JP2017014155A - Composition for protection of retina - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide compositions for protecting retina which can protect retinal pigment epithelial cells from the damage of active oxygen more effectively and inhibit retinopathy.SOLUTION: A composition for protecting retina contains as active ingredients astaxanthin and at least one ingredient selected from the group consisting of Ipomoea batatas young-leaf processed materials, Pueraria montana flower processed materials, Vaccinium myrtillus processed materials, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, vitamin C, vitamin D3, vitamin K, a calcium compound, a zinc compound, an iron compound, a molybdenum compound, a chromium compound, and shellac.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a retinal protective composition, and more particularly to a retinal protective composition containing astaxanthin and specific other components as active ingredients.

  It is known that the function of the retinal pigment epithelium, which is involved in photoreceptor repair and regeneration, decreases with age. When you are young, there are enough SOD enzymes in your body to prevent the negative effects of active oxygen, but with the aging, the function of the SOD enzyme decreases and the function of the retinal pigment epithelium also decreases.

  On the other hand, astaxanthin has a high antioxidant capacity and an antioxidant capacity about 1000 times that of vitamin E, and therefore has been proposed for applications such as in vivo antioxidants and improving eye regulation. For example, a health food mainly composed of anthocyanin and astaxanthin has been proposed (see Patent Document 1).

JP 2008-035714 A

  An object of the present invention is to provide a retinal protective composition capable of effectively protecting retinal pigment epithelial cells from active oxygen damage and suppressing retinal damage.

  As a result of diligent research and research on the protection of retinal pigment epithelial cells, which play an important role in the retina, from active oxygen, the combination of astaxanthin and certain other components has resulted in high retinal pigment epithelial cell protection. It has been found that an effect can be obtained, and the present invention has been completed. In other words, it has been found that the combination of astaxanthin and other specific components that have little or no ability to protect retinal pigment epithelial cells can more effectively suppress damage to retinal pigment epithelial cells due to active oxygen. It was.

  That is, the present invention relates to astaxanthin, sweet potato young leaf processed product, kuzu flower processed product, bilberry processed product, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, vitamin C, vitamin D3. And an active ingredient containing at least one component selected from the group consisting of vitamin K, calcium compound, zinc compound, iron compound, molybdenum compound, chromium compound, and shellac.

  The retinal protective composition of the present invention can be used as an eye function maintaining agent having a retinal protective action.

  The retinal protective composition of the present invention may be obtained by adding an active ingredient.

  The retinal protective composition of the present invention is preferably for oral use, and is preferably a tablet, capsule, powder, granule, or liquid.

  The present invention also relates to a method for maintaining an eye function (except for medical practice), characterized in that the retinal protective composition of the present invention is ingested.

  According to the retinal protective composition of the present invention, retinal pigment epithelial cells can be effectively protected from active oxygen damage, and retinal damage can be suppressed.

It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + plant material) of this invention to a human retinal pigment epithelial cell (ARPE-19), and an upper left figure as plant material Shown using processed sweet potato leaves, upper right figure using kuzu flower processed plant material, lower left figure using bilberry processed plant material, lower right figure plant material As shown below, gallic acid is used. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + amino acid) of this invention to a human retinal pigment epithelial cell (ARPE-19), The upper left figure is glutamic acid as an amino acid. The upper right figure shows the one using histidine as the amino acid, the lower left figure shows the one using methionine as the amino acid, and the lower right figure shows the one using serine as the amino acid. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + amino acid) of this invention to a human retinal pigment epithelial cell (ARPE-19), and the left figure shows tyrosine as an amino acid. The right figure shows what used valine as an amino acid. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + vitamins) of this invention to a human retinal pigment epithelial cell (ARPE-19), and an upper left figure is vitamins The figure using vitamin B6 is shown, the upper right figure shows using vitamin B12 as vitamins, the lower left figure shows using vitamin C as vitamins, and the lower right figure uses vitamin D3 as vitamins Show things. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + vitamins) of this invention to a human retinal pigment epithelial cell (ARPE-19), and vitamin K is used as vitamins. Shows what was used. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + minerals) of this invention to a human retinal pigment epithelial cell (ARPE-19), and an upper left figure as minerals The figure using the calcium compound is shown, the upper right figure shows the one using the zinc compound as the minerals, the lower left figure shows the one using the iron compound as the minerals, the lower right figure uses the molybdenum compound as the minerals Show things. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + minerals) of this invention to a human retinal pigment epithelial cell (ARPE-19), and uses chromium compound as minerals. Shows what was used. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker at the time of applying the retinal protective composition (astaxanthin + shellac) of this invention to a human retinal pigment epithelial cell (ARPE-19).

