino acids are used except cell walls of bacteria, and therefore, it is considered that only L-amino acids exist and only L-amino acids are used in mammals including humans. (Gentianzhong Chengyao et al, protein nuclease, 50: 453-460(2005), Lehninger's new biochemistry [ supra ] pp 2-147(1993) Guanchuan bookstore, Huppe. Biochemical original paperwork pp 22 21-30(1991) Wanshan). Therefore, conventionally, the L-amino acid has been used mainly as an amino acid, and the L-amino acid has been used industrially.

As examples of the exceptional use of D-amino acids, there are cases where they are used as a raw material for producing antibiotics in bacteria; and an example of a dietary supplement in which D-amino acid is directly used in the form of DL-amino acid mixture in order to save the cost of separating and extracting only L-amino acid from a mixture of L-amino acid and D-amino acid obtained in the same amount as when chemically synthesizing amino acid. However, there has never been an example of using only D-amino acids as a substance having physiological activity in the industry.

D-serine and D-aspartic acid have been studied more rapidly because of the high proportion of D-bodies. D-serine is locally present in the brain, hippocampus, and is known to be a regulator of NMDA receptors in the brain. It was confirmed that D-aspartic acid is localized in the testis and pineal gland and is involved in controlling the secretion of hormones (Japanese patent laid-open No. 2005-3558). The physiological role of D-aspartic acid and D-alanine in the skin is not clear.

As shown in the following examples, the collagen production-increasing effects of D-aspartic acid and D-alanine have not been known so far. Therefore, the collagen production promoting composition of the present invention containing D-aspartic acid and/or D-alanine is a novel invention.

In recent years, it has been reported that, when ddY mice were allowed to freely take 10mM aqueous solution of D-amino acid for two weeks and then the concentration of D-amino acid in each organ was measured, the results were as follows: in the pineal body, the concentration of D-amino acid is 3 to 1000pmol relative to 1 pineal body as a secretion organ; in brain tissue, the D Amino acid concentration is 2-500nmol relative to 1 gram wet mass (Morikawa, A. et al, Amino Acids, 32: 13-20 (2007)). Based on this, the lower limit of the daily intake of D-aspartic acid and D-alanine contained in the composition of the present invention described below was calculated.

As shown in the following examples, D-aspartic acid of the present invention has an effect of promoting collagen production in cultured human fibroblasts at a concentration ranging from 0.01 to 320. mu.M. Therefore, the amount of D-aspartic acid contained in the composition of the present invention may be any amount as long as the D-aspartic acid in the concentration range satisfies the condition that D-aspartic acid can be delivered to fibroblasts of the skin tissue of the living body. When the composition of the present invention is an external preparation, the content of D-aspartic acid may be 0.0000001 to 50 mass% or may be contained in the maximum mass concentration range, based on the total amount of the composition of the present invention. That is, the content of D-aspartic acid in the composition for external use is preferably 0.000001 to 30% by mass, and most preferably 0.00001 to 3% by mass. When the composition of the present invention is an oral preparation, the content of D-aspartic acid may be in the range of 0.0000001 to 100% by mass. The content of D-aspartic acid in the composition of the present invention is preferably 0.0000002 to 80% by mass, and most preferably 0.000001 to 60% by mass, when the composition is an oral preparation. The lower limit of the daily intake of D-aspartic acid contained in the composition of the present invention may be 0.01ng, preferably 0.1ng, more preferably 1ng, per 1kg body weight.

As shown in the following examples, D-alanine of the present invention has an effect of promoting collagen production in cultured human fibroblasts at a concentration ranging from 0.01 to 1000. mu.M. Therefore, the amount of D-alanine contained in the dermatological disease remedy, the external skin preparation, and the food composition of the present invention may be any amount as long as the D-alanine in the concentration range satisfies the condition that D-alanine can reach fibroblasts in living skin tissues. When the composition of the present invention is an external preparation, the content of D-alanine in the total amount of the composition of the present invention may be 0.000001 mass% to 50 mass% or may be contained in the maximum mass concentration range. That is, the content of D-alanine in the composition for external use is preferably 0.00001 to 30% by mass, and most preferably 0.0001 to 10% by mass. The daily content of D-alanine in the case of an oral preparation of the composition of the present invention may be in the range of 0.000001 to 100% by mass. The content of D-alanine in the composition of the present invention is preferably 0.00001 to 80% by mass, and most preferably 0.0001 to 60% by mass when the composition is an oral preparation. The lower limit of the daily intake of D-alanine contained in the composition of the present invention may be 0.01ng, preferably 0.1ng, more preferably 1ng, per 1kg body weight.

