WO2020139057A2 - Composition for preventing and treating muscle disease or improving muscle function, comprising illicium verum extract or shikimic acid - Google Patents

Composition for preventing and treating muscle disease or improving muscle function, comprising illicium verum extract or shikimic acid Download PDF

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Publication number
WO2020139057A2
WO2020139057A2 PCT/KR2019/018726 KR2019018726W WO2020139057A2 WO 2020139057 A2 WO2020139057 A2 WO 2020139057A2 KR 2019018726 W KR2019018726 W KR 2019018726W WO 2020139057 A2 WO2020139057 A2 WO 2020139057A2
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Prior art keywords
muscle
extract
octagonal
acid
octagonal fennel
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PCT/KR2019/018726
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French (fr)
Korean (ko)
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WO2020139057A3 (en
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황재관
김창희
유지희
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연세대학교 산학협력단
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Priority claimed from KR1020190076670A external-priority patent/KR20200083146A/en
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Publication of WO2020139057A2 publication Critical patent/WO2020139057A2/en
Publication of WO2020139057A3 publication Critical patent/WO2020139057A3/en

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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)
    • A61K36/79—Schisandraceae (Schisandra family)
    • 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
    • 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]
    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/316—Foods, ingredients or supplements having a functional effect on health having an effect on regeneration or building of ligaments or muscles

Definitions

  • the present invention relates to a composition for preventing and treating muscle disease or improving muscle function, comprising an octagonal fennel extract or shikimic acid.
  • Muscle atrophy is caused by a gradual decrease in muscle mass and refers to muscle weakness and degeneration (Cell, 119(7): 907-910, 2004). Muscle atrophy is promoted by inactivity, oxidative stress, and chronic inflammation and weakens muscle function and motor capacity (Clinical Nutrition, 26(5): 524-534, 2007). The most important factor in determining muscle function is muscle mass, which is maintained by a balance of protein synthesis and degradation. Muscular dystrophy occurs when proteolysis 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 anabolic or catabolism in the muscle, and when there are more signaling reactions that induce synthesis rather than degradation of muscle proteins Muscle protein synthesis is increased, which results in an increase in muscle size (hypertrophy) or an increase in muscle fiber number (hyperplasia) as muscle protein increases (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 gain 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 cell differentiation and muscle formation are regulated by a variety of muscle regulatory factors. Among them, MyoD initiates the expression of muscle-specific genes, and induces differentiation of muscle satellite cells into myoblasts. Induction of myogenin expression by the activity of MyoD is the most important factor in fusion of myoblasts and is involved in the formation of myotubes. The muscle fibers formed through this process form a bundle and finally form muscles (Cellular and Molecular Life Sciences, 70: 4117-4130, 2013).
  • the octagonal fennel ( Illiumium verum ) refers to the fruits of magnolia and evergreen, and is called star anise because eight pods wrapped in a hard shell are shaped like a star. It is widely used as a spice, mainly after drying and in powder form.
  • Anti-cancer Journal of Medicinal Plants Research, 4(24): 2666-2673. 2016
  • Antioxidant Food Chemistry, 104(1): 59-66, 2007
  • Anti-inflammatory activity (Journal of Ethnopharmacology, 144(1): 182-189, 2012) has been reported.
  • Shikimic acid is one of the components included in octagonal fennel, and is an important biochemical intermediate involved in the biosynthesis of alkaloids, flavonoids, and lignin, as well as amino acids.
  • Sikkim acid is an antithrombotic (Journal of Natural Medicine, 70(3): 492-501, 2016), antibacterial (Bioengineered, 4(4): 244-248, 2013), neuroprotection (Journal of Molecular Neuroscience, 56(4) : 956-965, 2015), antiviral (Journal of Agricultural and Food Chemistry, 52(17): 5391-5395, 2004).
  • the present invention provides a muscle disease prevention, for the improvement or treatment compositions comprising octagonal anise (Illicium verum) extract or fractions thereof as an active ingredient.
  • octagonal anise Illicium verum
  • the present invention provides a composition for preventing, improving or treating muscle diseases comprising shikimic acid represented by Formula 1 below or a salt thereof as an active ingredient.
  • Illicium verum extract or shikimic acid increases mTOR activity involved in muscle protein synthesis, decreases mRNA expression of MuRF-1 and atrogin-1 involved in muscle protein degradation, increases muscle mass And it has an excellent effect on improving muscle function.
