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WO2025110266A1 - Utilisation d'un extrait de stewartia pseudocamellia pour protéger la vue ou soulager, prévenir ou traiter les maladies rétiniennes - Google Patents

Utilisation d'un extrait de stewartia pseudocamellia pour protéger la vue ou soulager, prévenir ou traiter les maladies rétiniennes Download PDF

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Publication number
WO2025110266A1
WO2025110266A1 PCT/KR2023/018795 KR2023018795W WO2025110266A1 WO 2025110266 A1 WO2025110266 A1 WO 2025110266A1 KR 2023018795 W KR2023018795 W KR 2023018795W WO 2025110266 A1 WO2025110266 A1 WO 2025110266A1
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Prior art keywords
retinal
extract
preventing
tree
retinal diseases
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English (en)
Korean (ko)
Inventor
남우현
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Purebio Co Ltd
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Purebio Co Ltd
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Priority to PCT/KR2023/018795 priority Critical patent/WO2025110266A1/fr
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/82—Theaceae (Tea family), e.g. camellia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents

Definitions

  • the present application relates to the use of extracts of the Chinese juniper tree for protecting eyesight or improving, preventing and treating retinal diseases.
  • the eyes are the most important sensory organ, as approximately 80% of the information we obtain in our daily lives is obtained through the eyes. Therefore, functional impairment or loss of the eyes is one of the biggest factors that reduces the quality of life.
  • Retinal pigment epithelial cells are single-layer 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.
  • Retinal disease is not a fatal disease that leads to death, but it has been rapidly increasing recently due to the increase in the elderly population, industrialization, and changes in eating habits. Therefore, in addition to surgical methods, it is urgent to develop a composition that can be supplied in the form of herbal medicines that we have been taking, rather than a treatment method that is synthesized artificially.
  • Stewartia pseudocamellia Maxim is a deciduous tree endemic to Korea that grows mainly in the southern region. It is about 7 to 15 m tall, and its bark is a dark reddish brown, but the bark peels off in circles or long stripes, revealing reddish yellow patterns.
  • the leaves are oval, alternate, and have small serrations on the leaf margins.
  • the flowers are white and bloom in June and July where the leaves grow.
  • the flowers are cymes with five petals and many stamens.
  • the hard fruits that ripen in October are covered with soft hair on the outside.
  • the leaves turn yellow in the fall, and the flowers look like camellia flowers, so they are commonly planted as ornamental trees in gardens and parks.
  • the wood is hard and is often used for furniture, and many people currently seek it out as a valuable medicine and tea.
  • the known effects of the Stewartia include anticancer and liver protection.
  • the problem that this application seeks to solve is to provide a food composition for protecting eyesight or improving and preventing retinal diseases, which contains an extract of the zelkova tree as an effective ingredient.
  • Another problem to be solved by the present application is to provide a pharmaceutical composition for preventing or treating retinal diseases containing an extract of the zelkova tree as an active ingredient.
  • Another problem to be solved by the present application is to provide a feed composition for protecting eyesight or improving and preventing retinal diseases, containing an extract of the zelkova tree as an effective ingredient.
  • 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, containing an extract of the zelkova tree as an active ingredient.
  • Another problem to be solved by the present application is to provide a method for preventing or treating retinal disease, comprising a step of administering a pharmaceutical composition containing an extract of the zelkova tree as an active ingredient to a human or non-human animal subject.
  • Another problem that the present application seeks to solve is to provide a use of the extract of the said zelkova tree for protecting eyesight or for improving, preventing and treating retinal diseases.
  • 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.
  • 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.
  • the extract of the zelkova tree may have an activity to proliferate retinal pigment epithelial cells.
  • the extract of the zelkova tree can inhibit the production of reactive oxygen species induced by oxidative stress in retinal pigment epithelial cells.
  • the extract of the zelkova tree can prevent the death of retinal pigment epithelial cells.
  • the composition may be formed in a dosage form selected from a tablet, a capsule, and a liquid.
  • One embodiment of the present application provides a pharmaceutical composition for preventing or treating retinal disease, comprising an extract of the zelkova tree as an active ingredient.
