WO2022030719A1 - Composition for improving, treating, or preventing muscle disease, or improving muscle function, containing rose hip as acitve ingredient - Google Patents
Composition for improving, treating, or preventing muscle disease, or improving muscle function, containing rose hip as acitve ingredient Download PDFInfo
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- WO2022030719A1 WO2022030719A1 PCT/KR2021/003055 KR2021003055W WO2022030719A1 WO 2022030719 A1 WO2022030719 A1 WO 2022030719A1 KR 2021003055 W KR2021003055 W KR 2021003055W WO 2022030719 A1 WO2022030719 A1 WO 2022030719A1
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- rosehip
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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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- 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
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- 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)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/738—Rosa (rose)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
Definitions
- the present invention relates to a composition for improving, treating or preventing muscle disease, or to a composition for improving muscle function, and more particularly, to improve or treat muscle disease containing rose hips such as pulverized rosehip or rosehip extract as an active ingredient Or for prevention, or relates to a composition for improving muscle function.
- Muscle atrophy is caused by a gradual decrease in muscle mass, and refers to muscle weakness and degeneration (Cell, 119(7):907-910, 2004). Muscular atrophy is promoted by inactivity, oxidative stress, and chronic inflammation, and weakens muscle function and athletic performance (Clinical Nutrition, 26(5):524-534, 2007). The most important factor that determines muscle function is muscle mass, which is maintained by a balance between protein synthesis and breakdown. Muscular dystrophy occurs when protein degradation occurs more than synthesis (The International Journal of Biochemistry and Cell Biology, 37(10):1985-1996, 2005).
- Muscle size is regulated by intracellular signaling pathways that induce anabolism or catabolism within the muscle.
- a signal transduction reaction that induces synthesis rather than degradation of muscle protein occurs, muscle protein synthesis is increased, which appears as an increase in muscle size (hypertrophy) or an increase in the number of muscle fibers (hyperplasia) according to an increase in muscle protein (The Korea Journal of Sports) Science, 20(3):1551-1561, 2011).
- PI3K phosphatidylinositol-3 kinase
- mTOR mammalian target of rapamycin
- mTOR contributes to muscle mass increase by inducing muscle protein synthesis by activating two factors that initiate mRNA translation, 4E-binding protein (4EBP1) and phosphorylated 70-kDa ribosomal S6 kinase (p70S6K) (The Korea Journal of Sports Science, 20(3):1551-1561, 2011; The International Journal of Biochemistry and Cell Biology, 43(9):1267-1276, 2011).
- 4EBP1 4E-binding protein
- p70S6K phosphorylated 70-kDa ribosomal S6 kinase
- muscle cells and muscle formation are regulated by various muscle regulatory factors. Among them, the induction of myogenin expression by the activity of MyoD is the most important factor in the fusion of myoblasts, and is involved in the formation of myotubes. The muscle fibers formed through this process are bundled to finally form a muscle (Cellular and Molecular Life Sciences, 70:4117-4130, 2013).
- Rose hip is a plant belonging to the family Rosaceae, and the scientific name is known as Rosa canina L. So far, in relation to rosehip, anti-aging effects (Clinical Interventions in Aging, 19(10): 1849-1856, 2015), antioxidant effects (Antioxidants, 8(4): 92, 2019), anti-cancer effects (Journal of Pharmaceutical Analysis 8 , 8(6): 394-399, 2018), antidiabetic effect (Journal of Evidence-Based Complementary & Alternative Medicine, 21(4): 25-30, 2016), anti-obesity effect (Preventive Nutrition and Food Science, 18) (2): 85-91, 2013) has been reported to have the activity.
- the present inventors have searched for natural substances that have excellent muscle function control activity and can be safely applied.
- rose hips such as pulverized rosehip or rosehip extract are used for improving, treating or preventing muscle disease, or improving muscle function.
- an object of the present invention is to provide a food composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient.
- Another object of the present invention is to provide a pharmaceutical composition for treating or preventing muscle disease containing rosehip as an active ingredient.
- Another object of the present invention is to provide a cosmetic composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient.
- Another object of the present invention is to provide a feed additive for improving or preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient containing rosehip as an active ingredient.
- the present invention provides a food composition for improving or preventing muscle disease, or for improving muscle function, containing rose hip, such as pulverized rosehip or rosehip extract, as an active ingredient.
- the present invention also provides a pharmaceutical composition for treating or preventing muscle disease containing rosehip as an active ingredient.
- the present invention provides a cosmetic composition for improving or preventing muscle disease or improving muscle function containing rosehip as an active ingredient.
- the present invention provides a feed additive for improving or preventing muscle disease or improving muscle function containing rosehip as an active ingredient.
- the present invention provides the use of a pharmaceutical composition comprising rosehip as an active ingredient for the manufacture of a medicament for the prevention or treatment of muscle disease.
- the present invention also provides a method for treating muscle disease, comprising administering to a patient with a muscle disease a pharmaceutical composition comprising rosehip as an active ingredient.
- Rosehip according to the present invention has an excellent effect in increasing the activity of mTOR involved in muscle protein synthesis and inhibiting the mRNA expression of MuRF1 and atrogin-1 involved in muscle protein degradation.
- the present invention is a natural product, it can be safely used without side effects and can be used as a pharmaceutical, food, cosmetic or feed additive.
- 5 is a result confirming the effect of increasing the protein expression level of p-mTOR, p-p70S6K and p-4EBP-1 according to the treatment with hot water extract of rosehip in muscle atrophy-induced mice.
- the present invention relates to a food composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip, such as a pulverized rosehip or a rosehip extract; a pharmaceutical composition for treating or preventing a muscle disease containing rosehip, such as a pulverized rosehip or a rosehip extract; Or a cosmetic composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip, such as a pulverized rosehip or a rosehip extract; or a feed additive for improving or preventing muscle disease or improving muscle function containing rosehip, such as rosehip pulverized or rosehip extract; Or it provides a method for treating a muscle disease comprising applying a rosehip, such as a pulverized rosehip or a rosehip extract, to a human or non-human mammal.
- 'rose hip' is a plant of the Rosaceae family, a fruit of Rosa canina L., and a crushed rosehip or a rosehip extract may be used.
- 'pulverized rosehip' refers to a method that can be easily performed by a person of ordinary skill in the art to which the present invention pertains to dried rosehip fruit, and ingested while being easily ingested by mammals, including humans.
- the active ingredient is easily released from the dry pulverized material in the intestine, and as a result, it can be prepared and used in a form that can be easily absorbed into the body of a mammal.
- the shape or size of the dry pulverized particles is not limited, but as the surface of the particles is maximized, it is easier to release the active ingredient described above, so it is preferable to prepare it in the form of a fine powder as much as possible.
- 'rosehip extract' is obtained by extracting dried rosehip fruit using a suitable solvent, and is obtained in the form of an extract, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a prepared or purified product thereof.
- the method for preparing the rosehip extract may be obtained by extracting from the rosehip with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and ultra-high pressure, subcritical or supercritical fluid, but is not limited thereto.
- the rosehip extract of the present invention can be prepared using conventional extraction methods in the art, such as heat extraction, cold-chill extraction, ultrasonic extraction, filtration, and reflux extraction. Cultivated ones can be used.
- the rosehip extract according to the present invention can be separated using a conventional solvent according to a conventional method known in the art for preparing an extract from a natural product, that is, under conditions of conventional temperature and pressure.
- the term 'fraction' refers to a result obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various components. In the present invention, it refers to a result obtained by a fractionation method for separating a specific component or a specific group from the rosehip extract prepared as described above.
- a conventional fractionation solvent known in the art, for example, a polar solvent such as anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms such as water, ethanol, and methanol, and hexane, butanol, ethyl
- a non-polar solvent such as acetate, chloroform, dichloromethane, or a mixed solvent thereof may be used, but is not limited thereto.
- the rosehip fraction of the present invention may include those obtained by additionally applying a purification process.
- 'muscle' refers to tendons, muscles, and tendons comprehensively
- 'muscular function' means the ability to exert force by contraction of the muscle, It includes muscle strength, which is the ability to perform, muscular endurance, which is the ability to repeat contractions and relaxations for how long or how many times a muscle can repeat contractions and relaxations with a given weight, and instantaneous power, which is the ability to exert strong force in a short period of time.
- the term 'improving muscle function' refers to improving muscle function by increasing muscle mass.
- the present invention relates to a food composition for improving and preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient, such as a pulverized rosehip or a rosehip extract; pharmaceutical compositions for preventing and treating muscle diseases; A cosmetic composition for improving and preventing muscle disease, or improving muscle function; It provides a feed additive for improving or preventing muscle disease, or for improving muscle function.
- the rosehip extract may be a hot water extract, an ethanol extract, an ethyl acetate extract, a hexane extract, an ultra-high pressure extract, a subcritical extract, or a supercritical extract, but is not limited thereto.
- the rosehip extract can be obtained by extracting the fruit of rosehip with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and subcritical and supercritical fluids. It can also be obtained by extracting rosehip under ultra-high pressure conditions of 100 MPa or more. If necessary, it may be prepared by additionally including filtration and concentration steps according to methods known in the art.
- the organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, It may be at least one selected from the group consisting of methylene chloride, hexane, cyclohexane, and petroleum ether.
- the present inventors repeatedly extract the dried rosehip fruit pulverized product using ethanol, ethyl acetate and hexane as solvents at room temperature, respectively, hot water extraction, ultra-high pressure extraction, subcritical fluid extraction, supercritical A rosehip extract was prepared using fluid extraction.
- the food composition for preventing and improving muscle disease or improving muscle function of the present invention can be used for preventing or improving muscle disease due to muscle wasting or degeneration.
- Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by a gradual loss of muscle mass, weakness and degeneration of muscles, particularly skeletal or voluntary muscles and cardiac muscles.
- Examples of related diseases include sarcopenia, muscular atrophy, muscular dystrophy, atony, muscle degeneration, myasthenia gravis, cachexia, and the like.
- the composition of the present invention has an effect of increasing muscle mass, and the type of muscle is not limited.
- the food composition of the present invention includes all forms such as functional food, nutritional supplement, health function food, food additives and feed, including humans or livestock Animals are intended for food.
- Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
- Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
- Common foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat and their processed foods (eg, ham, sausages) Corned beef, etc.), breads and noodles (eg udon noodles, soba noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (eg butter, cheese, etc.), edible vegetable oils and fats, margarine , vegetable protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.) can be prepared by adding crushed rosehip or rosehip extract.
- the nutritional supplement is not limited thereto, but may be prepared by adding crushed rosehip or rosehip extract to capsules, tablets, pills, and the like.
- the pulverized rosehip or rosehip extract itself is prepared in the form of tea, juice, and drink, and liquefied, granulated, encapsulated and powdered so that it can be consumed (health drink). can be ingested.
- the pulverized rosehip or rosehip extract in the form of a food additive, it may be prepared and used in the form of a powder or concentrate.
- it can be prepared in the form of a composition by mixing the pulverized rosehip or rosehip extract with an active ingredient known to be effective in preventing and improving muscle disease, or improving muscle function.
- the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional drink.
- the above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol.
- Sweeteners include natural sweeteners such as thaumatin, stevia extract; A synthetic sweetener such as saccharin or aspartame may be used.
- the proportion of the natural carbohydrate is generally about 0.01 to 5.0 g, preferably about 0.1 to 1.0 g per 100 mL of the composition of the present invention.
- the pulverized rosehip or rosehip extract may be contained as an active ingredient in a food composition for preventing and improving muscle disease or improving muscle function, the amount of which is effective to achieve the action for preventing and improving muscle disease or improving muscle function.
- the amount is not particularly limited, but is preferably 0.01 to 100% by weight based on the total weight of the total composition.
- the food composition of the present invention may be prepared by mixing with other active ingredients known to be effective in preventing and improving muscle disease or improving muscle function together with pulverized rosehip or rosehip extract.
- the composition for preventing and treating muscle disease of the present invention may also be a pharmaceutical composition.
- the composition for preventing and treating muscle disease of the present invention is a pharmaceutical composition, it may be used for the prevention or treatment of muscle disease caused by muscle wasting or degeneration.
- Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by a gradual loss of muscle mass, weakness and degeneration of muscles, particularly skeletal or voluntary muscles and cardiac muscles.
- related diseases include sarcopenia, muscular atrophy, muscular dystrophy, atony, muscle degeneration, myasthenia gravis, cachexia, and the like.
- the composition of the present invention has an effect of increasing muscle mass, and the type of muscle is not limited.
- the 'pharmaceutical composition for preventing and treating muscle disease of the present invention may include a pharmaceutically acceptable carrier.
- the pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration.
- Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like.
- Carriers for parenteral administration may also include water, suitable oils, saline, aqueous glucose and glycols, and the like.
- it may further include a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid.
- Suitable preservatives are benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol.
- reference may be made to those described in the following literature Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).
- the pharmaceutical composition of the present invention may be administered to mammals including humans by any method.
- it may be administered orally or parenterally, and parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal. , intranasal, enteral, topical, sublingual or rectal administration.
- the pharmaceutical composition of the present invention may be formulated as a formulation for oral administration or parenteral administration according to the administration route as described above.
- one or more buffers eg, saline or PBS
- antioxidants e.g, bacteriostatic agents, chelating agents (eg, EDTA or glutathione), fillers, bulking agents, binders, adjuvants (eg, aluminum hydroxide) side), suspending agents, thickening agents, wetting agents, disintegrating agents or surfactants, diluents or excipients.
- bacteriostatic agents eg, EDTA or glutathione
- fillers eg, bulking agents, binders, adjuvants (eg, aluminum hydroxide) side), suspending agents, thickening agents, wetting agents, disintegrating agents or surfactants, diluents or excipients.
- Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions or capsules, and such solid preparations include at least one excipient in the pharmaceutical composition of the present invention, for example , starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose , methyl cellulose, sodium carboxymethyl cellulose, and hydroxypropylmethyl-cellulose or gelatin may be mixed and prepared.
- tablets or sugar tablets can be obtained by blending the active ingredient with a solid excipient, grinding it, adding a suitable adjuvant, and processing it into a granule mixture.
- Liquid formulations for oral use include suspensions, solutions, emulsions, or syrups.
- various excipients for example, wetting agents, sweeteners, flavoring agents, or preservatives may be included. .
- cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, and an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifier, and a preservative may be additionally included. .
- the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations and nasal inhalants together with suitable parenteral carriers.
- suitable parenteral carriers include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and/or vegetable oils.
- suitable carriers include Hanks' solution, Ringer's solution, PBS (phosphate buffered saline) with triethanolamine or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. etc. can be used.
- PBS phosphate buffered saline
- isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. etc.
- it may further include various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
- the injection may further contain an isotonic agent such as sugar or sodium chloride.
- transdermal administration forms such as ointment, cream, lotion, gel, external solution, pasta, liniment, and air are included.
- 'transdermal administration' means that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin by topically administering the pharmaceutical composition to the skin.
- the compounds for use according to the invention may be administered in pressurized packs or using a suitable propellant, for example, dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered in the form of an aerosol spray from the nebulizer.
- the dosage unit may be determined by providing a valve to deliver a metered amount.
- gelatin capsules and cartridges for use in inhalers or insufflators may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a recipe commonly known to all pharmaceutical chemistry.
- the pharmaceutical composition for preventing and treating muscle disease of the present invention contains an effective amount of a rosehip extract, it is possible to provide a desirable preventive and therapeutic effect on muscle disease.
- the term 'effective amount' refers to an amount that exhibits a higher response than that of the negative control group, and preferably refers to an amount sufficient to improve muscle function.
- the pharmaceutical composition of the present invention may contain 0.01 to 99.99% of the rosehip extract, and the remaining amount may be occupied by a pharmaceutically acceptable carrier.
- the effective amount of the rosehip extract contained in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.
- the total effective amount of the pharmaceutical composition of the present invention may be administered to a patient as a single dose, or may be administered by a fractionated treatment protocol in which multiple doses are administered for a long period of time.
- the pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease. When administered parenterally, it is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per 1 kg of body weight per day based on the rosehip extract, and when administered orally, the body weight per day based on the rosehip extract It may be administered in 1 to several divided doses so as to be administered in an amount of preferably 0.01 to 100 mg, more preferably 0.1 to 50 mg per 1 kg.
- the dosage of the rosehip extract is determined by considering various factors such as the age, weight, health status, sex, severity of the disease, diet and excretion rate of the patient, as well as the administration route and number of treatments of the pharmaceutical composition, and the effective dosage for the patient is determined. Therefore, in consideration of this point, those of ordinary skill in the art will be able to determine an appropriate effective dosage according to the specific use of the rosehip extract for the prevention and treatment of muscle diseases.
- the pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route and administration method as long as the effect of the present invention is exhibited.
- the pharmaceutical composition for preventing and treating muscle disease of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, or biological response modifiers.
- the pharmaceutical composition for preventing and treating muscle disease of the present invention may also be provided in the form of an external preparation containing a rosehip extract as an active ingredient.
- the pharmaceutical composition for the prevention and treatment of muscle diseases of the present invention is used as an external preparation for skin, additionally, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, and a foaming agent (foaming agent) ), fragrance, surfactant, water, ionic emulsifier, nonionic emulsifier, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic active agent, lipophilic active agent Or it may contain adjuvants commonly used in the field of dermatology, such as any other ingredients commonly used in external preparations for skin, such as lipid vesicles. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.
- the pharmaceutical composition for preventing and treating muscle disease of the present invention is provided as an external preparation for skin, it may be in the form of an ointment, patch, gel, cream or spray, but is not limited thereto.
- the composition for preventing and improving muscle disease or for improving muscle function of the present invention may also be a cosmetic composition.
- the cosmetic composition of the present invention contains rosehip extract as an active ingredient, and a basic cosmetic composition (toilet water, cream, essence, face wash such as lotion, cream, essence, cleansing foam and cleansing water, pack, body oil), color cosmetic composition together with a dermatologically acceptable excipient (foundation, lipstick, mascara, makeup base), hair product compositions (shampoo, conditioner, hair conditioner, hair gel), and soap.
- a basic cosmetic composition toilet water, cream, essence, face wash such as lotion, cream, essence, cleansing foam and cleansing water, pack, body oil
- color cosmetic composition together with a dermatologically acceptable excipient (foundation, lipstick, mascara, makeup base), hair product compositions (shampoo, conditioner, hair conditioner, hair gel), and soap.
- the excipient is not limited thereto, but may include, for example, an emollient, a skin penetration enhancer, a colorant, a fragrance, an emulsifier, a thickening agent, and a solvent.
- fragrances, dyes, bactericides, antioxidants, preservatives and moisturizing agents may be additionally included, and for the purpose of improving physical properties, thickeners, inorganic salts, synthetic polymer materials, and the like may be included.
- a face wash and soap with the cosmetic composition of the present invention it can be easily prepared by adding a rosehip extract to a conventional face wash and soap base.
- a cream it can be prepared by adding rosehip extract to a general oil-in-water type (O/W) cream base.
- synthetic or natural materials such as proteins, minerals and vitamins for the purpose of improving physical properties such as fragrances, chelating agents, pigments, antioxidants, and preservatives may be additionally added.
- the content of the rosehip extract contained in the cosmetic composition of the present invention is not limited thereto, but is preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, based on the total weight of the composition. If the content is less than 0.001% by weight, the desired effect cannot be expected, and if it exceeds 10% by weight, there may be difficulties in safety or formulation.
- composition of the present invention may be added to a feed additive or a feed composition comprising the same for the purpose of preventing or improving muscle disease.
- the term 'feed additive' refers to a substance added to feed for various effects such as nutrient supplementation and weight loss prevention, enhancement of digestibility of fiber in feed, oil quality improvement, reproductive disorder prevention and fertility improvement, and prevention of high temperature stress in summer includes
- the feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act, and mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, and trace minerals such as zinc, copper, cobalt, selenium, and kerotene.
- vitamins AD, E, nicotinic acid, vitamin B complex, etc. protective amino acids such as methionine and lysine, protective fatty acids such as fatty acid calcium salts, probiotics (lactic acid bacteria), yeast cultures, live bacteria such as mold fermented products, Yeast agents and the like may be further included.
- the term 'feed' refers to any natural or artificial diet, one-meal meal, etc. or a component of the one-meal meal for animals to eat, ingest and digest, or to prevent or improve muscle disease according to the present invention.
