Use of stewartia pseudocamellia extract for protecting eyesight or alleviating, preventing or treating retinal diseases

Abstract

The present application relates to a use of a Stewartia pseudocamellia extract to be used for protecting eyesight and alleviating, preventing or treating retinal diseases. The Stewartia pseudocamellia extract of the present application exhibits activity of proliferating retinal epithelial cells, thereby having the excellent effects of eyesight protection, and alleviation and prevention or treatment of retinal diseases.

Description

Use of extracts of the aralia elata for the protection of vision or for the improvement, prevention or treatment of retinal diseases

[0001] The present application relates to the use of extracts of the Korean yam tree for protecting eyesight or improving, preventing and treating retinal diseases.

[0002]

[0003] The eyes are our most important sensory organ, providing approximately 80% of the information we receive in our daily lives. Therefore, functional impairment or loss of the eyes is one of the biggest factors affecting our quality of life.

[0004] Retinal pigment epithelial cells (RPE) are a single layer of cells located at the interface between photoreceptors and the choroidal capillary layer. They perform biochemical functions such as light reception, material transport, visual circuit regulation, phagocytosis, and immune regulation, and play a key role in maintaining normal vision.

[0005] Although retinal disease is not a fatal condition, its incidence has been rapidly increasing in recent years due to the aging population, industrialization, and changes in dietary habits. Therefore, beyond surgical methods, there is an urgent need to develop a composition that can be supplied in the form of traditional herbal medicines, rather than synthetic treatments.

[0006] Meanwhile, the Stewartia pseudocamellia Maxima is a deciduous tree native to Korea, growing primarily in the southern regions. It grows to a height of 7 to 15 meters, and its bark is a dark reddish-brown, but here and there, the bark peels off in circular or long stripes, revealing reddish-yellow patterns. The leaves are oval, alternate, and have fine serrations along the leaf margins. The white flowers bloom in June and July, where the leaves emerge. The flowers are cymes, with five petals and numerous stamens. The hard fruit, which ripens in October, is covered with soft hairs. The leaves turn yellow in autumn, and the flowers resemble camellia flowers, making it a common ornamental tree in gardens and parks. The hard wood is often used for furniture, and today, it is sought after as a valuable medicinal herb and tea. Known health benefits of the Stewartia pseudocamellia include anti-cancer and liver-protective properties.

[0007] Recently, there has been active development of eye health maintenance medications and health functional foods, including those that prevent oxidative damage to the eye. In particular, a vision-enhancing agent using lutein, derived from marigold extract, as its main ingredient has been developed. However, no studies have been reported on eye health using the quince tree, creating a need for the development of eye-health-related foods and pharmaceuticals.

[0008]

[0009] The problem to be solved by the present application is to provide a food composition for protecting eyesight or improving and preventing retinal diseases, which contains an extract of the Korean fir tree as an active ingredient.

[0010] Another problem to be solved by the present application is to provide a pharmaceutical composition for preventing or treating retinal diseases, which contains an extract of the zelkova tree as an active ingredient.

[0011] Another problem to be solved by the present application is to provide a feed composition for protecting eyesight or improving and preventing retinal diseases, which contains an extract of the Korean fir tree as an active ingredient.

[0012] Another problem to be solved by the present application is to provide an over-the-counter pharmaceutical composition for eye washing, vision protection, or improvement and prevention of retinal diseases, which contains an extract of the Korean angelica tree as an active ingredient.

[0013] Another problem to be solved by the present application is to provide a method for preventing or treating retinal diseases, which comprises a step of administering to a human or non-human animal a pharmaceutical composition containing an extract of the zelkova tree as an active ingredient.

[0014] Another problem to be solved by the present application is to provide a use of the extract of the said zelkova tree for protecting eyesight or for improving, preventing and treating retinal diseases.

[0015]

[0016] One embodiment of the present application provides a food composition for protecting eyesight or improving and preventing retinal diseases, comprising an extract of the zelkova tree as an active ingredient.

[0017] In one embodiment of the present application, the retinal disease may be selected from the group consisting of retinal degeneration, macular degeneration, diabetic retinopathy, hypertensive retinopathy, retinitis pigmentosa, Leber congenital amaurosis, retinal detachment, dry eye syndrome, glaucoma, Stargardt disease, choroideremia, gyrate-atrophy, dry eye syndrome, eye strain, retinopathy of prematurity, macular hole, macular telangiectasia (MacTel), and optic neuropathy.

[0018] In one embodiment of the present application, the extract of the zelkova tree may have an activity to proliferate retinal pigment epithelial cells.

[0019] In one embodiment of the present application, the extract of the zelkova tree can inhibit the production of reactive oxygen species induced by oxidative stress in retinal pigment epithelial cells.

[0020] In one embodiment of the present application, the extract of the zelkova tree can prevent the death of retinal pigment epithelial cells.

[0021] In one embodiment of the present application, the composition may be in a dosage form selected from among tablets, capsules, and liquids.

[0022] One embodiment of the present application provides a pharmaceutical composition for preventing or treating retinal diseases, comprising an extract of the zelkova tree as an active ingredient.

[0023] One embodiment of the present application provides a feed composition for protecting eyesight or improving and preventing retinal diseases, comprising an extract of the zelkova tree as an active ingredient.

[0024] One embodiment of the present application provides an over-the-counter pharmaceutical composition for eye washing, vision protection, or improvement and prevention of retinal diseases, comprising an extract of the zelkova tree as an active ingredient.

[0025] One embodiment of the present application provides a method for preventing or treating a retinal disease, comprising administering to a human or non-human animal a pharmaceutical composition comprising an extract of the zelkova tree as an active ingredient.

[0026] One embodiment of the present application provides a use of the extract of the above-mentioned tree for protecting eyesight or for improving, preventing and treating retinal diseases.

[0027]

[0028] The extract of the present invention has the activity of promoting the proliferation of retinal epithelial cells, and thus has the effect of protecting the eyes, protecting vision, and improving, preventing, or treating retinal diseases.