  • 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 effective ingredient.
  • 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, containing an extract of the zelkova tree as an active ingredient.
  • One embodiment of the present application provides a method for preventing or treating a retinal disease, comprising the step of administering to a human or non-human animal subject a pharmaceutical composition comprising an extract of the zelkova tree as an active ingredient.
  • One embodiment of the present application provides a use of the extract of the zelkova tree for protecting eyesight or for improving, preventing and treating retinal diseases.
  • 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.
  • the extract of the zelkova tree 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.
  • Figure 1 shows the toxicity of the extract of the Chinese juniper tree against ARPE-19 cells.
  • Figure 2 shows the proliferation activity of extracts of the Chinese juniper tree and extracts of marigold on ARPE-19 cells.
  • Figure 3 shows the ARPE-19 cell death concentration in response to H 2 O 2 cytotoxicity.
  • Figure 4 shows the ARPE-19 cell protective effect of extracts of the Chinese juniper tree and extracts of marigold against H2O2 cytotoxicity .
  • Figure 5 shows the ROS production inhibition activity of the extract of the Chinese juniper tree and the extract of the marigold.
  • One embodiment of the present application provides a food composition for protecting the eyes, protecting vision, or improving and/or preventing retinal diseases, containing an extract of the ginkgo tree as an effective ingredient.
  • the composition can be variously commercialized, and can be formulated as, for example, a pharmaceutical composition, a food composition including a health functional food, a feed additive, an over-the-counter drug composition, etc.
  • the term "including as an active ingredient” means including an amount sufficient to effect a desired biological effect, such as a beneficial result, including but not limited to preventing, reducing, alleviating or eliminating the signs or symptoms of a disease or disorder, and this amount is referred to as an "effective amount.” Accordingly, the total amount of each active ingredient of the composition or additive is sufficient to exhibit a significant individual benefit. Accordingly, the effective amount will vary depending on the circumstances under which it is administered.
  • improvement in this application means any action that reduces a parameter related to the condition being treated by ingestion of the composition, for example, the severity of a symptom.
  • prevention in this application refers to blocking the appearance of a disease or a symptom associated with a disease in an asymptomatic subject at risk of developing a disease due to exposure to a disease cause. It also refers to any act of suppressing or delaying the onset of a disease by administration of the composition of this application.
  • treatment refers to a therapeutic intervention that improves the signs or symptoms of a disease or pathological condition after it has been onset.
  • “Amelioration” 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 of the 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.
  • 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 dryness of the eye.
  • Stewartia pseudocamellia Maxim is a deciduous tree in the Theaceae family, and is also called silk tree or brocade tree because of its mottled bark. Its leaves turn yellow in the fall, and its flowers resemble camellia flowers, so it is commonly planted as an ornamental tree in gardens and parks.
  • the wood is hard and is often used for furniture, and is currently sought after by many people as a valuable medicine and as a drinkable tea.
  • the bark falls off in thin pieces, forming mottled reddish brown, grayish white, and grayish brown patterns.
  • the axils grow at the bases of new branches, and the fruit is a 5-sided capsule. This plant is also known by the synonyms Stewartia koreana Nakai or Stewartia pseudocamellia Maxim.
  • var. koreana (Nakai) Kim, and is considered a Korean endemic species because of its cup-shaped flowers and straight, round branches.
  • Another synonym is Stewartia ptero-petiolata Cheng var. Koreana (Rehd.) Sealy is recorded.
  • the flowers of the Korean fir tree are large and showy, so they are used for ornamental purposes.
  • the wood is hard and has a beautiful pattern, so it is used for furniture and decorations.
  • the bark and root bark are called peony ( ⁇ ), and it has the effect of relaxing muscles and promoting blood circulation, so it relieves blood stasis caused by bruises, and related activities due to rheumatism have also been studied.
  • the above-mentioned extract of the hornbeam tree can be extracted from various organs of the natural, hybrid or mutant plant of the hornbeam tree, for example, the seeds, roots, aerial parts, stems, leaves, flowers, body of the fruit, peel of the fruit, as well as plant tissue cultures.
  • the extract of the hornbeam tree can be at least one of an extract of the stem of the hornbeam tree and an extract of the leaf of the hornbeam tree.
