HK1171382A - Antioxidant composition - Google Patents

Description

Antioxidant composition

Technical Field

The present invention relates to an antioxidant composition comprising 1 or 2 or more compounds selected from the group consisting of D-aspartic acid and derivatives and/or salts thereof; and a method for improving skin conditions comprising the step of administering the compound; and methods of treating and/or preventing cataracts comprising the step of administering said compounds.

Background

Reactive Oxygen Species (ROS) non-selectively oxidize biological components such as nucleic acids, proteins, and lipids, and thus damage functions of the living body and structures of tissues and organs. ROS in the skin is referred to as a cause of skin diseases such as skin cancer, skin allergy, skin inflammation, and photoallergic skin diseases (non-patent document 1). ROS are known to act on the epidermis of the skin and cause the formation of skin conditions such as fine wrinkles, rough skin, and dryness.

Documents of the prior art

Non-patent document

Non-patent document 1: bickers, D.R and Athar, m., j., invest. 2565(2006)

Disclosure of Invention

Problems to be solved by the invention

Conventionally, ascorbic acid (vitamin C), α -tocopherol (vitamin E), and the like have been used as antioxidant compositions in cosmetics and pharmaceuticals, but their stability is insufficient. Accordingly, there is a need to develop an antioxidant composition that can be used daily, is stable, and is safe.

Means for solving the problems

The present invention provides an antioxidant composition comprising 1 or 2 or more compounds selected from the group consisting of D-and/or L-aspartic acid and derivatives and/or salts thereof.

The antioxidant composition of the present invention may be used for the purpose of inhibiting and/or improving skin conditions.

In the antioxidant composition of the present invention, the skin conditions include, but are not limited to, fine wrinkles, rough skin, dryness, skin cancer, skin allergy, skin inflammation, and photoallergic skin diseases.

The antioxidant composition of the present invention can be used as a skin external agent.

The antioxidant composition of the present invention can be used as food.

The antioxidant composition of the invention can be used as a medicine for cataract.

The drug for cataract in the antioxidant composition of the present invention may be a therapeutic agent for cataract or a prophylactic agent for cataract.

The antioxidant composition of the present invention can be suitably used as an eye drop.

The cataract may be senile cataract.

The present invention provides a method for improving skin conditions, comprising the step of administering an antioxidant composition containing 1 or 2 or more compounds selected from the group consisting of D-and/or L-aspartic acid and derivatives and/or salts thereof.

Skin conditions that are inhibited and/or improved by the methods of the present invention include, but are not limited to, fine wrinkles, rough skin, dryness, skin cancer, skin allergy, skin inflammation, and photoallergic skin diseases.

In the method of the present invention, the antioxidant composition may be a skin external preparation.

In the method of the present invention, the antioxidant composition may be a food composition.

The present invention provides a method for treating and/or preventing cataract, which comprises the step of administering a composition containing 1 or 2 or more compounds selected from the group consisting of D-and/or L-aspartic acid and derivatives and/or salts thereof.

In the method for treating and/or preventing cataract according to the present invention, the drug for cataract may be an eye drop.

In the method for treating and/or preventing cataract of the present invention, the cataract may be senile cataract.

In the present specification, "salt" of aspartic acid means: any salt including metal salts, amine salts and the like, without impairing the effect of aspartic acid on resistance to oxidative damage. 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 aspartic acid refer to: a compound in which an aspartic acid molecule is covalently bonded to an arbitrary atomic group at an amino group, a carboxyl group, or a side chain without impairing the effect of aspartic acid on resistance to oxidative damage. 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. According to (Gentianzhong Chengyao et al, protein nuclease, 50: 453-460(2005), Lehninger's new biochemistry [ supra ] pp 2-147(1993) Guanchuan bookstore, Hupo. Biochemical original document 22 pp21-30(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 raw materials for antibiotics produced 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 been no example in which only D-amino acids are industrially used as physiologically active substances.