[0029] In addition, the extract of the present invention inhibits the production of reactive oxygen species induced by oxidative stress in retinal epithelial cells, thereby preventing the death of retinal epithelial cells, and thus has effects of protecting the eyes, protecting vision, and improving, preventing, or treating retinal diseases.

[0030]

[0031] Figure 1 shows the toxicity of the extract of the Korean fir tree against ARPE-19 cells.

[0032] Figure 2 shows the proliferation activity of extracts of the Chinese juniper tree and extracts of marigold on ARPE-19 cells.

[0033] Figure 3 shows the ARPE-19 cell death concentration for H2O2 cytotoxicity.

[0034] Figure 4 shows the ARPE-19 cell protective effect of extracts of the magnolia tree and extracts of marigold against H2O2 cytotoxicity.

[0035] Figure 5 shows the ROS production inhibitory activity of the extract of the Chinese juniper tree and the extract of the marigold.

[0036]

[0037] Hereinafter, the present invention will be described in more detail.

[0038] The specific functional descriptions below are merely illustrative for the purpose of explaining embodiments according to the concept of the present invention, and embodiments according to the concept of the present invention may be implemented in various forms and should not be construed as being limited to the embodiments described in this specification.

[0039] Embodiments according to the concept of the present invention may be subject to various modifications and take various forms, and thus specific embodiments are described in detail herein. However, this is not intended to limit the embodiments according to the concept of the present invention to a specific disclosed form, and it should be understood that all modifications, equivalents, and alternatives included within the spirit and technical scope of the present invention are included.

[0040] The terminology used herein is merely used to describe specific embodiments and is not intended to limit the invention. Singular expressions include plural expressions unless the context clearly dictates otherwise.

[0041] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and will not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0042] One embodiment of the present application provides a food composition for eye protection, vision protection, or the improvement and / or prevention of retinal diseases, comprising an extract of the Korean zelkova tree as an active ingredient. The composition can be formulated into various products, such as pharmaceutical compositions, food compositions including health functional foods, feed additives, and quasi-drug compositions.

[0043] As used herein, the term "including as an active ingredient" means including an amount sufficient to effect a desired biological effect, such as a beneficial outcome, including but not limited to the prevention, reduction, alleviation, or elimination of signs or symptoms of a disease or disorder. This amount is referred to as an "effective amount." Accordingly, the total amount of each active ingredient of a composition or additive is sufficient to produce a significant individual benefit. Therefore, the effective amount will vary depending on the circumstances under which it is administered.

[0044] The term "improvement" in this application means any action that reduces a parameter associated with the condition being treated by ingestion of the composition, for example, the severity of a symptom.

[0045] The term "prevention" in this application refers to blocking the appearance of a disease or symptoms associated with a disease in an asymptomatic subject at risk of developing the disease due to exposure to a disease agent. It also refers to any action that suppresses or delays the onset of a disease by administering the composition of this application.

[0046] The term "treatment" in this application refers to a therapeutic intervention that improves the signs or symptoms of a disease or pathological condition after it has developed. "Improvement" and "palliation" with respect to the signs or symptoms of the disease or pathological condition refer to any observable beneficial effect of the treatment. Such beneficial effects may be evidenced by a delayed onset of clinical symptoms of the disease in a susceptible subject, a reduction in the severity of some or all clinical symptoms of the disease, a slower progression of the disease, an improvement in the overall health or well-being of the subject, or other variables well known in the art to be specific to a particular disease. For example, treatment of a retinal disease may be evidenced by an improvement in visual acuity, a reduction in clinical symptoms due to the retinal disease, or other variables well known in the art to be specific to the retinal disease, such as the degree of dry eye.

[0047] Stewartia pseudocamellia Maxim. is a deciduous tree in the Theaceae family. Its tree bark has a mottled pattern, so it is also called the satin tree or the brocade tree. Its leaves turn yellow in the fall, and its flowers resemble camellia flowers, so it is often planted as an ornamental tree in gardens, parks, etc. Its wood is hard and is widely used for furniture. Currently, it is sought after by many people as a precious medicinal material and a drinkable tea. Its bark falls off in thin pieces, with purplish-red, gray-white, and gray-brown patterns, looking mottled. It exudes a liquid at the base of new branches, and its fruits are capsules with a pentagonal shape. This plant is also known as Stewartia koreana Nakai or Stewartia pseudocamellia Maxim. var. koreana (Nakai) Kim, and is regarded as a native species of Korea because its flowers are cup-shaped and its branches are straight and round. Another scientific name recorded is Stewartia ptero-petiolata Cheng var. koreana (Rehd.) Sealy. Stewartia pseudocamellia Maxim. has large and showy flowers and is used for ornamental purposes. Its wood is hard and has beautiful patterns, so it is used for furniture materials, decorative materials, etc. Its tree bark and root bark are called molan, which has the effect of relaxing tendons and activating blood circulation. It has been studied for relieving stasis caused by bruises and related activities caused by rheumatism.

[0048] The extract of Stewartia pseudocamellia Maxim. can be extracted from various organs of the natural, hybrid or variant plants of Stewartia pseudocamellia Maxim., for example, from the seeds, roots, above-ground parts, stems, leaves, flowers, fruit bodies, fruit skins of Stewartia pseudocamellia Maxim., as well as from plant tissue cultures. Without being limited thereto, the extract of Stewartia pseudocamellia Maxim. can be one or more of the stem extract and leaf extract of Stewartia pseudocamellia Maxim.

[0049] The term "extract" in this application has the meaning commonly used in the art as a crude extract, but in a broad sense, it also includes fractions obtained by further fractionating the extract. That is, the extract refers to the results such as a liquid component obtained by immersing a target substance in various solvents and then extracting it for a certain period of time at room temperature, low temperature, or heated conditions, and a solid component obtained by removing the solvent from the liquid component. In addition, it can be comprehensively interpreted to include all of the diluted solution of the result, the concentrate thereof, the adjusted preparation thereof, the purified product thereof, etc. in addition to the resultant product. The extract of the zelkova tree is preferably included in the composition at a concentration of 0.1 μg / ml to 5 μg / ml. If the concentration is less than 0.1 μg / ml, the effect is minimal, and if it is included at a concentration exceeding 5 μg / ml, it may exhibit toxicity to retinal pigment epithelial cells.