The composition of the present invention may contain 1 or 2 or more pharmaceutically acceptable additives without impairing the collagen production-promoting effect of D-aspartic acid and D-alanine, in addition to D-aspartic acid and D-alanine, and salts of D-aspartic acid and D-alanine, and/or derivatives capable of releasing D-aspartic acid and D-alanine in vivo by a drug metabolizing enzyme or the like. The additives include, but are not limited to, diluents and bulking agents, binders and binders, lubricants, glidants, plasticizers, disintegrants, carrier solvents, buffers, coloring materials, flavors, sweeteners, preservatives and stabilizers, adsorbents, and other pharmaceutical additives known to those skilled in the art.

The composition of the present invention may be prepared by using only D-aspartic acid and D-alanine, a salt of D-aspartic acid and D-alanine, and/or a derivative capable of releasing D-aspartic acid and D-alanine in vivo by a drug metabolizing enzyme or the like as an active ingredient, but other ingredients used in cosmetics, external skin preparations such as drugs including quasi drugs, and the like may be appropriately blended as necessary within a range not impairing the effect of the present invention. Examples of the other components (optional components) include oils, surfactants, powders, coloring materials, water, alcohols, thickeners, chelating agents, silicones, antioxidants, ultraviolet absorbers, humectants, perfumes, various medicinal components, preservatives, pH regulators, and neutralizers.

The dosage form of the collagen production promoting composition (hereinafter referred to as "skin condition improving agent") used for the purpose of inhibiting and/or improving the skin condition of the present invention may be any one of conventional quasi-pharmaceutical compositions and pharmaceutical compositions, and includes, for example, external preparations such as ointments, creams, lotions, masks, gels, and patches, oral preparations such as powders, granules, soft capsules, and tablets, and nasal preparations such as nasal spray, and injections.

The dosage form of the external preparation for skin of the present invention may be any one of conventional external preparations for skin, and includes, for example, ointment, cream, milky lotion, face toilet, pack, gel, patch and the like.

The food composition of the present invention may contain, as food-acceptable ingredients, seasonings, coloring materials, preservatives and the like, in addition to the D-aspartic acid and D-alanine, salts of D-aspartic acid and D-alanine, and/or derivatives capable of releasing D-aspartic acid and D-alanine in vivo by a drug metabolizing enzyme or the like, without impairing the collagen production promoting effect of D-aspartic acid and D-alanine.

The food composition of the present invention may be used in conventional food compositions such as candies, cookies, miso, French salad dressing, mayonnaise, French bread, soy sauce, yogurt, powdery food spread on rice, seasoning of seasoning natto, and moromi black vinegar, but is not limited to the above examples.

Drawings

FIG. 1 is a graph showing the effect of D-aspartic acid on type I collagen production in normal human dermal fibroblasts.

FIG. 2 is a graph showing the effect of D-alanine on type I collagen production in normal human dermal fibroblasts.

FIG. 3 is a graph showing the effect of L-and D-aspartic acid on type I collagen production in normal human dermal fibroblasts.

FIG. 4 is a graph showing the effect of L-and D-alanine on type I collagen production in normal human dermal fibroblasts.

FIG. 5 is a graph showing the effect of D-aspartic acid and D-alanine on type I atelocollagen production in normal human dermal fibroblasts.

FIG. 6 is a graph showing the effect of D-alanine on type I atelocollagen production in normal human dermal fibroblasts.

Detailed Description

The following examples of the present invention are given for illustrative purposes only and do not limit the scope of the present invention. The scope of the present invention is defined only by the description of the claims.

All documents referred to in this specification are incorporated in their entirety by reference.

Example 1

Effect of D-aspartic acid on promoting type I collagen production

Method of producing a composite material

Cell culture

The cells used were commercially available human neonatal dermal fibroblasts (CryoNHDF-Neo, Sanko pure chemical). The cells were 2X 10 per well⁵The cells were inoculated in a commercially available 24-well cell culture plate, and cultured in a medium (hereinafter referred to as "normal medium") containing 10% fetal bovine serum added to a cell culture medium (D-MEM (1g/L glucose) or Wako pure chemical industries). The cells were incubated at 37 ℃ with 5% CO₂And culturing under saturated steam condition for 4 hours.