- the feed containing the composition for use as an active ingredient can be prepared in various types of feed known in the art, and preferably, a concentrated feed, roughage and/or special feed may be included, but is not limited thereto.
- seed fruits containing grains such as wheat, oats, and corn, bran containing rice bran, bran, barley bran, etc.
- Fish Soluble is a concentrated fish meal, fish waste, and fresh liquid obtained from fish, fish meal, fish waste, and residual starch, which is the main component of starch residue, which is a by-product obtained by subtracting starch from sweet potatoes and potatoes.
- animal feeds such as dried whey, yeast, chlorella, and seaweed, which are dried whey, which is the remainder of the production of soluble), meat meal, blood meal, cow meal, powdered skim milk, cheese from skim milk, and casein from skim milk. It is not limited thereto.
- raw grass feed such as wild grasses, grasses, and green cuttings, turnips for feed, beets for feed, root vegetables such as rutherbearers, a type of turnip, raw herbs, green crops, grains, etc.
- silage which is stored feed fermented with lactic acid after filling, wild grass, hay obtained by cutting and drying grass, straw of crops for breeding purposes, and leaves of legumes.
- Special feed includes mineral feed such as oyster shells and rock salt, urea feed such as urea or its derivative diureide isobutane, supplementing ingredients that are easily lacking when only natural feed raw materials are mixed, or added to formulated feed to increase feed storage.
- feed additives and dietary supplements which are substances added in trace amounts, but are not limited thereto.
- the feed additive for preventing or improving muscle disease according to the present invention can be prepared by adding ground rosehip or rosehip extract in an appropriate effective concentration range according to various feed preparation methods known in the art.
- the feed additive according to the present invention can be applied without limitation as long as it is an individual for the purpose of preventing or improving muscle disease.
- non-human animals such as cattle, horses, pigs, goats, sheep, dogs, cats, rabbits, etc., birds and fish can be applied to any object.
- the dried rosehip fruit was crushed with a mixer, and 10 g of the crushed rosehip fruit was charged into a sample cartridge and extracted using a supercritical fluid extraction device (SFX 3560, Isco Inc., Lincoln, NE, USA).
- the supercritical extraction conditions were an extraction pressure of 300 bar, an extraction temperature of 50° C., a flow rate of supercritical carbon dioxide of 60 mL/min, and an extraction time of 2 hours.
- the supercritical fluid state was released by lowering the pressure of the extraction device to obtain a rosehip fruit supercritical extract.
- Example 2 Effect of increasing mTOR activity, a biomarker for muscle protein synthesis of rosehip hot water extract and ethanol extract
- mTOR protein when mTOR protein is phosphorylated and activated, it can induce activation of proteins involved in muscle protein synthesis and increase in muscle mass in the PI3K/Akt signaling pathway in muscle cells. Therefore, in order to confirm the muscle-generating activity of the rosehip extract, the activity of mTOR was confirmed using the mTOR Sandwich ELISA kit (Cell Signaling Technology, Beverly, MA, USA).
- L6 myoblasts ATCC; Manassas, VA, USA
- DMEM Dulbecco's modified Eagle's media
- FBS fetal bovine serum
- HS horse serum
- the hot water extract of rosehip of Example 1-1 was dissolved in DMEM (Hyclone) at a concentration of 40 ⁇ g/mL
- the ethanol extract of rosehip of Example 1-2 was dissolved in DMEM (Hyclone) at a concentration of 5.0 ⁇ g/mL
- the cells were treated and Incubated for 12 hours.
- cells were lysed by treatment with cell lysis buffer. Proteins in the obtained cell lysate were quantified at a concentration of 1 mg/mL using Bradford (Bio-Rad Laboratories Inc., Hercules, CA, USA). 50 ⁇ L of cell lysate was dispensed into microwells to which anti-mTOR antibody was attached and left at 37° C.
- the experiment was repeated a total of three times, and the measured values obtained for each were calculated as a percentage (%) relative to the control and expressed as the mean ⁇ standard deviation. Differences between groups were confirmed through Duncan multi-range analysis by one-way analysis of variance using the SPSS25.0 statistical package (SPSS Inc., Chicago, IL, USA). At this time, when the p value was less than 5%, it was marked as statistically significant.
- mTOR activity was measured in the same manner as in Example 2, except that 5.0 ⁇ g/mL of each of the rosehip extracts of Examples 1-3 to 1-7 was treated.
- the rosehip extract of the present invention has an excellent ability to promote muscle protein synthesis in muscle cells.
- L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at a concentration of 1 ⁇ 10 5 cell/mL.
- DMEM Hyclone
- FBS Hyclone
- the medium in the well was removed and the cells were treated with DMEM (Hyclone) containing 2% HS (Hyclone) to induce myotube differentiation. Differentiation was performed for a total of 6 days by replacing the medium with a new medium once every 2 days.
- the rosehip hot water extract prepared in Example 1-1 was added to DMEM (Hyclone) containing 50 ng/mL tumor necrosis factor alpha (TNF- ⁇ ; PeproTech, Rocky Hills, NJ, USA). After dissolving at a concentration of 80 ⁇ g/mL, the cells were treated and incubated for 12 hours. After incubation, the cells were lysed with a NP-40 buffer (ELPIS-Biotech, Daejeon, Korea) containing a protease inhibitor cocktail (Sigma-Aldrich St. Louis, MO, USA) to obtain a cell lysate. The obtained cell lysate was centrifuged at 13,000 rpm for 10 minutes to obtain a supernatant.
- DMEM Hyclone
- TNF- ⁇ tumor necrosis factor alpha
- the protein concentration in the supernatant was quantified with a Bradford (Bio-Rad Laboratories Inc.), and then the protein was heated for 5 minutes, and then the protein was separated by running on a 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gel. The separated protein was transferred to a nitrocellulose membrane.
- p-mTOR, mTOR, p-p70S6K, p70S6K, p-4EBP1, 4EBP1, or ⁇ -tubulin antibody (Cell Signaling Technology) was diluted 1:1000 in 2.5% bovine serum albumin (BSA) at a ratio of 1:1000 to nitro
- BSA bovine serum albumin
- the protein transferred to the cellulose membrane was reacted at room temperature for 20 hours. After reacting with the primary antibody, the nitrocellulose membrane was washed 3 times for 10 minutes using Tris-buffer Saline Tween20 (TBST).
- a secondary antibody (Bethyl Laboratories, Inc., Montgomery, TA, USA) conjugated with horseradish peroxidase (HRP), which recognizes the primary antibody, was diluted 1:5000 in 2.5% BSA, and then placed with a nitrocellulose membrane at room temperature for 2 hours. , and washed 3 times for 10 minutes each using TBST.
- the protein band was developed using ECL Western Blot detection reagent (GE Healthcare, Piscataway, NJ, USA), and the colored protein band was confirmed using the G;BOX EF imaging system (Syngene, Cambridge, UK).
- the protein expression levels of p-mTOR, p-p70S6K and p-4EBP-1 involved in muscle protein synthesis by TNF- ⁇ were significantly ( ## p ⁇ 0.01) decreased.
- the protein expression levels of p-mTOR, p-p70S6K, and p-4EBP-1 were significantly increased ( ** p ⁇ 0.01) as the rosehip hot water extract was treated. This means that the hot water extract of rosehip of the present invention has excellent ability to increase the expression of biomarkers involved in muscle protein synthesis in muscle cells.
- L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at a concentration of 1 ⁇ 10 5 cell/mL.
- DMEM Hyclone
- FBS Hyclone
- the medium in the well was removed and the cells were treated with DMEM (Hyclone) containing 2% HS (Hyclone) to induce myotube differentiation. Differentiation was performed for a total of 6 days by replacing the medium with a new medium once every 2 days.
- Example 1-1 After differentiation, the hot water extract of Example 1-1 was mixed with 40 ⁇ g/mL of the rosehip hot water extract of Example 1-1 in DMEM (Hyclone) containing 50 ng/mL tumor necrosis factor alpha (TNF- ⁇ ; PeproTech, Rocky Hills, NJ, USA). After dissolving in DMEM (Hyclone) at a concentration of 80 ⁇ g/mL, the cells were treated. After 12 hours, total RNA was isolated using TRIzol reagent (Takara, Otsu, Japan). The isolated total RNA was quantified using a nanodrop (NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, MA, USA).
- RNA 16 ⁇ L of quantified RNA was analyzed using Reverse Transcriptase Premix (ELPIS-Biotech) and a PCR machine (Gene Amp PCR System 2700; Applied Biosystems, Foster City, CA, USA) at 42°C, 55 minutes and 70°C, for 15 minutes. It was synthesized as cDNA under conditions. Out of 16 ⁇ L of the generated cDNA, 1 ⁇ L of cDNA, the following specific primers (Bioneer, Daejeon, Korea) and PCR premix (ELPIS-Biotech) were used for 30 seconds at 95°C, 1 minute at 60°C, and 1 minute at 72°C. PCR was performed by repeating 30 times.
- EPIS-Biotech Reverse Transcriptase Premix
- Reverse primer 5'-AGCCTGGAAGATGTCGTTGG-3' (SEQ ID NO: 2)
- Reverse primer 5'-CCGTATGAGTCTTATGTTTTGCTGG-3' (SEQ ID NO: 4)
- Reverse primer 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6)
- the amplified cDNA was separated by electrophoresis on 1.5% agarose gel, and the cDNA band was confirmed using the G;BOX EF imaging system (Syngene).
- the hot water extract of rosehip of the present invention has excellent ability to suppress the expression of biomarkers involved in muscle protein degradation in muscle cells.
- Example 5 In the same manner as in Example 5, except that 5 ⁇ g/mL of the rosehip ethanol, ethyl acetate, hexane, ultra-high pressure, subcritical, and supercritical extracts of Examples 1-2 to 1-7 were treated, respectively. This was repeated 3 times in total. Next, the band density of MuRF1 and atrogin-1 mRNA was measured using the Image J program (National Institute of Health, Bethesda, MD, USA), and then each measured value was calculated as a percentage (%) for the control group and the mean ⁇ It is expressed as standard deviation.
- mice 7-week-old male mice (C57BL/6J; DBL, Korea) were purchased and the experiment was carried out.
- the breeding room environment of all animals was maintained at a temperature of 23 ⁇ 2 °C and a relative humidity of 55 ⁇ 10%.
- a total of 20 rats were randomly divided into 5 rats per group.
- anesthesia was induced by intraperitoneal injection of 325 mg/kg of tribromoethanol (Sigma-Aldrich).
- the right hindlimb gastrocnemius muscle and right sole of the rats in the muscle atrophy group and the sample administration group were cut with a stapler seam using a skin stapler (Unidus, Chungcheongbuk-do, Korea).
- the damage resulted in immobilization of the right hind leg, which was maintained for a week.
- the stapler shim fixed to the calf muscles and the soles of the feet was removed, and dried rosehips 500 mg/kg for another week; 200 mg/kg of hot water extract of rosehip of Example 1-1; 200 mg/kg of rosehip ethanol extract of Example 1-2; 200 mg/kg of the rosehip supercritical extract of Example 1-7 was orally administered daily for 7 days.
- the normal group and the muscular atrophy group were orally administered with saline instead of the sample.
- the muscle strength of the rats was measured using a muscle strength meter (Panlab, Barcelona, Spain).
- the rat's tail was pulled with a constant force until the rat released the youngest member of the muscle strength meter, and a total of five consecutive tests were performed per rat.
- muscle strength was significantly ( # p ⁇ 0.05) decreased in the muscle atrophy group compared to the normal group, but when treated with the crushed rosehip powder and rosehip extract, the muscle strength was significantly reduced compared to the muscle atrophy group ( * * p ⁇ 0.01) was confirmed.
- mice were respiratory anesthetized with isoflurane and the volume and density of the right hind limb muscle were measured using positron emission tomography/computed tomography/single photon emission tomography (microPET/CT/SPECT; Siemens Inveon, Knoxville, TN, USA). did
- the muscle volume of the muscle atrophy group was significantly reduced ( ## p ⁇ 0.01) compared to the normal group. It was confirmed that the muscle volume increased significantly ( ** p ⁇ 0.01). This means that the pulverized rosehip and the rosehip extract of the present invention are excellent in increasing the muscle volume reduced due to muscle atrophy.
- the experimental animals were sacrificed through deep blood sampling after anesthesia by intraperitoneal injection of 325 mg/kg of tribromethanol (Sigma-Aldrich). After confirming that the heartbeat has stopped, the uninjured tibialis anterior muscle was removed from the right hind leg and the weight was measured.
- the weight of the tibialis anterior muscle in the muscular atrophy group was significantly ( ## p ⁇ 0.01) decreased compared to the normal group, but when treated with the crushed rosehip and rosehip extract, the weight was increased compared to the muscle atrophy group. It was confirmed that it increased significantly ( ** p ⁇ 0.01). This means that the pulverized rosehip and the rosehip extract of the present invention are excellent in increasing the muscle weight reduced due to muscle atrophy.
- the tissue was crushed with NP-40 buffer (ELPIS-Biotech, Daejeon, Korea) containing a protease inhibitor cocktail (Sigma-Aldrich St. Louis, MO, USA) to the tibialis anterior muscle extracted in Example 7-3. After obtaining a muscle tissue lysate, centrifuged at 13,000 rpm for 10 minutes to obtain a supernatant. Next, the expression level of the muscle protein synthesis biomarker was measured in the same manner as in Example 4 using the supernatant.
- the capsules were prepared by filling in gelatin capsules according to a conventional capsule preparation method.
- vitamin A acetate 70 ug 1000 mg of crushed rosehip or rosehip extract of the present invention, vitamin A acetate 70 ug, vitamin E 1.0 mg, vitamin B1 0.13 mg, vitamin B2 0.15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 ug, vitamin C 10 mg, biotin 10 ug, nicotinic acid amide 1.7 mg, folic acid 50 ug, calcium pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monobasic 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, and magnesium chloride 24.8 mg can be mixed to prepare the mixture, and the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to the usual health food manufacturing method, and then granules are prepared, , it can be used to prepare a health food composition according to a conventional method.
- the above ingredients After adding purified water to 1000 mg of rosehip extract of the present invention, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, and 1 g of taurine, the above ingredients are mixed according to the general method for preparing a total 900 mL health drink, and then about 1 hour After stirring and heating at 85 for a while, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and then stored in a refrigerator, and then it can be used in the manufacture of health beverage compositions.
- Chewing gum was prepared in a conventional manner by mixing 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of flavoring and 2% by weight of water and 0.1% by weight of rosehip extract of the present invention.
- Nutrient lotion was prepared according to a conventional method by using the rosehip extract of the present invention according to the formulation ratio of the nutrient lotion in Table 3 below.
- Softening lotion (skin lotion) A softening lotion was prepared according to a conventional method by using the rosehip extract of the present invention according to the ratio of the softening lotion formulation in Table 4 below.
- Nutrient Cream A nourishing cream was prepared according to a conventional method by using the rosehip extract of the present invention according to the formulation ratio of the nourishing cream in Table 5 below.
- a massage cream was prepared according to a conventional method by using the crushed rosehip powder or rosehip extract of the present invention according to the massage cream formulation ratio shown in Table 6 below.
- Pack A pack was prepared according to a conventional method by using the pulverized rosehip powder or rosehip extract of the present invention according to the pack formulation ratio in Table 7 below.
- 'appropriate amount' means that an appropriate amount can be added according to preference, and 'to 100' is the weight% remaining after subtracting the sum of the weight% of the remaining ingredients except for purified water from 100% by weight of purified water. It means that it can be done in weight %.
- the present invention provides a composition for improving, treating or preventing muscle disease, or for improving muscle function, containing rose hips such as pulverized rosehips or rosehip extracts. More specifically, the rose hip of the present invention improves muscle mass and muscle strength by increasing the activity of mTOR and reducing the mRNA expression levels of MuRF1 and atrogin-1, thereby improving, treating or preventing muscle diseases, or improving muscle function. indicates. Accordingly, the present invention has high industrial applicability because it can provide a composition showing an excellent effect for improving, treating or preventing muscle disease, or improving muscle function.
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Abstract
The present invention relates to a novel use of rose hip as ground rose hip or a rose hip extract, and to a composition for improving, treating, or preventing muscle disease, or improving muscle function, containing rose hip as an active ingredient. The composition of the present invention has the effect of increasing the activity of mTOR involved in muscle protein synthesis and reducing the mRNA expression of MuRF-1 and atrogin-1 involved in muscle protein degradation, and increases muscle mass and enhances muscle function such that the composition of the present invention can be helpfully used for preventing, improving, or treating various muscle diseases such as sarcopenia, muscular atrophy, muscular dystrophy, atony, muscle degeneration, myasthenia, cachexia, etc.
Description
본 발명은 근육 질환 개선, 치료 또는 예방용, 또는 근 기능 개선용 조성물에 관한 것으로, 보다 상세하게는 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙(rose hip)을 유효성분으로 함유하는 근육 질환 개선, 치료 또는 예방용, 또는 근 기능 개선용 조성물에 관한 것이다.The present invention relates to a composition for improving, treating or preventing muscle disease, or to a composition for improving muscle function, and more particularly, to improve or treat muscle disease containing rose hips such as pulverized rosehip or rosehip extract as an active ingredient Or for prevention, or relates to a composition for improving muscle function.
근위축(Muscle atrophy)이란 근육량의 점진적 감소에 의하여 발생하는 것으로, 근육의 약화 및 퇴행을 일컫는다(Cell, 119(7):907-910, 2004). 근 위축은 비활동, 산화적 스트레스, 만성 염증에 의해 촉진되며 근육 기능과 운동 능력을 약화시킨다(Clinical Nutrition, 26(5):524-534, 2007). 근 기능을 결정짓는 가장 중요한 요소는 근육량이며, 이는 단백질 합성과 분해의 균형에 의해 유지된다. 근 위축증은 단백질 분해가 합성보다 더 일어날 때 발생한다(The International Journal of Biochemistry and Cell Biology, 37(10):1985-1996, 2005).Muscle atrophy is caused by a gradual decrease in muscle mass, and refers to muscle weakness and degeneration (Cell, 119(7):907-910, 2004). Muscular atrophy is promoted by inactivity, oxidative stress, and chronic inflammation, and weakens muscle function and athletic performance (Clinical Nutrition, 26(5):524-534, 2007). The most important factor that determines muscle function is muscle mass, which is maintained by a balance between protein synthesis and breakdown. Muscular dystrophy occurs when protein degradation occurs more than synthesis (The International Journal of Biochemistry and Cell Biology, 37(10):1985-1996, 2005).
근육 크기는 근육 내에서 일어나는 동화작용(anabolism)이나 이화작용(catabolism)을 유도하는 세포 내 신호전달 과정(signaling pathways)에 의해 조절된다. 근 단백질의 분해보다 합성을 유도하는 신호전달 반응이 많이 일어날 경우 근 단백질 합성이 증가되는데, 이는 근 단백질 증가에 따른 근육 크기 증가(hypertrophy)나 근섬유 수 증가(hyperplasia)로 나타난다(The Korea Journal of Sports Science, 20(3):1551-1561, 2011).Muscle size is regulated by intracellular signaling pathways that induce anabolism or catabolism within the muscle. When a signal transduction reaction that induces synthesis rather than degradation of muscle protein occurs, muscle protein synthesis is increased, which appears as an increase in muscle size (hypertrophy) or an increase in the number of muscle fibers (hyperplasia) according to an increase in muscle protein (The Korea Journal of Sports) Science, 20(3):1551-1561, 2011).