  • the present invention since the present invention is a natural product, it can be safely used without side effects, and thus can be used as a pharmaceutical, food, or cosmetic material.
  • 1 is a result of measuring the activity of mTOR according to the treatment of octagonal fennel extract in L6 muscle cells.
  • 3 is a result of measuring mRNA expression levels of atrogin-1 and MuRF-1 according to treatment of octagonal fennel hot water extract, 50% ethanol extract, and ethanol extract in L6 muscle cells.
  • the present invention provides a octagonal fennel (Illicium verum) extracts or fractions muscle disorders or for the prevention and treatment of muscle function improving composition comprising as an active ingredient thereof.
  • the present invention provides a composition for preventing and treating muscle disease or improving muscle function, including shikimic acid or a salt thereof as an active ingredient.
  • the present inventors tried to develop a natural substance that improves muscle function, and found that the octagonal fennel extract or its active ingredient, shikimic acid, has an effect of preventing or treating muscle diseases by increasing the expression and activity of genes involved in muscle function. Did.
  • composition of the present invention may include an octagonal fennel extract as an active ingredient.
  • The'octagonal fennel' used in the composition of the present invention belongs to the family Illiciaceae, of course, including its subspecies and varieties.
  • the octagonal fennel of the present invention is self-evident in the art and includes a homogenous drug that can be used for the same or similar purposes as the prevention and treatment purposes of the present invention.
  • extract' means that a substance is dissolved in a solvent to separate its active ingredient or characteristic ingredient.
  • the extract includes an extract extracted by adding an extractant to a plant and a fraction fractionated by adding a fractional solvent to the extract prepared by extracting with an extractant.
  • the octagonal fennel extract of the present invention is meant to include both the extract extracted by adding a solvent to the octagonal fennel and the fractions fractionated by adding a fractional solvent thereto.
  • the octagonal fennel extract contains octagonal fennel water, an organic solvent having 1 to 6 carbon atoms, or a mixture thereof, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether, subcritical fluid and supercritical It may be extracted with any one or more solvents selected from the group consisting of fluids.
  • the organic solvent alcohol having 1 to 6 carbon atoms (acecohol), acetone (acetone), ether (ether), benzene (benzene), chloroform (chloroform), ethyl acetate (ethyl acetate), methylene chloride (methylene chloride), hexane It may include at least one solvent selected from the group consisting of (hexane), cyclohexane and petroleum ether.
  • the present invention may include an extraction method of the octagonal fennel extract and an octagonal extract extracted from the extraction method.
  • composition of the present invention may contain sikkim acid as an active ingredient.
  • the "shimkimic acid” is a compound extracted or purified from natural products or synthesized, and is represented by the following Chemical Formula 1.
  • the shikimic acid may be derived from octagonal fennel. At this time, it may be prepared by a method of layer separation by adding a solvent to the octagonal fennel extract.
  • the shikimic acid of the present invention may include hydrates, derivatives, and the like within a range having the same efficacy, and may also include solvent compounds or stereoisomers thereof.
  • the method for obtaining the shikimic acid is not particularly limited, and a chemically synthesized or commercially available product may be used using a known manufacturing method.
  • the octagonal fennel extract can be obtained by extracting octagonal fennel under ultra high pressure conditions of 100 MPa or more.
  • muscle refers to the tendon, muscle, tendon, and'muscle function' refers to the ability to exert power by contraction of the muscle, and the muscle exerts the greatest contractile force to overcome resistance. Includes muscle power, the ability to perform, muscle endurance, the ability to indicate how long or how many times a muscle can repeat contractions and relaxations at a given weight, and wits, the ability to exert strong power in a short period of time.
  • This muscle function is subject to the liver and is proportional to muscle mass.
  • the term'muscle function improvement' refers to improving muscle function better.
  • the octagonal fennel extract or sickimic acid which is an active ingredient according to the present invention, has an excellent ability to increase the synthesis of muscle protein from increasing the activity of mTOR in muscle cells.
  • the octagonal fennel extract or shikimic acid which is an active ingredient according to the present invention, has an excellent ability to increase muscle from increasing anterior muscle weight and improving muscle strength in musculoskeletal induced mice.
  • the composition of the present invention may be a pharmaceutical composition for preventing and treating muscle disease.