[0050] For example, fractions obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, fractions obtained through various additional purification methods such as separation by chromatography, etc. are also included in the extract of the present application.

[0051] The method for extracting the extract of the zelkova tree in this application is not particularly limited, and extraction can be performed according to methods commonly used in the relevant technical field. These include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, and these can be performed alone or in combination of two or more methods, but are not limited thereto.

[0052] The type of extraction solvent used for extraction in the present application is not particularly limited, and any solvent known in the art can be used. As the extraction solvent, water (or distilled water); alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol, butyl alcohol, etc.; polyhydric alcohols such as glycerin, butylene glycol, propylene glycol, etc.; and hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, dichloromethane, etc.; or mixtures thereof can be used, but are not limited thereto. In addition, a solvent extract can be prepared by extracting the branches one or more times using the solvent, and a dried extract can be prepared by distilling the solvent extract under reduced pressure and then freeze-drying or spray-drying the obtained solvent extract.

[0053] In the present application, it is preferable to use 10 to 95% (v / v) of ethanol as a solvent for the production of the extract. More preferably, it is preferable to use 50 to 95% (v / v) of ethanol, and even more preferably, 70 to 95% (v / v) of ethanol. In addition, the amount of the solvent as described above is preferably 1 to 25 times the weight of the extraction raw material, and more preferably 5 to 20 times. The extraction temperature is preferably 50 to 90°C, and more preferably 60 to 80°C. The extraction time is preferably 1 to 12 hours, and more preferably 5 to 7 hours. According to these extraction conditions, the component exhibiting the effect targeted in the present invention can be efficiently extracted from the raw material.

[0054] The above-mentioned prepared extract can then be filtered, concentrated, or dried to remove the solvent. Filtration, concentration, and drying can all be performed. For example, filtration can be performed using filter paper or a vacuum filter, concentration can be performed using a vacuum concentrator, and drying can be performed using a freeze-drying method, but the present invention is not limited thereto.

[0055] The term "fraction" in this application means a result obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various components.

[0056] In the present application, the fractions can be interpreted as fractions obtained by applying the extract of the zelkova tree to various fractionation methods. The fractions of the extract can be obtained by applying the extract to various fractionation methods, and the fractionation methods are not particularly limited thereto, but may include, but are not limited to, solvent fractionation methods performed by treating various solvents, ultrafiltration fractionation methods performed by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, and chromatographic fractionation methods performed by performing various chromatographies (designed for separation according to size, charge, hydrophobicity, or affinity).

[0057] In particular, the solvent used in the solvent fractionation method is not particularly limited thereto, but a polar solvent or a non-polar solvent may be used, and specifically, a non-polar solvent may be used. The solvent fractionation method may be performed in a manner of sequentially fractionating the extract using a solvent with a high degree of non-polarity to a solvent with a low degree of non-polarity. For example, a method of sequentially fractionating the extract using hexane or ethyl acetate may be used. The fractionation solvent may be a polar solvent such as water or alcohol; or a non-polar solvent such as hexane, ethyl acetate, chloroform, or dichloromethane, and these may be used alone or in a mixture of two or more thereof, but is not limited thereto.

[0058] The term "retinal disease" in this application refers to a disease caused by abnormalities or damage to the retina due to exogenous, intrinsic, or genetic causes. The retinal disease may be caused by oxidative stress.

[0059] In one embodiment of the present application, the retinal disease may be a disease selected from the group consisting of retinal degeneration, macular degeneration, diabetic retinopathy, hypertensive retinopathy, retinitis pigmentosa, Leber congenital amaurosis, retinal detachment, dry eye syndrome, glaucoma, Stargardt disease, choroideremia, gyrate-atrophy, dry eye syndrome, eye strain, retinopathy of prematurity, macular hole, macular telangiectasia (MacTel), and optic neuropathy.

[0060] The retina is one of the most metabolically active tissues in the body, has a high oxygen demand, is continuously exposed to light, and contains a high concentration of unsaturated fatty acids, which are known to be involved in the occurrence of oxidative damage.

[0061] In this application, the term "retinal pigment epithelium cell (RPE)" refers to a single layer of pigment cells that form part of the outer retinal blood barrier. These cells are characterized by playing an important role in maintaining the function of the retina, and are known to absorb light energy, interact with photoreceptors to maintain the visual cycle, secrete various growth factors that maintain the joint structure of photoreceptors and the choriocapillaris, transport ions, water, and metabolites from the subretinal space to the blood, and perform phagocytosis of the outer segment of photoreceptors. RPE cells are cells that form a single layer between the retina and the choriocapillaris, and physiologically play an important role in maintaining vision, such as supplying nutrients to photoreceptors, excreting metabolites, and phagocytosing cell debris. They also consume a lot of oxygen and are exposed to oxidative stress.

[0062] Oxidative stress causes structural defects and functional impairment of the RPE. Reactive oxygen species (ROS) induced by oxidative stress include free radicals, hydrogen peroxide, and oxygen ions, which are byproducts of oxygen metabolism, and induce cell damage and apoptosis in various cells. In particular, cells with a high metabolic rate, such as the RPE, are more vulnerable to oxidative damage, and sustained exposure to high-concentration oxidative stress induces damage to mitochondrial DNA in the RPE, disrupting mitochondrial function and ultimately leading to RPE apoptosis. Retinal damage-induced visual impairment is one of the leading causes of blindness, and oxidative stress is believed to be a contributing factor to retinal diseases.

[0063] In the present application, the retinal pigment epithelial cells may be ARPE-19 cells, but are not limited thereto, and may be used without limitation as long as they correspond to the type of human retinal pigment epithelial cells. The extract of the zelkova tree may have an activity to proliferate retinal pigment epithelial cells. The extract of the zelkova tree has the advantage of excellent cell proliferation activity when included in the composition at a concentration of 0.1 μg / ml to 5 μg / ml. If the concentration is less than 0.1 μg / ml, the effect is minimal, and if it is included at a concentration exceeding 5 μg / ml, it may exhibit toxicity to retinal pigment epithelial cells.