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). However, the physiological role of D-aspartic acid in the skin is not clear.

As shown in the examples below, the effect of L-or D-aspartic acid for inhibiting oxidative damage has not been known so far. Accordingly, the oxidizing composition of the present invention containing L-and/or D-aspartic acid 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; the D-Amino acid concentration in brain tissue is 2-500nmol relative to 1 gram wet weight (Morikawa, A. et al, Amino Acids, 32: 13-20 (2007)). Based on this, the lower limit of the daily intake of L-and D-aspartic acid contained in the composition of the present invention described below is calculated.

As shown in the following examples, aspartic acid of the present invention has an effect of inhibiting oxidative damage to cultured human fibroblasts at a concentration of 0.1 to 10. mu.M. Therefore, the amount of aspartic acid contained in the skin disorder-ameliorating agent, the external skin preparation and the food composition of the present invention may be any amount as long as the aspartic acid in the concentration range satisfies the condition that the aspartic acid can reach fibroblasts of the living skin tissue. When the composition of the present invention is an external preparation, the aspartic acid content in the total amount of the composition of the present invention may be 0.000015 wt% to 50 wt% or may be contained in the maximum concentration range by weight. That is, the content of aspartic acid in the composition for external use is preferably 0.00003 to 30% by weight, and most preferably 0.0003 to 3% by weight. In addition, the content of aspartic acid in the composition of the present invention may be in the range of 0.00001 to 100% by weight when the composition is an oral preparation. The content of aspartic acid in the composition of the present invention in the case of an oral preparation is preferably 0.00002 to 80% by weight, and most preferably 0.0002 to 60% by weight. The lower limit of the daily intake of D-aspartic acid contained in the composition of the present invention may be 0.01ng per 1kg body weight, preferably 0.1ng, and more preferably 1 ng. The lower limit of the daily intake of L-aspartic acid contained in the composition of the present invention is less than the usual dose (2 mg per 1kg body weight) of a commercially available health drug, and it is sufficient if the dose is 0.01mg per 1kg body weight, preferably 0.1mg, more preferably 1 mg.

The composition of the present invention may contain 1 or 2 or more pharmaceutically acceptable additives without impairing the effect of aspartic acid on the resistance to oxidative damage, in addition to a single substance of aspartic acid, a salt of aspartic acid, and/or a derivative capable of releasing aspartic acid 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 aspartic acid, a salt of aspartic acid, and/or a derivative capable of releasing aspartic acid 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 pharmaceuticals including quasi drugs, and the like may be appropriately blended as necessary within a range not impairing the effects 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 antioxidant composition (hereinafter referred to as "skin condition improving agent") used for the purpose of suppressing and/or improving the skin condition of the present invention may be any one of conventional quasi-drug 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 aspartic acid, salts of aspartic acid and/or derivatives capable of releasing aspartic acid in vivo by a drug metabolizing enzyme or the like, without impairing the effect of aspartic acid against oxidative damage.

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.

ROS is considered not only as a skin disease but also as one of the causes of cataracts. It is considered that lipid peroxides generated in the interior of the crystals by reduction reaction of hydrogen peroxide, radical chain reaction of polyvalent unsaturated fatty acids, or the like cause not only disturbance of lipid composition but also denaturation of proteins to break down membrane functions, thereby causing turbidity of the crystals. (Shimaduojiamei and Shitian, structure of crystalline and its biochemistry, page 318 & 323, Shitian, editions of Shitian, medical sunflower publications, Tokyo (1986)). Therefore, according to these findings and the examples below, D-and/or L-aspartic acid having an antioxidant effect is effective for the prevention or treatment of cataract.

Drawings

Fig. 1 is a graph showing the effect of administration of carnosine on oxidative damage caused by hydrogen peroxide in normal human dermal fibroblasts.

FIG. 2 is a graph showing the effect of L-aspartic acid administration on oxidative damage caused by hydrogen peroxide in normal human dermal fibroblasts.

