Composition for protection of retina

Abstract

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

[Technical Field]

[0001] The present invention relates to a retinal protective composition, and more particularly to a retinal protective composition containing astaxanthin and other specific ingredients as active ingredients. [Background technology]

[0002] It is known that the function of the retinal pigment epithelium, which is involved in the repair and regeneration of photoreceptors, declines with age. When we are young, our bodies have sufficient SOD enzymes, which prevent the harmful effects of reactive oxygen species. However, as we age, the activity of SOD enzymes declines, and the function of the retinal pigment epithelium also declines.

[0003] On the other hand, astaxanthin has a high antioxidant capacity, approximately 1,000 times that of vitamin E, and so its use as an antioxidant in the body and for improving the accommodative function of the eyes has been proposed. For example, health foods containing anthocyanins and astaxanthin as their main ingredients have been proposed. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Laid-Open No. 2008-035714 Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a retinal protective composition that can effectively protect retinal pigment epithelial cells from damage caused by active oxygen and suppress retinal disorders. [Means for solving the problem]

[0006] The present inventors have conducted extensive research and studies into the protection of retinal pigment epithelial cells, which play an important role in the retina, from reactive oxygen species. As a result, they have discovered that a high level of protective effect on retinal pigment epithelial cells can be achieved by combining astaxanthin with specific other components, leading to the completion of the present invention. In other words, they have discovered that damage to retinal pigment epithelial cells caused by reactive oxygen species can be more effectively suppressed by combining astaxanthin with specific other components that have little or no protective ability on retinal pigment epithelial cells.

[0007] Specifically, the present invention relates to a retinal protective composition characterized by containing astaxanthin and at least one component selected from the group consisting of processed young sweet potato leaves, processed kudzu flowers, processed bilberry, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, vitamin C, vitamin D3, vitamin K, calcium compounds, zinc compounds, iron compounds, molybdenum compounds, chromium compounds, and shellac as active ingredients.

[0008] The retinal protective composition of the present invention can be used as an agent for maintaining ocular function having a retinal protective effect.

[0009] The retinal protective composition of the present invention may be obtained by adding an active ingredient.

[0010] The retinal protective composition of the present invention is preferably for oral administration, and is preferably in the form of tablets, capsules, powders, granules, or liquid.

[0011] The present invention also relates to a method for maintaining ocular function (excluding medical procedures) which comprises ingesting the retinal protective composition of the present invention. [Effects of the Invention]

[0012] The retinal protective composition of the present invention can effectively protect retinal pigment epithelial cells from damage caused by active oxygen and suppress retinal disorders. [Brief explanation of the drawings]

[0013] [Figure 1] This figure shows the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin + plant material) is applied to human retinal pigment epithelial cells (ARPE-19). The upper left figure shows the result when processed young sweet potato leaves are used as the plant material, the upper right figure shows the result when processed kudzu flowers are used as the plant material, the lower left figure shows the result when processed bilberry is used as the plant material, and the lower right figure shows the result when gallic acid is used as the plant material. [Figure 2] FIG. 1 shows the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin + amino acid) was applied to human retinal pigment epithelial cells (ARPE-19). The upper left figure shows the result when glutamic acid was used as the amino acid, the upper right figure shows the result when histidine was used as the amino acid, the lower left figure shows the result when methionine was used as the amino acid, and the lower right figure shows the result when serine was used as the amino acid. [Figure 3] FIG. 1 shows the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin + amino acid) was applied to human retinal pigment epithelial cells (ARPE-19). The left figure shows the result when tyrosine was used as the amino acid, and the right figure shows the result when valine was used as the amino acid. [Figure 4] FIG. 1 shows the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin + vitamins) is applied to human retinal pigment epithelial cells (ARPE-19). The upper left figure shows the result when vitamin B6 is used as the vitamin, the upper right figure shows the result when vitamin B12 is used as the vitamin, the lower left figure shows the result when vitamin C is used as the vitamin, and the lower right figure shows the result when vitamin D3 is used as the vitamin. [Figure 5] FIG. 1 shows the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin + vitamins) was applied to human retinal pigment epithelial cells (ARPE-19), and shows the case where vitamin K was used as the vitamin. [Figure 6] This figure shows the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin + minerals) was applied to human retinal pigment epithelial cells (ARPE-19). The upper left figure shows the result when a calcium compound was used as the mineral, the upper right figure shows the result when a zinc compound was used as the mineral, the lower left figure shows the result when an iron compound was used as the mineral, and the lower right figure shows the result when a molybdenum compound was used as the mineral. [Figure 7] FIG. 1 shows the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin + minerals) was applied to human retinal pigment epithelial cells (ARPE-19), and shows the case where a chromium compound was used as the mineral. [Figure 8] FIG. 1 is a graph showing the expression level of TGFβ1, an oxidative stress marker, when the retinal protective composition of the present invention (astaxanthin+shellac) was applied to human retinal pigment epithelial cells (ARPE-19). DETAILED DESCRIPTION OF THE INVENTION