[0064] In addition, the extract of the zelkova tree can inhibit the production of reactive oxygen species induced by oxidative stress in retinal pigment epithelial cells, and specifically, when included in a concentration of 1 μg / ml to 5 μg / ml, the effect of inhibiting the production of reactive oxygen species becomes more excellent as the concentration increases. When the extract of the zelkova tree is included in a concentration exceeding 5 μg / ml, it may exhibit toxicity to retinal pigment epithelial cells.

[0065] The above-mentioned extract of the Korean zelkova tree has the effect of preventing the death of retinal pigment epithelial cells against H2O2 toxicity. Specifically, when included at a concentration of 1 μg / ml to 5 μg / ml, the cell protection effect is even more excellent. However, when the extract of the Korean zelkova tree is included at a concentration exceeding 5 μg / ml, it may exhibit toxicity to retinal pigment epithelial cells.

[0066] Excessive production of reactive oxygen species (ROS) due to oxidative stress oxidizes major cellular components of retinal pigment epithelial cells and activates the intrinsic apoptosis pathway, which is linked to mitochondrial dysfunction and DNA damage. This is a major cause of various retinal diseases, including age-related macular degeneration (AMD), diabetic retinopathy, and proliferative retinal diseases. Therefore, protecting retinal pigment epithelial cells from oxidative stress can be used to prevent and treat various retinal diseases, including AMD.

[0067] The present application provides a food composition for eye protection, vision protection, or the prevention and improvement of retinal diseases, comprising a zelkova tree extract as an active ingredient. The food composition of the present application may include a zelkova tree extract or a fraction thereof.

[0068] In this application, the term "improvement" refers to any act that improves or benefits the symptoms of a subject suspected of or diagnosed with a retinal disease that is prevented or treated using a composition containing an extract of the zelkova tree or a fraction thereof as an active ingredient.

[0069] The food composition of the present application is very useful because it can be consumed on a daily basis and is expected to have effects of protecting the eyes, protecting vision, reducing eye fatigue, and preventing and improving retinal diseases.

[0070] In this application, the term "food" means a natural product or processed product containing one or more nutrients, preferably a product that has gone through a certain degree of processing to become directly edible, and in its conventional sense includes all foods, food additives, functional foods, and beverages.

[0071] Examples of foods to which the composition for eye protection, vision protection, or prevention and improvement of retinal disease symptoms according to the present application can be added include various foods, beverages, gums, tea, vitamin complexes, functional foods, etc. In addition, foods in the present application include, but are not limited to, special nutritional foods (e.g., formulated milk, infant and baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g., ramen, noodles, etc.), bread, health supplements, seasoned foods (e.g., soy sauce, soybean paste, red pepper paste, mixed paste, etc.), sauces, confectionery (e.g., snacks), candies, chocolates, gums, ice cream, dairy products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi, pickled vegetables, etc.), beverages (e.g., fruit drinks, vegetable drinks, soy milk, fermented drinks, etc.), and natural seasonings (e.g., ramen soup, etc.). The above food, beverage or food additive can be manufactured by a conventional manufacturing method.

[0072] In addition, the above "functional food" refers to a food group or food composition that has been designed and processed to sufficiently express the body's regulatory function for regulating the biological defense rhythm, disease prevention, and recovery, etc., by using physical, biochemical, or bioengineering techniques to provide added value to the food so that the food's function can be performed and expressed for a specific purpose, and may be specifically a health functional food. The above functional food may include food additives that are acceptable in terms of food science, and may further include appropriate carriers, excipients, and diluents that are commonly used in the manufacture of functional foods.

[0073] In this application, the term "health functional food" refers to a food manufactured (including processed) using raw materials or ingredients with functionality useful to the human body according to the Health Functional Food Act, and "functionality" means obtaining a useful effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological action. Meanwhile, health food refers to a food that has a more active health maintenance or promotion effect than general food, and health supplement refers to a food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement may be used interchangeably. The health functional food of this application can be manufactured by a method commonly used in the art. It can be manufactured in various forms, and unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs because it uses food as a raw material, and can be highly portable.

[0074] Additionally, the term "beverage" in this application refers to a general term for anything consumed to quench thirst or enjoy a flavor, and includes functional beverages. The beverage, in addition to including the composition for eye protection, vision protection, or prevention and improvement of retinal disease symptoms in the indicated proportions as essential ingredients, has no particular limitations on other ingredients, and, like conventional beverages, may contain various flavorings or natural carbohydrates as additional ingredients.

[0075] Furthermore, in addition to those described above, a food containing the composition of the present application for eye protection, vision protection, or prevention and improvement of retinal disease symptoms may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and fillers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., and the above components may be used independently or in combination.

[0076] In the food containing the composition for eye protection, vision protection or prevention and improvement of retinal disease symptoms of the present application, the amount of the composition according to the present invention may be comprised in an amount of 0.001 wt% to 90 wt% of the total food weight, preferably 0.1 wt% to 40 wt%, and in the case of a beverage, it may be comprised in an amount of 0.001 g to 2 g, preferably 0.01 g to 0.1 g, based on 100 ml. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since the active ingredient does not have any problems in terms of safety, it may be used in an amount greater than the above range, and thus is not limited to the above range.

[0077] The food composition of the present application may contain conventional food additives, and its suitability as a food additive is determined by the specifications and standards for the relevant item in accordance with the general provisions and general test methods of the Food Additive Code approved by the Ministry of Food and Drug Safety, unless otherwise specified.

[0078] The above food additive refers to a food composition intentionally added to food, usually in small quantities, for the purpose of improving the appearance, flavor, texture, or storability. It refers to the use for the purpose of improving the quality of food, enhancing its storability or palatability, and enhancing its nutritional value and practical value. As defined in Article 2, Paragraph 2 of the Food Sanitation Act, it may be a substance added to food, mixed, infiltrated, or used by other means during the manufacturing, processing, or preservation of food.