FIG. 3 is a graph showing the effect of administration of D-aspartic acid on oxidative damage caused by hydrogen peroxide in normal human dermal fibroblasts.

Fig. 4 is a graph showing the effect of administration of carnosine on oxidative damage due to AAPH in normal human dermal fibroblasts.

FIG. 5 is a graph showing the effect of administration of L-and D-aspartic acid on oxidative damage due to AAPH 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.

Example 1

Evaluation test of antioxidant Effect

1. Purpose(s) to

ROS include the narrow sense of reactive oxygen species consisting of superoxide anion, hydroxyl radical, hydrogen peroxide, and singlet oxygen, and the broad sense of reactive oxygen species including alkyl oxygen radicals, peroxy hydroxyl radical, peroxy radicals, hydroperoxides, transition metal ion oxygen complexes, and the like. Although the hydroxyl radical has the highest oxidizing power in ROS, it has a very short life, and thus only living components such as nucleic acids, proteins, and lipids in the vicinity of the site of production are oxidized non-selectively. The peroxy radical oxidation force is weak relative to the hydroxyl radical, but since it is relatively stable, it diffuses and causes cell membrane damage through radical chain reaction of polyvalent unsaturated fatty acids. Hydroxyl radicals can produce peroxy radicals, but peroxy radicals cannot. Thus, effective antioxidants may differ due to the different mechanisms of action of hydroxyl and peroxy radicals. In this example, the antioxidant effect of hydrogen peroxide, a representative compound generating hydroxyl radicals, and 2, 2' -azobis (2-amidinopropane) dihydrochloride (hereinafter referred to as "AAPH"), a representative compound generating peroxy radicals, were evaluated. As a positive control, carnosine known to have an antioxidant effect was used.

2. Materials and methods

2-1. cells

In the evaluation of the antioxidant effect of hydrogen peroxide, human neonatal dermal fibroblasts (Cryo NHDF-Neo, sanguang pure drug) were used as cells. The cells were 1X 10 per well⁵Each of the cells was inoculated into a 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. In the evaluation of the antioxidant effect of AAPH, antibiotics (penicillin, streptomycin, amphotericin B (fungizone)) were added to the ordinary medium, and the cells were cultured for 1 day.

2-2. culture medium for evaluating antioxidant effect

Then, the cells were transferred to a medium containing 0.01% or 0.05% carnosine or 0.1. mu.M or 10. mu.M D-or L-aspartic acid in a medium for cell culture (D-MEM (1g/L glucose) and Wako pure chemical industries, all of which were supplemented with 0.5% fetal bovine serum) (hereinafter referred to as "low serum medium"), and the cells were cultured at 37 ℃ in 5% CO₂And culturing for 2 days under saturated water vapor condition. In the AAPH induced oxidative damage evaluation experiment, the cells were transferred to a medium further supplemented with 5ppm or 100ppm of carnosine or 10. mu.M of D-or L-aspartic acid, and cultured for 2 days. Make itThe low serum medium was used as a negative control with neither carnosine nor aspartic acid added.

2-3, adding an oxidizing agent

After 2 days of culture, 1mM or 4mM hydrogen peroxide or 50mM or 100mM AAPH was added to the medium for evaluation of antioxidant effect, and the antioxidant effect of carnosine or aspartic acid was investigated. As a control for evaluation of toxicity of the antioxidant when no oxidizing agent was added, a low serum medium to which neither hydrogen peroxide nor AAPH was added was used.

2-4. quantification of oxidative damage

After 2 hours from the addition of hydrogen peroxide or AAPH, alamarBlue (trademark, Biosource, Biosource International) was added to the medium so that the final concentration reached 10%, and after 2 to 3 hours, the fluorescence intensity of the supernatant was measured at an excitation wavelength of 544nm and a fluorescence wavelength of 590nm according to Ahmed S.A. et al (J.Immunol. method.170, 211-224(1994)) and the manufacturer's instructions.