  • extract in this application has a 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 means a result such as a liquid component obtained by immersing a target substance in various solvents and then extracting it at room temperature, low temperature, or heated state for a certain period of time, and a solid component obtained by removing the solvent from the liquid component.
  • the extract 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 included in the composition at a concentration of 0.1 ⁇ g/ml to 5 ⁇ g/ml. If the concentration is less than 0.1 ⁇ g/ml, the effect is minimal, and if it is included at a concentration exceeding 5 ⁇ g/ml, it may exhibit toxicity to retinal pigment epithelial cells.
  • 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.
  • the method for extracting the extract of the zelkova tree in this application is not particularly limited, and extraction can be performed according to a method commonly used in the relevant technical field. These include a hot water extraction method, an ultrasonic extraction method, a filtration method, a reflux extraction method, etc., and these can be performed alone or in combination of two or more methods, but are not limited thereto.
  • 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.
  • 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.
  • a solvent extract can be prepared by extracting a branch one or more times using the solvent, and a dried extract obtained by distilling the solvent extract under reduced pressure and then freeze-drying or spray-drying can be prepared.
  • the amount of the solvent as described above is 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.
  • the above-mentioned manufactured extract can then be filtered, concentrated, or dried to remove the solvent, and filtration, concentration, and drying can all be performed.
  • filtration can be performed using filter paper or a vacuum filter
  • concentration can be performed using a vacuum concentrator
  • drying can be performed using a freeze-drying method, but is not limited thereto.
  • 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.
  • the fraction can be interpreted as a fraction obtained by applying the extract of the zelkova tree to various fractionation methods.
  • the fraction of the extract can be obtained by applying the extract to various fractionation methods, and the fractionation methods are not particularly limited thereto, but can be a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, a chromatography fractionation method performed by performing various chromatography (designed for separation according to size, charge, hydrophobicity, or affinity), etc.
  • the solvent used in the solvent fractionation method is not particularly limited thereto, but a polar solvent or a nonpolar solvent can be used, and specifically, a nonpolar solvent can be used.
  • the solvent fractionation method can be performed in a manner of sequentially fractionating the extract using a solvent having a high degree of nonpolarity to a solvent having a low degree of nonpolarity, and for example, a method of sequentially fractionating the extract using hexane or ethyl acetate can be used.
  • a polar solvent such as water or alcohol
  • a nonpolar solvent such as hexane, ethyl acetate, chloroform, or dichloromethane
  • these can be used alone or in a mixture of two or more thereof, but are not limited thereto.
  • retinal disease in this application means a disease caused by abnormality or damage to the retina due to extrinsic, intrinsic or genetic causes.
  • the retinal disease may be caused by oxidative stress.
  • 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.
  • 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,
  • 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.
  • retinal pigment epithelium cell refers to a single layer of pigment cells forming a part of the external retinal blood vessel 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 for maintaining the visual cycle, secrete various growth factors that maintain the bonding structure of photoreceptors and the choroidal layer, transport ions, water, and metabolites from the subretinal space to the blood, and perform phagocytosis of the outer segment of photoreceptors.
  • RPE retina pigment epithelium cell
  • Retinal pigment epithelium cells are cells forming a single layer between the retina and the choroidal layer, and physiologically play an important role in maintaining vision, such as providing nutrition to photoreceptors, excreting metabolites, and phagocytosis of cell debris, and are exposed to oxidative stress due to their high oxygen consumption.
  • Oxidative stress causes structural defects and functional damage of the RPE.
  • Reactive oxygen species (ROS) induced by oxidative stress include free radicals, hydrogen peroxide, and oxygen ions from byproducts of oxygen metabolism, and induce cell damage and apoptosis in various cells.
  • ROS reactive oxygen species
  • cells with high metabolic rates such as RPE are more vulnerable to oxidative damage, and continuous exposure to high-concentration oxidative stress not only destroys mitochondrial function by inducing mitochondrial DNA damage in RPE, but also ultimately leads to apoptosis of RPE.
  • Visual impairment due to retinal damage is one of the most important causes of blindness, and oxidative stress is considered to be the cause of retinal diseases.