  • prophylaxis refers to all actions that inhibit or delay the onset of the administration of the composition.
  • “improvement” or “treatment” refers to all actions in which the symptoms of the disease are improved or beneficially altered by administration of the composition.
  • the composition of the present invention can be used for the prevention or treatment of muscle diseases caused by muscle wasting or degeneration.
  • Muscle wasting and degeneration are caused by genetic factors, acquired factors, aging, and the like, and muscle wasting is characterized by gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or veterinary muscles and cardiac muscles.
  • muscle diseases due to muscle wasting or degeneration are not limited thereto, but atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and muscular dystrophy ( sarcopenia), and these diseases can be used to treat or prevent various diseases in which natural recovery of muscles is difficult.
  • the composition of the present invention has an effect of increasing muscle mass, and the muscle does not limit its type.
  • the pharmaceutical composition of the present invention may include shikimic acid or a pharmaceutically acceptable salt thereof.
  • pharmaceutically acceptable means physiologically acceptable and does not generally cause an allergic reaction or similar reaction when administered to a human being, and the salt may include a pharmaceutically acceptable free acid (free). acid) is preferred.
  • the above-mentioned acid addition salts are a) general salts that directly mix sickimic acid, or b) dissolve and mix it in a solvent or hydrous solvent, or c) place skimic acid in an acid in a solvent or a water-based solvent and mix them. It is manufactured by the manufacturing method.
  • possible salt forms include gaba salt, gabapentin salt, pregabalin salt, nicotinate salt, adipate salt, hemimalonate salt, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt or And aspartates.
  • composition for preventing and treating muscle diseases of the present invention may further include a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers 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 include water, suitable oils, saline, aqueous glucose and glycols, and the like.
  • stabilizers and preservatives may be further included. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-parabens and chlorobutanol.
  • the pharmaceutical composition of the present invention can be administered by any method to mammals, including humans.
  • it can be administered orally or parenterally, and the parenteral administration method is not limited thereto, but intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal , Intranasal, intestinal, 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 e.g. saline or PBS
  • antioxidants e.g. saline or PBS
  • bacteriostatic agents e.g. EDTA or glutathione
  • fillers e.g. EDTA or glutathione
  • extenders e.g. EDTA or glutathione
  • binders e.g. aluminum hydroxide
  • adjuvants e.g. aluminum hydroxide
  • suspension thickener wetting agent, disintegrant or surfactant, diluent or excipient.
  • Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions or capsules, etc. These solid preparations include at least one excipient in the pharmaceutical composition of the present invention. , 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 hydroxypropyl methyl-cellulose or gelatin, and the like. For example, tablets or tablets of sugar can be obtained by mixing the active ingredient with a solid excipient and then grinding it and adding a suitable adjuvant to the granular mixture.
  • Liquid preparations for oral use include suspensions, intravenous solutions, emulsions or syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances or preservatives, in addition to water or liquid paraffin, a simple diluent commonly used. .
  • cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and may further include an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, and a preservative. .
  • compositions of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations and nasal inhalants, along with suitable parenteral carriers.
  • the injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi.
  • suitable carriers include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and/or vegetable oils.
  • solvents or dispersion media including 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, phosphate buffered saline (PBS) containing triethanol amine or sterile water for injection, isotonic solutions such as 10% ethanol, 40% propylene glycol and 5% dextrose. Etc. can be used.
  • PBS phosphate buffered saline
  • isotonic solutions such as 10% ethanol, 40% propylene glycol and 5% dextrose. Etc. can be used.
  • it may further include various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • the injection may further include isotonic agents such as sugars or sodium chloride in most cases.
  • the compounds used in accordance with the present invention may be pressurized packs or, using suitable propellants, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered in the form of an aerosol spray from a nebulizer.
  • the dosage unit can be determined by providing a valve that delivers a metered amount.
  • gelatin capsules and cartridges used in inhalers or insufflators can be formulated to contain a powder mixture of a 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 prescription generally known to all pharmaceutical chemistries.
  • the pharmaceutical composition for preventing and treating muscle diseases of the present invention can provide a desirable muscle disease prevention and treatment effect when an octagonal fennel extract or shikimic acid is included in an effective amount.
  • the term'effective amount' refers to an amount that exhibits a higher response than a negative control, and preferably refers to an amount sufficient to improve muscle function.
