JP6736137B2 - Retinal protection composition - Google Patents

Description

The present invention relates to a retinal protection composition, and more particularly, to a retinal protection composition containing lutein and certain other components as active ingredients.

It is known that the function of the retinal pigment epithelium, which is involved in repair and regeneration of photoreceptor cells, decreases with age. When I was younger, there were enough SOD enzymes in my body to decompose active oxygen generated in cells, which prevented the adverse effects of active oxygen, but the function of SOD enzyme decreased with age, and the function of retinal pigment epithelium decreased. Will also decrease.

On the other hand, lutein exists along with zeaxanthin in the macula and the lens of the retina, has an SOD-like action, and is said to remove active oxygen generated in the eye, and maintains the function of the eye, cataract, and age-related macula. It is known to be effective in preventing and treating degeneration and the like (see, for example, Patent Documents 1 and 2).

JP, 2003-238442, A JP 2005-287376 A

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

As a result of diligent investigations and studies on protection of retinal pigment epithelial cells that play an important role in the retina from active oxygen, the present inventors have found that by combining lutein with a specific other component, high retinal pigment epithelial cell protection can be achieved. It was found that the effect was obtained, and the present invention was completed. That is, it was found that the damage of retinal pigment epithelial cells due to active oxygen can be more effectively suppressed by combining lutein with other specific components having little or little ability to protect retinal pigment epithelial cells. It was

That is, the present invention relates to a retinal protection composition containing lutein and at least one component selected from the group consisting of the following (a) to (d) as active ingredients.
(A) at least one plant material selected from kudzu, rice, and coconut (b) at least one amino acid selected from arginine, phenylalanine, serine, threonine, tryptophan, and tyrosine (c) gallic acid and chlorogenic acid At least one polyphenol selected from (d) vitamin B1, vitamin D3, biotin, vitamin K, at least one vitamin/mineral selected from calcium, manganese, zinc, iron, molybdenum, and chromium

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

The retinal protection composition of the present invention may be obtained by adding an active ingredient comprising lutein and at least one component selected from the group consisting of (a) to (d).

The retinal protection 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 eye function (however, excluding medical practice), which comprises ingesting the above-mentioned retinal protection composition of the present invention.

The retinal protective composition of the present invention can effectively protect retinal pigment epithelial cells from damage of active oxygen and suppress retinal damage.

It is a figure which shows the expression level of TGFβ1 which is an oxidative stress marker when the retinal protection composition (lutein+plant material) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19), and the upper left figure shows the plant material. The one using kudzu is shown, the one on the upper right shows the one using rice (rice bran) as the plant material, and the one on the lower left shows the one using coconut (coconut) as the plant material. It is a figure which shows the expression level of TGFβ1 which is an oxidative stress marker when the retinal protective composition (lutein+amino acid) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19), and the upper left figure shows arginine as an amino acid. The upper right figure shows the one using phenylalanine as the amino acid, the lower left figure shows the one using serine as the amino acid, and the lower right figure shows the one using threonine as the amino acid. It is a figure which shows the expression level of TGFβ1 which is an oxidative stress marker when the retinal protection composition (lutein+amino acid) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19), and the left figure shows tryptophan as an amino acid. The one used is shown, and the right figure shows the one using tyrosine as an amino acid. It is a figure which shows the expression level of TGFβ1 which is an oxidative stress marker when the retinal protective composition (lutein+polyphenol) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19), and the left figure shows gallic acid as polyphenol. Is shown, and the figure on the right shows the one using chlorogenic acid as the polyphenol. It is a figure which shows the expression level of TGFβ1 which is an oxidative stress marker when the retinal protective composition (lutein+vitamins) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19), and the upper left figure shows vitamins. The one using vitamin B1, the one using the vitamin D3 as the vitamins in the upper right figure, the one using biotin as the vitamins in the lower left figure, and the one using vitamin K as the vitamins in the lower right figure Indicates. It is a figure which shows the expression level of TGFβ1 which is an oxidative stress marker when the retinal protective composition (lutein+minerals) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19), and the upper left figure shows the minerals. The figure using calcium is shown, the figure on the upper right shows the one using manganese as the minerals, the figure on the lower left shows the one using zinc as the minerals, and the lower right figure shows the one using iron as the minerals. It is a figure which shows the expression level of TGFβ1 which is an oxidative stress marker when the retinal protection composition (lutein+minerals) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19), and the left figure shows the minerals. The one using molybdenum is shown, and the one on the right shows the one using chromium as a mineral.

The retinal protection composition of the present invention contains lutein and at least one component selected from the group consisting of the following (a) to (d) (hereinafter sometimes referred to as other components) as active ingredients: Characterize. The following components (a) to (d) used together with lutein may be used alone (for example, the component (a)), or may be used in combination of two or more kinds. When two or more other components are used in combination, it is preferable to combine the other components having a high synergistic effect with lutein.