Addition of amino acids

Then, the medium used for culturing the cells was replaced with a medium prepared by adding 0.5% fetal bovine serum to a commercially available cell culture medium (D-MEM (1g/L glucose) and Wako pure chemical industries, Inc. (hereinafter referred to as "low serum medium") at 37 ℃ and 5% CO₂And culturing under saturated steam condition for about 1 day. Here, D-aspartic acid (and Wako pure chemical industries, 018-04821) was added to the low-serum medium at concentrations of 0.01. mu.M, 0.1. mu.M, 10. mu.M, 100. mu.M and 320. mu.M, respectively. Magnesium L-ascorbyl Phosphate (L-Ascorbic Acid Phosphate Magnesium salt n-Hydrate, hereinafter referred to as "APM" and Wako pure chemical industries, 013-19641) as a precursor of vitamin C was added to the low serum medium at a concentration of 150. mu.M, 250. mu.M or 500. mu.M, respectively, as a positive control. In addition, the low serum medium without the addition of APM and D-aspartic acid served as a negative control.

Quantification of type I collagen production

After 2 days of culture, the culture supernatant was collected, and the concentration of the C-terminal peptide of type I Procollagen (hereinafter referred to as "PIP") produced from human neonatal dermal fibroblasts was measured using Procollagen type I C-peptide EIA kit (manufactured by Takara Bio Inc.) according to the manufacturer's instructions.

Quantitative results

FIG. 1 shows the results of an experiment for examining the effect of D-aspartic acid addition on type I collagen production in human neonatal dermal fibroblasts. Error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 4-6 times under the same condition. In addition, in the Bonferroni/Dunn test, asterisks (. + -.) indicate that the p value was less than 1%.

The PIP concentration of the negative control was 583 ng/mL. The PIP concentrations increased to 1183ng/mL, 1666ng/mL or 1416ng/mL with the addition of APM at 150. mu.M, 250. mu.M and 500. mu.M concentrations (positive control), respectively. PIP concentrations at the addition of D-aspartic acid at concentrations of 0.01. mu.M, 0.1. mu.M, 10. mu.M, 100. mu.M or 320. mu.M were 1286ng/mL, 1159ng/mL, 1117ng/mL, 1119ng/mL or 1007ng/mL, respectively. The effect of promoting collagen I production in the medium supplemented with APM and D-aspartic acid was statistically significantly different at all concentrations compared to the negative control. Furthermore, the effect of D-aspartic acid at a concentration of 0.01 to 100. mu.M on the promotion of type I collagen production was the same as that at the minimum concentration of 150. mu.M of APM, and D-aspartic acid was shown to have a far stronger type I collagen production promoting effect than APM.

Example 2

Collagen production-promoting effect of D-alanine

Method of producing a composite material

The cell culture and the amounts of amino acids and type I collagen produced were determined in the same manner as in example 1. D-alanine was used as the amino acid at a concentration of 0.01. mu.M, 0.1. mu.M, 10. mu.M, 1000. mu.M, 17400. mu.M (peptide research institute, 2801). Furthermore, the low serum medium without addition of APM and D-alanine was used as a negative control.

Quantitative results

FIG. 2 shows the results of experiments to investigate the effect of D-alanine addition on type I collagen production in human neonatal dermal fibroblasts. Error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 4-6 times under the same condition. In addition, in the Bonferroni/Dunn test, asterisks (. + -.) indicate that the p-value was less than 5%. Asterisks (—) indicate p values less than 1%.

The PIP concentration of the negative control was 551 ng/mL. The PIP concentration increased to 1183ng/mL, 1666ng/mL or 1416ng/mL when APM was added at a concentration of 150. mu.M, 250. mu.M or 500. mu.M (positive control), respectively, and collagen type I production was promoted. PIP concentrations when D-alanine was added at a concentration of 0.01. mu.M, 0.1. mu.M, 10. mu.M, 1000. mu.M or 17400. mu.M were 750ng/mL, 789ng/mL, 876ng/mL, 823ng/mL or 799ng/mL, respectively. The effect of promoting collagen I production in the medium supplemented with APM and D-alanine was statistically significantly different at all concentrations compared to the negative control.

Example 3

Collagen production-promoting effect of L-and D-aspartic acid

Method of producing a composite material

The cell culture and the amounts of amino acids and type I collagen produced were determined in the same manner as in example 1. The amino acids used were D-aspartic acid at a concentration of 0.1. mu.M (Wako pure chemical industries, 018-. Furthermore, the low serum medium without addition of L-and D-aspartic acid was used as a negative control.