  • the pharmaceutical composition of the present invention may include octagonal fennel extract or shikimic acid in an amount of 0.01 to 99.99%, and the remaining amount may be occupied by a pharmaceutically acceptable carrier.
  • the effective amount of the octagonal fennel extract or shikimic acid included 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 can be administered to a patient in a single dose, and can be administered by a fractionated treatment protocol that is administered for a long time in multiple doses.
  • the pharmaceutical composition of the present invention can vary the content of the active ingredient according to the degree of 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 kg of body weight per day based on the octagonal extract or shikimic acid, and when administered orally, the octagonal extract or It may be administered in divided portions of 1 to several times to be administered in an amount of preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per kg of body weight per day, based on sikimic acid.
  • the pharmaceutical composition for preventing and treating muscle diseases of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormonal therapy, chemotherapy, or biological response modifiers.
  • composition for preventing and treating muscle diseases or improving muscle function of the present invention may contain octagonal fennel extract or shikimic acid alone.
  • additional components may further enhance the muscle function improving effect of the composition of the present invention.
  • the pharmaceutical composition of the present invention may be provided as a formulation of an external preparation containing octagonal fennel extract or shikimic acid as an active ingredient.
  • the pharmaceutical composition for preventing and treating muscle diseases of the present invention is provided as an external preparation for skin, it is not limited thereto, and may be a formulation such as ointment, patch, gel, cream or spray.
  • the composition of the present invention may be prepared as a food composition for preventing muscle disease or improving muscle function.
  • the food composition for preventing muscle disease and improving muscle function of the present invention is a food composition, it can be used to prevent or improve muscle disease due to muscle wasting or degeneration.
  • Muscle wasting and degeneration are caused by genetic factors, acquired factors, aging, and the like, and muscle wasting is characterized by gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or veterinary muscles and cardiac muscles. Examples of related diseases include atony, muscle atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and sarcopenia.
  • the composition of the present invention has an effect of increasing muscle mass, and the muscle does not limit its type.
  • the food composition of the present invention includes all forms of functional foods, nutritional supplements, health foods, food additives and feeds, and includes animals including humans or livestock. Subject to eating. 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.
  • Beverages including alcoholic beverages
  • fruits and processed foods thereof e.g. canned fruits, bottled foods, jams, marmalades, etc.
  • fish meat and processed foods
  • breads and noodles e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.
  • juice various drinks
  • cookies syrup
  • dairy products e.g. butter, cheese, etc.
  • edible plant maintenance margarine
  • Plant protein, retort food, frozen food, various seasonings eg, miso, soy sauce, sauce, etc.
  • seasonings eg, miso, soy sauce, sauce, etc.
  • the nutritional supplement it is not limited to the nutritional supplement, but may be prepared by adding octagonal fennel extract to capsules, tablets, pills, and the like.
  • the health functional food is not limited to this, for example, the octagonal fennel extract itself is prepared in the form of tea, juice, and drink to be consumed by liquefaction, granulation, encapsulation, and powdering for drinking (health drink). Can.
  • the octagonal fennel extract in order to use the octagonal fennel extract as a food additive, it can be prepared and used in the form of a powder or a concentrate.
  • the octagonal fennel extract can be prepared in the form of a composition by mixing with a known active ingredient known to have an effect of preventing muscle disease and improving muscle function.
  • the food composition of the present invention may be added to a feed additive or feed composition for the purpose of preventing muscle disease or improving muscle function.
  • the feed additive of the present invention corresponds to supplementary feeds under the feed management method, and mineral preparations such as sodium hydrogen carbonate, bentonite, magnesium oxide, and complex minerals, mineral preparations such as zinc, copper, cobalt, and selenium, and keratin , Vitamin A, D, E, nicotinic acid, vitamins such as vitamin B complex, protective amino acids such as methionine and lysine, protective fatty acids such as fatty acid calcium salts, probiotics (lactic acid bacteria), yeast culture, mold fermentation, etc. Probiotics, yeasts, etc. may be further included.