The retinal protective composition of the present invention more effectively protects retinal pigment epithelial cells from damage of active oxygen, can suppress retinal disorders, delay aging of the eye, and maintain the function of the eye. it can. Further, it is possible to suppress the development of retinal diseases such as age-related macular degeneration, retinitis pigmentosa, central retinochoroidopathy, macular retinal epithelium, and macular hole. Therefore, the retinal protection composition of the present invention can be used as an eye function maintenance agent, a retinal protection agent, a retinal disorder inhibitor, a retinal disease preventive agent, and the like.

[Lutein]
Lutein used in the present invention is a kind of carotenoid existing in leaves, flowers, fruits and the like of plants, and plant-derived ones can be used. For example, one derived from marigold containing a large amount of lutein can be used. Moreover, the thing manufactured by synthesis can also be used.

[Other ingredients]
(A) Plant Material In the retinal protection composition of the present invention, it is preferable to use at least one plant material selected from kudzu, rice, and coconut as an active ingredient together with lutein.

These plant materials may be any part of the plant such as leaves, stems, roots, flowers, fruits, trunks, branches, etc., in addition to the plant materials themselves (including dried products), crushed products and squeezed juices thereof. It is possible to use a processed plant material such as an extract. Examples of the crushed product include powder and granules. The squeezed juice or the extract may be liquid, but may be used as a paste or a dry powder. The extract can be obtained by extraction with a suitable solvent, and as the solvent, for example, water (hot water), ethanol, or hydrous ethanol can be used. As these plant materials, commercially available products can be used.

Kuzu is a climbing perennial plant of the genus Leguminosae. As the part used as the plant material of the present invention, flowers and roots are preferable, and flowers are particularly preferable. As Katsura flowers, those collected in any process from buds to fully-opened flowers may be used, and those collected in each process may be mixed and used. The type of kudzu is not particularly limited, but examples thereof include Pueraria thomsonii, Pueraria lobata, and Pueraria thunbergiana.

Rice is a plant belonging to the genus Poaceae. As the part used as the plant material of the present invention, the hulls and embryos of brown rice are preferable, and specifically, rice bran obtained as a by-product when whitening brown rice is preferably used. The type of rice is not particularly limited, and examples thereof include Japonica variety and Indica variety.

Coconut is a plant of the palm family, and the part used as the plant material of the present invention is preferably fruit (endosperm).

(B) Amino Acid In the retinal protection composition of the present invention, it is preferable to use at least one amino acid (including salt) selected from arginine, phenylalanine, serine, threonine, tryptophan, and tyrosine as an active ingredient together with lutein. preferable. Examples of the salt include sodium salt and hydrochloride.

(C) Polyphenol In the retinal protective composition of the present invention, it is preferable to use at least one polyphenol selected from gallic acid and chlorogenic acid as an active ingredient together with lutein.

(D) Vitamin/Minerals In the retinal protective composition of the present invention, the active ingredient is selected from vitamin B1, vitamin D3, biotin, vitamin K, calcium, manganese, zinc, iron, molybdenum, and chromium together with lutein. It is preferable to use at least one vitamin/mineral. In addition, calcium, manganese, zinc, iron, molybdenum, and chromium as minerals of the present invention include forms of compounds containing these metals.

The retinal protective composition of the present invention can be used as an eye function maintaining agent having a retinal protecting effect, and such an eye function maintaining agent contains lutein and a predetermined other component, and is used for maintaining eye function. The product is not particularly limited as long as it can be distinguished from other products. For example, the body of the product according to the present invention, the packaging, instructions, or any of the advertised items that indicate that there is a function of maintaining the function of the eye, such as a retinal protective effect, a retinal disorder suppressing effect, a retinal disease preventing effect, etc. Within the scope of the invention. For example, so-called health foods such as pharmaceuticals (including quasi-drugs), cosmetics, foods for specified health uses, foods with nutritional function, foods with functional claims, etc. Can be mentioned. In so-called health foods, "maintain the pigment amount in the macula of the eye (eye)," "worry of the eye (eye) due to aging", "protect the eye (eye) from light stimulation", " "For blue light measures", "For liquid crystal light measures", "If you are tired of reading, PC, smartphone", "For people who are uncomfortable", "For improving contrast sensitivity", "For tired eyes (eyes)" , "Tones eyes", "removes eyes", etc. can be exemplified. The retinal protection composition of the present invention can be ingested by any person who is concerned about the above effects regardless of sex or age, but the effects of the retinal protection composition of the present invention can be more effectively enjoyed. Therefore, it is desirable that healthy people except sick people should take it. In addition, it is desirable that it is taken by a person who works for overworking the eyes for a long time, and particularly it is desirable that it is taken by a deskworker who works for a long time on a personal computer or the like.