[0014] The retinal protective composition of the present invention is characterized by containing astaxanthin and at least one component (hereinafter sometimes referred to as "other component") selected from the group consisting of processed young sweet potato leaves, processed kudzu flowers, processed bilberry, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, vitamin C, vitamin D3, vitamin K, calcium compounds, zinc compounds, iron compounds, molybdenum compounds, chromium compounds, and shellac as active ingredients. The other components used together with astaxanthin may be used alone or in combination of two or more. When two or more other components are used in combination, it is preferable to combine other components that have a high synergistic effect with astaxanthin.

[0015] The retinal protective composition of the present invention can more effectively protect retinal pigment epithelial cells from damage caused by reactive oxygen species, suppress retinal disorders, delay ocular aging, and maintain ocular function. It can also suppress the occurrence of retinal diseases such as age-related macular degeneration, retinitis pigmentosa, central chorioretinopathy, macular epithelial dystrophy, and macular hole. Therefore, the retinal protective composition of the present invention can be used as an agent for maintaining ocular function, a retinal protective agent, an agent for suppressing retinal disorders, an agent for preventing retinal diseases, etc.

[0016] [Astaxanthin] The astaxanthin used in the present invention is a type of carotenoid, said to have antioxidant activity 1,000 times stronger than vitamin E. It may be derived from natural products or obtained synthetically. Examples of naturally derived astaxanthin include astaxanthin obtained from algae such as Haematococcus algae, crustaceans such as shrimp and crab, and fish such as salmon and sea bream. Extracts derived from natural products and chemically synthesized products are commercially available, and these commercially available products can be used.

[0017] [Other ingredients] (Processed young sweet potato leaves) Sweet potato refers to a plant belonging to the Convolvulaceae family, commonly known as Satsuma Imo (sweet potato). The variety of sweet potato is not particularly limited, and examples include Suio, Joy White, Koganesengan, Shiroyutaka, Satsuma Starch, and Ayamurasaki. Examples of processed sweet potato young leaves of the present invention include processed products such as crushed, squeezed, and extracts of sweet potato young leaves. Examples of crushed products include powder and granules. The squeezed juice and extract may be in liquid form, but can also be used as a paste or dry powder. The extract can be obtained by extraction using an appropriate solvent, such as water (hot water), ethanol, or aqueous ethanol.

[0018] (Processed kudzu flowers) Kudzu is a perennial climbing plant of the genus Pueraria in the family Fabaceae. Kudzu flowers may be harvested at any stage, from bud to fully opened flower, or a mixture of flowers harvested at each stage may be used. The type of kudzu is not particularly limited, but examples include Pueraria thomsonii, Pueraria lobata, and Pueraria thunbergiana. Examples of processed kudzu flower products of the present invention include crushed kudzu flowers, juice, and extracts. Crushed products include powders and granules. Juice and extracts may be in liquid form, but can also be used as pastes or dried powders. Extracts can be obtained by extraction using an appropriate solvent, such as water (hot water), ethanol, or aqueous ethanol.