[0079] One embodiment of the present application provides a pharmaceutical composition for preventing or treating retinal diseases, comprising an extract of the zelkova tree as an active ingredient.

[0080] The composition according to the present application may contain a pharmaceutically effective amount of a zelkova tree extract alone or may contain one or more pharmaceutically acceptable carriers, excipients or diluents.

[0081] In addition, the term "pharmaceutically acceptable" as used above refers to a composition that is physiologically acceptable and does not typically cause allergic reactions such as gastrointestinal upset or dizziness or similar reactions when administered to humans. Examples of the carrier, excipient, and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, and preservatives may be additionally included.

[0082] The pharmaceutical composition according to the present application may further include a pharmaceutically acceptable carrier in addition to the active ingredient. When formulating the pharmaceutical composition, the pharmaceutical preparation may be prepared by mixing it with pharmaceutically acceptable excipients, adjuvants, analgesics, isotonic agents, preservatives, and other adjuvants and formulating it into a pharmaceutically acceptable formulation, but is not limited thereto.

[0083] In the form of such pharmaceutical preparations, the present application can be administered in various oral and parenteral dosage forms during actual clinical administration, and the most preferred route of administration is oral administration. Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc. In addition, liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and preparations for parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, or topical application) include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories, but can be appropriately selected by those skilled in the art.

[0084] The pharmaceutically effective amount as described above refers to an amount sufficient to prevent, improve and / or achieve a therapeutic effect on retinal disease symptoms.

[0085] The preferred dosage of the pharmaceutical composition above varies depending on the patient's condition and weight, the severity of the disease, the drug form, the route of administration, and the duration of administration, but can be appropriately selected by those skilled in the art. However, for desirable effects, the pharmaceutical composition is preferably administered at 0.01 mg / kg to 10 g / kg per day, preferably 0.5 mg / kg to 1 g / kg. Administration may be administered once a day or divided into several doses. Therefore, the above dosage does not limit the scope of the present invention in any way.

[0086] When the above pharmaceutical composition is used clinically, it can be formulated into various preparations such as oral preparations such as tablets, capsules, powders, granules, pills, liquids, and suspensions by mixing it with a conventional carrier in the pharmaceutical field; injectable preparations in the form of a ready-to-use injectable dry powder that can be prepared with distilled water for injection at the time of injection; or ointments. Pharmaceutical preparations prepared using a conventional carrier can be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally, or topically. Therefore, the pharmaceutical composition of the present invention may further include appropriate carriers, excipients, and diluents commonly used in the manufacture of pharmaceuticals.

[0087] Additionally, the composition of the present application can be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to a mammal. The formulation can be in the form of a powder, granule, tablet, emulsion, syrup, aerosol, soft or hard gelatin capsule, sterile injectable solution, or sterile powder.

[0088] The pharmaceutical composition of the present application can be administered orally or parenterally (e.g., applied to the mucosa, intravenously, or subcutaneously) depending on the intended method.

[0089] One embodiment of the present application provides a feed composition for protecting the eyes, protecting vision, or improving and preventing retinal diseases, comprising an extract of the zelkova tree as an active ingredient.

[0090] The above feed composition may include feed additives. The feed additives of the present application fall under supplementary feed under the Feed Management Act.

[0091] In this application, the term "feed" means any natural or artificial diet, meal, etc. or a component of said meal, intended for or suitable for eating, ingesting and digesting by an animal.

[0092] The type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include plant feed such as grains, roots, food processing by-products, algae, fiber, pharmaceutical by-products, oils, starches, meal, or grain by-products; and animal feed such as proteins, inorganic substances, oils, minerals, oils, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0093] The term "feed additive" in this application includes substances added to feed for various purposes such as nutrient supplementation and weight loss prevention, increasing the digestibility and availability of fiber in feed, improving milk quality, preventing reproductive disorders and improving conception rate, and preventing summer heat stress. The feed additive of this application corresponds to supplementary feed under the Feed Management Act, and may additionally include mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals; mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium; vitamins such as carotene, vitamin A, vitamin D, vitamin E, nicotinic acid, and vitamin B complex; protected amino acids such as methionine and lysine; protected fatty acids such as fatty acid calcium salts; probiotics (lactic acid bacteria), yeast cultures, and mold fermentations; and yeast agents.

[0094] One embodiment of the present application provides a method for preventing or treating a retinal disease, comprising administering to a human or non-human animal subject a pharmaceutical composition comprising an extract of the zelkova tree as an active ingredient.

[0095] One embodiment of the present application provides a use of the composition for improving vision or for improving, preventing and treating retinal diseases.

[0096] The term "subject" in this application means any animal that is susceptible to or has developed a retinal disease.

[0097] The feed composition according to the present application may be applied to any non-human animal for the purpose of improving, preventing, or alleviating retinal disease, without limitation. For example, it may be applied to any non-human animal, such as dogs, cats, monkeys, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds, and fish.

[0098] One embodiment of the present application provides an external pharmaceutical composition for eye washing, eye protection, vision protection, and improvement and / or prevention of retinal diseases, comprising an extract of the zelkova tree as an active ingredient.

[0099] In the specification of this application, the term "quasi-drug" means a product that has a milder effect than a pharmaceutical product among products used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases of humans or animals. For example, according to the Pharmaceutical Affairs Act, a quasi-drug means a product excluding products used for the purpose of pharmaceutical products.

[0100] Quasi-drugs are fibers, rubber products, or similar products used for the purpose of treating, alleviating, managing, or preventing diseases of humans or animals; products that have a weak effect on the human body or do not directly affect the human body and are not devices or machines or similar products; products that are one of the preparations used for sterilization, insecticide, or similar purposes to prevent infection; products used for the purpose of diagnosing, treating, alleviating, managing, or preventing diseases of humans or animals, excluding products that are not devices, machines, or devices; and products used for the purpose of exerting a pharmacological effect on the structure and function of humans or animals, excluding products that are not devices, machines, or devices; and also include external preparations and personal hygiene products. Specifically, these may be eye drops, eye wash products, or ointments.

