JP2020054396A - Composition for protection of retina - Google Patents

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

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

  It is known that the function of the retinal pigment epithelium involved in the repair and regeneration of photoreceptors decreases with age. When young, the body has enough SOD enzyme to prevent the adverse effects of active oxygen, but with age, the activity of SOD enzyme decreases and the function of the retinal pigment epithelium also decreases.

  On the other hand, astaxanthin has a high antioxidant ability and has an antioxidant ability approximately 1000 times that of vitamin E, and thus has been proposed for uses such as an antioxidant in a living body and an improvement in the accommodation function of eyes. For example, health foods containing anthocyanins and astaxanthin as main components have been proposed.

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 damage by active oxygen and suppressing retinal damage.

  The present inventors have conducted intensive studies on the protection of retinal pigment epithelial cells that play an important role in the retina from reactive oxygen, and as a result, by combining astaxanthin with certain other components, high retinal pigment epithelial cell protection was achieved. The inventors have found that an effect can be obtained, and have completed the present invention. That is, it has been found that by combining astaxanthin with certain other components having little or no protective ability for retinal pigment epithelial cells, damage to retinal pigment epithelial cells by active oxygen can be more effectively suppressed. Was.

  That is, the present invention relates to astaxanthin, processed sweetpotato young leaves, processed kudzu flowers, processed bilberries, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, vitamin C, and vitamin D3. A vitamin C, a calcium compound, a zinc compound, an iron compound, a molybdenum compound, a chromium compound, and at least one component selected from the group consisting of shellac as an active ingredient.

  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 ocular function (excluding medical practice), comprising ingesting the retinal protective composition of the present invention.

  ADVANTAGE OF THE INVENTION According to the retinal protective composition of this invention, a retinal pigment epithelium cell can be protected effectively from damage of an active oxygen, and retinal disorder can be suppressed.

It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker when the retinal protective composition (astaxanthin + plant material) of this invention is applied to human retinal pigment epithelial cells (ARPE-19), and the upper left figure is a plant material. The one using the processed sweet potato young leaves is shown, the upper right figure shows the one using processed kudzu flowers as the plant material, the lower left figure shows the one using processed bilberry as the plant material, and the lower right figure is the plant material The one using gallic acid is shown. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker when the retinal protective composition (astaxanthin + amino acid) of this invention is applied to human retinal pigment epithelial cell (ARPE-19), and the upper left figure is glutamic acid as an amino acid The upper right panel shows the one using histidine as the amino acid, the lower left panel shows the one using methionine as the amino acid, and the lower right panel 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 when the retinal protective composition (astaxanthin + amino acid) of this invention is applied to human retinal pigment epithelial cell (ARPE-19), and the left figure is tyrosine as an amino acid. The figure on the right shows that using valine as the amino acid. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker when the retinal protective composition (astaxanthin + vitamins) of this invention is applied to human retinal pigment epithelial cells (ARPE-19), and the upper left figure is as a vitamin. The one using vitamin B6 is shown, the upper right figure shows the one using vitamin B12 as the vitamins, the lower left figure shows the one using vitamin C as the vitamins, and the lower right figure uses vitamin D3 as the vitamins Show things. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker when the retinal protective composition (astaxanthin + vitamins) of this invention is applied to human retinal pigment epithelial cells (ARPE-19). The ones used are shown. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker when the retinal protective composition (astaxanthin + minerals) of this invention is applied to human retinal pigment epithelial cells (ARPE-19), and the upper left figure is minerals. The one 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, and the lower right figure uses the molybdenum compound as the minerals Show things. It is a figure showing the expression level of TGFβ1, which is an oxidative stress marker, when the retinal protective composition (astaxanthin + minerals) of the present invention is applied to human retinal pigment epithelial cells (ARPE-19). The ones used are shown. It is a figure which shows the expression level of TGF (beta) 1 which is an oxidative stress marker when the retinal protective composition (astaxanthin + shellac) of this invention is applied to human retinal pigment epithelial cells (ARPE-19).