Quantitative results

FIG. 3 shows the results of experiments investigating the effect of L-and D-aspartic acid addition on type I collagen production in human neonatal dermal fibroblasts. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 6-12 times under the same condition. In addition, in the Bonferroni/Dunn test, asterisks (. + -.) indicate that the p value was less than 1%.

The negative control had a PIP concentration of 361 ng/mL. PIP concentrations at the addition of 0.1. mu.M L-and D-aspartic acid were 406ng/mL and 456ng/mL, respectively. The above results show that the promotion of type I collagen production by the addition of 0.1. mu.M D-aspartic acid is statistically significantly different. On the other hand, addition of 0.1. mu.M of L-aspartic acid was not effective in promoting collagen I production.

Example 4

Collagen production-promoting effects of L-and D-alanine

Method of producing a composite material

The cell culture and the amounts of amino acids and type I collagen produced were determined in the same manner as in example 1. As amino acids, 0.1. mu.M and 150. mu.M of D-alanine (peptide research institute, 2801) and 0.1. mu.M and 150. mu.M of L-alanine (peptide research institute, 2701) were used. Furthermore, the low serum medium without addition of L-and D-alanine was used as a negative control.

Quantitative results

FIG. 4 shows the results of experiments investigating the effect of addition of L-and D-alanine to type I collagen production in human neonatal dermal fibroblasts. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 6-12 times under the same condition. In the Bonferroni/Dunn test, asterisks (. + -) and asterisks (. + -) indicate p values of less than 5% and 1%, respectively.

The negative control had a PIP concentration of 361 ng/mL. PIP concentrations were 502ng/mL and 450ng/mL with the addition of 0.1. mu.M and 150. mu.M D-alanine, respectively. PIP concentrations at the addition of 0.1. mu.M and 150. mu.M L-alanine were 405ng/mL and 413ng/mL, respectively. The above results show that there is a statistically significant difference in the promotion of type I collagen production by the addition of 0.1. mu.M and 150. mu.M D-alanine. On the other hand, addition of 0.1. mu.M and 150. mu.M L-alanine was not effective in promoting type I collagen production.

Example 5

Collagen production-promoting effects of D-aspartic acid and D-alanine

Method of producing a composite material

Cell culture and amino acid addition were performed in the same manner as in example 1. As the amino acids, 0.1. mu.M of D-aspartic acid (and Wako pure chemical industries, 018. quadrature., 04821), 0.1. mu.M and 0.001. mu.M of D-alanine (peptide research institute, 2801) were used. Furthermore, the low serum medium without addition of D-aspartic acid and D-alanine was used as a negative control. APM was adjusted to 250 μ M and added to the low serum medium as a positive control. To evaluate the amount of type I collagen produced, type I procollagen and procollagen produced by human neonatal dermal fibroblasts were treated with pepsin (800-.

Quantitative results (1)

FIG. 5 shows the results of experiments to investigate the effect of the addition of D-aspartic acid and D-alanine to type I atelocollagen production in human neonatal dermal fibroblasts. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 5-6 times under the same condition. In the Bonferroni/Dunn test, asterisks (. + -) and asterisks (. + -) indicate p values of less than 5% and 1%, respectively.

The negative control had a type I atelopeptide collagen concentration of 1.8. mu.g/mL. When 250mM APM was added (positive control), the type I atelocollagen concentration increased to 3.9. mu.g/mL, and the type I collagen production was promoted. The concentration of type I atelocollagen when 0.1. mu.M D-alanine was added was 4.0. mu.g/mL. The concentration of type I atelocollagen when 0.1. mu.M of D-aspartic acid was added was 3.8. mu.g/mL. The above results show that the promotion of type I collagen production by the addition of 0.1. mu.M of D-aspartic acid and D-alanine is statistically significantly different.

Quantitative results (2)

FIG. 6 shows the results of experiments to investigate the effect of D-alanine addition to human neonatal dermal fibroblasts on type I atelocollagen production. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 2 times under the same condition.

The negative control had a type I atelopeptide collagen concentration of 1.7. mu.g/mL. When 250mM APM was added (positive control), the type I atelocollagen concentration increased to 2.6. mu.g/mL, and the type I collagen production was promoted. The concentration of type I atelocollagen when 0.001. mu.M D-alanine was added was 2.1. mu.g/mL.