[0019] (Processed bilberry) Bilberry is the fruit of a low shrub species of the genus Vaccinium in the family Ericaceae, and is also called water berry, wiener berry, braeberry, or European blueberry. Examples of processed bilberry products of the present invention include pulverized bilberry, juice, extract, and other processed products. Examples of pulverized products include powder and granules. The juice and extract may be in liquid form, but can also be used as a paste or dry powder. The extract can be obtained by extraction using an appropriate solvent, and examples of the solvent that can be used include water (hot water), ethanol, and aqueous ethanol.

[0020] (others) Gallic acid may be derived from plants, but is not limited to this. Amino acids such as glutamic acid, histidine, methionine, serine, tyrosine, and valine may be in the form of salts. Examples of salts include sodium salts and hydrochlorides.

[0021] The retinal protective composition of the present invention can be used as an ocular function maintenance agent with retinal protective activity. Such an ocular function maintenance agent contains astaxanthin and other specified ingredients and is not particularly limited as long as it can be distinguished from other products in terms of its use in maintaining ocular function. For example, the scope of the present invention includes products that display on the product itself, packaging, instructions, or promotional materials an ocular function maintenance effect, such as retinal protection, retinal damage suppression, or retinal disease prevention. Examples of such products include so-called health foods, such as pharmaceuticals (including quasi-drugs), cosmetics, and functional foods with efficacy claims approved by designated organizations, such as foods for specified health uses, foods with nutrient functions, and foods with functional claims. Examples of so-called health foods include those that "maintain pigment levels in the macula of the eye," "relieve eye problems caused by aging," "protect eyes from light irritation," "protect against blue light," "protect against LCD light," "relieve fatigue from reading, using a computer, or using a smartphone," "for those who are concerned about blurred vision," "improve contrast sensitivity," "for tired eyes," "regulate eye condition," and "relieve eye fatigue." The retinal protective composition of the present invention can be taken by anyone concerned about the above-mentioned effects, regardless of gender or age, but it is preferably taken by healthy individuals excluding those with illnesses, as this allows the effects of the retinal protective composition of the present invention to be enjoyed more effectively. It is also preferably taken by individuals who perform work that strains the eyes for long periods of time, and is particularly desirable for desk workers who work long periods of time on a computer, etc.

[0022] The retinal protective composition of the present invention may be in the form of, for example, a tablet, capsule, powder, granule, liquid, granule, bar, plate, block, solid, pill, paste, cream, caplet, gel, chewable, or stick. Among these, tablets, capsules, powder, granule, and liquid are particularly preferred. Specific examples include supplements, packaged beverages filled in PET bottles, cans, or bottles, and instant powdered or granular beverages that can be dissolved in water (hot water), milk, fruit juice, green juice, or the like. These are preferred because they are easy to consume during meals and can enhance palatability.

[0023] The retinal protective composition of the present invention may contain astaxanthin and other ingredients (active ingredients) in an amount appropriate to the extent that the desired effect is achieved.

[0024] Generally, when the retinal protective composition of the present invention is a pharmaceutical or supplement (tablet, capsule), the active ingredient is preferably contained in an amount of 0.01 to 100% by mass, more preferably 0.1 to 85% by mass, and even more preferably 0.5 to 70% by mass, of the total mass, calculated on a dry mass basis.

[0025] 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 mass% of the total, calculated on a dry mass basis, more preferably 0.03 to 12.5 mass%, and even more preferably 0.05 to 10 mass%.

[0026] Furthermore, when the retinal protective composition of the present invention is an instant powdered beverage (powder) or an instant granular beverage (granules), the active ingredient is preferably contained in an amount of 0.1 to 80 mass% of the total, calculated on a dry mass basis, more preferably 0.5 to 70 mass%, and even more preferably 1 to 60 mass%.

[0027] In order to more effectively exert the effects of the present invention, the active ingredient is preferably contained in an amount of 80% by mass or more of the total retinal protective composition of the present invention (excluding water) in dry mass terms, more preferably 90% by mass or more, even more preferably 95% by mass or more, and particularly preferably 100% by mass.

[0028] There is no particular limit to the amount of intake of the retinal protective composition of the present invention, but from the viewpoint of more significantly exerting the effects of the present invention, it is preferable to take so that the daily intake of the active ingredient is 0.1 mg / day or more, more preferably 1 mg / day or more, and particularly preferably 2 mg / day or more. The upper limit is not particularly limited, but is, for example, 4 g / day, preferably 2 g / day.