  The retinal protective composition of the present invention comprises astaxanthin, sweet potato young leaf processed product, kuzu flower processed product, bilberry processed product, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, vitamin C. , Vitamin D3, vitamin K, calcium compound, zinc compound, iron compound, molybdenum compound, chromium compound, and at least one component selected from the group consisting of shellac (hereinafter sometimes referred to as other components) as an active ingredient It is characterized by containing. The other component used with astaxanthin may be used individually by 1 type, and may be used in combination of 2 or more type. When using 2 or more types of other components in combination, it is preferable to combine other components having high synergistic effect with astaxanthin.

  The retinal protective composition of the present invention can more effectively protect retinal pigment epithelial cells from active oxygen damage, suppress retinal damage, delay eye aging, and maintain eye function. it can. In addition, it is possible to suppress the occurrence of retinal diseases such as age-related macular degeneration, retinitis pigmentosa, central choroidal disease, macular retinal epitheliogenesis, and macular hole. Therefore, the retinal protective composition of the present invention can be used as an eye function maintaining agent, retinal protective agent, retinal disorder inhibitor, retinal disease preventive agent, and the like.

[Astaxanthin]
Astaxanthin used in the present invention is a kind of carotenoid, and is said to have an antioxidant action 1000 times that of vitamin E, and may be derived from natural products or obtained by synthesis. Examples of those derived from natural products include astaxanthin obtained from algae such as Haematococcus algae, crustaceans such as shrimp and crab, and fishes such as salmon and Thailand. Naturally derived extracts and chemically synthesized products are commercially available, and these commercially available products can be used.

[Other ingredients]
(Processed sweet potato leaves)
Sweet potato is a plant belonging to the convolvulaceae family and is generally called sweet potato. The sweet potato varieties are not particularly limited, and examples include suiou, joy white, koganesengan, white yutaka, sweet potato, and ayamurasaki. Examples of the processed sweet potato leaves of the present invention include processed products such as crushed, squeezed, and extract of sweet potato young leaves. Examples of the pulverized product include powder and granules. The juice or extract may be liquid, but can also be used as a paste or dry powder. The extract can be obtained by extraction using an appropriate solvent. As the solvent, for example, water (hot water), ethanol, or hydrous ethanol can be used.

(Kuzu's flower processed product)
Kuzu is a perennial plant of the leguminous genus Kudzu. As the kuzu flower, those collected in any process from the bud to the fully opened flower may be used, and those collected in each process may be used in combination. The type of kuzu is not particularly limited, and examples thereof include Pueraria thomsonii, Pueraria lobata, Pueraria thunbergiana, and the like. Examples of the kuzu flower processed product of the present invention include processed products such as a kuzu flower pulverized product, juice, and extract. Examples of the pulverized product include powder and granules. The juice or extract may be liquid, but can also be used as a paste or dry powder. The extract can be obtained by extraction using an appropriate solvent. As the solvent, for example, water (hot water), ethanol, or hydrous ethanol can be used.

(Bilberry processed product)
Bilberry is a fruit of a low shrub species of the genus Vaccinaceae, and is sometimes referred to as white berry, winberry, braeberry, and European blueberry. Examples of the processed bilberry product of the present invention include processed products such as a pulverized product, juice, and extract of bilberry. Examples of the pulverized product include powder and granules. The juice or extract may be liquid, but can also be used as a paste or dry powder. The extract can be obtained by extraction using an appropriate solvent. As the solvent, for example, water (hot water), ethanol, or hydrous ethanol can be used.

(Other)
The gallic acid may be derived from a plant, but is not limited thereto. Amino acids such as glutamic acid, histidine, methionine, serine, tyrosine, and valine include salt forms. Examples of the salt include sodium salt and hydrochloride.