Example 6

Examples of formulations of the emulsion preparation, patch, tablet, soft capsule, granule, beverage, candy, biscuit, miso, french salad dressing, mayonnaise, french bread, soy sauce, yogurt, powdery food spread on rice, dressing of seasoning natto, fermented black vinegar, cream, body cream, gel, peel-off pack, impregnated pack, emulsion, lotion, and aerosol according to the present invention are shown below. These examples are given for the purpose of illustration and are not intended to limit the scope of the present invention.

COMPARATIVE EXAMPLE 1 (emulsion preparation)

COMPARATIVE EXAMPLE 2 (emulsion preparation)

COMPARATIVE EXAMPLE 3 (Patch)

COMPARATIVE EXAMPLE 4 (Patch)

COMPARATIVE EXAMPLE 5 (TABLET)

COMPARATIVE EXAMPLE 6 (TABLET)

COMPARATIVE EXAMPLE 7 (Soft Capsule)

COMPARATIVE EXAMPLE 8 (Soft Capsule)

COMPARATIVE EXAMPLE 9 (PARTICLES)

COMPARATIVE EXAMPLE 10 (DRINK)

COMPARATIVE EXAMPLE 11 (beverage)

COMPARATIVE EXAMPLE 12 (candy)

COMPARATIVE EXAMPLE 13 (biscuit)

Preparation of blending example 13 (biscuit)

While stirring butter, slowly adding fine granulated sugar, and stirring with egg, D-aspartic acid and/or D-alanine and spice. After mixing thoroughly, add the shaken whole low gluten flour, stir at low speed, store in a refrigerator in blocks. Then, the mixture was molded and baked at 170 ℃ for 15 minutes to make a biscuit.

COMPATIVE EXAMPLE 14 (miso)

Preparation example 14 (miso) production method

The rice koji and the salt are mixed well. Soaking cleaned semen glycines in 3 times of water overnight, removing water, adding new water, cooking, and sieving. The boiled broth (raw juice) was collected and dissolved to a concentration of 10% w/v D-aspartic acid and/or D-alanine. Immediately mashing the cooked beans, adding rice koji mixed with salt, and adding the raw juice dissolved with D-aspartic acid and/or D-alanine while uniformly mixing to have hardness similar to that of clay. The kneaded dough is tightly packed into a barrel without a gap until each corner is filled up, and after the surface is flattened, the dough is covered with a wrapper and sealed. After 3 months, the container was replaced, the surface was leveled and covered with a wrap. In addition, instead of adding D-aspartic acid and/or D-alanine to the raw juice, rice koji which produces a large amount of D-aspartic acid and/or D-alanine may be used. The rice koji can be obtained by quantifying D-aspartic acid and/or D-alanine by the method described in Japanese patent application laid-open No. 2008-185558. Further, D-aspartic acid and/or D-alanine or a salt thereof may be added to commercially available miso.

COMPARATIVE EXAMPLE 15 (French salad dressing)

COMPARATIVE EXAMPLE 16 (French salad dressing)

Preparation of blend examples 15 and 16 (French salad dressing)

After adding sodium chloride and D-aspartic acid or D-alanine to vinegar, the mixture was sufficiently stirred and dissolved. Salad oil was added, stirred well and pepper was added.

COMPATIVE EXAMPLE 17 (mayonnaise)

COMPATIVE EXAMPLE 18 (mayonnaise)

Preparation of blending examples 17 and 18 (mayonnaise)

Adding vinegar, sodium chloride, D-aspartic acid or D-alanine and pepper into egg yolk (at room temperature), and stirring with a whisk. Then, while adding salad oil little by little, stirring was continued until an emulsion was prepared. Finally adding granulated sugar and stirring.

COMPATIVE EXAMPLE 19 (French bread)

COMPATIVE EXAMPLE 20 (French bread)

Method for producing blend example 19 and blend example 20 (French bread)

Add 1g granulated sugar and dry yeast to warm water for pre-fermentation. The hard flour, the weak flour, sodium chloride, 5g granulated sugar and D-aspartic acid or D-alanine were put into a bowl, and the previously fermented yeast was added thereto. After sufficient kneading, the pellets were formed, and the first fermentation in the main fermentation was conducted at 30 ℃. The dough was kneaded again, later shaped to an appropriate shape, and post-fermented using an electronic fermenter. After scoring the dough surface, it was baked in an oven at 220 ℃ for 30 minutes.