[0029] The retinal protective composition of the present invention can be stored as a daily dose in a single container or divided into, for example, 2 to 3 containers so that the daily intake amount is the above-mentioned intake amount.

[0030] The blending mass ratio of astaxanthin to other components, calculated on a dry mass basis, is preferably in the range of 0.5:1 to 70:1, more preferably in the range of 0.75:1 to 60:1, even more preferably in the range of 1:1 to 60:1, and particularly preferably in the range of 1:1 to 50:1. When the blending ratio of astaxanthin to other components is in the above range, the effects of the present invention can be more effectively exhibited.

[0031] The retinal protective composition of the present invention can be produced by known methods, by adding ingredients other than the active ingredient that are orally acceptable, as needed.

[0032] Furthermore, examples of the retinal protective composition as an oral preparation of the present invention include retinal protective compositions containing an active ingredient, as well as retinal protective compositions (retinal protective foods) obtained by adding an active ingredient to a food, such as foods in which the content of the active ingredient of the present invention is increased compared to ordinary foods (including natural foods), and foods in which the active ingredient of the present invention has been added to foods that do not normally contain the active ingredient. The active ingredients may be added separately or simultaneously, or together with other ingredients other than the active ingredient.

[0033] Examples of the retinal protective food of the present invention include beverages such as carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, smoothies, and green juice; frozen desserts such as ice cream, iced sherbet, and shaved ice; noodles such as soba, udon, harusame, Chinese noodles, and instant noodles; sweets such as candy, candy, gum, chocolate, tablet sweets, snacks, biscuits, jelly, jam, cream, baked goods, and bread; processed seafood and livestock foods such as kamaboko, ham, and sausage; processed milk, fermented milk, Examples include dairy products such as yogurt; oils and fats such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing, and processed foods thereof; seasonings such as sauces and soy sauce; and retort pouch foods such as curry, stew, oyakodon, rice porridge, porridge, Chinese rice bowl, katsudon, tempura bowl, beef bowl, hayashi rice, omelet rice, oden, mapo dolphin, gyoza, shumai, hamburger steak, meatballs, various sauces, and various soups.

[0034] The method for maintaining ocular function of the present invention is characterized by ingestion of the retinal protective composition of the present invention as described above, but does not include medical procedures. The retinal protective composition of the present invention is preferably ingested by healthy individuals, excluding those with illnesses. It is also preferably ingested by individuals who perform tasks that strain the eyes for long periods of time, particularly desk workers who work long hours at a computer, etc. An example of the method of the present invention is a method for achieving the effect of maintaining ocular function by providing the retinal protective food of the present invention in a restaurant or other food service establishment. [Example]

[0035] The present invention will be described below based on examples. [Example 1] 1.Cell culture (1) In a 37°C, 5% CO2 incubator, 75 cm 2 Human retinal pigment epithelial cells (ARPE-19) were cultured in flasks using a medium prepared by mixing DMEM and F-12HAM medium at a 1:1 ratio, with 10% FBS. (2) Cells suspended by trypsin treatment were collected at 75 cm 2 Transfer 1.0 x 10 cells from the flask to each well of a 96-well plate. 4 The cells were seeded at a density of 100 cells / well. (3) The cells were pre-cultured for 72 hours in a 37°C, 5% CO2 incubator. (4) After removing the medium from each well, 200 μL of medium containing the test substance adjusted to a predetermined concentration was added to each well, and the cells were cultured in a CO 2 incubator for 20 hours.

[0036] For samples containing astaxanthin in combination with other ingredients, a 2x concentration of the test substance was prepared and 100 μL of each was added to achieve the desired concentration. Controls were prepared using 0.5% DMSO-containing medium for test substances dissolved in DMSO, and 0.25% DMSO-containing medium for other test substances.

[0037] Commercially available astaxanthin was used. As other components used together with astaxanthin, the substances shown in Tables 1 to 4 were used.