근 단백질 합성에 관여하는 인자들은 근 세포 내에서 phosphatidylinositol-3 kinase (PI3K)/Akt pathway의 자극을 기점으로 다운스트림 단백질(downstream proteins)을 인산화시킴으로써 단백질 합성을 유도한다. PI3K/Akt 신호전달에 의한 mammalian target of rapamycin (mTOR)의 활성은 세포 내에서 다양한 성장 신호를 통합하는 중심 성장 신호전달 인자로 인정되고 있다. mTOR는 mRNA translation을 개시하는 두 인자, 4E-binding protein (4EBP1)과 phosphorylated 70-kDa ribosomal S6 kinase (p70S6K)를 활성화시킴으로써 근 단백질 합성을 유도하여 근육량 증가에 기여한다(The Korea Journal of Sports Science, 20(3):1551-1561, 2011; The International Journal of Biochemistry and Cell Biology, 43(9):1267-1276, 2011). 반면에 전사 인자(transcription factor)인 forhead box (FoxO)가 세포질에서 핵 내로 이동하면 단백질 분해에 관여하는 E3 ubiquitin ligase인자 atrogin-1과 MuRF1의 발현을 증가시킨다(Disease Models and Mechanisms, 6:25-39, 2013). 이들의 발현량이 증가하면 근육 내의 단백질 분해가 촉진되어 근육량이 줄어들게 된다. 따라서 mTOR의 활성 촉진과 atrogin-1과 MuRF1 발현 억제는 근육 단백질의 양을 증가시켜 근육량을 증가시킨다. Factors involved in muscle protein synthesis induce protein synthesis by phosphorylating downstream proteins starting from stimulation of phosphatidylinositol-3 kinase (PI3K)/Akt pathway in muscle cells. The activity of the mammalian target of rapamycin (mTOR) by PI3K/Akt signaling is recognized as a central growth signaling factor that integrates various growth signals in cells. mTOR contributes to muscle mass increase by inducing muscle protein synthesis by activating two factors that initiate mRNA translation, 4E-binding protein (4EBP1) and phosphorylated 70-kDa ribosomal S6 kinase (p70S6K) (The Korea Journal of Sports Science, 20(3):1551-1561, 2011; The International Journal of Biochemistry and Cell Biology, 43(9):1267-1276, 2011). On the other hand, when the forward box (FoxO), a transcription factor, moves from the cytoplasm to the nucleus, it increases the expression of atrogin-1 and MuRF1, the E3 ubiquitin ligase factors involved in protein degradation (Disease Models and Mechanisms, 6:25- 39, 2013). When their expression level increases, protein breakdown in the muscle is accelerated, resulting in a decrease in muscle mass. Therefore, promotion of mTOR activity and suppression of atrogin-1 and MuRF1 expression increases the amount of muscle protein and increases muscle mass.
근육세포의 분화와 근육 형성은 다양한 근육 조절 인자(muscle regulatory factors)에 의해 조절된다. 그 중, MyoD의 활성에 의한 myogenin 발현의 유도는 근원세포의 결합(fusion)에 가장 중요한 요소로, 근관세포(myotube)의 형성에 관여한다. 이와 같은 과정을 통해 형성된 근섬유는 다발을 이루어 최종적으로 근육을 형성하게 된다(Cellular and Molecular Life Sciences, 70:4117-4130, 2013).Differentiation of muscle cells and muscle formation are regulated by various muscle regulatory factors. Among them, the induction of myogenin expression by the activity of MyoD is the most important factor in the fusion of myoblasts, and is involved in the formation of myotubes. The muscle fibers formed through this process are bundled to finally form a muscle (Cellular and Molecular Life Sciences, 70:4117-4130, 2013).
로즈힙(Rose hip)은 장미과(Rosaceae)에 속는 식물로서 학명은 로사 카니나(Rosa canina L.)로 알려져 있다. 지금까지 로즈힙과 관련하여 항노화 효과(Clinical Interventions in Aging, 19(10): 1849-1856, 2015), 항산화 효과(Antioxidants, 8(4): 92, 2019), 항암 효과(Journal of Pharmaceutical Analysis 8, 8(6): 394-399, 2018), 항당뇨 효과(Journal of Evidence-Based Complementary & Alternative Medicine, 21(4): 25-30, 2016), 항비만 효과(Preventive Nutrition and Food Science, 18(2): 85-91, 2013) 등의 활성이 보고된 바 있다. Rose hip is a plant belonging to the family Rosaceae, and the scientific name is known as Rosa canina L. So far, in relation to rosehip, anti-aging effects (Clinical Interventions in Aging, 19(10): 1849-1856, 2015), antioxidant effects (Antioxidants, 8(4): 92, 2019), anti-cancer effects (Journal of Pharmaceutical Analysis 8 , 8(6): 394-399, 2018), antidiabetic effect (Journal of Evidence-Based Complementary & Alternative Medicine, 21(4): 25-30, 2016), anti-obesity effect (Preventive Nutrition and Food Science, 18) (2): 85-91, 2013) has been reported to have the activity.
그러나, 본 발명의 이전에는 로즈힙의 근육 질환 예방 및 치료, 또는 근 기능 개선에 관해서는 알려진 바 없었다.However, prior to the present invention, it was not known about the prevention and treatment of muscle disease or improvement of muscle function of rosehip.
[선행기술문헌][Prior art literature]
[특허문헌][Patent Literature]
한국공개특허 제2006-7008605호Korean Patent Publication No. 2006-7008605
한국공개특허 제2016-0008200호Korea Patent Publication No. 2016-0008200
이에 본 발명자들은 우수한 근 기능 조절 활성을 가지며 안전하게 적용될 수 있는 천연물질을 탐색한 결과, 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙(rose hip)이 근육 질환 개선, 치료 또는 예방용, 또는 근 기능 개선 활성이 있음을 확인하여 본 발명을 완성하였다.Accordingly, the present inventors have searched for natural substances that have excellent muscle function control activity and can be safely applied. As a result, rose hips such as pulverized rosehip or rosehip extract are used for improving, treating or preventing muscle disease, or improving muscle function. By confirming that this exists, the present invention was completed.
따라서, 본 발명의 목적은 로즈힙을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물을 제공하는 것이다Accordingly, an object of the present invention is to provide a food composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient.
본 발명의 또 다른 목적은 로즈힙을 유효성분으로 함유하는 근육 질환 치료 또는 예방용 약학 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for treating or preventing muscle disease containing rosehip as an active ingredient.
본 발명의 또 다른 목적은 로즈힙을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 화장료 조성물을 제공하는 것이다.Another object of the present invention is to provide a cosmetic composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient.
본 발명의 또 다른 목적은 로즈힙을 유효성분으로 함유하는 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 사료 첨가제를 제공하는 것이다.Another object of the present invention is to provide a feed additive for improving or preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient containing rosehip as an active ingredient.
상기 목적을 달성하기 위해, 본 발명은 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙(rose hip)을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a food composition for improving or preventing muscle disease, or for improving muscle function, containing rose hip, such as pulverized rosehip or rosehip extract, as an active ingredient.
또한 본 발명은 로즈힙을 유효성분으로 함유하는 근육 질환 치료 또는 예방용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for treating or preventing muscle disease containing rosehip as an active ingredient.
또한 본 발명은 로즈힙을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 화장료 조성물을 제공한다.In addition, the present invention provides a cosmetic composition for improving or preventing muscle disease or improving muscle function containing rosehip as an active ingredient.
또한 본 발명은 로즈힙을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 사료 첨가제를 제공한다.In addition, the present invention provides a feed additive for improving or preventing muscle disease or improving muscle function containing rosehip as an active ingredient.
또한 본 발명은 근육 질환 예방 또는 치료용 의약 제조를 위한 로즈힙을 유효성분으로 포함하는 약학 조성물의 용도를 제공한다.In addition, the present invention provides the use of a pharmaceutical composition comprising rosehip as an active ingredient for the manufacture of a medicament for the prevention or treatment of muscle disease.
또한 본 발명은 근육 질환 환자에게 로즈힙을 유효성분으로 포함하는 약학 조성물을 투여하는 것을 포함하는 근육 질환의 치료방법을 제공한다.The present invention also provides a method for treating muscle disease, comprising administering to a patient with a muscle disease a pharmaceutical composition comprising rosehip as an active ingredient.
본 발명에 따른 로즈힙은 근 단백질 합성에 관여하는 mTOR의 활성을 증가시키고, 근 단백질 분해에 관여하는 MuRF1과 atrogin-1의 mRNA 발현을 억제시키는데 우수한 효과가 있다. 또한, 근육량을 증가시키고, 근기능을 향상시켜 각종 질병에 의하여 유발되는 근육의 기능 저하, 근육의 손실 등을 예방, 치료 또는 개선 할 수 있다. 본 발명은 천연물이므로 부작용 없이 안전하게 사용될 수 있어 의약품, 식품, 화장품 또는 사료첨가제로 사용될 수 있다.Rosehip according to the present invention has an excellent effect in increasing the activity of mTOR involved in muscle protein synthesis and inhibiting the mRNA expression of MuRF1 and atrogin-1 involved in muscle protein degradation. In addition, by increasing muscle mass and improving muscle function, it is possible to prevent, treat, or improve muscle function deterioration and muscle loss caused by various diseases. Since the present invention is a natural product, it can be safely used without side effects and can be used as a pharmaceutical, food, cosmetic or feed additive.
도 1은 L6 근육세포에서, 로즈힙 열수 추출물 및 에탄올 추출물 처리에 따른 mTOR의 활성 증가 효과를 확인한 결과이다.1 is a result confirming the effect of increasing the activity of mTOR in L6 muscle cells according to the treatment with hot water extract of rosehip and ethanol extract.
도 2는 L6 근육세포에서, 로즈힙 열수 추출물 처리에 따른 p-mTOR, p-p70S6K 및 p-4EBP-1의 단백질 발현량 증가 효과를 확인한 결과이다.2 is a result confirming the effect of increasing the protein expression level of p-mTOR, p-p70S6K, and p-4EBP-1 in L6 muscle cells according to the treatment with hot water extract of rosehip.
도 3은 L6 근육세포에서, 로즈힙 열수 추출물 처리에 따른 atrogin-1 및 MuRF1의 mRNA 발현량 감소 효과를 확인한 결과이다.3 is a result confirming the effect of reducing the mRNA expression levels of atrogin-1 and MuRF1 in L6 muscle cells according to the treatment with hot water extract of rosehip.
도 4는 근위축 유도 쥐에서, 로즈힙 처리에 따른 근육 부피 증가 효과를 확인한 결과이다.4 is a result confirming the effect of increasing muscle volume according to the treatment of rosehip in muscle atrophy-induced rats.
도 5는 근위축 유도 쥐에서, 로즈힙 열수 추출물 처리에 따른 p-mTOR, p-p70S6K 및 p-4EBP-1의 단백질 발현량 증가 효과를 확인한 결과이다.5 is a result confirming the effect of increasing the protein expression level of p-mTOR, p-p70S6K and p-4EBP-1 according to the treatment with hot water extract of rosehip in muscle atrophy-induced mice.
이하, 본 발명의 구성을 구체적으로 설명한다.Hereinafter, the configuration of the present invention will be described in detail.
본 발명은 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙을 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물; 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙을 함유하는 근육 질환 치료 또는 예방용 약학 조성물; 또는 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙을 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 화장료 조성물; 또는 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙을 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 사료 첨가제; 또는 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙을 인간, 또는 인간을 제외한 포유동물에 적용하는 것을 함유하는 근육 질환 치료방법을 제공한다.The present invention relates to a food composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip, such as a pulverized rosehip or a rosehip extract; a pharmaceutical composition for treating or preventing a muscle disease containing rosehip, such as a pulverized rosehip or a rosehip extract; Or a cosmetic composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip, such as a pulverized rosehip or a rosehip extract; or a feed additive for improving or preventing muscle disease or improving muscle function containing rosehip, such as rosehip pulverized or rosehip extract; Or it provides a method for treating a muscle disease comprising applying a rosehip, such as a pulverized rosehip or a rosehip extract, to a human or non-human mammal.
본 명세서에서 '로즈힙(rose hip)'은 장미과(Rosaceae) 식물, 로사 카니나(Rosa canina L.)의 열매이며, 로즈힙 분쇄물 또는 로즈힙 추출물을 사용할 수 있다. In the present specification, 'rose hip' is a plant of the Rosaceae family, a fruit of Rosa canina L., and a crushed rosehip or a rosehip extract may be used.
본 명세서에서 '로즈힙 분쇄물'은 건조된 로즈힙 열매를 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 인간을 포함한 포유동물이 용이하게 섭취할 수 있으면서 섭취 후 장에서 유효성분이 건조 분쇄물로부터 쉽게 유리되어 결과적으로 포유동물 체내로 용이하게 흡수될 수 있는 형태로 준비되어 사용될 수 있다. 이를 위한 건조 분쇄물 입자의 모양이나 크기는 제한되지 않으나, 입자의 표면이 극대화 될수록 상기에 기재한 유효성분의 유리가 용이하므로 가급적 고운 가루형태로 제조함이 바람직하다. In the present specification, 'pulverized rosehip' refers to a method that can be easily performed by a person of ordinary skill in the art to which the present invention pertains to dried rosehip fruit, and ingested while being easily ingested by mammals, including humans. Afterwards, the active ingredient is easily released from the dry pulverized material in the intestine, and as a result, it can be prepared and used in a form that can be easily absorbed into the body of a mammal. For this purpose, the shape or size of the dry pulverized particles is not limited, but as the surface of the particles is maximized, it is easier to release the active ingredient described above, so it is preferable to prepare it in the form of a fine powder as much as possible.
본 명세서에서 '로즈힙 추출물'은 건조된 로즈힙 열매를 적합한 용매를 사용하여 추출하여 수득한 것으로, 추출액, 추출액의 희석액 또는 농축액, 추출액을 건조하여 얻어지는 건조물, 또는 이들의 조정제물 또는 정제물의 형태를 모두 포함한다. 로즈힙 추출물의 제조방법은 로즈힙으로부터 물, 탄소수 1 내지 6의 유기용매 및 초고압, 아임계 또는 초임계 유체로 이루어진 군에서 선택된 하나 이상의 용매로 추출하여 수득할 수 있으나, 이에 한정되지 않는다.In the present specification, 'rosehip extract' is obtained by extracting dried rosehip fruit using a suitable solvent, and is obtained in the form of an extract, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a prepared or purified product thereof. include The method for preparing the rosehip extract may be obtained by extracting from the rosehip with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and ultra-high pressure, subcritical or supercritical fluid, but is not limited thereto.
본 발명의 로즈힙 추출물은 가열 추출, 냉침 추출 초음파 추출법, 여과법 및 환류추출법 등 당 업계의 통상적인 추출방법을 사용하여 제조될 수 있으며, 로즈힙은 상업적으로 판매되는 것을 구입하여 사용하거나, 자연에서 채취 또는 재배된 것을 사용할 수 있다.The rosehip extract of the present invention can be prepared using conventional extraction methods in the art, such as heat extraction, cold-chill extraction, ultrasonic extraction, filtration, and reflux extraction. Cultivated ones can be used.
본 발명에 따른 로즈힙 추출물은 천연물로부터 추출물을 제조하는 당업계에 공지된 통상적인 방법에 따라, 즉 통상적인 온도, 압력의 조건 하에서 통상적인 용매를 사용하여 분리할 수 있다.The rosehip extract according to the present invention can be separated using a conventional solvent according to a conventional method known in the art for preparing an extract from a natural product, that is, under conditions of conventional temperature and pressure.
본 명세서에서 사용된 용어 '분획물'은 다양한 구성성분을 포함하는 혼합물로부터 특정 성분 또는 특정 그룹을 분리하는 분획방법에 의하여 얻어진 결과물을 의미한다. 본 발명에서는 상기와 같이 제조된 로즈힙 추출물로부터 특정 성분 또는 특정 그룹을 분리하는 분획방법에 의하여 얻어진 결과물을 의미한다.As used herein, the term 'fraction' refers to a result obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various components. In the present invention, it refers to a result obtained by a fractionation method for separating a specific component or a specific group from the rosehip extract prepared as described above.
본 발명에 따른 로즈힙 분획물을 얻기 위하여 당업계에서 공지된 통상적인 분획 용매, 예를 들어, 물, 에탄올, 메탄올과 같은 탄소수 1 내지 4의 무수 또는 함수 저급 알코올 등의 극성 용매 및 헥산, 부탄올, 에틸아세테이트, 클로로포름, 디클로로메탄 등의 비극성 용매, 또는 이들의 혼합 용매가 사용될 수 있으나, 이에 제한되지 않는다.In order to obtain the rosehip fraction according to the present invention, a conventional fractionation solvent known in the art, for example, a polar solvent such as anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms such as water, ethanol, and methanol, and hexane, butanol, ethyl A non-polar solvent such as acetate, chloroform, dichloromethane, or a mixed solvent thereof may be used, but is not limited thereto.
본 발명의 로즈힙 분획물은 정제과정을 추가적으로 적용하여 얻은 것도 포함될 수 있다. 예를 들어, 본 발명에 따른 로즈힙 추출물을 일정한 분자량 컷-오프(cut-off) 값을 갖는 한외 여과막(ultrafiltration membrane)을 통과시켜 얻은 분획물, 다양한 크로마토그래피(크기, 전하, 소수성 또는 친화성에 따른 분리를 위해 제작된 것)에 의한 분리 등으로 추가적으로 실시된 다양한 정제 방법을 통해 얻어진 분획물도 본 발명의 로즈힙 추출물에 포함된다.The rosehip fraction of the present invention may include those obtained by additionally applying a purification process. For example, a fraction obtained by passing a rosehip extract according to the present invention through an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (separation according to size, charge, hydrophobicity or affinity) Fractions obtained through various purification methods additionally carried out such as separation by ) are also included in the rosehip extract of the present invention.
본 명세서에서, '근'은 심줄, 근육, 건을 포괄적으로 지칭하고, '근 기능'은 근육의 수축에 의해 힘을 발휘하는 능력을 의미하며, 근육이 저항을 이겨내기 위하여 최대한으로 수축력을 발휘할 수 있는 능력인 근력, 근육이 주어진 중량에 얼마나 오랫동안 또는 얼마나 여러 번 수축과 이완을 반복할 수 있는지를 나타내는 능력인 근지구력, 단시간 내에 강한 힘을 발휘하는 능력인 순발력을 포함한다. 본 명세서의 용어 '근 기능 개선'은 근육량을 증가시켜 근 기능을 더 좋게 향상시키는 것을 말한다.In this specification, 'muscle' refers to tendons, muscles, and tendons comprehensively, and 'muscular function' means the ability to exert force by contraction of the muscle, It includes muscle strength, which is the ability to perform, muscular endurance, which is the ability to repeat contractions and relaxations for how long or how many times a muscle can repeat contractions and relaxations with a given weight, and instantaneous power, which is the ability to exert strong force in a short period of time. As used herein, the term 'improving muscle function' refers to improving muscle function by increasing muscle mass.
본 발명은 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙을 유효성분으로 함유하는 근육 질환 개선 및 예방용, 또는 근 기능 개선용 식품 조성물; 근육 질환 예방 및 치료용 약학적 조성물; 근육 질환 개선 및 예방용, 또는 근 기능 개선용 화장료 조성물; 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 사료 첨가제를 제공한다.The present invention relates to a food composition for improving and preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient, such as a pulverized rosehip or a rosehip extract; pharmaceutical compositions for preventing and treating muscle diseases; A cosmetic composition for improving and preventing muscle disease, or improving muscle function; It provides a feed additive for improving or preventing muscle disease, or for improving muscle function.
한 구체 예에서, 로즈힙 추출물은 열수 추출물, 에탄올 추출물, 에틸아세테이트 추출물, 헥산 추출물, 초고압 추출물, 아임계 추출물, 초임계 추출물일 수 있으나, 이로 한정되지 않는다.In one embodiment, the rosehip extract may be a hot water extract, an ethanol extract, an ethyl acetate extract, a hexane extract, an ultra-high pressure extract, a subcritical extract, or a supercritical extract, but is not limited thereto.
한 구체 예에서, 로즈힙 추출물은 물, 탄소수 1 내지 6의 유기용매, 아임계 및 초임계 유체로 이루어진 군으로부터 선택되는 하나 이상의 용매로 로즈힙의 열매를 추출하여 수득할 수 있다. 로즈힙을 100 MPa 이상의 초고압 조건 하에서 추출하여 수득할 수도 있다. 필요한 경우에는 당업계에 공지된 방법에 따라 여과 및 농축 단계를 추가적으로 포함하여 제조할 수 있다.In one embodiment, the rosehip extract can be obtained by extracting the fruit of rosehip with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and subcritical and supercritical fluids. It can also be obtained by extracting rosehip under ultra-high pressure conditions of 100 MPa or more. If necessary, it may be prepared by additionally including filtration and concentration steps according to methods known in the art.