  • 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 an advantage in that it has excellent cell proliferation activity when included in a concentration of 0.1 ⁇ g/ml to 5 ⁇ g/ml in the composition. If it is included in a concentration of less than 0.1 ⁇ g/ml, the effect is minimal, and if it is included in a concentration exceeding 5 ⁇ g/ml, it may exhibit toxicity to retinal pigment epithelial cells.
  • 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 ⁇ g/ml to 5 ⁇ g/ml, the effect of inhibiting the production of reactive oxygen species becomes more excellent as the concentration increases.
  • the extract of the zelkova tree is included in a concentration exceeding 5 ⁇ g/ml, it can exhibit toxicity to retinal pigment epithelial cells.
  • the above extract of the hornbeam tree has the effect of preventing the death of retinal pigment epithelial cells against H2O2 toxicity. Specifically, when included in a concentration of 1 ⁇ g/ml to 5 ⁇ g/ml, it has the advantage of having a more excellent cell protection effect. When the extract of the hornbeam tree is included in a concentration exceeding 5 ⁇ g/ml, it may exhibit toxicity to retinal pigment epithelial cells.
  • ROS reactive oxygen species
  • the present application provides a food composition for protecting the eyes, protecting vision, or preventing and improving retinal diseases, containing an extract of the zelkova tree as an effective ingredient.
  • the food composition of the present application may contain an extract of the zelkova tree or a fraction thereof.
  • improvement refers to any act of improving or benefiting the symptoms of a subject suspected of or suffering from a retinal disease that is prevented or treated by using a composition containing an extract of the zelkova tree or a fraction thereof as an effective ingredient.
  • the food composition of the present application is very useful because it can be consumed on a daily basis and can be expected to have effects such as protecting the eyes, protecting vision, reducing eye fatigue, and preventing and improving retinal diseases.
  • food in this application means a natural product or processed product containing one or more nutrients, preferably one that has undergone a certain degree of processing to become directly edible, and in its usual sense includes all of food, food additives, functional foods, and beverages.
  • Examples of foods to which the composition for protecting the eyes, protecting vision, or preventing and improving symptoms of retinal diseases according to the present application can be added include various foods, beverages, gum, tea, vitamin complexes, functional foods, etc.
  • the 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, gum, ice cream, processed dairy products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi types, pickled vegetables, etc.), beverages (e.g., fruit drinks, vegetable drinks, soy milk, fermented beverages, etc.), and natural seasonings (e.g
  • the above “functional food” means a food group or food composition that has been processed to sufficiently express the body's regulatory functions 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 specifically, it may be a health functional food.
  • the above functional food may include a food additive acceptable from a food science perspective, and may further include appropriate carriers, excipients, and diluents commonly used in the manufacture of functional foods.
  • health functional food means 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 effects.
  • health food means a food that has a more active health maintenance or promotion effect than general food
  • health supplement means a food for the purpose of health supplementation.
  • the terms health functional food, health food, and health supplement may be used interchangeably.
  • the health functional food of the present application can be manufactured by a method commonly used in the art. It can be manufactured in various forms of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long period of time because it uses food as a raw material, and can be highly portable.
  • the "beverage” in the present application means a general term for something that is consumed to quench thirst or enjoy the taste, and includes a functional beverage.
  • the beverage has no special restrictions on other ingredients except that it contains the composition for eye protection, vision protection, or prevention and improvement of retinal disease symptoms as an essential ingredient in the indicated proportion, and may contain various flavoring agents or natural carbohydrates as additional ingredients like a typical beverage.
  • 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, colorants 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.
  • various nutrients vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, colorants 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 amount of the composition according to the present invention may be comprised in an amount of 0.001 to 90 wt% of the total food weight, preferably 0.1 to 40 wt%, and in the case of a beverage, may be comprised in an amount of 0.001 to 2 g, preferably 0.01 to 0.1 g, based on 100 ml.
  • 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 therefore is not limited to the above range.
  • 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 according to the general provisions and general test methods of the Food Additive Code approved by the Ministry of Food and Drug Safety, unless otherwise specified.