[0101] When the composition of the present application is used as a quasi-drug composition, the extract of the zelkova tree can be added directly or used appropriately in combination with other quasi-drug ingredients according to conventional methods. The amount of active ingredients mixed can be appropriately determined depending on the intended use.

[0102] The contents of the pharmaceutical composition or health functional food composition may be applied to the quasi-drug composition of the present application. The eye wash or quasi-drug composition for eye wash according to the present application may further include a conventional therapeutic anti-allergic agent or anti-inflammatory agent for the prevention or improvement of retinal diseases, but is not limited thereto.

[0103] In addition, the composition may further include additives such as fragrances, pigments, sterilizers, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, emulsifiers, and synthetic polymers to improve physical properties, but is not limited thereto.

[0104] Furthermore, the composition may be administered in a spray, squeeze, or electric machine form, and may be used in any formulation form, but is preferably in a liquid form for ease of washing. It may also be prepared and used in the form of a paste, ointment, spray formulation, eye drop formulation, etc., but is not limited thereto.

[0105] Hereinafter, the present invention will be described in detail through examples, comparative examples, and experimental examples. However, the following examples and comparative examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0106]

[0107] <Example> Preparation of extract of the zelkova tree

[0108] The leaves and stems of the Korean yam tree were dried in a shaded area, then ground and powdered. The powder was used to prepare an extract, and 100% of the leaf and stem powder was used.

[0109] After mixing the powder with 70% ethanol at a ratio of 1:20, the mixture was refluxed and extracted at 65°C for 6 hours, and the ethanol extracts of Examples 1 to 5 were prepared by varying the concentration.

[0110]

[0111] <Comparative Example> Preparation of Marigold Extract

[0112] Commercially available eye protection marigold extract (manufactured by Nutraceuticals International Group, weight 100 g) was purchased, dissolved in 60% ethanol, and used as a comparative example.

[0113]

[0114] <Experimental Example 1> Measurement of toxicity in retinal pigment epithelial cells (ARPE-19)

[0115] 1) Culture of retinal pigment epithelial cells (ARPE-19)

[0116] ARPE-19 cells, human retinal pigmented epithelial cells, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

[0117] The medium used was DMEM High glucose growth medium containing 10% Feral Bovine Serum (FBS). ARPE-19 cells were cultured in an incubator controlled at 95% humidity, 5% CO2, and 37°C, and antibiotics for the medium (Penicillin streptomycin, Gibco, CA, USA) were used to suppress microbial contamination or proliferation. When the ARPE-19 cells covered approximately 80% of the dish, they were washed with phosphate-buffered saline-EDTA (PBS-EDTA, Thermo Fisher Scientific, Massachusetts, USA), and then treated with 0.25% Trypsin-EDTA (Gibco, Thermo Fisher Scientific, Massachusetts, USA) to detach the cells attached to the bottom of the dish and subculture them. The medium was changed every 48 to 72 hours to culture ARPE-19 cells.

[0118]

[0119] 2) Toxicity measurement

[0120] ARPE-19 cell line 1Х10 4 Cells were seeded into a 96-well plate at a concentration of 10 cells / well and cultured in a CO2 incubator controlled at 37°C, 95% humidity, and 5% CO2 for 24 hours. Each sample was treated to cells in a fresh medium at an appropriate concentration range that did not show cytotoxicity, and cultured for 24 hours. To measure the viability of the cell line, MTS (CellTiter 96 ®Absorbance was measured at 490 nm using a microplate reader (Molecular Devices, Versa Max ELISA Microplate Reader, USA) using the AQueous One Solution Cell Proliferation Assay (Promega, USA). Cell toxicity was expressed as the absorbance of the extract-treated group compared to the control group that was not treated with the extract.

[0121] In order to measure the toxicity to the ARPE-19 cells, the ethanol extract of the zelkova tree of Example 1 was treated to the cells at concentrations ranging from 0.1 to 10 μg / ml, and the results are shown in Fig. 1 and Table 1.

[0122] Extract concentration (㎍ / ml)ARPE-19 cell viability (%)Control-100.0 ± 13.8Example 10.1101.7 ± 4.7Example 20.5106.9 ± 1.8Example 31108.3 ± 4.6Example 45109.9 ± 4.7Example 51086.7 ± 2.0*

[0123] * p<0.05 indicates a significant difference as compared to the control group.

[0124] The above-mentioned ethanol extract of the Korean zelkova tree exhibited cytotoxicity at a treatment concentration of 10 μg / ml, whereas no cytotoxicity was observed at treatment concentrations below 5 μg / ml. Therefore, the efficacy of the Korean zelkova tree extract for improving, preventing, or treating retinal pigment epithelial cells was confirmed in the concentration range of 0.1 to 5 μg / ml.

[0125]

[0126] <Experimental Example 2> Proliferation activity of retinal pigment epithelial cells (ARPE-19)

[0127] ARPE-19 cell line 1Х10 4Cells were seeded into 96-well plates at a concentration of 10 cells / well and cultured in a CO2 incubator controlled at 37°C, 95% humidity, and 5% CO2 for 24 hours. Each sample (extract) was treated to the cells in a fresh medium at an appropriate concentration range that did not show cytotoxicity and cultured for 2 days. To measure the viability of the cell line, the absorbance was measured at 450 nm with a microplate reader (Molecular Devices, Versa Max ELISA Microplate Reader, USA) using a Cell Counting Kit-8 (CCK-8, Dojindo Mol. Tech., Rockville., MD, USA). The cell proliferation activity was expressed as the absorbance of the sample-treated group compared to the control group that was not treated with the sample (extract).

[0128] In order to measure the proliferation activity of the ARPE-19 cells, ARPE-19 cells were treated with the ethanol extract of the example tree and the marigold extract of the comparative example at concentrations of 0.1 to 5 μg / ml, and the results are as shown in Fig. 2 and Table 2.