한 구체 예에서, 탄소수 1 내지 6의 유기용매는 탄소수 1 내지 6의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌 클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane) 및 석유에테르(petroleum ether)로 이루어진 군중에서 선택되는 하나 이상일 수 있다.In one embodiment, the organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, It may be at least one selected from the group consisting of methylene chloride, hexane, cyclohexane, and petroleum ether.
본 발명의 구체적인 실시예에 있어서, 본 발명자들은 건조된 로즈힙 열매의 분쇄물을 에탄올, 에틸아세테이트 및 헥산을 용매로 하여 각각 상온에서 반복 추출하거나, 열수 추출, 초고압 추출, 아임계 유체 추출, 초임계 유체 추출을 이용하여 로즈힙 추출물을 제조하였다.In a specific embodiment of the present invention, the present inventors repeatedly extract the dried rosehip fruit pulverized product using ethanol, ethyl acetate and hexane as solvents at room temperature, respectively, hot water extraction, ultra-high pressure extraction, subcritical fluid extraction, supercritical A rosehip extract was prepared using fluid extraction.
로즈힙 추출물을 L6 근육세포에 처리한 결과, 근 단백질 합성에 관여하는 p-mTOR의 활성과 근 단백질 합성 생체지표인 p-mTOR, p-p70S6K, p-4EBP-1의 단백질 발현량의 발현이 유의적으로 증가되는 것을 확인하였다(도 1, 도 2, 표 1). 또한, 로즈힙 추출물을 L6 근육세포에 처리한 결과, 근 단백질 분해에 관여하는 MuRF-1과 atrogin-1의 mRNA 발현을 유의적으로 억제시키는 것을 확인하였다(도 3, 표 2).As a result of treatment with rosehip extract on L6 muscle cells, the activity of p-mTOR involved in muscle protein synthesis and the expression of protein expression levels of p-mTOR, p-p70S6K, and p-4EBP-1, which are biomarkers for muscle protein synthesis, were significant. It was confirmed that there is a negative increase (Fig. 1, Fig. 2, Table 1). In addition, as a result of treating the rosehip extract in L6 muscle cells, it was confirmed that the mRNA expression of MuRF-1 and atrogin-1, which are involved in muscle protein degradation, was significantly suppressed (FIG. 3, Table 2).
피부 스테이플러(Skin stapler)를 이용한 부동화(immobilization) 마우스 동물 모델을 이용하여 로즈힙 분쇄믈, 로즈힙 열수 추출물, 로즈힙 에탄올 추출물, 로즈힙 초임계 추출물을 처리한 결과, 근위축군에 비하여 근력, 근육부피 및 근육무게가 각각 유의적으로 증가하였다(도4, 표 3, 표 4). 또한, 로즈힙 열수 추출물 처리에 의해 근 단백질 합성 생체지표인 p-mTOR, p-p70S6K, p-4EBP-1의 단백질 발현량이 증가하였다(도 5).As a result of processing rosehip crushed water, rosehip hot water extract, rosehip ethanol extract, and rosehip supercritical extract using an immobilization mouse animal model using a skin stapler, muscle strength, muscle volume and muscle weight compared to the muscle atrophy group each significantly increased (Fig. 4, Table 3, Table 4). In addition, protein expression levels of p-mTOR, p-p70S6K, and p-4EBP-1, which are biomarkers for muscle protein synthesis, were increased by treatment with hot water extract of rosehip (FIG. 5).
본 발명의 근육 질환 예방 및 개선, 또는 근 기능 개선용이 식품 조성물인 경우, 근육 소모 또는 퇴화로 인한 근육 질환의 예방 또는 개선에 사용될 수 있다. 근육 소모 및 퇴화는 유전적 요인, 후천적 요인, 노화 등을 원인으로 발생하며, 근육 소모는 근육량의 점진적 손실, 근육, 특히 골격근 또는 수의근 및 심장근육의 약화 및 퇴행을 특징으로 한다. 이와 관련된 질환의 예로는 근감소증(sarcopenia), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 긴장감퇴증(atony), 근육 퇴화, 근무력증, 악액질(cachexia) 등을 들 수 있다. 본 발명의 조성물은 근육량 증대 효과가 있으며, 근육은 그 종류를 제한하지 않는다.In the case of the food composition for preventing and improving muscle disease or improving muscle function of the present invention, it can be used for preventing or improving muscle disease due to muscle wasting or degeneration. Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by a gradual loss of muscle mass, weakness and degeneration of muscles, particularly skeletal or voluntary muscles and cardiac muscles. Examples of related diseases include sarcopenia, muscular atrophy, muscular dystrophy, atony, muscle degeneration, myasthenia gravis, cachexia, and the like. The composition of the present invention has an effect of increasing muscle mass, and the type of muscle is not limited.
본 발명의 식품 조성물은 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강기능식품(health function food), 식품 첨가제(food additives) 및 사료 등의 모든 형태를 포함하며, 인간 또는 가축을 비롯한 동물을 취식대상으로 한다. 상기 유형의 식품 조성물은 당 업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다.The food composition of the present invention includes all forms such as functional food, nutritional supplement, health function food, food additives and feed, including humans or livestock Animals are intended for food. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
상기 유형의 식품 조성물은 당업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다. 일반 식품으로는 이에 한정되지 않지만 음료(알콜성 음료 포함), 과실 및 그의 가공식품(예: 과일통조림, 병조림, 잼, 마아말레이드 등), 어류, 육류 및 그 가공식품(예: 햄, 소시지 콘비이프 등), 빵류 및 면류(예: 우동, 메밀국수, 라면, 스파게이트, 마카로니 등), 과즙, 각종 드링크, 쿠키, 엿, 유제품(예: 버터, 치이즈 등), 식용식물 유지, 마아가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등) 등에 로즈힙 분쇄물 또는 로즈힙 추출물을 첨가하여 제조할 수 있다. 또한, 영양보조제로는 이에 한정되지 않지만 캡슐, 타블렛, 환 등에 로즈힙 분쇄물 또는 로즈힙 추출물물을 첨가하여 제조할 수 있다. 또한, 건강기능식품으로는 이에 한정되지 않지만 예를 들면, 로즈힙 분쇄물 또는 로즈힙 추출물 자체를 차, 쥬스 및 드링크의 형태로 제조하여 음용(건강음료)할 수 있도록 액상화, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다. 또한, 로즈힙, 로즈힙 분쇄물 또는 로즈힙 추출물을 식품 첨가제의 형태로 사용하기 위해서는 분말 또는 농축액 형태로 제조하여 사용할 수 있다. 또한, 로즈힙 분쇄물 또는 로즈힙 추출물과 근육 질환 예방 및 개선, 또는 근 기능 개선 효과가 있다고 알려진 공지의 활성 성분과 함께 혼합하여 조성물의 형태로 제조할 수 있다.Food compositions of this type can be prepared in various forms according to conventional methods known in the art. Common foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat and their processed foods (eg, ham, sausages) Corned beef, etc.), breads and noodles (eg udon noodles, soba noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (eg butter, cheese, etc.), edible vegetable oils and fats, margarine , vegetable protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.) can be prepared by adding crushed rosehip or rosehip extract. In addition, the nutritional supplement is not limited thereto, but may be prepared by adding crushed rosehip or rosehip extract to capsules, tablets, pills, and the like. In addition, although not limited to this as a health functional food, for example, the pulverized rosehip or rosehip extract itself is prepared in the form of tea, juice, and drink, and liquefied, granulated, encapsulated and powdered so that it can be consumed (health drink). can be ingested. In addition, in order to use rosehip, pulverized rosehip or rosehip extract in the form of a food additive, it may be prepared and used in the form of a powder or concentrate. In addition, it can be prepared in the form of a composition by mixing the pulverized rosehip or rosehip extract with an active ingredient known to be effective in preventing and improving muscle disease, or improving muscle function.
본 발명의 근육 질환 개선 및 예방, 또는 근 기능 개선용 조성물이 건강음료 조성물로 이용되는 경우, 상기 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드(monosaccharide); 말토스, 슈크로스와 같은 디사카라이드(disaccharide); 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드(polysaccharide); 자일리톨, 소르비톨, 에리트리톨 등의 당알콜(sugar alcohol)일 수 있다. 감미제는 타우마틴(thaumatin), 스테비아(stevia) 추출물과 같은 천연 감미제; 사카린(saccharin), 아스파탐(aspartame)과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 mL 당 일반적으로 약 0.01~5.0g, 바람직하게는 약 0.1~1.0g이다.When the composition for improving and preventing muscle disease or improving muscle function of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional drink. . The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as thaumatin, stevia extract; A synthetic sweetener such as saccharin or aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01 to 5.0 g, preferably about 0.1 to 1.0 g per 100 mL of the composition of the present invention.
로즈힙 분쇄물 또는 로즈힙 추출물은 근육 질환 예방 및 개선용, 또는 근 기능 개선용 식품 조성물의 유효성분으로 함유될 수 있는데, 그 양은 근육 질환 예방 및 개선용, 또는 근 기능 개선용 작용을 달성하기에 유효한 양으로 특별히 한정되는 것은 아니나, 전체 조성물 총 중량에 대하여 0.01 내지 100 중량%인 것이 바람직하다. 본 발명의 식품 조성물은 로즈힙 분쇄물 또는 로즈힙 추출물과 함께 근육 질환 예방 및 개선, 또는 근 기능 개선용 조성물에 효과가 있는 것으로 알려진 다른 활성 성분과 함께 혼합하여 제조될 수 있다.The pulverized rosehip or rosehip extract may be contained as an active ingredient in a food composition for preventing and improving muscle disease or improving muscle function, the amount of which is effective to achieve the action for preventing and improving muscle disease or improving muscle function. The amount is not particularly limited, but is preferably 0.01 to 100% by weight based on the total weight of the total composition. The food composition of the present invention may be prepared by mixing with other active ingredients known to be effective in preventing and improving muscle disease or improving muscle function together with pulverized rosehip or rosehip extract.
본 발명의 근육 질환 예방 및 치료용 조성물은 또한 약학 조성물일 수 있다. 본 발명의 근육 질환 예방 및 치료용 조성물이 약학 조성물인 경우, 근육 소모 또는 퇴화로 인한 근육 질환의 예방 또는 치료에 사용될 수 있다. 근육 소모 및 퇴화는 유전적 요인, 후천적 요인, 노화 등을 원인으로 발생하며, 근육 소모는 근육량의 점진적 손실, 근육, 특히 골격근 또는 수의근 및 심장근육의 약화 및 퇴행을 특징으로 한다. 이와 관련된 질환의 예로는 근감소증(sarcopenia), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 긴장감퇴증(atony), 근육 퇴화, 근무력증, 악액질(cachexia) 등을 들 수 있다. 본 발명의 조성물은 근육량 증대 효과가 있으며, 근육은 그 종류를 제한하지 않는다.The composition for preventing and treating muscle disease of the present invention may also be a pharmaceutical composition. When the composition for preventing and treating muscle disease of the present invention is a pharmaceutical composition, it may be used for the prevention or treatment of muscle disease caused by muscle wasting or degeneration. Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by a gradual loss of muscle mass, weakness and degeneration of muscles, particularly skeletal or voluntary muscles and cardiac muscles. Examples of related diseases include sarcopenia, muscular atrophy, muscular dystrophy, atony, muscle degeneration, myasthenia gravis, cachexia, and the like. The composition of the present invention has an effect of increasing muscle mass, and the type of muscle is not limited.
본 발명의 '근육 질환 예방 및 치료용 약학 조성물은 약학적으로 허용 가능한 담체를 포함할 수 있다.The 'pharmaceutical composition for preventing and treating muscle disease of the present invention may include a pharmaceutically acceptable carrier.
약학적으로 허용되는 담체로는 예컨대, 경구 투여용 담체 또는 비경구 투여용 담체를 추가로 포함할 수 있다. 경구 투여용 담체는 락토스, 전분, 셀룰로스 유도체, 마그네슘 스테아레이트, 스테아르산 등을 포함할 수 있다. 또한 비경구 투여용 담체는 물, 적합한 오일, 식염수, 수성 글루코스 및 글리콜 등을 포함할 수 있다. 또한, 안정화제 및 보존제를 추가로 포함할 수 있다. 적합한 안정화제로는 아황산수소나트륨, 아황산나트륨 또는 아스코르브산과 같은 항산화제가 있다. 적합한 보존제로는 벤즈알코늄 클로라이드, 메틸- 또는 프로필-파라벤 및 클로로부탄올이 있다. 그 밖의 약학적으로 허용되는 담체로는 다음의 문헌에 기재되어 있는 것을 참고로 할 수 있다(Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).The pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Carriers for parenteral administration may also include water, suitable oils, saline, aqueous glucose and glycols, and the like. In addition, it may further include a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives are benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. As other pharmaceutically acceptable carriers, reference may be made to those described in the following literature (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).
본 발명의 약학 조성물은 인간을 비롯한 포유동물에 어떠한 방법으로도 투여할 수 있다. 예를 들어, 경구 또는 비경구로 투여할 수 있으며, 비경구적인 투여방법으로는 이에 제한되는 것은 아니나, 정맥내, 근육내, 동맥내, 골수내, 경막내, 심장내, 경피, 피하, 복강내, 비강내, 장관, 국소, 설하 또는 직장내 투여일 수 있다.The pharmaceutical composition of the present invention may be administered to mammals including humans by any method. For example, it may be administered orally or parenterally, and parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal. , intranasal, enteral, topical, sublingual or rectal administration.
본 발명의 약학 조성물은 상술한 바와 같은 투여 경로에 따라 경구 투여용 또는 비경구 투여용 제제로 제형화 할 수 있다. 제형화할 경우에는 하나 이상의 완충제(예를 들어, 식염수 또는 PBS), 항산화제, 정균제, 킬레이트화제(예를 들어, EDTA 또는 글루타치온), 충진제, 증량제, 결합제, 아쥬반트(예를 들어, 알루미늄 하이드록사이드), 현탁제, 농후제 습윤제, 붕해제 또는 계면활성제, 희석제 또는 부형제를 사용하여 조제될 수 있다.The pharmaceutical composition of the present invention may be formulated as a formulation for oral administration or parenteral administration according to the administration route as described above. When formulated, one or more buffers (eg, saline or PBS), antioxidants, bacteriostatic agents, chelating agents (eg, EDTA or glutathione), fillers, bulking agents, binders, adjuvants (eg, aluminum hydroxide) side), suspending agents, thickening agents, wetting agents, disintegrating agents or surfactants, diluents or excipients.
경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 액제, 겔제, 시럽제, 슬러리제, 현탁액 또는 캡슐제 등이 포함되며, 이러한 고형제제는 본 발명의 약학 조성물에 적어도 하나 이상의 부형제 예를 들면, 전분(옥수수 전분, 밀 전분, 쌀 전분, 감자 전분 등 포함), 칼슘카보네이트(calcium carbonate), 수크로스(sucrose), 락토오스(lactose), 덱스트로오스, 솔비톨, 만니톨, 자일리톨, 에리스리톨 말티톨, 셀룰로즈, 메틸 셀룰로즈, 나트륨 카르복시메틸셀룰로오즈 및 하이드록시프로필메틸-셀룰로즈 또는 젤라틴 등을 섞어 조제될 수 있다. 예컨대, 활성성분을 고체 부형제와 배합한 다음 이를 분쇄하고 적합한 보조제를 첨가한 후 과립 혼합물로 가공함으로써 정제 또는 당의 정제를 수득할 수 있다.Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions or capsules, and such solid preparations include at least one excipient in the pharmaceutical composition of the present invention, for example , starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose , methyl cellulose, sodium carboxymethyl cellulose, and hydroxypropylmethyl-cellulose or gelatin may be mixed and prepared. For example, tablets or sugar tablets can be obtained by blending the active ingredient with a solid excipient, grinding it, adding a suitable adjuvant, and processing it into a granule mixture.
단순한 부형제 이외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제 또는 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물 또는 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제 또는 보존제 등이 포함될 수 있다.In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, or syrups. In addition to water or liquid paraffin, which are commonly used simple diluents, various excipients, for example, wetting agents, sweeteners, flavoring agents, or preservatives may be included. .
또한, 경우에 따라 가교결합 폴리비닐피롤리돈, 한천, 알긴산 또는 나트륨 알기네이트 등을 붕해제로 첨가할 수 있으며, 항응집제, 윤활제, 습윤제, 향료, 유화제 및 방부제 등을 추가로 포함할 수 있다.In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, and an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifier, and a preservative may be additionally included. .
비경구적으로 투여하는 경우 본 발명의 약학 조성물은 적합한 비경구용 담체와 함께 주사제, 경피 투여제 및 비강 흡입제의 형태로 당 업계에 공지된 방법에 따라 제형화될 수 있다. 상기 주사제의 경우에는 반드시 멸균되어야 하며 박테리아 및 진균과 같은 미생물의 오염으로부터 보호되어야 한다. 주사제의 경우 적합한 담체의 예로는 이에 한정되지는 않으나, 물, 에탄올, 폴리올(예를 들어, 글리세롤, 프로필렌 글리콜 및 액체 폴리에틸렌 글리콜 등), 이들의 혼합물 및/또는 식물유를 함유하는 용매 또는 분산매질일 수 있다. 보다 바람직하게는, 적합한 담체로는 행크스 용액, 링거 용액, 트리에탄올 아민이 함유된 PBS (phosphate buffered saline) 또는 주사용 멸균수, 10% 에탄올, 40% 프로필렌 글리콜 및 5% 덱스트로즈와 같은 등장 용액 등을 사용할 수 있다. 상기 주사제를 미생물 오염으로부터 보호하기 위해서는 파라벤, 클로로부탄올, 페놀, 소르빈산, 티메로살 등과 같은 다양한 항균제 및 항진균제를 추가로 포함할 수 있다. 또한, 상기 주사제는 대부분의 경우 당 또는 나트륨 클로라이드와 같은 등장화제를 추가로 포함할 수 있다.When administered parenterally, the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations and nasal inhalants together with suitable parenteral carriers. In the case of the injection, it must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. For injection, examples of suitable carriers include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and/or vegetable oils. can More preferably, suitable carriers include Hanks' solution, Ringer's solution, PBS (phosphate buffered saline) with triethanolamine or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. etc. can be used. In order to protect the injection from microbial contamination, it may further include various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, in most cases, the injection may further contain an isotonic agent such as sugar or sodium chloride.
경피 투여제의 경우 연고제, 크림제, 로션제, 겔제, 외용액제, 파스타제, 리니멘트제, 에어롤제 등의 형태가 포함된다. 상기에서 '경피 투여'는 약학 조성물을 국소적으로 피부에 투여하여 약학 조성물에 함유된 유효한 양의 활성성분이 피부 내로 전달되는 것을 의미한다. In the case of transdermal administration, forms such as ointment, cream, lotion, gel, external solution, pasta, liniment, and air are included. In the above, 'transdermal administration' means that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin by topically administering the pharmaceutical composition to the skin.
흡입 투여제의 경우, 본 발명에 따라 사용되는 화합물은 적합한 추진제, 예를 들면, 디클로로플루오로메탄, 트리클로로플루오로메탄, 디클로로테트라플루오로에탄, 이산화탄소 또는 다른 적합한 기체를 사용하여, 가압 팩 또는 연무기로부터 에어로졸 스프레이 형태로 편리하게 전달 할 수 있다. 가압 에어로졸의 경우, 투약 단위는 계량된 양을 전달하는 밸브를 제공하여 결정할 수 있다. 예를 들면, 흡입기 또는 취입기에 사용되는 젤라틴 캡슐 및 카트리지는 화합물, 및 락토즈 또는 전분과 같은 적합한 분말 기제의 분말 혼합물을 함유하도록 제형화할 수 있다. 비경구 투여용 제형은 모든 제약 화학에 일반적으로 공지된 처방서인 문헌(Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour)에 기재되어 있다.In the case of administration by inhalation, the compounds for use according to the invention may be administered in pressurized packs or using a suitable propellant, for example, dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered in the form of an aerosol spray from the nebulizer. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a recipe commonly known to all pharmaceutical chemistry.