  • the above food additives refer to food compositions that are usually intentionally added to food in small quantities for the purpose of improving the appearance, flavor, texture, or storability, and are used for the purpose of improving the quality of food, enhancing the storability or palatability, and increasing the nutritional value and actual value of food.
  • this may be a substance used by adding, mixing, infiltrating, or other methods to food when manufacturing, processing, or preserving food.
  • One embodiment of the present application provides a pharmaceutical composition for preventing or treating retinal disease, comprising an extract of the zelkova tree as an active ingredient.
  • composition according to the present application may contain a pharmaceutically effective amount of a zelkova extract alone or may contain one or more pharmaceutically acceptable carriers, excipients or diluents.
  • the term "pharmaceutically acceptable” as used herein 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.
  • 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.
  • fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be additionally included.
  • the pharmaceutical composition according to the present application may further include a pharmaceutically acceptable carrier in addition to the active ingredient.
  • the pharmaceutical preparation may be prepared by mixing it with pharmaceutically acceptable excipients, adjuvants, analgesics, isotonic agents, preservatives, and other adjuvants and formulating it in a pharmaceutically acceptable formulation form, but is not limited thereto.
  • Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc.
  • liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc.
  • preparations for parenteral administration e.g., intravenous, subcutaneous, intraperitoneal, or topical application
  • parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories, but can be appropriately selected by those skilled in the art.
  • the pharmaceutically effective amount as described above refers to an amount sufficient to prevent, improve and/or treat retinal disease symptoms.
  • the preferred dosage of the pharmaceutical composition above varies depending on the patient's condition and weight, the degree of the disease, the drug form, the route of administration, and the period of administration, but can be appropriately selected by those skilled in the art.
  • 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.
  • the administration may be administered once a day or divided into several times. Therefore, the above dosage does not limit the scope of the present invention in any way.
  • the pharmaceutical composition When 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 ready-to-use injectable dry powders 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 can further include appropriate carriers, excipients, and diluents commonly used in the manufacture of pharmaceuticals.
  • compositions of the present application can be formulated using methods known in the art so as 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.
  • 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.
  • 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.
  • the above feed composition may include a feed additive.
  • the feed additive of the present application corresponds to a supplementary feed under the Feed Management Act.
  • feed means any natural or artificial diet, meal, etc., or a component of said meal, intended for eating, ingesting, or digesting by an animal or suitable therefor.
  • 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, fibers, pharmaceutical by-products, fats, starches, meal, or grain by-products; and animal feed such as proteins, inorganic substances, fats, minerals, fats, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.
  • feed additive in the present application includes substances added to feed for various purposes such as nutrient supplementation and prevention of weight loss, promotion of digestibility and utilization of fiber in feed, improvement of milk quality, prevention of reproductive disorders and improvement of conception rate, and prevention of high temperature stress in summer.
  • the feed additive of the present 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 fermented molds; and yeast agents.
  • 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 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
  • 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.
  • One embodiment of the present application provides a use of the composition for improving vision or for improving, preventing and treating retinal diseases.
  • subject in this application means any animal that is susceptible to or has developed a retinal disease.
  • the feed composition according to the present application can be applied without limitation to any entity other than a human for the purpose of improving, preventing, or alleviating retinal diseases.
  • entity including non-human animals such as dogs, cats, monkeys, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds, and fish.
  • 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.
  • a quasi-drug in the specification of the present application 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.
  • a quasi-drug means a product excluding products used for the purpose of pharmaceutical products.
  • Non-medicinal products 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 instruments or machines and similar products; products that are one of the preparations used for sterilization, insecticide and 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 instruments, 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 instruments, machines or devices; and also includes external preparations and personal hygiene products. Specifically, they can be eye drops, eye wash preparations and ointments.
  • the extract of the zelkova tree may be added as is or may be used appropriately according to a conventional method together with other over-the-counter drug ingredients.
  • the mixing amount of the effective ingredients may be appropriately determined depending on the intended use.
  • the contents of the pharmaceutical composition or health functional food composition may be applied to the pharmaceutical composition of the present application.
  • the eye wash or eye wash pharmaceutical composition 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.