  The retinal protective composition of the present invention comprises astaxanthin, sweet potato young leaf treatment, kuzu flower treatment, bilberry treatment, gallic acid, glutamic acid, histidine, methionine, serine, tyrosine, valine, vitamin B6, vitamin B12, and vitamin C. , Vitamin D3, vitamin K, a calcium compound, a zinc compound, an iron compound, a molybdenum compound, a 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. 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 astaxanthin with another component having a high synergistic effect.

  The retinal protective composition of the present invention can more effectively protect retinal pigment epithelial cells from damage by active oxygen, can suppress retinal damage, delay eye aging, and maintain eye function. it can. In addition, the occurrence of retinal diseases such as age-related macular degeneration, retinitis pigmentosa, central retinochoroidosis, macular retinal epithelial dysplasia, and macular hole can be suppressed. Therefore, the retinal protective composition of the present invention can be used as an eye function maintaining agent, a retinal protective agent, a retinal disorder inhibitor, a retinal disease preventive, 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 natural substances include astaxanthin obtained from algae such as Haematococcus algae, crustaceans such as shrimp and crab, and fish such as salmon and Thai. Extracts and chemically synthesized products derived from natural products are commercially available, and these commercially available products can be used.

[Other components]
(Processed sweet potato leaves)
Sweet potato refers to a plant belonging to the convolvulaceae family and is generally called sweet potato. The variety of sweet potatoes is not particularly limited, and examples thereof include sweet potato, joy white, koganesengan, shiroyutaka, satsu masterch, ayamurasaki and the like. Examples of the processed sweet potato young leaves according to the present invention include processed products such as pulverized sweet potato leaves, squeezed juices, and extracts. Examples of the pulverized product include powder, granules, and the like. The squeezed juice and the extract may be liquid, but may be used as a paste or a dry powder. The extract can be obtained by extraction using an appropriate solvent, and as the solvent, for example, water (hot water), ethanol, or hydrated ethanol can be used.

(Processed kudzu flower)
Kuzu is a vine perennial plant of the genus Kudzu. As the kudzu flowers, 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. Although there is no particular limitation on the type of kudzu, Pueraria thomsonii, Pueraria lobata, Pueraria thunbergiana and the like can be exemplified. Examples of the processed material of kudzu flower of the present invention include processed products of crushed kuzu flower, squeezed juice, extract, and the like. Examples of the pulverized product include powder, granules, and the like. The squeezed juice and the extract may be liquid, but may be used as a paste or a dry powder. The extract can be obtained by extraction using an appropriate solvent, and as the solvent, for example, water (hot water), ethanol, or hydrated ethanol can be used.

(Bilberry processed product)
Bilberry is a fruit of a low-shrub species of the ericaceae genus Vaccinium, and is sometimes referred to as white berries, winberries, blueberries, and European blueberries. Examples of the processed bilberry product of the present invention include processed products such as ground bilberry, squeezed juice, and extract. Examples of the pulverized product include powder, granules, and the like. The squeezed juice and the extract may be liquid, but may be used as a paste or a dry powder. The extract can be obtained by extraction using an appropriate solvent, and as the solvent, for example, water (hot water), ethanol, or hydrated ethanol can be used.

(Other)
Gallic acid may be of plant origin, but is not limited thereto. Amino acids such as glutamic acid, histidine, methionine, serine, tyrosine, and valine include salts. Examples of the salt include a sodium salt, a hydrochloride, and the like.