본 발명의 근육 질환 예방 및 치료용 약학 조성물은 로즈힙 추출물을 유효량으로 포함할 때 바람직한 근육 질환 예방 및 치료 효과를 제공할 수 있다. 본 명세서에서, '유효량'이라 함은 음성 대조군에 비해 그 이상의 반응을 나타내는 양을 말하며 바람직하게는 근 기능을 향상시키기에 충분한 양을 말한다. 본 발명의 약학 조성물에 로즈힙 추출물이 0.01 내지 99.99% 포함될 수 있으며, 잔량은 약학적으로 허용 가능한 담체가 차지할 수 있다. 본 발명의 약학 조성물에 포함되는 로즈힙 추출물의 유효량은 조성물이 제품화되는 형태 등에 따라 달라질 것이다.When the pharmaceutical composition for preventing and treating muscle disease of the present invention contains an effective amount of a rosehip extract, it is possible to provide a desirable preventive and therapeutic effect on muscle disease. As used herein, the term 'effective amount' refers to an amount that exhibits a higher response than that of the negative control group, and preferably refers to an amount sufficient to improve muscle function. The pharmaceutical composition of the present invention may contain 0.01 to 99.99% of the rosehip extract, and the remaining amount may be occupied by a pharmaceutically acceptable carrier. The effective amount of the rosehip extract contained in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.
본 발명의 약학 조성물의 총 유효량은 단일 투여량(single dose)으로 환자에게 투여될 수 있으며, 다중 투여량(multiple dose)으로 장기간 투여되는 분할 치료 방법(fractionated treatment protocol)에 의해 투여될 수 있다. 본 발명의 약학 조성물은 질환의 정도에 따라 유효성분의 함량을 달리할 수 있다. 비경구 투여시는 로즈힙 추출물을 기준으로 하루에 체중 1 kg당 바람직하게 0.01 내지 50 mg, 더 바람직하게는 0.1 내지 30 mg의 양으로 투여되도록, 그리고 경구 투여시는 로즈힙 추출물을 기준으로 하루에 체중 1 kg당 바람직하게 0.01 내지 100 mg, 더 바람직하게는 0.1 내지 50 mg의 양으로 투여되도록 1 내지 수회에 나누어 투여할 수 있다. 그러나 상기 로즈힙 추출물의 용량은 약학 조성물의 투여 경로 및 치료 횟수뿐만 아니라 환자의 연령, 체중, 건강 상태, 성별, 질환의 중증도, 식이 및 배설율 등 다양한 요인들을 고려하여 환자에 대한 유효 투여량이 결정되는 것이므로, 이러한 점을 고려할 때 당 분야의 통상적인 지식을 가진 자라면 상기 로즈힙 추출물을 근육 질환 예방 및 치료를 위한 특정한 용도에 따른 적절한 유효 투여량을 결정할 수 있을 것이다. 본 발명에 따른 약학 조성물은 본 발명의 효과를 보이는 한 그 제형, 투여 경로 및 투여 방법에 특별히 제한되지 아니한다.The total effective amount of the pharmaceutical composition of the present invention may be administered to a patient as a single dose, or may be administered by a fractionated treatment protocol in which multiple doses are administered for a long period of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease. When administered parenterally, it is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per 1 kg of body weight per day based on the rosehip extract, and when administered orally, the body weight per day based on the rosehip extract It may be administered in 1 to several divided doses so as to be administered in an amount of preferably 0.01 to 100 mg, more preferably 0.1 to 50 mg per 1 kg. However, the dosage of the rosehip extract is determined by considering various factors such as the age, weight, health status, sex, severity of the disease, diet and excretion rate of the patient, as well as the administration route and number of treatments of the pharmaceutical composition, and the effective dosage for the patient is determined. Therefore, in consideration of this point, those of ordinary skill in the art will be able to determine an appropriate effective dosage according to the specific use of the rosehip extract for the prevention and treatment of muscle diseases. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route and administration method as long as the effect of the present invention is exhibited.
본 발명의 근육 질환 예방 및 치료용 약학 조성물은 단독으로, 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 또는 생물학적 반응조절제를 사용하는 방법들과 병용하여 사용할 수 있다.The pharmaceutical composition for preventing and treating muscle disease of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, or biological response modifiers.
본 발명의 근육 질환 예방 및 치료용 약학 조성물은 또한 로즈힙 추출물을 유효성분으로 함유하는 외용제의 제형으로 제공할 수 있다.The pharmaceutical composition for preventing and treating muscle disease of the present invention may also be provided in the form of an external preparation containing a rosehip extract as an active ingredient.
본 발명의 근육 질환 예방 및 치료용 약학 조성물을 피부외용제로 사용하는 경우, 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 유화제, 비이온형 유화제, 충전제, 금속이온봉쇄제, 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 활성제, 친유성 활성제 또는 지질 소낭 등 피부 외용제에 통상적으로 사용되는 임의의 다른 성분과 같은 피부 과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 또한 상기 성분들은 피부 과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다.When the pharmaceutical composition for the prevention and treatment of muscle diseases of the present invention is used as an external preparation for skin, additionally, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, and a foaming agent (foaming agent) ), fragrance, surfactant, water, ionic emulsifier, nonionic emulsifier, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic active agent, lipophilic active agent Or it may contain adjuvants commonly used in the field of dermatology, such as any other ingredients commonly used in external preparations for skin, such as lipid vesicles. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.
본 발명의 근육 질환 예방 및 치료용 약학 조성물이 피부 외용제로 제공될 경우, 이에 제한되는 것은 아니나, 연고, 패취, 겔, 크림 또는 분무제 등의 제형일 수 있다.When the pharmaceutical composition for preventing and treating muscle disease of the present invention is provided as an external preparation for skin, it may be in the form of an ointment, patch, gel, cream or spray, but is not limited thereto.
본 발명의 근육 질환 예방 및 개선용, 또는 근 기능 개선용 조성물은 또한 화장료 조성물일 수 있다. 본 발명의 화장료 조성물은 로즈힙 추출물을 유효성분으로 함유하며 피부학적으로 허용 가능한 부형제와 함께 기초 화장품 조성물(화장수, 크림, 에센스, 클렌징 폼 및 클렌징 워터와 같은 세안제, 팩, 보디오일), 색조 화장품 조성물(화운데이션, 립스틱, 마스카라, 메이크업 베이스), 두발 제품 조성물(샴푸, 린스, 헤어컨디셔너, 헤어젤) 및 비누 등의 형태로 제조될 수 있다.The composition for preventing and improving muscle disease or for improving muscle function of the present invention may also be a cosmetic composition. The cosmetic composition of the present invention contains rosehip extract as an active ingredient, and a basic cosmetic composition (toilet water, cream, essence, face wash such as lotion, cream, essence, cleansing foam and cleansing water, pack, body oil), color cosmetic composition together with a dermatologically acceptable excipient (foundation, lipstick, mascara, makeup base), hair product compositions (shampoo, conditioner, hair conditioner, hair gel), and soap.
상기 부형제로는 이에 한정되지는 않으나 예를 들어, 피부연화제, 피부 침투 증강제, 착색제, 방향제, 유화제, 농화제 및 용매를 포함할 수 있다. 또한, 향료, 색소, 살균제, 산화방지제, 방부제 및 보습제 등을 추가로 포함할 수 있으며, 물성개선을 목적으로 점증제, 무기염류, 합성 고분자 물질 등을 포함할 수 있다. 예를 들면, 본 발명의 화장료 조성물로 세안제 및 비누를 제조하는 경우에는 통상의 세안제 및 비누 베이스에 로즈힙 추출물을 첨가하여 용이하게 제조할 수 있다. 크림을 제조하는 경우에는 일반적인 수중유적형(O/W)의 크림베이스에 로즈힙 추출물을 첨가하여 제조할 수 있다. 여기에 향료, 킬레이트제, 색소, 산화방지제, 방부제 등과 물성개선을 목적으로 한 단백질, 미네랄, 비타민 등 합성 또는 천연소재를 추가로 첨가할 수 있다.The excipient is not limited thereto, but may include, for example, an emollient, a skin penetration enhancer, a colorant, a fragrance, an emulsifier, a thickening agent, and a solvent. In addition, fragrances, dyes, bactericides, antioxidants, preservatives and moisturizing agents may be additionally included, and for the purpose of improving physical properties, thickeners, inorganic salts, synthetic polymer materials, and the like may be included. For example, when preparing a face wash and soap with the cosmetic composition of the present invention, it can be easily prepared by adding a rosehip extract to a conventional face wash and soap base. In the case of preparing a cream, it can be prepared by adding rosehip extract to a general oil-in-water type (O/W) cream base. Here, synthetic or natural materials such as proteins, minerals and vitamins for the purpose of improving physical properties such as fragrances, chelating agents, pigments, antioxidants, and preservatives may be additionally added.
본 발명의 화장료 조성물에 함유되는 로즈힙 추출물의 함량은 이에 한정되지 않지만 전체 조성물 총중량에 대하여 0.001 내지 10 중량%인 것이 바람직하고, 0.01 내지 5중량%인 것이 더욱 바람직하다. 상기 함량이 0.001중량% 미만에서는 목적하는 효과를 기대할 수 없고, 10중량% 초과에서는 안전성 또는 제형상의 제조에 어려움이 있을 수 있다.The content of the rosehip extract contained in the cosmetic composition of the present invention is not limited thereto, but is preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, based on the total weight of the composition. If the content is less than 0.001% by weight, the desired effect cannot be expected, and if it exceeds 10% by weight, there may be difficulties in safety or formulation.
본 발명의 조성물은 근육 질환 예방 또는 개선을 목적으로 사료첨가제 또는 이를 포함하는 사료 조성물에 첨가할 수 있다.The composition of the present invention may be added to a feed additive or a feed composition comprising the same for the purpose of preventing or improving muscle disease.
본 발명에서 용어, '사료첨가제'는 영양소 보충 및 체중감소 예방, 사료 내 섬유소의 소화 이용성 증진, 유질개선, 번식장애 예방 및 수태율 향상, 하절기 고온 스트레스 예방 등 다양한 효과를 목적으로 사료에 첨가하는 물질을 포함한다. 본 발명의 사료첨가제는 사료관리법상의 보조사료에 해당하며, 탄산수소나트륨, 벤토나이트(bentonite), 산화마그네슘, 복합광물질 등의 광물질제제, 아연, 구리, 코발트, 셀레늄 등의 미량 광물질인 미네랄제제, 케로틴, 비타민 A D, E, 니코틴산, 비타민 B 복합체 등의 비타민제, 메티오닌, 라이신 등의 보호아미노산제, 지방산 칼슘염 등의 보호지방산제, 생균제(유산균제), 효모배양물, 곰팡이 발효물 등의 생균, 효모제 등이 추가로 포함될 수 있다.As used herein, the term 'feed additive' refers to a substance added to feed for various effects such as nutrient supplementation and weight loss prevention, enhancement of digestibility of fiber in feed, oil quality improvement, reproductive disorder prevention and fertility improvement, and prevention of high temperature stress in summer includes The feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act, and mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, and trace minerals such as zinc, copper, cobalt, selenium, and kerotene. , vitamins AD, E, nicotinic acid, vitamin B complex, etc., protective amino acids such as methionine and lysine, protective fatty acids such as fatty acid calcium salts, probiotics (lactic acid bacteria), yeast cultures, live bacteria such as mold fermented products, Yeast agents and the like may be further included.
본 발명에서 용어 '사료'는, 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분으로, 본 발명에 따른 근육 질환 예방 또는 개선용 조성물을 유효성분으로 포함하는 사료는 당업계에 공지된 다양한 형태의 사료로 제조가능하며, 바람직하게는 농후 사료, 조사료 및/또는 특수사료가 포함될 수 있으나, 이로 한정되지 않는다.In the present invention, the term 'feed' refers to any natural or artificial diet, one-meal meal, etc. or a component of the one-meal meal for animals to eat, ingest and digest, or to prevent or improve muscle disease according to the present invention. The feed containing the composition for use as an active ingredient can be prepared in various types of feed known in the art, and preferably, a concentrated feed, roughage and/or special feed may be included, but is not limited thereto.
농후사료에는 밀, 귀리, 옥수수 등의 곡류를 포함하는 종자열매류, 곡물을 정제하고 얻는 부산물로서 쌀겨, 밀기울, 보릿겨 등을 포함하는 겨류, 콩, 유체, 깨, 아마인, 코코야자 등을 채유하고 얻는 부산물인 깻묵류와 고구마, 감자 등에서 녹말을 뺀 나머지인 녹말찌꺼기의 주성분인 잔존녹말질류 등의 찌꺼기류, 어분, 물고기찌꺼기, 어류에서 얻은 신선한 액상물을 농축시킨 것인 피시솔루블(fish soluble), 육분(肉粉), 혈분, 우모분, 탈지분유, 우유에서 치즈, 탈지유에서 카제인을 제조할 때의 잔액인 훼이(whey)를 건조한 건조훼이 등의 동물질사료, 효모, 클로렐라, 해조류가 있으나 이에 제한되지 않는다.In the enriched feed, seed fruits containing grains such as wheat, oats, and corn, bran containing rice bran, bran, barley bran, etc. Fish Soluble is a concentrated fish meal, fish waste, and fresh liquid obtained from fish, fish meal, fish waste, and residual starch, which is the main component of starch residue, which is a by-product obtained by subtracting starch from sweet potatoes and potatoes. There are animal feeds such as dried whey, yeast, chlorella, and seaweed, which are dried whey, which is the remainder of the production of soluble), meat meal, blood meal, cow meal, powdered skim milk, cheese from skim milk, and casein from skim milk. It is not limited thereto.
조사료에는 야초, 목초, 풋베기 등의 생초(生草)사료, 사료용 순무, 사료용 비트, 순무의 일종인 루터베어거 등의 뿌리채소류, 생초, 풋베기작물, 곡실(穀實) 등을 사일로에 채워 놓고 젖산발효시킨 저장사료인 사일리지(silage), 야초, 목초를 베어 건조시킨 건초, 종축용(種畜用) 작물의 짚, 콩과 식물의 나뭇잎이 있으며, 이에 제한되지 않는다. 특수사료에는 굴껍데기, 암염 등의 미네랄 사료, 요소나 그 유도체인 디우레이드이소부탄 등의 요소사료, 천연사료원료만을 배합했을 때 부족하기 쉬운 성분을 보충하거나, 사료의 저장성을 높이기 위해서 배합사료에 미량으로 첨가하는 물질인 사료첨가물, 식이보조제가 있으나 이에 제한되지 않는다.For roughage, raw grass feed such as wild grasses, grasses, and green cuttings, turnips for feed, beets for feed, root vegetables such as rutherbearers, a type of turnip, raw herbs, green crops, grains, etc. are placed in a silo. Examples include, but are not limited to, silage, which is stored feed fermented with lactic acid after filling, wild grass, hay obtained by cutting and drying grass, straw of crops for breeding purposes, and leaves of legumes. Special feed includes mineral feed such as oyster shells and rock salt, urea feed such as urea or its derivative diureide isobutane, supplementing ingredients that are easily lacking when only natural feed raw materials are mixed, or added to formulated feed to increase feed storage. There are feed additives and dietary supplements, which are substances added in trace amounts, but are not limited thereto.
본 발명에 따른 상기 근육 질환의 예방 또는 개선용 사료 첨가제는 당업계에 공지된 다양한 사료 제조방법에 따라 적절한 유효 농도 범위에서 로즈힙 분쇄물 또는 로즈힙 추출물을 첨가하여 제조 가능하다.The feed additive for preventing or improving muscle disease according to the present invention can be prepared by adding ground rosehip or rosehip extract in an appropriate effective concentration range according to various feed preparation methods known in the art.
본 발명에 따른 사료 첨가제는 근육 질환의 예방 또는 개선을 목적으로 하는 개체이면 제한 없이 적용가능하다. 예를 들면, 소, 말, 돼지, 염소, 양, 개, 고양이, 토끼 등과 같은 비인간동물, 조류 및 어류 등 어느 개체에도 적용이 가능하다.The feed additive according to the present invention can be applied without limitation as long as it is an individual for the purpose of preventing or improving muscle disease. For example, non-human animals such as cattle, horses, pigs, goats, sheep, dogs, cats, rabbits, etc., birds and fish can be applied to any object.
이하 본 발명을 실시 예를 통하여 보다 상세하게 설명한다. Hereinafter, the present invention will be described in more detail through examples.
단, 하기 실시 예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시 예에 한정되는 것은 아니다. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.
[실시예 1] 로즈힙 추출물의 제조[Example 1] Preparation of rosehip extract
<1-1> 로즈힙 열수 추출물의 제조<1-1> Preparation of hot water extract of rosehip
건조된 로즈힙 열매를 믹서로 분쇄한 다음, 분쇄한 로즈힙 열매 시료 10 g을 물 100 mL에 넣고 80℃에서 3시간 동안 추출하였다. 추출된 시료는 와트만(Whatman) 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 로즈힙 열수 추출물을 얻었다.After grinding the dried rosehip fruit with a mixer, 10 g of the crushed rosehip fruit sample was added to 100 mL of water and extracted at 80° C. for 3 hours. The extracted sample was filtered under reduced pressure with Whatman No. 1 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove solvent components, and then hot water extract of rosehip was obtained.
<1-2> 로즈힙 에탄올 추출물의 제조<1-2> Preparation of rosehip ethanol extract
건조된 로즈힙 열매를 믹서로 분쇄한 다음, 분쇄한 로즈힙 열매 시료 10 g을 100% 에탄올 100 mL에 넣고 40℃에서 3시간 동안 추출하였다. 추출된 시료는 와트만 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 로즈힙 에탄올 추출물을 얻었다.After crushing the dried rosehip fruit with a mixer, 10 g of the crushed rosehip fruit sample was added to 100 mL of 100% ethanol and extracted at 40° C. for 3 hours. The extracted sample was filtered under reduced pressure with Whatman No. 1 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove solvent components, and then a rosehip ethanol extract was obtained.
<1-3> 로즈힙 에틸아세테이트 추출물의 제조<1-3> Preparation of rosehip ethyl acetate extract
건조된 로즈힙 열매를 믹서로 분쇄한 다음, 분쇄한 로즈힙 열매 시료 10 g을 에틸아세테이트 100 mL에 넣고 40℃에서 3시간 동안 추출하였다. 추출된 시료는 와트만 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 로즈힙 에틸아세테이트 추출물을 얻었다.After grinding the dried rosehip fruit with a mixer, 10 g of the crushed rosehip fruit sample was added to 100 mL of ethyl acetate and extracted at 40°C for 3 hours. The extracted sample was filtered under reduced pressure with Whatman No. 1 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove solvent components, and then a rosehip ethyl acetate extract was obtained.
<1-4> 로즈힙 헥산 추출물의 제조<1-4> Preparation of rosehip hexane extract
건조된 로즈힙 열매를 믹서로 분쇄한 다음, 분쇄한 로즈힙 열매 시료 10 g을 헥산 100 mL에 넣고 40℃에서 3시간 동안 추출하였다. 추출된 시료는 와트만 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 로즈힙 헥산 추출물을 얻었다.After grinding the dried rosehip fruit with a mixer, 10 g of the crushed rosehip fruit sample was added to 100 mL of hexane and extracted at 40° C. for 3 hours. The extracted sample was filtered under reduced pressure with Whatman No. 1 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove solvent components, and then a rosehip hexane extract was obtained.
<1-5> 로즈힙 초고압 추출물의 제조<1-5> Preparation of ultra-high pressure rosehip extract
건조된 로즈힙 열매를 믹서로 분쇄한 다음, 분쇄한 로즈힙 열매 1g과 18% 에탄올 76 mL을 폴리에틸렌(polyethylene) 팩에 넣고 밀봉한 후 초고압 추출장치(Frescal MFP-7000; Mitsubishi Heavy Industries)를 이용하여 추출하였다. 초고압 추출 조건은 추출압력이 320 MPa, 추출시간은 5 min 이였다. 추출된 시료는 와트만(Whatman) 2번 여과지로 여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거함으로써 로즈힙 초고압 추출물을 얻었다. After crushing the dried rosehip fruit with a mixer, 1 g of crushed rosehip fruit and 76 mL of 18% ethanol are put in a polyethylene pack, sealed, and extracted using an ultra-high pressure extraction device (Frescal MFP-7000; Mitsubishi Heavy Industries) did Under the ultra-high pressure extraction conditions, the extraction pressure was 320 MPa and the extraction time was 5 min. The extracted sample was filtered with Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove solvent components to obtain a rosehip ultra-high pressure extract.