  • composition may further include additives such as fragrances, pigments, bactericides, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, emulsifiers, and synthetic polymers to improve physical properties, but is not limited thereto.
  • additives such as fragrances, pigments, bactericides, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, emulsifiers, and synthetic polymers to improve physical properties, but is not limited thereto.
  • 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 easy washing.
  • the composition may also be prepared and used in the form of a paste, ointment, spray, or eye drop, but is not limited thereto.
  • the leaves and stems of the zelkova tree were dried in a shaded place, then ground and powdered.
  • the powder was prepared to produce an extract, and 100% of the leaf and stem powder was used.
  • a commercially available eye-protecting marigold extract (manufactured by Nutraceuticals International Group, weight 100 g) was purchased, dissolved in 60% ethanol, and used as a comparative example.
  • ARPE-19 cells human retinal pigmented epithelial cells, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).
  • 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 medium antibiotics (Penicillin streptomycin, Gibco, CA, USA) were used to suppress microbial contamination or proliferation.
  • the dish was washed with phosphated-buffered saline-EDTA (PBS-EDTA, Thermo Fisher Scientific, Massachusetts, USA), and then the cells attached to the bottom of the dish were detached by treating with 0.25% Trypsin-EDTA (Gibco, Thermo Fisher Scientific, Massachusetts, USA) and subcultured.
  • the medium was changed every 48 to 72 hours and ARPE-19 cells were cultured.
  • ARPE-19 cells were seeded in 96-well plates at a concentration of 1 ⁇ 10 4 cells/well and cultured in a CO 2 incubator controlled at 37°C, 95% humidity, and 5% CO 2 for 24 h. Each sample was treated to the cells in a fresh medium at an appropriate concentration range that did not show cytotoxicity and cultured for 24 h. To measure the viability of the cell line, the absorbance was measured at 490 nm with a microplate reader (Molecular Devices, Versa Max ELISA Microplate Reader, USA) using MTS (CellTiter 96 ® 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.
  • Example 1 which is a zelkova tree, at concentrations ranging from 0.1 to 10 ⁇ g/ml, and the results are shown in Fig. 1 and Table 1.
  • * p ⁇ 0.05 indicates a significant difference as compared to the control group.
  • the above-mentioned ethanol extract of the hornbeam tree showed cytotoxicity at a treatment concentration of 10 ⁇ g/ml, whereas no cytotoxicity was observed at treatment concentrations below 5 ⁇ g/ml. Therefore, the efficacy of the hornbeam tree extract for improvement, prevention, or treatment of retinal pigment epithelial cells was confirmed in the concentration range of 0.1 to 5 ⁇ g/ml.
  • ARPE-19 cells were seeded in 96-well plates at a concentration of 1 ⁇ 10 4 cells/well and cultured in a CO 2 incubator controlled at 37 °C, 95% humidity, and 5% CO 2 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).
  • the ARPE-19 cells were treated with the ethanol extract of the example of the dogwood tree and the marigold extract of the comparative example at concentrations of 0.1 to 5 ⁇ g/ml, and the results are as shown in Fig. 2 and Table 2.
  • * p ⁇ 0.05 indicates a significant difference as compared to the control group.
  • Examples 1 to 4 which are the ethanol extracts of the above-mentioned hornbeam, showed an increase in the proliferation activity of ARPE-19 cells as the concentration increased.
  • the proliferation activity of the hornbeam ethanol extract was 142.8% at a concentration of 1 ⁇ g/ml, which was about 1.43 times better than that of the control.
  • the comparative examples 1 to 4 which are the marigold extracts, showed a very small difference compared to the control group.
  • ARPE-19 cells were damaged by H2O2 , and the protective efficacy against ARPE-19 cell damage was measured by cell viability.
  • the ARPE-19 cell line was seeded at a concentration of 1 ⁇ 10 4 cells/well in a 96-well plate, and cultured at 37°C, 5% CO 2 and 95% humidity for 24 hours to stabilize. After removing the medium, 10 ⁇ l of H 2 O 2 diluted in new medium was treated at each concentration, and the reaction was performed at 37°C, 5% CO 2 and 95% humidity for 24 hours.