  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 astaxanthin and certain other components, 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 product of the present invention, a package, an instruction, or any of the advertisements indicates that the product has a function of maintaining eye functions such as a retinal protective effect, a retinal disorder suppressing effect, and a retinal disease preventing effect. Are included in the scope of the present invention. For example, so-called health foods such as pharmaceuticals (including quasi-drugs), cosmetics, and functional foods that have been approved for their indications by specified organizations such as foods for specified health use, nutritional functional foods, and functionally labeled foods Can be mentioned. In so-called health foods, "maintain the amount of pigment in the macula of the eye (eye)", "worry the eye (eye) due to aging", "protect the eye (eye) from light stimulation", " For measures against blue light, for measures against liquid crystal lights, for reading, using PCs and smartphones, for those who are worried about blurring, for improving contrast sensitivity, and for tired eyes (eye) , "Conditioning the eyes (eye)", "relieving eyes (eye) fatigue" and the like can be exemplified. The retinal protective composition of the present invention can be taken regardless of gender or age if the above-mentioned effects are a concern for a person, but it is possible to more effectively enjoy the effects of the retinal protective composition of the present invention. Because it is possible, it is desirable that healthy people except the sick take it. In addition, it is desirable for a person who performs a task that abuses the eyes for a long time to take it, and particularly 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, 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, the forms of tablets, capsules, powders, granules and liquids are particularly preferred. Specifically, packaged beverages filled in supplements, PET bottles, cans, bottles, etc., instant powdered beverages to be dissolved in water (hot water), milk, fruit juice, green juice, etc., and instant granular beverages Can be exemplified. These are preferred in that they are easily drinkable at the time of meals and the like, and can enhance palatability.

  The content of astaxanthin and other components (active ingredients) in the retinal protective composition of the present invention may be appropriately contained as long as the effects are exhibited.

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

  When the retinal protective composition of the present invention is a packaged beverage (solution), the active ingredient is preferably contained in an amount of 0.01 to 15% by mass in terms of dry mass, and preferably from 0.03 to 12%. The content is more preferably 5% by mass, and even more preferably 0.05 to 10% by mass.

  When the retinal protective composition of the present invention is an instant powdered beverage (powder) or an instant granulated beverage (granule), the active ingredient is contained in an amount of 0.1 to 80% by mass in terms of dry mass. The content is preferably 0.5 to 70% by mass, more preferably 1 to 60% by mass.

  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 entire retinal protective composition (excluding water) of the present invention in terms of dry mass, and more preferably 90% by mass. More preferably, the content is 95% by mass or more, and particularly preferably 100% by mass.

  The amount of the retinal protective composition of the present invention is not particularly limited, but from the viewpoint of more remarkably exhibiting the effects of the present invention, the daily intake of the active ingredient is 0.1 mg / day or more. , Preferably 1 mg / day or more, more preferably 2 mg / day or more. The upper limit is not particularly limited, but is, for example, 4 g / day, preferably 2 g / day.

  The retinal protective composition of the present invention can be contained in one container or, for example, divided into a plurality of two or three containers so that the daily intake is the above intake, and accommodated as a daily supply. .

  The blending mass ratio of astaxanthin and other components is preferably in the range of 0.5: 1 to 70: 1, more preferably 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 mixing ratio of astaxanthin and other components is within the above range, the effects of the present invention can be more effectively exerted.

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

  In addition, as the retinal protective composition as an oral preparation of the present invention, in addition to the retinal protective composition containing an active ingredient, a retinal protective composition obtained by adding an active ingredient to a food (retinal protective food). Examples thereof include, for example, a food in which the content of the active ingredient of the present invention is increased as compared with ordinary foods (including natural foods), and an active ingredient for a food that does not normally contain the active ingredient of the present invention. Can be mentioned. As for the addition of the active ingredients, the respective ingredients may be added separately, simultaneously, or may be added together with other ingredients other than the active ingredients.

  Examples of the retinal protective food of the present invention include drinks such as carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, smoothies, green juices, etc .; ice creams, ice sorbets, shaved ice, etc .; buckwheat, udon, harusame, Chinese noodles Noodles such as instant noodles; sweets such as candy, candy, gum, chocolate, tablets, snacks, biscuits, jellies, jams, creams, baked goods, bread; processed fishery and livestock products such as kamaboko, ham, sausage; Dairy products such as processed milk, fermented milk, yogurt, etc .; fats and oils such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing and processed foods; sauces, soy sauce, etc .; curry, stew, oyakodon, Porridge, porridge, Chinese rice bowl, katsudon, tendon, gyudon, hayashi rice, omelet, oden, marbodeau , Mention may be made of dumplings, Shumai, hamburger, meatballs, various sources, retort pouch foods such as various soups and the like.