<1-6> 로즈힙 아임계 추출물의 제조<1-6> Preparation of rosehip subcritical extract
건조된 로즈힙 열매를 믹서로 분쇄한 다음, 분쇄한 로즈힙 열매 50 g을 1 L 물과 함께 아임계 추출장치(Biovan, Gyeonggi, Korea)의 아임계수 반응기에 넣고 밀폐하였다. 밀폐 후, 반응기의 온도를 200℃로 상승시켰으며, 반응기의 온도가 200℃에 도달하면 가열을 중단하고, 상기 온도를 20분간 유지하여 추출을 하였다. 20분 후 추출물을 냉각수가 공급되는 저장탱크로 이송하여 30℃까지 급속 냉각시킨 후, 부유 잔사를 분리하기 위해 3,600 rpm으로 30분 동안 원심분리하여 상등액만 취하였다. 동결건조기(Ilshin Lab Co. Ltd., Seoul, Korea)를 이용하여 용매를 전부 제거함으로써 로즈힙 열매 아임계 추출물을 얻었다.After crushing the dried rosehip fruit with a mixer, 50 g of the crushed rosehip fruit was put in a subcritical water reactor of a subcritical extraction device (Biovan, Gyeonggi, Korea) with 1 L water and sealed. After sealing, the temperature of the reactor was raised to 200°C, and when the temperature of the reactor reached 200°C, heating was stopped, and the temperature was maintained for 20 minutes for extraction. After 20 minutes, the extract was transferred to a storage tank supplied with cooling water and rapidly cooled to 30° C., and then centrifuged at 3,600 rpm for 30 minutes to separate floating residues, and only the supernatant was taken. A subcritical extract of rosehip fruit was obtained by removing all solvents using a freeze dryer (Ilshin Lab Co. Ltd., Seoul, Korea).
<1-7> 로즈힙 초임계 추출물의 제조<1-7> Preparation of rosehip supercritical extract
건조된 로즈힙 열매를 믹서로 분쇄한 다음, 분쇄한 로즈힙 열매 10 g을 시료 카트리지에 충전하고 초임계 유체 추출 장치(SFX 3560, Isco Inc., Lincoln, NE, USA)를 이용하여 추출하였다. 초임계 추출 조건은 추출 압력이 300 bar, 추출 온도는 50℃, 초임계 이산화탄소의 유속은 60 mL/min, 추출시간은 2시간으로 하였다. 초임계 유체 추출이 완료되면, 추출 장치의 압력을 낮춰 초임계 유체 상태를 해제하여 로즈힙 열매 초임계 추출물을 얻었다.The dried rosehip fruit was crushed with a mixer, and 10 g of the crushed rosehip fruit was charged into a sample cartridge and extracted using a supercritical fluid extraction device (SFX 3560, Isco Inc., Lincoln, NE, USA). The supercritical extraction conditions were an extraction pressure of 300 bar, an extraction temperature of 50° C., a flow rate of supercritical carbon dioxide of 60 mL/min, and an extraction time of 2 hours. When the supercritical fluid extraction was completed, the supercritical fluid state was released by lowering the pressure of the extraction device to obtain a rosehip fruit supercritical extract.
[실시예 2] 로즈힙 열수 추출물 및 에탄올 추출물의 근 단백질 합성 생체지표 mTOR 활성 증가 효과[Example 2] Effect of increasing mTOR activity, a biomarker for muscle protein synthesis of rosehip hot water extract and ethanol extract
mTOR 단백질은 인산화되어 활성화되었을 때, 근육 세포 내의 PI3K/Akt 신호전달경로에서 근단백질 합성 및 근육량 증가에 관여하는 단백질의 활성화를 유도할 수 있음이 알려져 있다. 이에, 로즈힙 추출물의 근육 생성 유도 활성을 확인하기 위해, mTOR Sandwich ELISA kit (Cell Signaling Technology, Beverly, MA, USA)를 이용하여 mTOR의 활성을 확인하였다.It is known that when mTOR protein is phosphorylated and activated, it can induce activation of proteins involved in muscle protein synthesis and increase in muscle mass in the PI3K/Akt signaling pathway in muscle cells. Therefore, in order to confirm the muscle-generating activity of the rosehip extract, the activity of mTOR was confirmed using the mTOR Sandwich ELISA kit (Cell Signaling Technology, Beverly, MA, USA).
L6 근육모세포(ATCC; Manassas, VA, USA)를 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA)이 함유된 Dulbecco's modified Eagle's media (DMEM; Hyclone)와 함께 6-웰 플레이트에 1 × 105 cell/well이 되도록 seeding 후 24시간 동안 배양하였다. 배양 후, 웰에 있는 배지를 제거하고 2% horse serum (HS; Hyclone)이 함유된 DMEM (Hyclone)로 교환하여 6일간 추가 배양하여 L6 세포를 근관세포(myotube)로 분화시켰다. 다음, 상기 실시예 1-1의 로즈힙 열수 추출물을 40 μg/mL의 농도로, 실시예 1-2의 로즈힙 에탄올 추출물을 5.0 μg/mL의 농도로 DMEM (Hyclone)에 녹인 후, 세포에 처리하고 12시간 동안 배양하였다. 배양 후, 세포 용해 완충용액(cell lysis buffer)을 처리하여 세포를 용해시켰다. 수득한 세포 용해물 내 단백질을 브래드포드(Bio-Rad Laboratories Inc., Hercules, CA, USA)를 이용하여 1 mg/mL 농도로 정량하였다. 항-mTOR 항체가 부착된 마이크로웰에 세포 용해물을 50 μL씩 분주하여 37℃에서 2시간 동안 방치하였다. 세척 완충용액(Washing buffer)으로 총 4회 씻은 후, 확인용 항체(detection antibody)를 처리하고 37℃에서 1시간 방치했으며, 다시 세척 완충용액으로 총 4회 씻은 후, 겨자무 과산화효소(horseradish peroxidase)가 접합된 2차 항체를 넣고 37℃에서 30분 동안 방치하였다. 마지막으로 세척 완충용액으로 총 4회 씻은 후, TMB 기질을 각 웰에 넣고 37℃에서 10분 동안 방치하고 정지 완충용액(stop solution)을 가하여 TMB의 반응을 멈추었다. 2분 뒤, 450 nm의 파장으로 흡광도를 측정하여 로즈힙 추출물을 처리한 근관 세포 내 mTOR 수준을 측정하였다. 실험은 총 3회 반복하여 진행하였으며, 각각 얻은 측정값은 대조군에 대한 백분율(%)을 계산하여 평균±표준편차로 나타내었다. 군 간의 차이는 SPSS25.0 통계 패키지(SPSS Inc., Chicago, IL, USA)를 이용하여 일원배치분산분석(one-way analysis of variance)에 의한 Duncan 다중범위 분석을 통해 확인하였다. 이 때, p 값이 5% 미만일 경우 통계적으로 유의성이 있다고 표기하였다. L6 myoblasts (ATCC; Manassas, VA, USA) were mixed with Dulbecco's modified Eagle's media (DMEM; Hyclone) containing 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) in a 6-well plate at 1 × It was cultured for 24 hours after seeding so as to become 10 5 cells/well. After culturing, the medium in the well was removed, exchanged with DMEM (Hyclone) containing 2% horse serum (HS; Hyclone), and further cultured for 6 days to differentiate L6 cells into myotubes. Next, the hot water extract of rosehip of Example 1-1 was dissolved in DMEM (Hyclone) at a concentration of 40 μg/mL, and the ethanol extract of rosehip of Example 1-2 was dissolved in DMEM (Hyclone) at a concentration of 5.0 μg/mL, and the cells were treated and Incubated for 12 hours. After incubation, cells were lysed by treatment with cell lysis buffer. Proteins in the obtained cell lysate were quantified at a concentration of 1 mg/mL using Bradford (Bio-Rad Laboratories Inc., Hercules, CA, USA). 50 μL of cell lysate was dispensed into microwells to which anti-mTOR antibody was attached and left at 37° C. for 2 hours. After washing a total of 4 times with washing buffer, the detection antibody was treated and left at 37°C for 1 hour, washed again with washing buffer a total of 4 times, and horseradish peroxidase ) was added to the conjugated secondary antibody and left at 37° C. for 30 minutes. Finally, after washing a total of 4 times with wash buffer, TMB substrate was put into each well, left at 37° C. for 10 minutes, and a stop solution was added to stop the TMB reaction. After 2 minutes, absorbance was measured with a wavelength of 450 nm to measure the level of mTOR in myotube cells treated with rosehip extract. The experiment was repeated a total of three times, and the measured values obtained for each were calculated as a percentage (%) relative to the control and expressed as the mean ± standard deviation. Differences between groups were confirmed through Duncan multi-range analysis by one-way analysis of variance using the SPSS25.0 statistical package (SPSS Inc., Chicago, IL, USA). At this time, when the p value was less than 5%, it was marked as statistically significant.
그 결과, 도 1에 나타난 바와 같이 로즈힙 열수 추출물을 40 μg/mL의 농도로 처리함에 따라 대조군에 비하여 mTOR의 활성이 유의적(*
p < 0.05)으로 증가하였으며, 로즈힙 에탄올 추출물을 5.0 μg/mL의 농도로 처리함에 따라 대조군에 비하여 mTOR의 활성이 유의적(**
p < 0.01)으로 증가한 것을 확인할 수 있었다. 이는 본 발명의 로즈힙 열수 추출물과 에탄올 추출물이 근육세포 내에서 근 단백질 합성을 촉진시키는 능력이 우수하다는 것을 의미한다.As a result, as shown in FIG. 1, when the rosehip hot water extract was treated at a concentration of 40 μg/mL, the activity of mTOR increased significantly ( * p < 0.05) compared to the control, and the rosehip ethanol extract was treated with 5.0 μg/mL. As compared to the control group, it was confirmed that the activity of mTOR increased significantly ( ** p < 0.01) by treatment with a concentration of . This means that the hot water extract of rosehip and the ethanol extract of the present invention have excellent ability to promote muscle protein synthesis in muscle cells.
[실시예 3] 로즈힙 추출물의 근 단백질 합성 생체지표 mTOR 활성 증가 효과[Example 3] Increase effect of mTOR activity, a biomarker for muscle protein synthesis of rosehip extract
실시예 1-3 내지 실시예 1-7의 로즈힙 추출물을 각각 5.0 μg/mL씩 처리하였다는 점을 제외하고, 상기 실시예 2와 동일한 방법으로 mTOR 활성을 측정하였다. mTOR activity was measured in the same manner as in Example 2, except that 5.0 μg/mL of each of the rosehip extracts of Examples 1-3 to 1-7 was treated.
그 결과, 표 1에 나타난 바와 같이 로즈힙 추출물을 처리함에 따라 추출물을 처리하지 않은 대조군에 비하여 mTOR의 활성이 모두 유의적(**
p < 0.01)으로 증가한 것을 확인할 수 있었다. 이는 본 발명의 로즈힙 추출물이 근육세포 내에서 근 단백질 합성을 촉진시키는 능력이 우수하다는 것을 의미한다.As a result, as shown in Table 1, as the rosehip extract was treated, it was confirmed that all of the mTOR activities increased significantly ( ** p < 0.01) compared to the control group that was not treated with the extract. This means that the rosehip extract of the present invention has an excellent ability to promote muscle protein synthesis in muscle cells.
실험군experimental group | 상대적 mTOR 활성(%)Relative mTOR activity (%) |
실시예 1-3Examples 1-3 | 117.7±3.8**117.7±3.8** |
실시예 1-4Examples 1-4 | 128.3±6.4**128.3±6.4** |
실시예 1-5Examples 1-5 | 121.9±6.0**121.9±6.0** |
실시예 1-6Examples 1-6 | 122.5±5.2**122.5±5.2** |
실시예 1-7Examples 1-7 | 130.6±6.5**130.6±6.5** |
[실시예 4] 로즈힙 열수 추출물의 근 단백질 합성 생체지표 발현량 증가 효과[Example 4] Effect of increasing the expression level of muscle protein synthesis biomarkers of hot water extract of rosehip
근육세포인 L6 myoblast (ATCC)를 10% FBS (Hyclone)가 함유된 DMEM (Hyclone)과 함께 6-웰 플레이트에 1 Х 105 cell/mL이 되도록 넣었다. 세포밀도가 약 80~85%가 되었을 때, 웰에 있는 배지를 제거하고 2% HS (Hyclone)가 함유된 DMEM (Hyclone)을 세포에 처리하여 myotube 분화를 유도하였다. 2일에 한 번씩 새로운 배지로 교체하여 총 6일 동안 분화를 진행하였다. 분화 후, 50 ng/mL tumor necrosis factor alpha (TNF-α; PeproTech, Rocky Hills, NJ, USA)가 함유된 DMEM (Hyclone)에 상기 실시예 1-1에서 제조한 로즈힙 열수 추출물을 DMEM에 40과 80 μg/mL의 농도로 녹인 후, 세포에 처리하고 12시간 동안 배양하였다. 배양 후, 단백질 가수분해효소 억제제 칵테일(Sigma-Aldrich St. Louis, MO, USA)이 포함된 NP-40 완충용액(ELPIS-Biotech, Daejeon, Korea)으로 세포를 용해시켜 세포 용해물을 수득하였다. 수득한 세포 용해물은 13,000 rpm으로 10분간 원심분리하여 상등액을 취하였다. 상등액 내 단백질 농도를 브래드포드(Bio-Rad Laboratories Inc.)로 정량한 다음, 단백질을 5분간 가열하고 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gel에 전개하여 단백질을 분리하였다. 분리된 단백질은 니트로셀룰로스 막으로 전달하였다. 그런 다음, p-mTOR, mTOR, p-p70S6K, p70S6K, p-4EBP1, 4EBP1, 또는 α-tubulin 항체(Cell Signaling Technology)를 각각 2.5% bovine serum albumin (BSA)에 1:1000 비율로 희석하여 니트로셀룰로스 막에 전달된 단백질과 20시간 동안 상온에서 반응시켰다. 1차 항체를 반응시킨 다음, Tris-buffer Saline Tween20 (TBST)를 이용하여 니트로셀룰로스 막을 10분간 3 회 세척하였다. 1차 항체를 인지하는 horseradish peroxidase (HRP)가 접합된 2차 항체(Bethyl Laboratories, Inc., Montgomery, TA, USA)를 2.5% BSA에 1:5000이 되도록 희석하여 니트로셀룰로스 막과 2시간 동안 상온에서 반응시켰으며, TBST를 이용하여 10분씩 3회에 걸쳐 세척하였다. 단백질 밴드는 ECL 웨스턴블럿 검출 시약(GE Healthcare, Piscataway, NJ, USA)을 사용하여 발색하였으며, G;BOX EF imaging system (Syngene, Cambridge, UK)을 이용하여 발색된 단백질 밴드를 확인하였다. L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at a concentration of 1 Х 10 5 cell/mL. When the cell density reached about 80-85%, the medium in the well was removed and the cells were treated with DMEM (Hyclone) containing 2% HS (Hyclone) to induce myotube differentiation. Differentiation was performed for a total of 6 days by replacing the medium with a new medium once every 2 days. After differentiation, the rosehip hot water extract prepared in Example 1-1 was added to DMEM (Hyclone) containing 50 ng/mL tumor necrosis factor alpha (TNF-α; PeproTech, Rocky Hills, NJ, USA). After dissolving at a concentration of 80 μg/mL, the cells were treated and incubated for 12 hours. After incubation, the cells were lysed with a NP-40 buffer (ELPIS-Biotech, Daejeon, Korea) containing a protease inhibitor cocktail (Sigma-Aldrich St. Louis, MO, USA) to obtain a cell lysate. The obtained cell lysate was centrifuged at 13,000 rpm for 10 minutes to obtain a supernatant. The protein concentration in the supernatant was quantified with a Bradford (Bio-Rad Laboratories Inc.), and then the protein was heated for 5 minutes, and then the protein was separated by running on a 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gel. The separated protein was transferred to a nitrocellulose membrane. Then, p-mTOR, mTOR, p-p70S6K, p70S6K, p-4EBP1, 4EBP1, or α-tubulin antibody (Cell Signaling Technology) was diluted 1:1000 in 2.5% bovine serum albumin (BSA) at a ratio of 1:1000 to nitro The protein transferred to the cellulose membrane was reacted at room temperature for 20 hours. After reacting with the primary antibody, the nitrocellulose membrane was washed 3 times for 10 minutes using Tris-buffer Saline Tween20 (TBST). A secondary antibody (Bethyl Laboratories, Inc., Montgomery, TA, USA) conjugated with horseradish peroxidase (HRP), which recognizes the primary antibody, was diluted 1:5000 in 2.5% BSA, and then placed with a nitrocellulose membrane at room temperature for 2 hours. , and washed 3 times for 10 minutes each using TBST. The protein band was developed using ECL Western Blot detection reagent (GE Healthcare, Piscataway, NJ, USA), and the colored protein band was confirmed using the G;BOX EF imaging system (Syngene, Cambridge, UK).
그 결과, 도 2에 나타난 바와 같이, TNF-α에 의해 근단백질 합성에 관여하는 p-mTOR, p-p70S6K 및 p-4EBP-1의 단백질 발현 수준이 유의적(##
p < 0.01)으로 감소한 반면, 로즈힙 열수 추출물을 처리함에 따라 p-mTOR, p-p70S6K 및 p-4EBP-1의 단백질 발현 수준이 유의적(**
p < 0.01)으로 증가한 것을 확인할 수 있었다. 이는 본 발명의 로즈힙 열수 추출물이 근육세포 내에서 근단백질 합성에 관여하는 생체지표의 발현을 증가시키는 능력이 우수한 것을 의미한다.As a result, as shown in Figure 2, the protein expression levels of p-mTOR, p-p70S6K and p-4EBP-1 involved in muscle protein synthesis by TNF-α were significantly ( ## p < 0.01) decreased. On the other hand, it was confirmed that the protein expression levels of p-mTOR, p-p70S6K, and p-4EBP-1 were significantly increased ( ** p < 0.01) as the rosehip hot water extract was treated. This means that the hot water extract of rosehip of the present invention has excellent ability to increase the expression of biomarkers involved in muscle protein synthesis in muscle cells.
[실시예 5] 로즈힙 열수 추출물의 근 단백질 분해 생체지표 발현량 억제 효과[Example 5] Effect of inhibiting the expression level of muscle proteolysis biomarkers of hot water extract of rosehip
근육세포인 L6 myoblast (ATCC)를 10% FBS (Hyclone)가 함유된 DMEM (Hyclone)과 함께 6-웰 플레이트에 1 × 105 cell/mL이 되도록 넣었다. 세포밀도가 약 80~85%가 되었을 때, 웰에 있는 배지를 제거하고 2% HS (Hyclone)가 함유된 DMEM (Hyclone)을 세포에 처리하여 myotube 분화를 유도하였다. 2일에 한 번씩 새로운 배지로 교체하여 총 6일 동안 분화를 진행하였다. 분화 후, 50 ng/mL tumor necrosis factor alpha (TNF-α; PeproTech, Rocky Hills, NJ, USA)가 함유된 DMEM (Hyclone)에 상기 상기 실시예 1-1의 로즈힙 열수 추출물을 40 μg/mL와 80 μg/mL의 농도로 DMEM (Hyclone)에 녹인 후, 세포에 처리하였다. 12시간 후, TRIzol시약(Takara, Otsu, Japan)을 사용하여 총 RNA를 분리하였다. 분리한 총 RNA는 나노드랍(NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, MA, USA)을 이용하여 정량하였다. 정량된 16 μL의 RNA를 Reverse Transcriptase Premix (ELPIS-Biotech)와 PCR 기계(Gene Amp PCR System 2700; Applied Biosystems, Foster City, CA, USA)를 이용하여 42℃, 55분 및 70℃, 15분의 조건에서 cDNA로 합성하였다. 16 μL의 생성된 cDNA 중 1 μL의 cDNA, 하기의 특정 프라이머(Bioneer, Daejeon, Korea) 그리고 PCR premix (ELPIS-Biotech)로 95℃에서 30초, 60℃에서 1분, 72℃에서 1분을 30번 반복하여 PCR을 수행하였다. L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at a concentration of 1 × 10 5 cell/mL. When the cell density reached about 80-85%, the medium in the well was removed and the cells were treated with DMEM (Hyclone) containing 2% HS (Hyclone) to induce myotube differentiation. Differentiation was performed for a total of 6 days by replacing the medium with a new medium once every 2 days. After differentiation, the hot water extract of Example 1-1 was mixed with 40 μg/mL of the rosehip hot water extract of Example 1-1 in DMEM (Hyclone) containing 50 ng/mL tumor necrosis factor alpha (TNF-α; PeproTech, Rocky Hills, NJ, USA). After dissolving in DMEM (Hyclone) at a concentration of 80 μg/mL, the cells were treated. After 12 hours, total RNA was isolated using TRIzol reagent (Takara, Otsu, Japan). The isolated total RNA was quantified using a nanodrop (NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, MA, USA). 16 μL of quantified RNA was analyzed using Reverse Transcriptase Premix (ELPIS-Biotech) and a PCR machine (Gene Amp PCR System 2700; Applied Biosystems, Foster City, CA, USA) at 42°C, 55 minutes and 70°C, for 15 minutes. It was synthesized as cDNA under conditions. Out of 16 μL of the generated cDNA, 1 μL of cDNA, the following specific primers (Bioneer, Daejeon, Korea) and PCR premix (ELPIS-Biotech) were used for 30 seconds at 95°C, 1 minute at 60°C, and 1 minute at 72°C. PCR was performed by repeating 30 times.