  The retinal protective composition of the present invention can be used as an eye function maintaining agent having a retinal protective action, and the eye function maintaining agent contains astaxanthin and a predetermined other component, and is used for maintaining the eye function. The product is not particularly limited as long as it can be distinguished from other products. For example, an indication that any of the main body, packaging, instructions, and promotional material of the product according to the present invention has a function of maintaining the function of the eye such as a retinal protective effect, a retinal disorder suppressing effect, and a retinal disease preventing effect. Is included in the scope of the present invention. For example, so-called health foods such as pharmaceuticals (including quasi-drugs), cosmetics, functional foods for which the indication of efficacy has been approved by prescribed organizations such as foods for specified health use, functional nutritional foods, functional indication foods Can be mentioned. In so-called health foods, “maintain the amount of pigment in the macula of the eyes (eyes)”, “to worry about eyes (eyes) due to aging”, “protect eyes (eyes) from light irritation”, “ "For blue light measures", "For liquid crystal light measures", "If you are tired from reading, computer, smartphone", "For those who are worried about being blurred", "For improving contrast sensitivity", "For tired eyes (eyes)" , “To adjust the condition of the eyes (eyes)”, “to remove the fatigue of the eyes (eyes)”, and the like can be exemplified. The retinal protective composition of the present invention can be taken regardless of gender and age as long as the above effect is concerned, but the effect of the retinal protective composition of the present invention can be enjoyed more effectively. Therefore, it is desirable for healthy people except sick people to take it. In addition, it is desirable to be taken by a person who performs work that overuses the eyes for a long time, and it is particularly desirable for a desk worker who works for a long time using a personal computer or the like.

  Examples of the form of the retinal protective composition of the present invention include tablets, capsules, powders, granules, liquids, granules, rods, plates, blocks, solids, and rounds. , Paste agents, cream agents, caplet agents, gel agents, chewable agents, stick agents and the like. Among these, the forms of tablets, capsules, powders, granules, and liquids are particularly preferable. Specifically, supplements, container-packed beverages filled in PET bottles, cans, bottles, etc., instant powder beverages and instant granule beverages for dissolving and drinking in water (hot water), milk, fruit juice, green juice, etc. Can be illustrated. These are preferable in that they can be easily taken at the time of eating and the like, and can enhance palatability.

  The contents of astaxanthin and other components (active ingredients) in the retinal protective composition of the present invention may be appropriately contained within the range where the effects are exerted.

  In general, when the retinal protective composition of the present invention is a pharmaceutical product or a supplement (tablet, capsule), the active ingredient is preferably contained in an amount of 0.01 to 100% by mass in terms of dry mass. 0.1 to 85% by mass is more preferable, and 0.5 to 70% by mass is further preferable.

  When the retinal protective composition of the present invention is a packaged beverage (liquid), the active ingredient is preferably contained in an amount of 0.01 to 15% by mass in terms of dry mass, and 0.03 to 12. It is more preferable that 5 mass% is contained, and it is further more preferable that 0.05-10 mass% is contained.

  When the retinal protective composition of the present invention is an instant powder beverage (powder) or an instant granule beverage (granule), the active ingredient is contained in an amount of 0.1 to 80% by mass in terms of dry mass. It is preferable that 0.5 to 70% by mass is included, and 1 to 60% by mass is more preferable.

  In order to exhibit the effects of the present invention more effectively, the active ingredient is preferably contained in an amount of 80% by mass or more of the entire retinal protective composition of the present invention (excluding moisture) in terms of dry mass, and 90% by mass. It is more preferably contained, more preferably 95% by mass or more, and particularly preferably 100% by mass.

  The intake amount of the retinal protective composition of the present invention is not particularly limited, but from the viewpoint of exerting the effects of the present invention more remarkably, the intake amount of the active ingredient per day is 0.1 mg / day or more. It is preferable to take 1 mg / day or more, more preferably 2 mg / day or more. Although the upper limit in particular is not restrict | limited, For example, it is 4 g / day, Preferably it is 2 g / day.

  The retinal protective composition of the present invention can be accommodated in one container or divided into a plurality of containers, for example, as one day so that the daily intake becomes the above-mentioned intake. .

  The blending mass ratio of astaxanthin and other components is preferably in the range of 0.5: 1 to 70: 1 and more preferably in the range of 0.75: 1 to 60: 1 in terms of dry mass. The range of 1: 1 to 60: 1 is more preferable, and the range of 1: 1 to 50: 1 is particularly preferable. The effect of this invention can be exhibited more effectively because the compounding ratio of astaxanthin and other components is in the above range.

  The retinal protective composition of the present invention can be produced by a known method by adding components other than the active ingredient acceptable as an oral preparation, if necessary.