3. Results

3-1 antioxidant effect of carnosine on Hydrogen peroxide

FIG. 1 shows the results of experiments investigating the antioxidant effect of carnosine on hydrogen peroxide in Cryo NHDF-Neo cells. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 3 times under the same condition. In the Bonferroni/Dunn test, an asterisk (. + -.) indicates that t is less than 5%.

The viable cell rate of the control for evaluation of toxicity of antioxidant without addition of oxidizing agent was 102% when carnosine was not added, 100% when carnosine concentration was 0.01%, and 97% when carnosine concentration was 0.05%. The viable cell rate at a hydrogen peroxide concentration of 1mM was 45% when no carnosine was added, 51% when the carnosine concentration was 0.01%, and 53% when the carnosine concentration was 0.05%. The viable cell rate was 14% when no carnosine was added, 21% when the carnosine concentration was 0.01%, and 45% when the carnosine concentration was 0.05% at 4 mM. When the concentration of hydrogen peroxide was 4mM, the ratio of viable cells was considered to be significantly different when the concentration of carnosine was 0.05% compared to the ratio of viable cells when carnosine was not added. From the above results, in the experimental series of the present example, the antioxidant effect of carnosine against hydrogen peroxide was confirmed.

Antioxidant effect of L-aspartic acid on hydrogen peroxide

FIG. 2 shows the results of experiments investigating the antioxidant effect of L-aspartic acid on hydrogen peroxide in Cryo NHDF-Neo cells. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 3 times under the same condition.

The viable cell rate of the control used for evaluation of the toxicity of the antioxidant when no oxidizing agent was added was 93% when no L-aspartic acid was added, 88% when the concentration of L-aspartic acid was 0.1. mu.M, and 97% when the concentration of L-aspartic acid was 10. mu.M. The viable cell rate was 61% at a hydrogen peroxide concentration of 1mM, 62% at an L-aspartic acid concentration of 0.1. mu.M, and 62% at an L-aspartic acid concentration of 10. mu.M. The viable cell rate was 36% when hydrogen peroxide was 4mM, 33% when the concentration of L-aspartic acid was 0.1. mu.M, and 32% when the concentration of L-aspartic acid was 10. mu.M. From the above results, L-aspartic acid was found to have no statistically significant antioxidant effect against hydrogen peroxide.

Antioxidant effect of D-aspartic acid on hydrogen peroxide

FIG. 3 shows the results of experiments investigating the antioxidant effect of D-aspartic acid on oxidative damage due to hydrogen peroxide in Cryo NHDF-Neo cells. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 3 times under the same condition. In addition, in the Bonferroni/Dunn test, asterisks (. + -.) indicate less than 5% of p, and asterisks (. + -.) indicate less than 1% of p.

The viable cell rate of the control used for evaluation of the toxicity of the antioxidant when no oxidizing agent was added was 97% when no D-aspartic acid was added, 86% when the concentration of D-aspartic acid was 0.1. mu.M, and 97% when the concentration of D-aspartic acid was 10. mu.M. The viable cell rate was 55% at a hydrogen peroxide concentration of 1mM, 62% at a D-aspartic acid concentration of 0.1. mu.M, and 63% at a D-aspartic acid concentration of 10. mu.M. The viable cell rate was 22% at a hydrogen peroxide concentration of 4mM, 29% at a D-aspartic acid concentration of 0.1. mu.M, and 34% at a D-aspartic acid concentration of 10. mu.M. The difference in the viable cell rate between the addition of 0.1. mu.M and 10. mu.M D-aspartic acid was considered significant when the hydrogen peroxide concentration was 4mM, compared with the viable cell rate in the absence of the addition of D-aspartic acid. From the above results, D-aspartic acid is considered to have a concentration-dependent antioxidant effect on hydrogen peroxide.