  • mineral preparations such as sodium hydrogen carbonate, bentonite, magnesium oxide, and complex minerals, mineral preparations such as zinc, copper, cobalt, and selenium, and keratin , Vitamin A, D, E, nicotinic acid, vitamins such as vitamin B complex, protective amino acids such as methionine and lysine, protective fatty acids such as fatty acid calcium salts, probiotics (lactic acid bacteria
  • feed in the present invention, any natural or artificial diet for the animal to eat, eat, digest or suitable for this, or a component of the meal, or the composition of the meal, bone formation promoting composition according to the present invention
  • Feed containing as an active ingredient can be prepared in various forms of feed known in the art, and preferably may include a rich feed, forage and/or special feed, but is not limited thereto.
  • Thick feed contains seeds, wheat, oats, corn and other grains, and by-products obtained by refining and harvesting grains such as rice bran, wheat bran, barley bran, etc., rice, soybean, fluid, sesame, flaxseed, coco palm, etc.
  • Fish-soluble (fish fish) which is a concentrate of fresh liquid obtained from fish, fish meal, fish residues, fish residues, etc. soluble), meat powder, blood powder, milk powder, skim milk powder, cheese from milk, whey, which is the balance when manufacturing casein from skim milk, and animal feed materials such as dry whey, yeast, chlorella, and algae, but are not limited thereto. Does not.
  • the feed additive according to the present invention can be prepared by adding octagonal fennel extract or sickimic acid in an appropriate effective concentration range according to various methods for preparing feeds known in the art.
  • the excipients include, but are not limited to, for example, skin emollients, skin penetration enhancers, colorants, fragrances, emulsifiers, thickening agents and solvents.
  • fragrances, pigments, fungicides, antioxidants, preservatives and moisturizers may be further included, and for the purpose of improving physical properties, may include thickeners, inorganic salts, synthetic polymer materials, and the like.
  • it can be easily prepared by adding an octagonal fennel extract to a conventional cleanser and soap base.
  • the use of the octagonal fennel extract or the composition containing shikimic acid represented by the formula (1) as an active ingredient may be provided for the prevention or treatment of muscle disease.
  • L6 myoblasts (ATCC; Manassas, VA, USA) 1 ⁇ in a 6-well plate with Dulbecco's modified Eagle's media (DMEM; Hyclone) with 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) After seeding to be 10 5 cells/well, the cells were cultured for 24 hours. After incubation, the medium in the well was removed and exchanged with DMEM (Hyclone) containing 2% horse serum (HS; Hyclone) for 6 days to incubate L6 cells to differentiate into myotubes.
  • DMEM Dulbecco's modified Eagle's media
  • FBS Hyclone
  • FBS fetal bovine serum
  • each of the octagonal fennel extract prepared in Example 1 was dissolved in DMEM (Hyclone) at 10 ⁇ g/mL, and then treated with cells and cultured for 12 hours. After incubation, cells were lysed by treatment with a cell lysis buffer. The protein in the obtained cell lysate was quantified at a concentration of 1 mg/mL using Bradford (Bio-Rad Laboratories Inc., Hercules, CA, USA). Cell lysate was dispensed in 50 ⁇ L of each microwell attached with anti-mTOR antibody and left at 37°C for 2 hours. After washing a total of 4 times with a washing buffer, treated with a detection antibody, and allowed to stand for 1 hour at 37°C.
  • DMEM Hyclone
  • the anti-rabbit secondary antibody (Bethyl Laboratories, Inc., Montgomery, TA, USA) conjugated with horseradish peroxidase recognizing the primary antibody was diluted to 1:5000 in 2.5% BSA with nitrocellulose membrane 2 The mixture was reacted at room temperature for an hour, and washed three times with 10 minutes using TBST. Protein bands were developed using ECL western blotting detection reagents (Amersham, Tokyo, Japan), and protein bands developed using a G;BOX EF imaging system (Syngene, Cambridge, UK) were identified. The results are shown in FIG. 2.
  • the muscle cell L6 myoblast was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) to 1 ⁇ 10 5 cells/mL.
  • DMEM Hyclone
  • FBS Hyclone
  • RNA isolated was quantified using a nanodrop (NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, MA, USA). Quantitative 16 ⁇ L of RNA using Reverse Transcriptase Premix (ELPIS-Biotech) and 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 with cDNA under conditions.
  • Reverse primer 5'-CCGTATGAGTCTTATGTTTTGCTGG-3' (SEQ ID NO: 2)
  • Reverse primer 5'-AGCCTGGAAGATGTCGTTGG-3' (SEQ ID NO: 4)
  • Reverse primer 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6)
  • the amplified cDNA was separated by electrophoresis with a 1.5% agarose gel, and the cDNA band was confirmed using a G;BOX EF imaging system (Syngene). The results are shown in FIG. 3.