  The retinal protective composition as an oral preparation of the present invention includes a retinal protective composition (retinal protective food) obtained by adding an active ingredient to a food in addition to a retinal protective composition containing an active ingredient. For example, an active ingredient with respect to a food in which the content of the active ingredient of the present invention is increased compared to a normal food (including a natural food) or a food that does not normally contain the active ingredient of the present invention. Can be mentioned. For the addition of the active ingredients, the respective ingredients may be added separately, may be added simultaneously, or may be added together with other ingredients other than the active ingredients.

  Examples of the retina-protecting food of the present invention include carbonated beverages, nutritional beverages, fruit beverages, lactic acid beverages, smoothies, green juice, and other frozen desserts; ice cream, ice sherbet, shaved ice, and so on; soba, udon, harusame, chinese noodles Noodles such as instant noodles; candy, candies, gum, chocolate, tablet confectionery, snack confectionery, biscuits, jelly, jam, cream, baked confectionery, bakery confectionery, etc .; Dairy products such as processed milk, fermented milk, yogurt; salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing and other fats and processed foods; seasonings such as sauces, soy sauce; curry, stew, parent and rice bowl, Porridge, miscellaneous cooking, Chinese rice bowl, bonito, tempura bowl, beef bowl, hayashi rice, omelet rice, oden, mabodo , Mention may be made of dumplings, Shumai, hamburger, meatballs, various sources, retort pouch foods such as various soups and the like.

  The eye function maintaining method of the present invention is characterized by ingesting the retinal protective composition of the present invention described above, but does not include medical practice. The retinal protective composition of the present invention is desirably taken by healthy persons excluding the sick. In addition, it is desirable for a person who performs work that overuses the eyes for a long time, and particularly for a desk worker who performs work for a long time on a personal computer or the like. Examples of the method of the present invention include a method of obtaining an eye function maintaining effect by providing the retinal protective food of the present invention in a restaurant such as a restaurant.

Hereinafter, the present invention is described based on examples.
[Example 1]
1. Cell Culture (1) Human retinal pigment epithelial cells (ARPE-19) were cultured in a 75 cm ² flask in a 37 ° C., 5% CO ₂ incubator. A medium prepared by mixing DMEM medium and F-12HAM medium at a ratio of 1: 1 so that FBS was 10% was used.
(2) Cells suspended by trypsin treatment were seeded from a 75 cm ² flask to each well of 96 well plate at a cell density of 1.0 × 10 ⁴ cells / well.
(3) Pre-cultured for 72 hours in a 37 ° C., 5% CO ₂ incubator.
(4) After removing the medium from each well, 200 μL each of the test substance-containing medium prepared to a predetermined concentration was added and cultured in a CO ₂ incubator for 20 hours.

  In addition, about the combined sample of an astaxanthin and another component, the test substance containing culture medium of 2 times concentration was prepared, respectively, and it was set as the target density | concentration by adding 100 microliters. In addition, the control was prepared using a medium containing 0.5% DMSO for the test substance dissolved in DMSO, and prepared using a medium containing 0.25% DMSO for the other test substances. .

  Regarding astaxanthin, commercially available astaxanthin was used. As other components used with astaxanthin, the substances shown in Tables 1 to 4 were used.

(5) After culturing for 20 hours, 50 μL of hydrogen peroxide prepared to 1.5 mM or 2.5 mM in a serum-free medium was added and cultured for 20 hours. When the other components are plant materials and vitamins, the final concentration of hydrogen peroxide solution is 300 μM, and when the other components are amino acids, minerals, and polyphenols, the final concentration of hydrogen peroxide solution is 500 μM. For control, the test area and the final concentration of hydrogen peroxide solution were combined.

2. Expression analysis of TGFβ1 (1) After culturing for 20 hours, the medium was removed from the cultured cells, and RNA was collected using CellAmp ^™ Direct RNA Prep Kit for RT-PCR (Real Time) (manufactured by Takara), N = Three samples were mixed into one RNA sample.
(2) The RNA sample was diluted 3 times using Rnase-free water.
(3) In order to prepare a calibration curve, one sample was diluted 3 times or 9 times. PCR was performed using One Step SYBR (registered trademark) PrimeScript ^™ RT-PCR Kit II (Perfect Real Time) (manufactured by Takara). TGFβ1 (manufactured by QIAGEN) and RPLP0 (manufactured by QIAGEN) were used as primers.
(4) The gene expression level in each sample was calculated using a calibration curve prepared from samples diluted 1-fold, 3-fold, and 9-fold. Analysis was performed by relative quantification, and the amount of mRNA was corrected using RPLP0 (housekeeping gene) as an internal standard.