MuRF1MuRF1
Forward primer: 5'-CCGGACGGAAATGCTATGGA-3'(서열번호 1)Forward primer: 5'-CCGGACGGAAATGCTATGGA-3' (SEQ ID NO: 1)
Reverse primer: 5'-AGCCTGGAAGATGTCGTTGG-3'(서열번호 2)Reverse primer: 5'-AGCCTGGAAGATGTCGTTGG-3' (SEQ ID NO: 2)
Atrogin-1Atrogin-1
Forward primer: 5'-GTTACTGCAACAAGGAGAATCTGTT-3'(서열번호 3)Forward primer: 5'-GTTACTGCAACAAGGAGAATCTGTT-3' (SEQ ID NO: 3)
Reverse primer: 5'-CCGTATGAGTCTTATGTTTTGCTGG-3'(서열번호 4)Reverse primer: 5'-CCGTATGAGTCTTATGTTTTGCTGG-3' (SEQ ID NO: 4)
β-Actin:β-Actin:
Forward primer: 5'-TGACAGGATGCAGAAGGAGATTAC-3'(서열번호 5)Forward primer: 5'-TGACAGGATGCAGAAGGAGATTAC-3' (SEQ ID NO: 5)
Reverse primer: 5'-TAAAACGCAGCTCAGTAACAGTC-3'(서열번호 6)Reverse primer: 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6)
PCR 결과 증폭된 cDNA를 1.5% agarose gel로 전기영동하여 분리하였으며, G;BOX EF imaging system (Syngene)을 이용하여 cDNA band를 확인하였다. As a result of PCR, the amplified cDNA was separated by electrophoresis on 1.5% agarose gel, and the cDNA band was confirmed using the G;BOX EF imaging system (Syngene).
그 결과, 도 3에 나타낸 바와 같이 TNF-α에 의해 근단백질 분해에 관여하는 atrogin-1과 MuRF1의 mRNA 발현 수준이 유의적(##
p < 0.01)으로 증가한 반면, 로즈힙 열수 추출물을 처리함에 따라 atrogin-1과 MuRF1의 mRNA 발현 수준이 유의적(**
p < 0.01)으로 감소한 것을 확인할 수 있었다. 이는 본 발명의 로즈힙 열수 추출물이 근육세포 내에서 근단백질 분해에 관여하는 생체지표의 발현을 억제시키는 능력이 우수한 것을 의미한다.As a result, as shown in FIG. 3 , the mRNA expression levels of atrogin-1 and MuRF1, which are involved in myoprotein degradation by TNF-α, increased significantly ( ## p < 0.01), whereas as the rosehip hot water extract was treated, It was confirmed that the mRNA expression levels of atrogin-1 and MuRF1 were significantly reduced ( ** p < 0.01). This means that the hot water extract of rosehip of the present invention has excellent ability to suppress the expression of biomarkers involved in muscle protein degradation in muscle cells.
[실시예 6] 로즈힙 추출물의 근 단백질 분해 억제 효과[Example 6] Inhibitory effect on muscle protein degradation of rosehip extract
실시예 1-2 내지 실시예 1-7의 로즈힙 에탄올, 에틸아세테이트, 헥산, 초고압, 아임계, 초임계 추출물을 각각 5 μg/mL 처리하였다는 점을 제외하고, 상기 실시예 5와 동일한 방법으로 총 3회 반복하여 진행하였다. 다음, Image J 프로그램(National Institute of Health, Bethesda, MD, USA)을 이용하여 MuRF1 및 atrogin-1 mRNA의 band density를 측정한 다음, 각 측정값은 대조군에 대한 백분율(%)을 계산하여 평균±표준편차로 나타내었다. In the same manner as in Example 5, except that 5 μg/mL of the rosehip ethanol, ethyl acetate, hexane, ultra-high pressure, subcritical, and supercritical extracts of Examples 1-2 to 1-7 were treated, respectively. This was repeated 3 times in total. Next, the band density of MuRF1 and atrogin-1 mRNA was measured using the Image J program (National Institute of Health, Bethesda, MD, USA), and then each measured value was calculated as a percentage (%) for the control group and the mean± It is expressed as standard deviation.
그 결과, 표 2에 나타난 바와 같이 TNF-α를 처리함에 따라 대조군에 비해 atrogin-1 및 MuRF1의 mRNA 발현량이 유의적으로(##
p < 0.01) 증가하였으나, 로즈힙 추출물을 처리한 경우 TNF-α 처리군에 비해 MuRF1 및 Atrogin-1의 mRNA 발현량이 모두 유의적(**
p < 0.01)으로 감소한 것을 확인할 수 있었다. 이는 본 발명의 로즈힙 추출물이 근육세포 내에서 근 단백질 분해를 억제하는 능력이 우수하다는 것을 의미한다.As a result, as shown in Table 2, when TNF-α was treated, the mRNA expression levels of atrogin-1 and MuRF1 were significantly increased ( ## p < 0.01) compared to the control, but when treated with rosehip extract, TNF-α It was confirmed that both the mRNA expression levels of MuRF1 and Atrogin-1 were significantly reduced ( ** p < 0.01) compared to the treatment group. This means that the rosehip extract of the present invention has an excellent ability to inhibit muscle protein degradation in muscle cells.
실험군experimental group |
상대적 MuRF-1 발현량(%)Relative MuRF-1 Expression (%) |
상대적 Atrogin-1 발현량(%)Relative Atrogin-1 Expression (%) |
TNF-αTNF-α | 375.2±41.1## 375.2±41.1 ## | 438.4±50.3## 438.4±50.3 ## |
실시예 1-2Example 1-2 | 187.0±37.1**187.0±37.1** | 196.4±38.1**196.4±38.1** |
실시예 1-3Examples 1-3 | 185.1±39.3**185.1±39.3** | 224.6±41.4**224.6±41.4** |
실시예 1-4Examples 1-4 | 155.4±46.3**155.4±46.3** | 174.8±37.3**174.8±37.3** |
실시예 1-5Examples 1-5 | 190.3±30.3**190.3±30.3** | 217.9±45.5*217.9±45.5* |
실시예 1-6Examples 1-6 | 173.2±49.2**173.2±49.2** | 226.4±39.4**226.4±39.4** |
실시예 1-7Examples 1-7 | 150.3±41.5**150.3±41.5** | 188.4±49.1**188.4±49.1** |
[실시예 7] 동물모델에서 로즈힙의 근 기능 및 근육량 증가 효과[Example 7] Effects of increasing muscle function and muscle mass of rose hips in animal models
<7-1> 동물의 사육 및 근위축 유도<7-1> Animal rearing and muscle atrophy induction
실험동물로 생후 7주된 수컷쥐(C57BL/6J; DBL, Korea)를 구입하여 실험을 진행하였다. 모든 동물의 사육실 환경은 온도 23 ± 2℃, 상대습도 55 ± 10%로 유지시켰다. 실험 시작 전, 총 20마리의 쥐를 무작위로 1군당 5마리가 되도록 나누었다. 1주일간 적응시킨 후, 325 mg/kg의 트리브로모메탄올(tribromoethanol, Sigma-Aldrich)을 복강 주사하여 마취를 유도하였다. 마취 후, 근위축군과 시료 투여군에 있는 쥐의 오른쪽 뒷다리(hindlimb) 장딴지근육(gastrocnemius muscle)과 오른쪽 발바닥을 피부 스테이플러(skin stapler)(Unidus, Chungcheongbuk-do, Korea)를 사용하여 스테이플러 심으로 근육을 손상시켜 오른쪽 뒷다리가 움직이지 못하게 하였으며, 이 상태를 일주일 간 유지시켰다. 일주일 후, 장딴지근과 발바닥에 고정되어 있던 스테이플러 심을 제거하고, 다시 일주일간 로즈힙 건조 분쇄물 500 mg/kg; 실시예 1-1의 로즈힙 열수 추출물 200 mg/kg; 실시예 1-2의 로즈힙 에탄올 추출물 200 mg/kg; 실시예 1-7의 로즈힙 초임계 추출물 200 mg/kg을 매일 7일간 경구투여하였다. 이 때, 정상군과 근위축군은 시료 대신에 식염수로 경구투여를 실시하였다.As experimental animals, 7-week-old male mice (C57BL/6J; DBL, Korea) were purchased and the experiment was carried out. The breeding room environment of all animals was maintained at a temperature of 23 ± 2 °C and a relative humidity of 55 ± 10%. Before the start of the experiment, a total of 20 rats were randomly divided into 5 rats per group. After adaptation for 1 week, anesthesia was induced by intraperitoneal injection of 325 mg/kg of tribromoethanol (Sigma-Aldrich). After anesthesia, the right hindlimb gastrocnemius muscle and right sole of the rats in the muscle atrophy group and the sample administration group were cut with a stapler seam using a skin stapler (Unidus, Chungcheongbuk-do, Korea). The damage resulted in immobilization of the right hind leg, which was maintained for a week. After one week, the stapler shim fixed to the calf muscles and the soles of the feet was removed, and dried rosehips 500 mg/kg for another week; 200 mg/kg of hot water extract of rosehip of Example 1-1; 200 mg/kg of rosehip ethanol extract of Example 1-2; 200 mg/kg of the rosehip supercritical extract of Example 1-7 was orally administered daily for 7 days. At this time, the normal group and the muscular atrophy group were orally administered with saline instead of the sample.
<7-2> 로즈힙의 근력 향상 효과 확인<7-2> Confirmation of the muscle strength improvement effect of rose hips
경구 투여 기간이 끝나고, 근력측정기(Panlab, Barcelona, Spain)를 이용하여 쥐의 근력을 측정하였다. 쥐가 근력측정기의 막내를 놓을 때까지 일정한 힘으로 쥐의 꼬리를 당겼으며, 한 마리당 총 5회 연속 테스트를 실시하였다.After the oral administration period was over, the muscle strength of the rats was measured using a muscle strength meter (Panlab, Barcelona, Spain). The rat's tail was pulled with a constant force until the rat released the youngest member of the muscle strength meter, and a total of five consecutive tests were performed per rat.
그 결과, 표 3에 나타낸 바와 같이 정상군에 비해 근위축군에서 근력이 유의적 (#
p < 0.05)으로 감소하였으나, 로즈힙 분쇄물 및 로즈힙 추출물을 처리했을 때 근위축군에 비해 근력이 유의적(**
p < 0.01)으로 증가한 것을 확인하였다. 이는 본 발명의 로즈힙 분쇄물 및 로즈힙 추출물이 근위축으로 인해 감소된 근력을 증가시키는 효과가 우수하다는 것을 의미한다.As a result, as shown in Table 3, muscle strength was significantly ( # p < 0.05) decreased in the muscle atrophy group compared to the normal group, but when treated with the crushed rosehip powder and rosehip extract, the muscle strength was significantly reduced compared to the muscle atrophy group ( * * p < 0.01) was confirmed. This means that the pulverized rosehip and the rosehip extract of the present invention are excellent in the effect of increasing muscle strength reduced due to muscle atrophy.
실험군experimental group | 근력(g)strength (g) |
정상군 normal group | 263.8±8.4263.8±8.4 |
근위축군muscle atrophy | 194.2±7.2## 194.2±7.2 ## |
로즈힙 분쇄물Rosehip Grind | 232.6±6.4**232.6±6.4** |
로즈힙 열수 추출물Rosehip Hot Water Extract | 235.2±7.9**235.2±7.9** |
로즈힙 에탄올 추출물Rosehip Ethanol Extract | 240.1±8.7**240.1±8.7** |
로즈힙 초임계 추출물Rosehip Supercritical Extract | 252.8±9.5**252.8±9.5** |
<7-3> 로즈힙의 근육 부피 증가 효과<7-3> The effect of increasing muscle volume of rose hips
희생하기 전, 마우스를 isoflurane으로 호흡 마취하고 positron emission tomography/computed tomography/single photon emission tomography (microPET/CT/SPECT; Siemens Inveon, Knoxville, TN, USA)를 이용하여 오른쪽 뒷다리 근육의 부피 및 밀도를 측정하였다.Before sacrifice, the mice were respiratory anesthetized with isoflurane and the volume and density of the right hind limb muscle were measured using positron emission tomography/computed tomography/single photon emission tomography (microPET/CT/SPECT; Siemens Inveon, Knoxville, TN, USA). did
그 결과, 도 4에 나타낸 바와 같이 정상군에 비해 근위축군의 근육 부피가 유의적(##
p < 0.01)으로 감소하였으나, 로즈힙 분쇄물 및 로즈힙 추출물을 처리했을 때 근위축군에 비해 근위축군에 비해 근육 부피가 유의적(**
p < 0.01)으로 증가한 것을 확인하였다. 이는 본 발명의 로즈힙 분쇄물 및 로즈힙 추출물이 근위축으로 인해 감소된 근육의 부피를 증가시키는 효과가 우수하다는 것을 의미한다.As a result, as shown in FIG. 4 , the muscle volume of the muscle atrophy group was significantly reduced ( ## p < 0.01) compared to the normal group. It was confirmed that the muscle volume increased significantly ( ** p < 0.01). This means that the pulverized rosehip and the rosehip extract of the present invention are excellent in increasing the muscle volume reduced due to muscle atrophy.
<7-3> 로즈힙의 근육 무게 증가 효과 확인<7-3> Confirmation of the effect of increasing muscle weight of rose hips
근력 측정이 끝난 후, 실험동물을 325 mg/kg의 트리브로모메탄올(Sigma-Aldrich)을 복강 주사하여 마취 후 심채혈을 통해 희생시켰다. 심장박동이 멈춘 것을 확인한 뒤, 오른쪽 뒷다리에서 손상을 입지 않은 전경골근(tibialis anterior muscle)을 적출하여 무게를 측정하였다.After measuring the muscle strength, the experimental animals were sacrificed through deep blood sampling after anesthesia by intraperitoneal injection of 325 mg/kg of tribromethanol (Sigma-Aldrich). After confirming that the heartbeat has stopped, the uninjured tibialis anterior muscle was removed from the right hind leg and the weight was measured.
그 결과, 표 4에 나타낸 바와 같이 정상군에 비해 근위축군의 전경골근의 무게가 유의적(##
p < 0.01)으로 감소하였으나, 로즈힙 분쇄물 및 로즈힙 추출물을 처리했을 때 근위축군에 비해 무게가 유의적(**
p < 0.01)으로 증가한 것을 확인하였다. 이는 본 발명의 로즈힙 분쇄물 및 로즈힙 추출물이 근위축으로 인해 감소된 근육의 무게를 증가시키는 효과가 우수하다는 것을 의미한다.As a result, as shown in Table 4, the weight of the tibialis anterior muscle in the muscular atrophy group was significantly ( ## p < 0.01) decreased compared to the normal group, but when treated with the crushed rosehip and rosehip extract, the weight was increased compared to the muscle atrophy group. It was confirmed that it increased significantly ( ** p < 0.01). This means that the pulverized rosehip and the rosehip extract of the present invention are excellent in increasing the muscle weight reduced due to muscle atrophy.
실험군experimental group | 근육무게(mg)muscle weight (mg) |
정상군 normal group | 60.2±3.660.2±3.6 |
근위축군muscle atrophy | 45.6±3.4## 45.6±3.4 ## |
로즈힙 분쇄물Rosehip Grind | 51.9±3.5**51.9±3.5** |
로즈힙 열수 추출물Rosehip Hot Water Extract | 53.0±4.1**53.0±4.1** |
로즈힙 에탄올 추출물Rosehip Ethanol Extract | 55.4±3.8**55.4±3.8** |
로즈힙 초임계 추출물Rosehip Supercritical Extract | 58.0±4.3**58.0±4.3** |
<7-4> 동물모델에서 로즈힙 열수 추출물의 근 단백질 합성 생체지표 발현량 증가 효과<7-4> Effect of increasing expression level of muscle protein synthesis biomarkers of hot water extract of rosehip in animal model
실시예 7-3에서 적출한 전경골근에 단백질 가수분해효소 억제제 칵테일(Sigma-Aldrich St. Louis, MO, USA)이 포함된 NP-40 완충용액(ELPIS-Biotech, Daejeon, Korea)으로 조직을 분쇄하여 근 조직 용해물을 수득한 후, 13,000 rpm으로 10분간 원심분리하여 상등액을 취하였다. 다음, 상등액을 이용하여 상기 실시예 4와 동일한 방법으로 근 단백질 합성 생체지표 발현량을 측정하였다. The tissue was crushed with NP-40 buffer (ELPIS-Biotech, Daejeon, Korea) containing a protease inhibitor cocktail (Sigma-Aldrich St. Louis, MO, USA) to the tibialis anterior muscle extracted in Example 7-3. After obtaining a muscle tissue lysate, centrifuged at 13,000 rpm for 10 minutes to obtain a supernatant. Next, the expression level of the muscle protein synthesis biomarker was measured in the same manner as in Example 4 using the supernatant.
그 결과, 도 5에 나타난 바와 같이, 부동화 동물모델에서 부동화에 의해 p-mTOR, p-p70S6K 및 p-4EBP-1의 단백질 발현 수준이 감소한 반면, 로즈힙 열수 추출물을 처리함에 따라 p-mTOR, p-p70S6K 및 p-4EBP-1의 단백질 발현 수준이 증가한 것을 확인할 수 있었다. 이는 본 발명의 로즈힙 열수 추출물이 동물모델에서 근단백질 합성에 관여하는 생체지표의 발현을 증가시키는 능력이 우수한 것을 의미한다.As a result, as shown in FIG. 5, protein expression levels of p-mTOR, p-p70S6K and p-4EBP-1 were decreased by immobilization in the immobilized animal model, whereas p-mTOR, p It was confirmed that the protein expression levels of -p70S6K and p-4EBP-1 were increased. This means that the hot water extract of rosehip of the present invention has excellent ability to increase the expression of biomarkers involved in muscle protein synthesis in animal models.
이하, 본 발명에 따른 로즈힙 분쇄물 또는 로즈힙 추출물을 유효성분으로 함유하는 의약품, 식품 또는 화장품의 제조예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다. 상기 근육 질환 개선, 치료 또는 예방, 또는 근 기능 개선 효과가 우수한 로즈힙 분쇄물 또는 로즈힙 추출물을 이용하여 하기와 같은 조성성분 및 조성비에 따라 제조예 1 내지 3의 의약품, 식품 또는 화장료 조성물을 통상적인 방법에 따라서 제조하였다.Hereinafter, a manufacturing example of a pharmaceutical, food or cosmetic containing the pulverized rosehip product or rosehip extract according to the present invention as an active ingredient will be described, but the present invention is not intended to limit the present invention, but merely to describe it in detail. The pharmaceutical, food or cosmetic composition of Preparation Examples 1 to 3 according to the following compositional components and composition ratio using the pulverized rosehip or rosehip extract having excellent effect of improving, treating or preventing muscle disease, or improving muscle function in a conventional method prepared according to
[제조예 1] 의약품[Production Example 1] Pharmaceuticals
<1-1> 산제<1-1> Powder
본 발명의 로즈힙 추출물 50 mg, 결정셀룰로오즈 2 g을 혼합한 후 통상의 산제 제조방법에 따라서 기밀포에 충진하여 산제를 제조하였다.After mixing 50 mg of the rosehip extract of the present invention and 2 g of crystalline cellulose, it was filled in an airtight bag according to a conventional powder preparation method to prepare a powder.