  • Reverse primer 5'-GGTGTAACAACCATACCCCACT-3' (SEQ ID NO: 8)
  • Reverse primer 5'-GGGAGTTTGCGTTCCTCTGA-3'
  • the amplified cDNA was separated by electrophoresis with a 1.5% agarose gel, and the cDNA band was confirmed using a G;BOX EF imaging system (Syngene). The results are shown in FIG. 4.
  • the muscle strength of the rats was measured using a muscle strength meter (Panlab, Barcelona, Spain).
  • the rat's tail was pulled with constant force until the rat placed the youngest of the muscular strength tester, and a total of 5 consecutive tests were performed per head.
  • the octagonal hot water extract of the present invention has an excellent effect of increasing the weight of muscles reduced due to muscular atrophy.
  • muscle strength was significantly decreased ( ## p ⁇ 0.01) in the atrophy group compared to the normal group, but muscle strength was significantly increased ( ** p ⁇ 0.01) as treated with shikimic acid. was confirmed.
  • a powder was prepared by filling in an airtight fabric according to a conventional powder manufacturing method.
  • tablets After mixing the octagonal fennel extract or 50 mg of sikkim acid, 400 mg of crystalline cellulose, and 5 mg of magnesium stearate, tablets were prepared by tableting according to a conventional tablet manufacturing method.
  • the present invention relates to a composition for preventing and treating muscle disease or improving muscle function, comprising an octagonal fennel extract and shikimic acid.
  • the octagonal fennel extract and shikimic acid according to the present invention have 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, increasing muscle mass and muscle function It is useful for preventing, treating or improving various muscle diseases such as atony, muscle atrophy, muscle dystrophy, muscle degeneration, myasthenia, cachexia, and sarcopenia. As it can be used, it has industrial applicability.

Abstract

The present invention relates to a composition for preventing and treating a muscle disease or improving muscle function, comprising an Illicium verum extract and shikimic acid. The Illicium verum extract and shikimic acid according to the present invention have the effect of increasing 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 increase muscle mass and enhance muscle function, thereby being useful for preventing, treating, or alleviating various muscle diseases such as atony, muscular atrophy, muscular dystrophy, muscular degeneration, myasthenia, cachexia, and sarcopenia.

Description

팔각회향 추출물 또는 시킴산을 함유하는 근육 질환 예방 및 치료용 또는 근 기능 개선용 조성물Composition for preventing and treating muscle disease or improving muscle function, containing octagonal fennel extract or shikimic acid
본 발명은 팔각회향 추출물 또는 시킴산을 함유하는 근육 질환 예방 및 치료용 또는 근 기능 개선용 조성물에 관한 것이다.The present invention relates to a composition for preventing and treating muscle disease or improving muscle function, comprising an octagonal fennel extract or shikimic acid.
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). Muscle atrophy is promoted by inactivity, oxidative stress, and chronic inflammation and weakens muscle function and motor capacity (Clinical Nutrition, 26(5): 524-534, 2007). The most important factor in determining muscle function is muscle mass, which is maintained by a balance of protein synthesis and degradation. Muscular dystrophy occurs when proteolysis 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 조절되며 근 단백질의 분해보다 합성을 유도하는 신호전달 반응이 많이 일어날 경우 근 단백질 합성이 증가되는데, 이는 근 단백질 증가에 따른 근육 크기 증가(hypertrophy, 근비대)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 anabolic or catabolism in the muscle, and when there are more signaling reactions that induce synthesis rather than degradation of muscle proteins Muscle protein synthesis is increased, which results in an increase in muscle size (hypertrophy) or an increase in muscle fiber number (hyperplasia) as muscle protein increases (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). 반대로 전사 인자(transcription factor)인 forhead box (FoxO)가 세포질에서 핵 내로 이동하면 단백질 분해에 관여하는 E3 ubiquitin ligase인자 atrogin-1과 MuRF-1의 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 MuRF-1 발현 억제는 근육 단백질의 양을 증가시켜 근육량을 증가시키게 된다. Factors involved in muscle protein synthesis induce protein synthesis by phosphorylating downstream proteins starting from stimulation of the phosphatidylinositol-3 kinase (PI3K)/Akt pathway in muscle cells. The activity of 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 gain 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). Conversely, when the transcription factor forhead box (FoxO) moves from the cytoplasm into the nucleus, the expression of the E3 ubiquitin ligase factors atrogin-1 and MuRF-1, which are involved in protein degradation, increases (Disease Models and Mechanisms, 6: 25). -39, 2013). When these expression levels increase, protein breakdown in muscles is promoted, and muscle mass decreases. Therefore, promoting mTOR activity and suppressing atrogin-1 and MuRF-1 expression increases muscle protein and increases muscle mass.