[0038] (5) After 20 hours of incubation, 50 μL of aqueous hydrogen peroxide solution prepared at 1.5 mM or 2.5 mM in serum-free medium was added and incubated for another 20 hours. For samples containing plant materials or vitamins, the final concentration of aqueous hydrogen peroxide was set to 300 μM. For samples containing amino acids, minerals, or polyphenols, the final concentration of aqueous hydrogen peroxide was set to 500 μM. For the control, the final concentration of aqueous hydrogen peroxide was the same as that of the test group.

[0039] 2. Expression analysis of TGFβ1 (1) After 20 hours of culture, the medium was removed from the cultured cells and the cells were incubated in CellAmp TM RNA was collected using the Direct RNA Prep Kit for RT-PCR (Real Time) (Takara), and N=3 samples were mixed to form one RNA sample. (2) The RNA sample was diluted 3-fold with RNase-free water. (3) To create a standard curve, one sample was diluted 3-fold and 9-fold. One Step SYBR® PrimeScript TM PCR was performed using RT-PCR Kit II (Perfect Real Time) (Takara) with primers TGFβ1 (QIAGEN) and RPLP0 (QIAGEN). (4) Gene expression levels in each sample were calculated using a standard curve created from 1x, 3x, and 9x diluted samples. Analysis was performed by relative quantification, and mRNA levels were corrected using RPLP0 (a housekeeping gene) as an internal standard.

[0040] The results are shown in Tables 1 to 4 and FIGS.

[0041] From left to right, each graph shows the results for "control (hydrogen peroxide added)," "astaxanthin alone added," "other components alone added," and "astaxanthin + other components added." The numbers in the legend below the graph indicate the sample concentration; for example, "AST_2.5" in the graph in the upper left of Figure 1 indicates astaxanthin 2.5 (μg / mL). The concentration units for added components are μg / mL in all graphs. The vertical axis indicates "% of control."

[0042] [Table 1]

[0043] For sweet potato, finely ground fresh young leaves were used. For kudzu, a hot water extract (dried powder) of the flowers was used. For bilberry, commercially available dried fruit powder was used. For blueberries, commercially available dried fruit powder was used.

[0044] [Table 2]

[0045] [Table 3]

[0046] [Table 4]

[0047] As shown in Tables 1 to 4 and Figures 1 to 8, the combination of astaxanthin with the specific other components of the present invention synergistically reduced the expression of TGFβ1 caused by oxidative stress in human retinal pigment epithelial cells (ARPE-19). Therefore, the retinal protective composition of the present invention can effectively protect retinal pigment epithelial cells.

[0048] [Example 2] (Production of tablets) Tablets (200 mg) were prepared containing the following ingredients:

[0049] Astaxanthin 4mg Bilberry extract 60mg Blueberry extract 50mg Sweet potato young leaf powder 50mg 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 Remaining

[0050] Take one tablet per day with water without chewing.

[0051] [Example 3] (Production of capsules) 300 ml of the following mixture was filled into a soft capsule to prepare a capsule preparation.

[0052] Astaxanthin 2% Safflower oil 50% Bilberry extract 20% Blueberry extract 15% Beeswax 10% Kudzu flower 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

[0053] The capsules are taken once a day with water without chewing.

[0054] [Example 4] (Production of Granules) The following ingredients were mixed and prepared into granules (3000 mg) in a conventional manner.

[0055] 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 remainder

[0056] Take one packet of the above granules daily with water.

[0057] [Example 6] (Production of liquid preparation) A liquid formulation (30 mL) consisting of the following components was prepared.

[0058] Astaxanthin 4mg Erythritol 250mg Bilberry extract 200mg Blackcurrant extract 200mg Vitamin C 1200mg Citric acid 150mg Valine 20mg Vitamin K 5mg Iron chloride 0.5mg water remainder

[0059] The above liquid should be taken one bottle per day. [Industrial Applicability]

[0060] The retinal protective composition of the present invention has a high retinal protective effect and can be used as an oral preparation, and therefore the present invention is highly industrially useful.

Claims

[Claim 1] A retinal protective composition containing astaxanthin as an active ingredient, characterized in that it contains at least one component selected from the group consisting of processed kudzu flowers and processed young sweet potato leaves.

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