<1-2> 정제<1-2> Tablet
본 발명의 로즈힙 추출물 50 mg, 결정셀룰로오즈 400 mg, 스테아린산 마그네슘 5 mg을 혼합한 후 통상의 정제 제조방법에 따라서 타정하여 정제를 제조하였다.After mixing 50 mg of the rosehip extract of the present invention, 400 mg of crystalline cellulose, and 5 mg of magnesium stearate, tablets were prepared by tableting according to a conventional tablet manufacturing method.
<1-3> 캡슐제<1-3> Capsules
본 발명의 로즈힙 추출물 30 mg, 유청단백질 100 mg, 결정셀룰로오즈 400 mg, 스테아린산 마그네슘 6 mg을 혼합한 후 통상의 캡슐제 제조방법에 따라서 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.After mixing 30 mg of the rosehip extract of the present invention, 100 mg of whey protein, 400 mg of crystalline cellulose, and 6 mg of magnesium stearate, the capsules were prepared by filling in gelatin capsules according to a conventional capsule preparation method.
[제조예 2] 식품[Production Example 2] Food
<2-1> 건강식품의 제조<2-1> Manufacturing of health food
본 발명의 로즈힙 분쇄물 또는 로즈힙 추출물 1000 mg, 비타민 A 아세테이트 70 ug, 비타민 E 1.0 mg, 비타민 B1 0.13 mg, 비타민 B2 0.15 mg, 비타민 B6 0.5 mg, 비타민 B12 0.2 ug, 비타민 C 10 mg, 비오틴 10 ug, 니코틴산아미드 1.7 mg, 엽산 50 ug, 판토텐산 칼슘 0.5 mg, 황산제1철 1.75 mg, 산화아연 0.82 mg, 탄산마그네슘 25.3 mg, 제1인산칼륨 15 mg, 제2인산칼슘 55 mg, 구연산칼륨 90 mg, 탄산칼슘 100 mg, 염화마그네슘 24.8 mg를 혼합하여 제조할 수 있으며, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 조성물 제조에 사용할 수 있다.1000 mg of crushed rosehip or rosehip extract of the present invention, vitamin A acetate 70 ug, vitamin E 1.0 mg, vitamin B1 0.13 mg, vitamin B2 0.15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 ug, vitamin C 10 mg, biotin 10 ug, nicotinic acid amide 1.7 mg, folic acid 50 ug, calcium pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monobasic 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, and magnesium chloride 24.8 mg can be mixed to prepare the mixture, and the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to the usual health food manufacturing method, and then granules are prepared, , it can be used to prepare a health food composition according to a conventional method.
<2-2> 건강음료의 제조<2-2> Manufacturing of health drinks
본 발명의 로즈힙 추출물 1000 mg, 구연산 1000 mg, 올리고당 100 g, 매실농축액 2 g, 타우린 1 g에 정제수를 가하여 전체 900 mL 통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간동안 85에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 L용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 건강음료 조성물 제조에 사용할 수 있다.After adding purified water to 1000 mg of rosehip extract of the present invention, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, and 1 g of taurine, the above ingredients are mixed according to the general method for preparing a total 900 mL health drink, and then about 1 hour After stirring and heating at 85 for a while, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and then stored in a refrigerator, and then it can be used in the manufacture of health beverage compositions.
<2-3> 츄잉껌<2-3> Chewing Gum
껌 베이스 20 중량%, 설탕 76.9 중량%, 향료 1 중량% 및 물 2 중량%와 본 발명의 로즈힙 추출물 0.1 중량%를 배합하여 통상의 방법으로 츄잉껌을 제조하였다.Chewing gum was prepared in a conventional manner by mixing 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of flavoring and 2% by weight of water and 0.1% by weight of rosehip extract of the present invention.
<2-4> 캔디<2-4> Candy
설탕 60 중량%, 물엿 39.8 중량% 및 향료 0.1 중량%와 본 발명의 로즈힙 추출물 0.1 중량%를 배합하여 통상의 방법으로 캔디를 제조하였다. Sugar 60% by weight, starch syrup 39.8% by weight, and flavoring 0.1% by weight and 0.1% by weight of the rosehip extract of the present invention were mixed to prepare a candy in a conventional manner.
<2-5> 비스켓<2-5> Biscuit
박력 1급 25.59 중량%, 중력 1급 22.22 중량%, 정백당 4.80 중량%, 식염 0.73 중량%, 포도당 0.78 중량%, 팜쇼트닝 11.78 중량%, 암모늄 1.54 중량%, 중조 0.17 중량%, 중아황산나트륨 0.16 중량%, 쌀가루 1.45 중량%, 비타민 B 0.0001 중량%, 밀크향 0.04 중량%, 물 20.6998 중량%, 전지분유 1.16 중량%, 대용분유 0.29 중량%, 제1인산칼슘 0.03 중량%, 살포염 0.29 중량% 및 분무유 7.27 중량%와 본 발명의 로즈힙 분쇄물 또는 로즈힙 추출물 0.8301 중량%를 배합하여 통상의 방법으로 비스켓을 제조하였다.1st class strength 25.59 wt%, 1st class gravity 22.22 wt%, refined sugar 4.80 wt%, salt 0.73 wt%, glucose 0.78 wt%, palm shortening 11.78 wt%, ammonium 1.54 wt%, sodium bicarbonate 0.17 wt%, sodium bisulfite 0.16 wt% , rice flour 1.45% by weight, vitamin B 0.0001% by weight, milk flavor 0.04% by weight, water 20.6998% by weight, whole milk powder 1.16% by weight, powder substitute 0.29% by weight, monobasic calcium phosphate 0.03% by weight, dusting salt 0.29% by weight and spray A biscuit was prepared in a conventional manner by mixing 7.27 wt% of oil and 0.8301 wt% of the pulverized rosehip or rosehip extract of the present invention.
[제조예 3] 화장품[Production Example 3] Cosmetics
<3-1> 영양화장수(밀크로션)<3-1> Nutrient lotion (milk lotion)
본 발명의 로즈힙 추출물을 하기 표 3의 영양화장수 제형 비율대로 하여 통상적인 방법에 따라 영양화장수를 제조하였다. Nutrient lotion was prepared according to a conventional method by using the rosehip extract of the present invention according to the formulation ratio of the nutrient lotion in Table 3 below.
배합성분 Ingredients |
제조예3-1 (중량%)Preparation Example 3-1 (weight%) |
로즈힙 추출물rosehip extract | 2.02.0 |
스쿠알란squalane | 5.05.0 |
밀납beeswax | 4.04.0 |
폴리솔베이트 60Polysorbate 60 | 1.51.5 |
솔비탄세스퀴올레이트Sorbitan sesquioleate | 1.51.5 |
유동파라핀liquid paraffin | 0.50.5 |
카프릴릭/카프릭트리글리세라이드Caprylic/Capric Triglycerides | 5.05.0 |
글리세린glycerin | 3.03.0 |
부틸렌글리콜butylene glycol | 3.03.0 |
프로필렌글리콜propylene glycol | 3.03.0 |
카르복시비닐폴리머Carboxyvinyl Polymer | 0.10.1 |
트리에탄올아민triethanolamine | 0.20.2 |
방부제, 색소, 향료Preservatives, Colors, Flavors | 적량appropriate amount |
정제수Purified water | to 100to 100 |
<3-2> 유연화장수(스킨로션)본 발명의 로즈힙 추출물을 하기 표 4의 유연화장수 제형 비율대로 하여 통상적인 방법에 따라 유연화장수를 제조하였다.<3-2> Softening lotion (skin lotion) A softening lotion was prepared according to a conventional method by using the rosehip extract of the present invention according to the ratio of the softening lotion formulation in Table 4 below.
배합성분 Ingredients |
제조예3-2 (중량%)Preparation 3-2 (weight%) |
로즈힙 추출물rosehip extract | 2.02.0 |
글리세린glycerin | 3.03.0 |
부틸렌글리콜butylene glycol | 2.02.0 |
프로필렌글리콜propylene glycol | 2.02.0 |
카르복시비닐폴리머Carboxyvinyl Polymer | 0.10.1 |
PEG 12 노닐페닐에테르PEG 12 nonylphenyl ether | 0.20.2 |
폴리솔베이트 80Polysorbate 80 | 0.40.4 |
에탄올ethanol | 10.010.0 |
트리에탄올아민triethanolamine | 0.10.1 |
방부제, 색소, 향료Preservatives, Colors, Flavors | 적량appropriate amount |
정제수Purified water | to 100to 100 |
<3-3> 영양크림본 발명의 로즈힙 추출물을 하기 표 5의 영양크림 제형 비율대로 하여 통상적인 방법에 따라 영양크림을 제조하였다.<3-3> Nutrient Cream A nourishing cream was prepared according to a conventional method by using the rosehip extract of the present invention according to the formulation ratio of the nourishing cream in Table 5 below.
배합성분 Ingredients |
제조예3-3 (중량%)Production Example 3-3 (weight%) |
로즈힙 추출물rosehip extract | 2.02.0 |
폴리솔베이트 60Polysorbate 60 | 1.51.5 |
솔비탄세스퀴올레이트Sorbitan sesquioleate | 0.50.5 |
PEG60 경화피마자유PEG60 hydrogenated castor oil | 2.02.0 |
유동파라핀liquid paraffin | 1010 |
스쿠알란squalane | 5.05.0 |
카프릴릭/카프릭트리글리세라이드Caprylic/Capric Triglycerides | 5.05.0 |
글리세린glycerin | 5.05.0 |
부틸렌글리콜butylene glycol | 3.03.0 |
프로필렌글리콜propylene glycol | 3.03.0 |
트리에탄올아민triethanolamine | 0.20.2 |
방부제antiseptic | 적량appropriate amount |
색소pigment | 적량appropriate amount |
향료Spices | 적량appropriate amount |
정제수Purified water | to 100to 100 |
<3-4> 마사지크림본 발명의 로즈힙 분쇄물 또는 로즈힙 추출물을 하기 표 6의 마사지크림 제형 비율대로 하여 통상적인 방법에 따라 마사지크림을 제조하였다.<3-4> Massage Cream A massage cream was prepared according to a conventional method by using the crushed rosehip powder or rosehip extract of the present invention according to the massage cream formulation ratio shown in Table 6 below.
배합성분 Ingredients |
제조예3-4 (중량%)Preparation 3-4 (weight%) |
로즈힙 분쇄물 또는 로즈힙 추출물Rosehip Grind or Rosehip Extract | 1.01.0 |
밀납beeswax | 10.010.0 |
폴리솔베이트 60Polysorbate 60 | 1.51.5 |
PEG 60 경화피마자유 |
2.02.0 |
솔비탄세스퀴올레이트Sorbitan sesquioleate | 0.80.8 |
유동파라핀liquid paraffin | 40.040.0 |
스쿠알란squalane | 5.05.0 |
카프릴릭/카프릭트리글리세라이드Caprylic/Capric Triglycerides | 4.04.0 |
글리세린glycerin | 5.05.0 |
부틸렌글리콜butylene glycol | 3.03.0 |
프로필렌글리콜propylene glycol | 3.03.0 |
트리에탄올아민triethanolamine | 0.20.2 |
방부제, 색소, 향료Preservatives, Colors, Flavors | 적량appropriate amount |
정제수Purified water | to 100to 100 |
<3-5> 팩본 발명의 로즈힙 분쇄물 또는 로즈힙 추출물을 하기 표 7의 팩 제형 비율대로 하여 통상적인 방법에 따라 팩을 제조하였다. <3-5> Pack A pack was prepared according to a conventional method by using the pulverized rosehip powder or rosehip extract of the present invention according to the pack formulation ratio in Table 7 below.
배합성분 Ingredients |
제조예3-5 (중량%)Production Example 3-5 (weight%) |
로즈힙 분쇄물 또는 로즈힙 추출물Rosehip Grind or Rosehip Extract | 1.01.0 |
폴리비닐알콜polyvinyl alcohol | 13.013.0 |
소듐카르복시메틸셀룰로오스Sodium Carboxymethyl Cellulose | 0.20.2 |
글리세린glycerin | 5.05.0 |
알란토인allantoin | 0.10.1 |
에탄올ethanol | 6.06.0 |
PEG 12 노닐페닐에테르PEG 12 nonylphenyl ether | 0.30.3 |
폴리솔베이트 60Polysorbate 60 | 0.30.3 |
방부제, 색소, 향료Preservatives, Colors, Flavors | 적량appropriate amount |
정제수Purified water | to 100to 100 |
<3-6> 젤본 발명의 로즈힙 추출물을 하기 표 8의 젤 제형 비율대로 하여 통상적인 방법에 따라 젤을 제조하였다.<3-6> Gel A gel was prepared according to a conventional method by using the rosehip extract of the present invention according to the gel formulation ratio of Table 8 below.
배합성분Ingredients |
제조예3-6 (중량%)Preparation 3-6 (weight%) |
로즈힙 추출물rosehip extract | 0.50.5 |
에틸렌디아민초산나트륨Sodium ethylenediamine acetate | 0.050.05 |
글리세린glycerin | 5.05.0 |
카르복시비닐폴리머Carboxyvinyl Polymer | 0.30.3 |
에탄올ethanol | 5.05.0 |
PEG 60 경화피마자유 |
0.50.5 |
트리에탄올아민triethanolamine | 0.30.3 |
방부제, 색소, 향료Preservatives, Colors, Flavors | 적량appropriate amount |
정제수Purified water | to 100to 100 |
상기 표에서 '적량'은 기호도에 따라 적정량이 첨가될 수 있음을 의미하고, 'to 100'은 총 100중량% 에서 정제수를 제외한 나머지 배합성분의 중량%의 합을 제한 후 남은 중량%를 정제수의 중량%로 할 수 있다는 것을 의미한다.In the above table, 'appropriate amount' means that an appropriate amount can be added according to preference, and 'to 100' is the weight% remaining after subtracting the sum of the weight% of the remaining ingredients except for purified water from 100% by weight of purified water. It means that it can be done in weight %.
이상 살펴본 바와 같이, 본 발명은 로즈힙 분쇄물 또는 로즈힙 추출물과 같은 로즈힙(rose hip)을 함유하는 근육 질환 개선, 치료 또는 예방용, 또는 근 기능 개선용 조성물을 제공한다. 보다 상세하게는 본 발명의 로즈힙은 mTOR의 활성을 증가시키고 MuRF1과 atrogin-1의 mRNA 발현량을 감소시킴으로써 근육량과 근력을 향상시켜 근육 질환 개선, 치료 또는 예방용, 또는 근 기능 개선에 우수한 효과를 나타낸다. 따라서, 본 발명은 근육 질환 개선, 치료 또는 예방용, 또는 근 기능 개선에 탁월한 효과를 보이는 조성물을 제공할 수 있으므로 산업상 이용가능성이 높다.As described above, the present invention provides a composition for improving, treating or preventing muscle disease, or for improving muscle function, containing rose hips such as pulverized rosehips or rosehip extracts. More specifically, the rose hip of the present invention improves muscle mass and muscle strength by increasing the activity of mTOR and reducing the mRNA expression levels of MuRF1 and atrogin-1, thereby improving, treating or preventing muscle diseases, or improving muscle function. indicates. Accordingly, the present invention has high industrial applicability because it can provide a composition showing an excellent effect for improving, treating or preventing muscle disease, or improving muscle function.
Claims (8)
- 로즈힙(Rose hip)을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물.A food composition for improving or preventing muscle disease, or for improving muscle function, containing rose hip as an active ingredient.
- 제 1항에 있어서,The method of claim 1,상기 로즈힙은 로즈힙 분쇄물 또는 로즈힙 추출물인 것을 특징으로 하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물.The rosehip is a food composition for improving or preventing muscle disease, or for improving muscle function, characterized in that the rosehip is a pulverized rosehip or a rosehip extract.
- 제 2 항에 있어서,3. The method of claim 2,상기 로즈힙 추출물은 물, 탄소수 1 내지 6의 유기용매, 아임계 유체 및 초임계 유체로 이루어진 군으로부터 선택되는 하나 이상의 용매로 로즈힙을 추출하여 수득한 것을 특징으로 하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물.The rosehip extract is for improving or preventing muscle disease, characterized in that it is obtained by extracting rosehip with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid and a supercritical fluid, or A food composition for improving function.
- 제 3 항에 있어서, 4. The method of claim 3,상기 유기용매는 탄소수 1 내지 6의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌 클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane) 및 석유에테르(petroleum ether)로 이루어진 군 중에서 선택되는 하나 이상의 용매인 것을 특징으로 하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물.The organic solvent is an alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane ( hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) for improving or preventing muscle disease, characterized in that at least one solvent selected from the group consisting of, or a food composition for improving muscle function.
- 제 1 항 내지 제 4 항 중 어느 한 항에 있어서, 5. The method according to any one of claims 1 to 4,상기 근육 질환은 근감소증(sarcopenia), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 긴장감퇴증(atony), 근육 퇴화, 근무력증 및 악액질(cachexia)로 이루어진 군에서 선택되는 하나 이상인 것을 특징으로 하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 식품 조성물.The muscle disease is characterized in that at least one selected from the group consisting of sarcopenia, muscular atrophy, muscular dystrophy, atony, muscle degeneration, myasthenia gravis and cachexia. A food composition for improving or preventing muscle disease, or for improving muscle function.
- 로즈힙을 유효성분으로 함유하는 근육 질환 치료 또는 예방용 약학 조성물.A pharmaceutical composition for treating or preventing muscle disease containing rosehip as an active ingredient.
- 로즈힙을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 화장료 조성물.A cosmetic composition for improving or preventing muscle disease, or for improving muscle function, containing rosehip as an active ingredient.
- 로즈힙을 유효성분으로 함유하는 근육 질환 개선 또는 예방용, 또는 근 기능 개선용 사료 첨가제.Feed additive for improving or preventing muscle disease or improving muscle function containing rosehip as an active ingredient.
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110097427A1 (en) * | 2009-10-27 | 2011-04-28 | Shyam Ramakrishnan | Molecular Targets and Dietary Modulators of Exercise-Induced Muscle Damage |
WO2014005597A1 (en) * | 2012-07-06 | 2014-01-09 | Axellus A/S | Rose hip composition, process for the manufacture of the rose hip composition and said composition for use in a method for the maintenance of flexible joints and decreased inflammation |
JP2019517470A (en) * | 2016-05-27 | 2019-06-24 | スマートフィッシュ・アーエス | Use of a composition comprising marine oil and juice to improve muscle performance |
KR20200071363A (en) * | 2018-12-11 | 2020-06-19 | 주식회사 엘리바이오 | Composition for Cartilage Damage Prevention and Regeneration Containing Rosa Canina Extract |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110097427A1 (en) * | 2009-10-27 | 2011-04-28 | Shyam Ramakrishnan | Molecular Targets and Dietary Modulators of Exercise-Induced Muscle Damage |
WO2014005597A1 (en) * | 2012-07-06 | 2014-01-09 | Axellus A/S | Rose hip composition, process for the manufacture of the rose hip composition and said composition for use in a method for the maintenance of flexible joints and decreased inflammation |
JP2019517470A (en) * | 2016-05-27 | 2019-06-24 | スマートフィッシュ・アーエス | Use of a composition comprising marine oil and juice to improve muscle performance |
KR20200071363A (en) * | 2018-12-11 | 2020-06-19 | 주식회사 엘리바이오 | Composition for Cartilage Damage Prevention and Regeneration Containing Rosa Canina Extract |
Non-Patent Citations (1)
Title |
---|
CHRUBASIK COSIMA ET AL: "A systematic review on the Rosa canina effect and efficacy profiles.", PHYSIOTHERAPY RESEARCH, JOHN WILEY & SONS LTD. CHICHESTER., GB, vol. 22, no. 6, 1 June 2008 (2008-06-01), GB , pages 725 - 733, XP002490524, ISSN: 0951-418X, DOI: 10.1002/ptr.2400 * |
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