근육세포의 분화와 근육 형성은 다양한 근육 조절 인자(muscle regulatory factors)에 의해 조절된다. 그 중, MyoD는 근육 특이 유전자의 발현을 개시하게 하고, 근육위성세포(muscle satellite cells)가 근원세포(myoblast)로 분화(differentiation)되는 것을 유도한다. 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).Muscle cell differentiation and muscle formation are regulated by a variety of muscle regulatory factors. Among them, MyoD initiates the expression of muscle-specific genes, and induces differentiation of muscle satellite cells into myoblasts. Induction of myogenin expression by the activity of MyoD is the most important factor in fusion of myoblasts and is involved in the formation of myotubes. The muscle fibers formed through this process form a bundle and finally form muscles (Cellular and Molecular Life Sciences, 70: 4117-4130, 2013).
팔각회향(Illicium verum)은 목련과 상록수의 열매를 말하는 것으로, 단단한 껍질로 싸인 꼬투리 여덟 개가 마치 별처럼 붙어있는 모양을 하고 있어 일반적으로 스타아니스(star anise)라고 불린다. 주로 건조한 후 분말형태로 만들어 향신료로 널리 사용되고 있으며, 항암(Journal of Medicinal Plants Research, 4(24): 2666-2673. 2016), 항산화(Food Chemistry, 104(1): 59-66, 2007), 항염(Journal of Ethnopharmacology, 144(1): 182-189, 2012) 등의 활성이 보고되어 있다. The octagonal fennel ( Illiumium verum ) refers to the fruits of magnolia and evergreen, and is called star anise because eight pods wrapped in a hard shell are shaped like a star. It is widely used as a spice, mainly after drying and in powder form. Anti-cancer (Journal of Medicinal Plants Research, 4(24): 2666-2673. 2016), Antioxidant (Food Chemistry, 104(1): 59-66, 2007), Anti-inflammatory activity (Journal of Ethnopharmacology, 144(1): 182-189, 2012) has been reported.
시킴산(Shikimic acid)은 팔각회향에 포함되어 있는 성분 중 하나로 알칼로이드, 플라보노이드, 리그닌 뿐만 아니라 아미노산의 생합성에 관여하는 중요한 생화학적 중간물질이다. 시킴산은 항혈전(Journal of Natural Medicine, 70(3): 492-501, 2016), 항균(Bioengineered, 4(4): 244-248, 2013), 신경보호(Journal of Molecular Neuroscience, 56(4): 956-965, 2015), 항바이러스(Journal of Agricultural and Food Chemistry, 52(17): 5391-5395, 2004) 등의 활성을 갖는 것으로 보고되어 있다.Shikimic acid is one of the components included in octagonal fennel, and is an important biochemical intermediate involved in the biosynthesis of alkaloids, flavonoids, and lignin, as well as amino acids. Sikkim acid is an antithrombotic (Journal of Natural Medicine, 70(3): 492-501, 2016), antibacterial (Bioengineered, 4(4): 244-248, 2013), neuroprotection (Journal of Molecular Neuroscience, 56(4) : 956-965, 2015), antiviral (Journal of Agricultural and Food Chemistry, 52(17): 5391-5395, 2004).
그러나, 본 발명의 이전에는 팔각회향 추출물 또는 시킴산의 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 diseases of octagonal fennel extract or shikimic acid or improvement of muscle function.
본 발명은 부작용 없이 근 단백질 합성에 관여하는 단백질 발현 증가 및 근 단백질 분해에 관여하는 단백질 발현 억제 활성을 가짐으로써 근육 질환의 예방 및 치료 또는 근 기능을 개선할 수 있는 기술을 제공하는 것을 목적으로 한다.It is an object of the present invention to provide a technique capable of preventing and treating muscle disease or improving muscle function by having protein expression inhibitory activity involved in muscle protein degradation and protein expression involved in muscle protein synthesis without side effects. .