The form of the retinal protection composition of the present invention includes, for example, tablets, capsules, powders, granules, liquids, granules, rods, plates, blocks, solids, and pills. , Pastes, creams, caplets, gels, chewables, sticks and the like. Among these, tablets, capsules, powders, granules, and liquids are particularly preferable. Specifically, supplements, bottled beverages filled in PET bottles, cans, bottles, etc., instant powdered beverages and instant granular beverages to be dissolved in water (hot water), milk, fruit juice, green juice, etc. Can be illustrated. These are preferable in that they can be easily taken at meals and the taste can be enhanced.

The content of lutein and other components (active ingredients) in the retinal protection composition of the present invention may be appropriately contained within the range where the effect is exhibited.

Generally, when the retinal protection composition of the present invention is a drug or supplement (tablet, capsule), it is preferable that the active ingredient is contained in an amount of 0.01 to 100% by mass based on the dry mass. 0.1 to 85 mass% is more preferable, and 0.5 to 70 mass% is further preferable.

When the retinal protection composition of the present invention is a packaged beverage (solution), it is preferable that the active ingredient is contained in an amount of 0.01 to 15% by mass based on the dry mass, and 0.03 to 12. The content is more preferably 5% by mass, further preferably 0.05 to 10% by mass.

Moreover, when the retina protective composition of the present invention is an instant powder drink (powder) or an instant granule drink (granule), the active ingredient is contained in an amount of 0.1 to 80% by mass based on the dry mass. It is preferable that the content is 0.5 to 70% by mass, more preferably 1 to 60% by mass.

In order to exert the effect of the present invention more effectively, it is preferable that the active ingredient is contained in an amount of 80% by mass or more based on the dry mass of the entire retinal protection composition (excluding water), and 90% by mass. The above content is more preferable, the content is more preferably 95% by mass or more, still more preferably 100% by mass. Furthermore, when the retinal protection composition of the present invention contains any of the components (a) to (d), it is preferable that the contained component is composed of only the active ingredient of the present invention. That is, when the retinal protection composition of the present invention contains, for example, the component (a) (plant material), it is preferable that the composition not contain any plant material other than kudzu, rice, and coconut.

The intake of the retinal protection composition of the present invention is not particularly limited, but from the viewpoint of more significantly exhibiting the effects of the present invention, the active ingredient is ingested at a dose of 1 mg/day or more. It is preferable to ingest at 5 mg/day or more, more preferably at 10 mg/day or more. The upper limit is not particularly limited, but is, for example, 4 g/day, preferably 2 g/day.

The retinal protection composition of the present invention can be stored as one day in one container or divided into, for example, a plurality of containers such as 2-3 so that the daily intake becomes the above intake. ..

The blending mass ratio of lutein 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. , 1:1 to 60:1, more preferably 1:1 to 50:1. When the compounding ratio of lutein and other components is within the above range, the effects of the present invention can be more effectively exhibited.

The retinal protection 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.

Further, as the retinal protection composition as an oral preparation of the present invention, in addition to the retinal protection composition containing the active ingredient, a retinal protection composition obtained by adding the active ingredient to food (retinal protection food) Examples thereof include, for example, foods in which the content of the active ingredient of the present invention is increased as compared with normal foods (including natural foods), and active ingredients for foods that do not usually contain the active ingredient of the present invention. Foods to which is added can be mentioned. The active ingredient may be added separately, simultaneously, or may be added together with other ingredients than the active ingredient.

Examples of the retina-preserving food of the present invention include carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, smoothies, drinks such as green juice; frozen desserts such as ice cream, ice sorbet, shaved ice; buckwheat, udon, harusame, Chinese noodle , Instant noodles, etc.; candy, candy, gum, chocolate, tablets, snacks, biscuits, jellies, jams, creams, baked goods, breads, etc.; processed fish and livestock products such as kamaboko, ham, sausages; Dairy products such as processed milk, fermented milk and yogurt; oils and fats such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing and processed foods; seasonings such as sauces and soy sauces; curry, stew, oyakodon, Retort pouch foods such as porridge, porridge, Chinese rice bowl, katsudon, tempura bowl, beef bowl, hayashi rice, omelet rice, oden, marbo doff, gyoza, shumai, hamburger, meatballs, various sauces, and various soups can be mentioned.

The method for maintaining eye function of the present invention is characterized by ingesting the retinal protection composition of the present invention described above, but does not include medical practice. The retinal protection composition of the present invention can be ingested regardless of gender or age if the person is concerned about the above effects, but the effects of the retinal protection composition of the present invention can be more effectively enjoyed. Because it is possible, it is desirable to have healthy people excluding sick people ingest it. In addition, it is desirable to ingest it by a person who works for overworking the eyes for a long time, and in particular, it is desirable that it be ingested by a deskworker who works for a long time on a personal computer or the like. Examples of the method of the invention include a method of obtaining an eye function maintaining effect by providing the retinal protection food of the present invention in a restaurant such as a restaurant.