Preparation example 21 (Soy sauce)

COMPATIVE EXAMPLE 22 (Soy sauce)

Preparation of formulation examples 21 and 22 (Soy sauce)

D-aspartic acid or D-alanine was added to commercially available soy sauce and stirred well. In addition, instead of adding D-aspartic acid or D-alanine or a salt thereof, rice koji which produces a large amount of D-aspartic acid or D-alanine may be used to brew soy sauce. The rice koji can be obtained by quantifying D-aspartic acid or D-alanine by the method described in Japanese patent application laid-open No. 2008-185558.

COMPARATIVE EXAMPLE 23 (yogurt)

COMPATIVE EXAMPLE 24 (yogurt)

Preparation of preparation examples 23 and 24 (yogurt)

Fermenting at 40-45 deg.C. Other commercially available species of bacteria may be used, and D-aspartic acid or D-alanine may be added to commercially available yogurt. In addition, instead of adding D-aspartic acid or D-alanine and its salt, can also be used to produce a large amount of D-aspartic acid or D-alanine bacteria. The bacterium can be selected by quantifying D-aspartic acid or D-alanine by the method described in Japanese patent application laid-open No. 2008-185558.

COMPARATIVE EXAMPLE 25 (powdery food sprinkled on rice)

COMPATIVE EXAMPLE 26 (seasoning Natto sauce)

Blend example 27 (seasoning Natto seasoning juice)

COMPATIVE EXAMPLE 28 (Natto)

Preparation of blend example 28 (inner Bean)

It is also possible to produce natto using a bacterium that produces a large amount of D-aspartic acid and/or D-alanine instead of adding D-aspartic acid and/or D-alanine and a salt thereof. In order to obtain the above-mentioned bacteria, D-aspartic acid and/or D-alanine can be quantitatively determined and selected by the method described in Japanese patent application laid-open No. 2008-185558.

COMPATIVE EXAMPLE 29 (fermented glutinous Black Vinegar)

Blend example 30 (fermented glutinous black vinegar)

Preparation of blending examples 29 and 30 (fermented glutinous Black Vinegar)

Instead of adding D-aspartic acid or D-alanine and salts thereof, vinegar, black vinegar, and mash may be prepared using a bacterium that produces a large amount of D-aspartic acid or D-alanine. The bacterium can be selected by quantifying D-aspartic acid or D-alanine by the method described in Japanese patent application laid-open No. 2008-185558.

COMPARATIVE EXAMPLE 31 (cream)

COMPATIVE EXAMPLE 32 (cream)

COMPARATIVE EXAMPLE 33 (cream for body)

COMPATIVE EXAMPLE 34 (cream for body)

COMPARATIVE EXAMPLE 35 (gelata)

COMPARATIVE EXAMPLE 36 (gelata)

Blending example 37 (Peel-off type mask)

Blending example 38 (Peel-off type mask)

COMPATIVE EXAMPLE 39 (impregnation type mask)

Blend example 40 (impregnated type mask)

COMPARATIVE EXAMPLE 41 (EMULSION)

COMPARATIVE EXAMPLE 42 (EMULSION)

COMPARATIVE EXAMPLE 43 (emulsion)

COMPARATIVE EXAMPLE 44 (emulsion)

COMPARATIVE EXAMPLE 45 (astringent)

COMPATIVE EXAMPLE 46 (astringent)

COMPARATIVE EXAMPLE 47 (astringent)

COMPARATIVE EXAMPLE 48 (astringent)

Blend example 49 (stock solution of urea aerosol for external use)

Blend example 50 (stock solution of urea aerosol for external use)

Blend example 51 (Aerosol urea spray)

Blending example 51 (urea aerosol spray) filling method

An aerosol was prepared by filling a stock solution of an aerosol urea external preparation and dimethyl ether in a pressure-resistant aerosol aluminum can whose inner surface was treated by coating with teflon (registered trademark).

Claims (5)

1. Use of 1 or 2 or more compounds selected from the group consisting of D-aspartic acid and D-alanine and salts thereof for the preparation of a collagen production-promoting composition.

2. Use according to claim 1, wherein the collagen production-promoting composition is used for the purpose of inhibiting and/or improving the skin condition.

3. Use according to claim 2, characterized in that the skin condition is photoaging and/or wrinkles.

4. Use according to any one of claims 1 to 3, characterized in that the collagen production-promoting composition is used as a skin external agent.

5. Use according to any one of claims 1 to 3, wherein the collagen production-promoting composition is used as a food product.