3-4 antioxidant effect of carnosine in evaluation experiment of oxidative damage due to AAPH

FIG. 4 shows the results of experiments investigating the antioxidant effect of carnosine on oxidative damage due to AAPH in Cryo NHDF-Neo cells. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 3 times under the same condition. In addition, in the Bonferroni/Dunn test, asterisks (. + -.) indicate that p is less than 1%.

The viable cell rate of the control for evaluating the toxicity of the antioxidant without adding the oxidizing agent was 100% when no carnosine was added, 93% when the carnosine concentration was 5ppm, and 103% when the carnosine concentration was 100 ppm. The viable cell rate at an AAPH concentration of 100mM was 31% without carnosine, 60% at a carnosine concentration of 5ppm, and 85% at a carnosine concentration of 100 ppm. When the concentration of AAPH was 100mM, the difference in the viable cell rate was considered to be significant when 100ppm of carnosine was added, compared with the viable cell rate when no carnosine was added. From the above results, in the experimental system of this example, the antioxidant effect of carnosine on AAPH was confirmed.

3-5 antioxidant Effect of L-and D-aspartic acids in evaluation of oxidative Damage due to AAPH

FIG. 5 shows the results of experiments investigating the antioxidant effect of L-and D-aspartic acid on oxidative damage due to AAPH in Cryo NHDF-Neo cells. The error bars for each experimental condition represent the standard deviation of the measured values of the experimental results repeated 3 times under the same condition. In the Bonferroni/Dunn test, an asterisk (x) indicates that p is less than 5%, and an asterisk (x) indicates that p is less than 0.1%.

The toxicity of the antioxidant was evaluated in the absence of the oxidizing agent by using the control for live cell rate, which was 95% when L-and D-aspartic acid were not added, 102% when the concentration of D-aspartic acid was 10. mu.M, and 80% when the concentration of L-aspartic acid was 10. mu.M. The viable cell rate was 51% at an AAPH concentration of 100mM, 96% at a D-aspartic acid concentration of 10. mu.M, and 69% at an L-aspartic acid concentration of 10. mu.M, without addition of L-and D-aspartic acids. At an AAPH concentration of 100mM, the difference in the viable cell rate was considered significant when 10. mu.M of L and D-aspartic acid was added, compared with the viable cell rate when L-and D-aspartic acid were not added. From the above results, it was confirmed that D-aspartic acid has a better antioxidant effect on AAPH than L-aspartic acid.

4 conclusion

From the experimental results of this example, it was confirmed that D-aspartic acid had an antioxidant effect on both hydrogen peroxide and AAPH, but L-aspartic acid was confirmed to have an antioxidant effect only on AAPH. Here, it is suggested that D-aspartic acid has an antioxidant effect on both hydroxyl radicals and peroxy radicals, but L-aspartic acid has an antioxidant effect only on peroxy radicals.

Example 2

Examples of the formulation of the aspartic acid-containing 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, lotion, toner, and aerosol according to the present invention will be described below. The aspartic acid in the following formulation examples is D-form and/or L-form. 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 (Patch)

COMPARATIVE EXAMPLE 3 (TABLET)

COMPARATIVE EXAMPLE 4 (TABLET)

COMPARATIVE EXAMPLE 5 (Soft Capsule)

COMPARATIVE EXAMPLE 6 (Soft Capsule)

COMPARATIVE EXAMPLE 7 (PARTICLES)

COMPARATIVE EXAMPLE 8 (DRINKS)

COMPARATIVE EXAMPLE 9 (candy)

COMPARATIVE EXAMPLE 10 (biscuit)

Preparation of blending example 10 (biscuit)

While stirring the butter, slowly adding fine granulated sugar, and adding egg, aspartic acid and spice and stirring. 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 11 (miso)

Preparation of blend example 11 (miso)

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 the aspartic acid was dissolved to a concentration of 10% w/v. Immediately mashing the cooked beans, adding rice koji mixed with salt, and adding the raw juice dissolved with aspartic acid while uniformly mixing to have a 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 aspartic acid to the raw juice, rice koji which produces a large amount of aspartic acid may be used. In order to obtain the rice koji, aspartic acid can be quantitatively determined and selected by the method described in Japanese patent application laid-open No. 2008-185558. Further, aspartic acid or a salt thereof may be added to commercially available miso.