  • the viability of ARPE-19 cell line was measured using MTS (CellTiter 96 ® AQueous One Solution Cell Proliferation Assay, Promega, USA) reagent. 10 ⁇ l of the MTS reagent was treated per well, and after reaction for 60 minutes at 37°C, 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 (Control).
  • MTS CellTiter 96 ® AQueous One Solution Cell Proliferation Assay, Promega, USA
  • H2O2 was treated at a concentration of 0.5 - 10 mM, and the results are as shown in Fig. 3 and Table 3.
  • * p ⁇ 0.05 indicates a significant difference as compared to the control group.
  • ARPE-19 cells were seeded at a concentration of 1 ⁇ 10 4 cells/well in a 96-well plate and cultured at 37°C, 5% CO2 , and 95% humidity for 24 hours to stabilize. After removing the medium, 2 mM H2O2 was treated with new medium , and 10 ⁇ l of the sample (extract) was treated at each concentration and reacted at 37°C, 5% CO2 , and 95% humidity for 24 hours.
  • the viability of ARPE-19 cell line was measured using MTS (CellTiter 96 ® AQueous One Solution Cell Proliferation Assay, Promega, USA) reagent. That is, 10 ⁇ l of the MTS reagent was treated per well, and after reaction for 60 minutes at 37 °C, 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 treatment group compared to the control group that was not treated with the sample (extract).
  • MTS CellTiter 96 ® AQueous One Solution Cell Proliferation Assay, Promega, USA
  • * p ⁇ 0.05 indicates a significant difference as compared to the (+)control group.
  • ROS reactive oxygen species
  • ARPE-19 cells were dispensed at a concentration of 1 ⁇ 10 4 cells/well (100 ⁇ l) into a 96-well plate and 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 cultured for 24 hours. After 24 hours, 100 ⁇ l of PBS was added and washed, and this process was repeated 3 times. 100 ⁇ l of 2',7'-dichlorofluorescin diacetate (DCF-DA) dissolved in DMSO was diluted to 20 ⁇ M (in PBS), and cultured for 20 minutes. 100 ⁇ l of 1 mM H 2 O 2 (in PBS) was added, reacted for 30 minutes, and the fluorescence intensity was measured at 486 nm and 535 nm using an ELISA reader.
  • DCF-DA 2',7'-dichlorofluorescin diacetate
  • the ethanol extract of the example tree and the marigold extract of the comparative example were treated at concentrations of 0.1 to 5 ⁇ g/ml, and the results are as shown in Fig. 5 and Table 5.
  • * p ⁇ 0.05 indicates a significant difference as compared to the (+)control group.

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Abstract

La présente invention concerne l'utilisation d'un extrait de Stewartia pseudocamellia à utiliser pour protéger la vue et soulager, prévenir ou traiter les maladies rétiniennes. L'extrait de Stewartia pseudocamellia de la présente invention manifeste une activité de prolifération de cellules épithéliales rétiniennes, ce qui permet d'obtenir d'excellents effets de protéger la vue et de soulager, prévenir ou traiter les maladies rétiniennes.
PCT/KR2023/018795 2023-11-21 2023-11-21 Utilisation d'un extrait de stewartia pseudocamellia pour protéger la vue ou soulager, prévenir ou traiter les maladies rétiniennes Pending WO2025110266A1 (fr)

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KR20120095251A (ko) * 2011-02-18 2012-08-28 한국과학기술연구원 벌개미취 추출물 또는 이의 분획물을 포함하는 망막질환 예방 및 치료용 조성물
JP2019048858A (ja) * 2013-10-29 2019-03-28 プレジデント アンド フェローズ オブ ハーバード カレッジ 酸化ストレスを阻害するための方法および組成物
KR20220085035A (ko) * 2020-12-14 2022-06-21 경상국립대학교산학협력단 노각나무 추출물을 포함한 구강 건강 개선용 조성물 및 이를 포함하는 제품
KR20220162654A (ko) * 2022-11-10 2022-12-08 주식회사 아이두젠 노각나무 추출물을 유효성분으로 포함하는 시력 보호용 또는 망막 질환의 개선 및 예방용 조성물

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