  The results are shown in Tables 1 to 4 and FIGS.

  Each figure represents the results of “control (addition of hydrogen peroxide)”, “single addition of astaxanthin”, “single addition of other components”, and “addition of astaxanthin + other components” from the left. Moreover, the numerical value in description of the graph lower part shows sample concentration, for example, "AST_2.5" of the upper left graph of FIG. 1 shows astaxanthin 2.5 (microgram / mL). The concentration unit of the additive component is μg / mL in all graphs. The vertical axis represents “% of control”.

For sweet potato, finely ground powder of fresh young leaves was used.
For kuzu, a hot water extract of flowers (dry powder) was used.
For bilberry, a commercially available fruit dry powder was used.
For blueberries, a commercially available fruit dry powder was used.

  As shown in Tables 1 to 4 and FIGS. 1 to 8, the expression level of TGFβ1 due to oxidative stress of human retinal pigment epithelial cells (ARPE-19) is obtained by combining astaxanthin and specific other components of the present invention. Decreased synergistically. Therefore, according to the retinal protective composition of the present invention, retinal pigment epithelial cells can be effectively protected.

[Example 2] (Manufacture of tablets)
A tablet (200 mg) comprising the following components was produced.

Astaxanthin 4mg
Bilberry extract 60mg
Blueberry extract 50mg
50mg sweet potato leaves
Calcium stearate 5mg
Arginine 1.5mg
Glutamic acid 1.5mg
Tryptophan 1mg
Shellac 5mg
Vitamin B6 0.5mg
Vitamin B12 0.5mg
Iron chloride 0.25mg
Zinc gluconate 0.5mg
Cellulose remainder

  Take one tablet with water without chewing one tablet a day.

[Example 3] (Manufacture of capsules)
300 ml of the following mixture was sealed in a soft capsule to produce a capsule.

Astaxanthin 2%
Safflower oil 50%
Bilberry extract 20%
Blueberry extract 15%
Beeswax 10%
Kuzunohana Extract 2%
Water-soluble β-carotene powder 0.2%
Vitamin B12 0.01%
Biotin 0.01%
Gallic acid 0.01%
Vitamin E 0.01%
Iron chloride 0.01%
Gelatin remaining

  The capsule is taken with water without chewing one capsule a day.

[Example 4] (Production of granules)
The following ingredients were mixed to produce granules (3000 mg) by a conventional method.

Astaxanthin 3mg
Gallic acid 50mg
Aspartame 500mg
Glutamic acid 50mg
Tyrosine 50mg
Valine 30mg
Vitamin C 1000mg
Vitamin D3 10mg
Calcium chloride 2mg
Molybdenum chloride 1mg
Starch 340mg
Maltose balance

  Take one granule with water each day.

[Example 6] (Manufacture of solution)
A liquid (30 mL) comprising the following components was produced.

Astaxanthin 4mg
Erythritol 250mg
Bilberry extract 200mg
Cassis extract 200mg
Vitamin C 1200mg
Citric acid 150mg
Valine 20mg
Vitamin K 5mg
Iron chloride 0.5mg
Water balance

  Take one bottle of the above solution a day.

Since the retinal protective composition of the present invention has a high retinal protective effect and can be used as an oral preparation, the industrial utility of the present invention is high.

Claims (6)

  Astaxanthin, processed sweet potato leaves, processed kuzu flower, processed bilberry, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, vitamin C, vitamin D3, vitamin K, calcium compound A retinal protective composition comprising, as an active ingredient, at least one component selected from the group consisting of: a zinc compound, an iron compound, a molybdenum compound, a chromium compound, and shellac.   The retinal protective composition according to claim 1, which is an eye function maintaining agent having a retinal protective action.   The retinal protective composition according to claim 1, which is obtained by adding an active ingredient.   The retinal protective composition according to any one of claims 1 to 3, which is for oral use.   The retinal protective composition according to claim 4, which is a tablet, capsule, powder, granule, or liquid. A method for maintaining an eye function, wherein the retinal protective composition according to any one of claims 1 to 5 is ingested (except for medical practice).