[0129] Extract concentration (㎍ / ml)ARPE-19 cell proliferation activity (%)(ARPE-19 cell Proliferation (%))Control-100.0 ± 4.9Example 10.1110.3 ± 13.6Example 20.5130.3 ± 9.7Example 31142.8 ± 9.1*Example 45135.2 ± 3.8*Control-100.0 ± 4.6Comparative Example 10.1103.2 ± 9.0Comparative Example 20.5108.1 ± 4.8Comparative Example 31105.8 ± 5.2Comparative Example 45115.4 ± 10.1

[0130] * p<0.05 indicates a significant difference as compared to the control group.

[0131] Examples 1 to 4, which are the ethanol extracts of the above-mentioned hornbeam tree, showed that the proliferation activity of ARPE-19 cells increased as the concentration increased. In addition, referring to the results of the ethanol extract of the above-mentioned hornbeam tree, the proliferation activity of the hornbeam tree ethanol extract was 142.8% at a concentration of 1 ㎍ / ml, which was about 1.43 times better than that of the control. On the other hand, it was confirmed that the comparative examples 1 to 4, which are the marigold extracts, showed a very small difference compared to the control group. Through the above results, it can be seen that the hornbeam tree extract according to the present application has excellent proliferation activity of retinal pigment epithelial cells.

[0132]

[0133] <Experimental Example 3> Protective effect of retinal pigment epithelial cells (ARPE-19) against H2O2 cytotoxicity

[0134] ARPE-19 cells were damaged with H2O2, and the protective effect against ARPE-19 cell damage was measured by cell viability.

[0135]

[0136] 1) ARPE-19 cell death concentration for H2O2 cytotoxicity

[0137] To measure the ARPE-19 killing concentration by H2O2, hydrogen peroxide 30% (DAEJUNG, 4104-4405) was purchased and used.

[0138] The above ARPE-19 cell line was seeded in a 96-well plate at 1X10 4 After dispensing at a concentration of 1 cell / well, the cells were cultured at 95% humidity, 5% CO2, and 37°C for 24 hours for stabilization. After removing the medium, 10 ㎕ of H2O2 diluted with new medium was treated at each concentration and reacted at 95% humidity, 5% CO2, and 37°C for 24 hours.

[0139] The viability of ARPE-19 cell line was measured using MTS (CellTiter 96 ®The viability of cells was measured using the MTS reagent (AQueous One Solution Cell Proliferation Assay, Promega, USA). 10 μl of the MTS reagent was added per well, and the cells were incubated at 37°C for 60 minutes. The absorbance was measured at 490 nm using a microplate reader (Molecular Devices, VersaMax ELISA Microplate Reader, USA). The viability of ARPE-19 cells was expressed as the absorbance of the sample-treated group compared to the untreated control group.

[0140] We attempted to find the concentration that resulted in approximately 50% cell viability by treating cells with various concentrations of H2O2. H2O2 was treated at concentrations of 0.5 - 10 mM, and the results are shown in Figure 3 and Table 3.

[0141] Classification H2O2 concentration (㎍ / ml)ARPE-19 cell viability (%)(ARPE-19 cell Viability (%))Control-100.0 ± 5.8H2O20.594.2 ± 15.6179.4 ± 5.4*264.7 ± 2.3*40.0 ± 0.4*60.0 ± 0.4*80.0 ± 0.4*100.0 ± 0.5*

[0142] * p<0.05 indicates a significant difference as compared to the control group.

[0143] As a result of the experiment, referring to Fig. 3 and Table 3, as the concentration of H2O2 increased, the cell viability decreased, and the cell viability was confirmed to be approximately 60% at a concentration of 2 mM. Therefore, the concentration of H2O2 for studying the ARPE-19 cell protective effect against H2O2 cytotoxicity was confirmed to have ARPE-19 cell protective effect at 2 mM.

[0144]

[0145] 2) Protective effect of ARPE-19 cells against H2O2 cytotoxicity

[0146] ARPE-19 cell line was seeded in 96 well plates at 1X10 4 After dispensing at a concentration of cells / well, the cells were cultured for 24 hours at 37°C, 5% CO2, and 95% humidity for stabilization. After removing the medium, 2 mM H2O2 was treated with new medium, and 10 ㎕ of the sample (extract) was treated at each concentration and reacted for 24 hours at 37°C, 5% CO2, and 95% humidity.

[0147] The viability of ARPE-19 cell line was measured using MTS (CellTiter 96 ® The viability of ARPE-19 cells was measured using the MTS reagent (AQueous One Solution Cell Proliferation Assay, Promega, USA). That is, 10 ㎕ of the MTS reagent was treated per well, and after reaction at 37°C for 60 minutes, the absorbance was measured at 490 nm using a microplate reader (Molecular Devices, VersaMax ELISA Microplate Reader, USA). The viability of ARPE-19 cells was expressed as the absorbance of the sample-treated group compared to the control group that was not treated with the sample (extract).

[0148] In order to measure the ARPE-19 cell protective effect by the above H2O2 treatment, the cell protective efficacy of the ethanol extract of the Korean magnolia tree as an example and the marigold extract as a comparative example were confirmed. The results are shown in Fig. 4 and Table 4.

[0149] Extract concentration (㎍ / ml)ARPE-19 cell viability (%)Control-100.0 ± 11.7+55.0 ± 3.5Example 10.159.0 ± 5.0Example 20.560.8 ± 5.1Example 3164.4 ± 4.7*Example 4562.6 ± 2.5*Comparative Example 10.158.1 ± 2.6Comparative Example 20.557.3 ± 3.2Comparative Example 3156.1 ± 1.9Comparative Example 4552.6 ± 3.1

[0150] * p<0.05 indicates a significant difference as compared to the (+)control group.