COMPARATIVE EXAMPLE 12 (French salad dressing)

Preparation of blend example 12 (French salad dressing)

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

Preparation example 13 (mayonnaise)

Preparation method of blending example 13 (mayonnaise)

Adding vinegar, sodium chloride, aspartic acid 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 14 (French bread)

Preparation of formulation example 14 (French bread)

Add 1g granulated sugar and dry yeast to warm water for pre-fermentation. Hard flour, soft flour, sodium chloride, 5g granulated sugar and aspartic acid were placed in a bowl, to which previously fermented yeast was added. 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.

COMPARATIVE EXAMPLE 15 (Soy sauce)

Preparation of formulation example 15 (Soy sauce)

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

COMPATIVE EXAMPLE 16 (yogurt)

Preparation method of formula 16 (yogurt)

Fermenting at 40-45 deg.C. Other commercially available species of bacteria may be used, and aspartic acid may be added to commercially available yogurt. In addition, instead of adding aspartic acid or its salt, bacteria producing a large amount of aspartic acid may be used. In order to obtain such bacteria, aspartic acid can be quantified and selected by the method described in Japanese patent application laid-open No. 2008-185558. Further, aspartic acid or a salt thereof may be added to commercially available yogurt.

COMPARATIVE EXAMPLE 17 (powdery food sprinkled on rice)

COMPARATIVE EXAMPLE 18 (seasoning Natto sauce)

COMPATIVE EXAMPLE 19 (Natto)

Preparation of blend example 19 (Natto)

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

Blending example 20 (fermented glutinous black vinegar)

Preparation of blending example 20 (fermented glutinous Black Vinegar)

Instead of adding aspartic acid or its salt, the bacteria producing a large amount of aspartic acid can be used to prepare vinegar, black vinegar, and mash. In order to obtain such bacteria, aspartic acid can be quantified and selected by the method described in Japanese patent application laid-open No. 2008-185558. Further, aspartic acid or a salt thereof may be added to commercially available moromi black vinegar.

COMPARATIVE EXAMPLE 21 (cream)

COMPARATIVE EXAMPLE 22 (cream for body)

COMPARATIVE EXAMPLE 23 (gelata)

Blending example 24 (peel-off type mask)

Blend example 25 (impregnated type mask)

COMPARATIVE EXAMPLE 26 (emulsion)

COMPARATIVE EXAMPLE 27 (emulsion)

COMPARATIVE EXAMPLE 28 (astringent)

COMPARATIVE EXAMPLE 29 (astringent)

Blend example 30 (stock solution of urea aerosol for external use)

Blend example 31 (Aerosol urea spray)

Blending example 31 (Aerosol Urea 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 (8)

1. An antioxidant composition comprising 1 or 2 or more compounds selected from the group consisting of D-aspartic acid and derivatives and/or salts thereof.

2. Composition according to claim 1, characterized in that it is used for the purpose of inhibiting and/or improving the condition of the skin.

3. The composition of claim 2, wherein the skin condition is selected from the group consisting of fine wrinkles, rough skin, dryness, skin cancer, skin allergy, skin inflammation, and photoallergic skin diseases.

4. Composition according to any one of claims 1 to 3, characterized in that it is used as a skin external agent.

5. Composition according to any one of claims 1 to 3, characterized in that it is used as a food product.

6. Composition according to claim 1, for use as a medicament for cataracts.

7. The composition according to claim 6, wherein the drug for cataract is a therapeutic agent for cataract or a prophylactic agent for cataract.

8. A composition according to claim 6 or 7, characterized in that it is suitable as an eye drop.