Hereinafter, the present invention will be described based on examples.
[Example 1]
1. Cell Culture (1) Human retinal pigment epithelial cells (ARPE-19) were cultured using a 75 cm ² flask in a 37° C., 5% CO ₂ incubator. The medium used was a mixture of DMEM medium and F-12HAM medium in a ratio of 1:1 and prepared so that FBS was 10%.
(2) The cells suspended by the trypsin treatment were inoculated from a 75 cm ² flask into 96 well plates of each well at a cell density of 1.0×10 ⁴ cells/well.
(3) Precultured at 37° C. in a 5% CO ₂ incubator for 72 hours.
(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 the cells were cultured in a CO ₂ incubator for 20 hours.

In addition, regarding the combined use sample of lutein and other components, a test substance-containing medium having a double concentration was prepared, and 100 μL of each medium was added to obtain a target concentration. In addition, control was prepared using a medium containing 0.5% DMSO for the test substance dissolved in DMSO, and 0.25% DMSO medium for the other test substances.

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

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

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

The results are shown in Tables 1 to 4 and FIGS. 1 to 7.

Each figure shows the results of "control (addition of hydrogen peroxide)", "single addition of lutein", "single addition of other component", and "addition of lutein+other component" from the left. The numerical value in the description below the graph indicates the sample concentration, and for example, “lutein_0.62” in the upper left graph of FIG. 1 indicates lutein 0.62 (μg/mL). The concentration unit of the added component is μg/mL in all graphs. The vertical axis represents “% of control”.

For kudzu, a hot water extract of flowers (dry powder) was used.
A commercial item was used for the rice bran.
Regarding the coconut, a commercially available coconut milk was dried and the oil was removed.
For mugwort, finely ground powder of fresh leaves was used.
For maki berries, a commercially available dried fruit powder was used.

As shown in Tables 1 to 4 and FIGS. 1 to 7, the expression level of TGFβ1 due to the oxidative stress of human retinal pigment epithelial cells (ARPE-19) was obtained by combining lutein with a specific other component of the present invention. Has decreased synergistically. Therefore, the retinal protective composition of the present invention can effectively protect retinal pigment epithelial cells.

[Example 2] (Production of tablets)
A tablet (200 mg) consisting of the following ingredients was produced.

Lutein (marigold extract) 10mg
Bilberry extract 60mg
Blueberry extract 50mg
Calcium stearate 5mg
Arginine 1.5mg
Phenylalanine 1.5 mg
Tryptophan 1 mg
Chlorogenic acid 1 mg
Vitamin B1 0.5 mg
Vitamin K 0.5mg
Calcium chloride 0.5mg
Zinc gluconate 0.5mg
Cellulose balance

The above tablets are taken with water without chewing 2 tablets a day.

[Example 3] (Production of capsule)
The following mixture was enclosed in a soft capsule to manufacture a capsule.

Weight ratio Lutein (marigold extract) 30% (20 mg as lutein)
Safflower oil 50%
Beeswax 10%
Kuzu flower extract 2%
Blueberry extract 1%
Water-soluble β-carotene powder 0.2%
Hematococcus algal pigment 0.2%
Vitamin B12 0.01%
Biotin 0.01%
Gallic acid 0.01%
Vitamin E 0.01%
Iron chloride 0.01%

The above capsules are taken with water without chewing one capsule per day.

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

Lutein (marigold extract) 15mg
Pine bark extract 50mg
Coconut milk 200mg
Aspartame 500mg
Tyrosine 50mg
Threonine 50 mg
Tryptophan 30mg
Chlorogenic acid 20mg
Vitamin B1 5 mg
Vitamin K 5mg
Calcium chloride 2mg
Copper gluconate 1mg
Dextrin balance

The above granules are taken once a day with water.

[Example 6] (Production of liquid agent)
A liquid agent (125 mL) composed of the following components was produced.

Lutein (marigold extract) 18mg
Bilberry extract 150mg
Cassis extract 50mg
Rice bran extract 25mg
Vitamin C 10mg
Citric acid 5mg
Arginine 1.5mg
Phenylalanine 1.5 mg
Tryptophan 1 mg
Vitamin B1 0.5 mg
Vitamin K 0.5mg
Calcium chloride 0.5mg
Zinc gluconate 0.5mg
Remaining water

The above liquid medicine is to be drunk one pack per day.

The retinal protective composition of the present invention has a high retinal protective effect and can be used as an oral preparation, so that the industrial usefulness of the present invention is high.

Claims (2)

A retinal protection composition comprising lutein and kuzuhana as active ingredients. The retinal protective composition according to claim 1, which is an eye function maintaining agent having a retinal protective effect.