[0151] Referring to Examples 3 and 4, which are the ethanol extracts of the above-mentioned zelkova tree, a protective effect on ARPE-19 cells was observed, and a more excellent ARPE-19 cell protective effect was observed at a concentration of 1 μg / ml or higher. On the other hand, referring to Comparative Examples 1 to 4, which are the marigold extracts, no significant ARPE-19 cell protective effect was observed at any concentration. Through the above results, it can be seen that the zelkova tree extract according to the present application has an excellent protective effect on retinal pigment epithelial cells against H2O2 cytotoxicity.

[0152]

[0153] <Experimental Example 4> Inhibition of reactive oxygen species (ROS) production in retinal pigment epithelial cells (ARPE-19)

[0154] The production of reactive oxygen species (ROS) is measured by the intensity of fluorescent material generation using DCF-DA. Reduced or acetylated form of 2',7'-Dichlorofluorescin (DCF) releases fluorescent material (FITC) when the acetate group is removed by the oxidation of esterase within the cell and ROS generated within the cell.

[0155] ARPE-19 cells 1Х10 4 After dispensing 100 ㎕ of the solution into a 96-well plate at a concentration of 10 cells / well, the solution was cultured for 24 hours. After culture, the medium was removed, and the sample (extract) was treated at an appropriate concentration using a medium that did not contain serum, and the solution was cultured for 24 hours. After 24 hours, 100 ㎕ of PBS was added and washed, and this process was repeated three times. 100 ㎕ of 2',7'-dichlorofluorescin diacetate (DCF-DA) dissolved in DMSO was diluted to 20 ㎕ (in PBS), and the solution was cultured for 20 minutes. After adding 100 ㎕ of H2O21 mM (in PBS), the solution was reacted for 30 minutes, and the fluorescence intensity was measured at 486 nm and 535 nm using an ELISA reader.

[0156] In order to measure the ROS production inhibition activity in the ARPE-19 cells, the ethanol extract of the example tree and the marigold extract of the comparative example were treated at concentrations of 0.1 to 5 μg / ml, and the results are as shown in Fig. 5 and Table 5.

[0157] Extract concentration (㎍ / ml) 1M H2O2 Reactive oxygen species activity (%) Control--20.1 ± 1.7*+100.0 ± 6.9 Example 10.1+97.1 ± 7.5 Example 20.5+93.5 ± 5.9 Example 31+68.5 ± 1.6* Example 45+65.1 ± 11.4* Comparative Example 10.1+95.0 ± 7.7 Comparative Example 20.5+90.7 ± 2.8 Comparative Example 31+92.5 ± 5.4 Comparative Example 45+99.3 ± 6.3

[0158] * p<0.05 indicates a significant difference as compared to the (+)control group.

[0159] Referring to Examples 1 to 4, which are the ethanol extracts of the above-mentioned zelkova tree, the ROS production inhibition activity significantly increased as the concentration increased, and in particular, an excellent ROS production inhibition activity of 34.9% was confirmed at a concentration of 5 μg / ml. On the other hand, referring to Comparative Examples 1 to 4, which are the marigold extracts, no significant ROS production inhibition effect was confirmed.

[0160] Through the above results, it can be seen that the extract of the zelkova tree according to the present application has excellent ROS generation inhibition effect.

[0161]

[0162] While specific portions of this application have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred embodiments and are not intended to limit the scope of this application. Therefore, the substantial scope of this application is defined by the appended claims and their equivalents. Furthermore, various modifications and improvements made by those skilled in the art utilizing the basic concepts of this application defined in the claims are also within the scope of this application.

Claims

1. A food composition for protecting eyesight or improving and preventing retinal diseases, containing an extract of the Chinese juniper tree as an effective ingredient.

2. In claim 1, The above retinal diseases are, A food composition for protecting eyesight or improving and preventing retinal diseases, wherein the food composition is selected from the group consisting of retinal degeneration, macular degeneration, diabetic retinopathy, hypertensive retinopathy, retinitis pigmentosa, Leber congenital amaurosis, retinal detachment, dry eye syndrome, glaucoma, Stargardt disease, choroideremia, gyrate-atrophy, dry eye syndrome, eye strain, retinopathy of prematurity, macular hole, macular telangiectasia (MacTel), and optic neuropathy.

3. In claim 1, The above extract of the zelkova tree, A food composition for protecting vision or improving and preventing retinal diseases, which has the activity of proliferating retinal pigment epithelial cells.

4. In claim 1, The above extract of the zelkova tree, A food composition for protecting vision or improving and preventing retinal diseases, which inhibits the production of reactive oxygen species induced by oxidative stress in retinal pigment epithelial cells and prevents the death of retinal pigment epithelial cells.

5. A pharmaceutical composition for preventing or treating retinal disease, comprising an extract of the Chinese juniper tree as an active ingredient.

6. In claim 5, The above retinal diseases are, A pharmaceutical composition for preventing or treating a retinal disease, selected from the group consisting of retinal degeneration, macular degeneration, diabetic retinopathy, hypertensive retinopathy, retinitis pigmentosa, Leber congenital amaurosis, retinal detachment, dry eye syndrome, glaucoma, Stargardt disease, choroideremia, gyrate-atrophy, dry eye syndrome, eye strain, retinopathy of prematurity, macular hole, macular telangiectasia (MacTel), and optic neuropathy.

7. In claim 5, The above extract of the zelkova tree, A pharmaceutical composition for preventing or treating retinal diseases, which has the activity of proliferating retinal pigment epithelial cells.

8. In claim 5, The above extract of the zelkova tree, A pharmaceutical composition for preventing or treating retinal diseases, which inhibits the production of reactive oxygen species induced by oxidative stress in retinal pigment epithelial cells and prevents death of retinal pigment epithelial cells.

9. A feed composition for protecting eyesight or improving and preventing retinal diseases, containing an extract of the Chinese juniper tree as an effective ingredient.

10. An external pharmaceutical composition for eye washing, vision protection, or improvement and prevention of retinal diseases, containing an extract of the Chinese juniper tree as an active ingredient.

11. A method for treating a retinal disease, comprising the step of administering the pharmaceutical composition of claim 5 to a human or non-human animal subject.

12. Use of extracts of the zelkova tree for the protection of vision or for the improvement, prevention and treatment of retinal diseases.

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