  The method for maintaining ocular function of the present invention is characterized by ingesting the above-described retinal protective composition of the present invention, but does not include a medical practice. It is desirable that the retinal protective composition of the present invention be taken by a healthy person other than a sick person. In addition, it is desirable to ingest it for a person who performs a task that abuses the eyes for a long time, and particularly for a desk worker who performs a task for a long time using a personal computer or the like. Examples of the method of the present invention include a method of providing 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 will be 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 at 37 ° C. in a 5% CO ₂ incubator. The medium used was prepared by mixing the DMEM medium and the F-12HAM medium at a ratio of 1: 1 and adjusting the FBS to 10%.
(2) The cells suspended by trypsin treatment were seeded from a 75 cm ² flask into each well of a 96-well plate 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 of a test substance-containing medium adjusted to a predetermined concentration was added thereto, and the cells were cultured in a CO ₂ incubator for 20 hours.

  In addition, about the combination sample of astaxanthin and another component, the test substance-containing medium was prepared at a double concentration, and 100 μL each was added to obtain a target concentration. The control was prepared using a medium containing 0.5% DMSO for a test substance dissolved in DMSO, and prepared using a medium containing 0.25% DMSO for other test substances. .

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

(5) After culturing for 20 hours, 50 μL of a hydrogen peroxide solution adjusted to 1.5 mM or 2.5 mM in a serum-free medium was added, and the cells were 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 plot 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), and N = The three samples were mixed to make one RNA sample.
(2) The RNA sample was three-fold diluted using Rnase-free water.
(3) One sample was diluted 3-fold and 9-fold to prepare a calibration curve. PCR was performed using One Step SYBR (registered trademark) PrimeScript ^™ RT-PCR Kit II (Perfect Real Time) (manufactured by Takara). As primers, TGFβ1 (manufactured by QIAGEN) and RPLP0 (manufactured by QIAGEN) were used.
(4) The amount of gene expression in each sample was calculated using a calibration curve prepared from samples diluted 1-, 3-, and 9-fold. 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.

  Each figure shows the results of "control (addition of hydrogen peroxide)", "addition of astaxanthin alone", "addition of other components alone", and "addition of astaxanthin and other components" from the left. The numerical values in the description below the graph indicate the sample concentration. For example, “AST_2.5” in the upper left graph of FIG. 1 indicates astaxanthin 2.5 (μg / mL). The concentration unit of the added component is μg / mL in all the graphs. The vertical axis indicates “% of control”.

For sweet potatoes, finely ground raw young leaves were used.
For the kudzu, a hot water extract of flowers (dry powder) was used.
For bilberry, a commercially available dried fruit powder was used.
For blueberries, a commercially available dried powder of fruit was used.

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

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

Astaxanthin 4mg
Bilberry extract 60mg
Blueberry extract 50mg
Sweet potato young leaf powder 50mg
5mg calcium stearate
Arginine 1.5mg
Glutamic acid 1.5mg
Tryptophan 1mg
Shellac 5mg
Vitamin B6 0.5mg
Vitamin B12 0.5mg
0.25mg iron chloride
0.5mg zinc gluconate
Cellulose balance

  The tablets are taken with water without chewing one tablet per day.

[Example 3] (Production of capsule)
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%
Kuzu 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 balance

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

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

Astaxanthin 3mg
Gallic acid 50mg
500mg aspartame
Glutamic acid 50mg
Tyrosine 50mg
Valine 30mg
Vitamin C 1000mg
Vitamin D3 10mg
2mg calcium chloride
Molybdenum chloride 1mg
340mg starch
Maltose balance

  The above granules are taken one packet daily with water.

[Example 6] (Production of liquid agent)
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

  One bottle of the above liquid is consumed daily.

  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 (1)

  A retinal protective composition comprising astaxanthin as an active ingredient, further comprising at least one component selected from the group consisting of glutamic acid, methionine, serine, tyrosine and valine.