그러나, 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.
상기 과제를 해결하기 위한 수단으로서, 본 발명은 팔각회향(Illicium verum) 추출물 또는 이의 분획물을 유효성분으로 포함하는 근육 질환 예방, 개선 또는 치료용 조성물을 제공한다.As a means for solving the above problems, the present invention provides a muscle disease prevention, for the improvement or treatment compositions comprising octagonal anise (Illicium verum) extract or fractions thereof as an active ingredient.
다른 측면에서, 본 발명은 하기 화학식 1로 표시되는 시킴산(shikimic acid) 또는 이의 염을 유효성분으로 포함하는 근육 질환 예방, 개선 또는 치료용 조성물을 제공한다.In another aspect, the present invention provides a composition for preventing, improving or treating muscle diseases comprising shikimic acid represented by Formula 1 below or a salt thereof as an active ingredient.
[화학식 1][Formula 1]
Figure PCTKR2019018726-appb-I000001
Figure PCTKR2019018726-appb-I000001
본 발명에 따른 팔각회향(Illicium verum) 추출물 또는 시킴산(shikimic acid)은 근 단백질 합성에 관여하는 mTOR의 활성 증가, 근 단백질 분해에 관여하는 MuRF-1과 atrogin-1의 mRNA 발현 감소, 근육량 증가 및 근기능 향상에 우수한 효과가 있다. 또한, 본 발명은 천연물이므로 부작용 없이 안전하게 사용될 수 있어, 의약품, 식품 또는 화장품 소재로 사용될 수 있다. Illicium verum extract or shikimic acid according to the present invention increases mTOR activity involved in muscle protein synthesis, decreases mRNA expression of MuRF-1 and atrogin-1 involved in muscle protein degradation, increases muscle mass And it has an excellent effect on improving muscle function. In addition, since the present invention is a natural product, it can be safely used without side effects, and thus can be used as a pharmaceutical, food, or cosmetic material.
도 1은 L6 근육세포에서, 팔각회향 추출물의 처리에 따른 mTOR의 활성을 측정한 결과이다.1 is a result of measuring the activity of mTOR according to the treatment of octagonal fennel extract in L6 muscle cells.
도 2는 L6 근육세포에서, 팔각회향 열수 추출물, 50% 에탄올 추출물 및 에탄올 추출물의 처리에 따른 p-mTOR의 단백질 발현량을 측정한 결과이다.2 is a result of measuring the protein expression level of p-mTOR according to the treatment of octagonal hot water extract, 50% ethanol extract and ethanol extract in L6 muscle cells.
도 3은 L6 근육세포에서, 팔각회향 열수 추출물, 50% 에탄올 추출물 및 에탄올 추출물의 처리에 따른 atrogin-1과 MuRF-1의 mRNA 발현량을 측정한 결과이다.3 is a result of measuring mRNA expression levels of atrogin-1 and MuRF-1 according to treatment of octagonal fennel hot water extract, 50% ethanol extract, and ethanol extract in L6 muscle cells.
도 4는 L6 근육세포에서, 팔각회향 열수 추출물의 처리에 따른 미오디(MyoD), 미오게닌(myogenin) 및 MRF4의 mRNA 발현량을 측정한 결과이다.4 is a result of measuring mRNA expression levels of MioD, myogenin and MRF4 according to the treatment of octagonal hot water extract in L6 muscle cells.
도 5는 근위축 유도 쥐에서, 팔각회향 열수 추출물 처리에 따른 근력 향상 효과를 확인한 결과이다.5 is a result of confirming the effect of improving muscle strength according to the treatment of octagonal hot water extract in muscle atrophy-induced rats.
도 6은 근위축 유도 쥐에서, 팔각회향 열수 추출물 처리에 따른 전경골근 무게 증가 효과를 확인한 결과이다.6 is a result of confirming the effect of increasing the weight of the tibialis anterior muscle according to the treatment of octagonal hot water extract in the atrophy-induced rat.
도 7은 L6 근육세포에서, 시킴산 처리에 따른 mTOR의 활성을 측정한 결과이다.7 is a result of measuring the activity of mTOR according to the treatment of sikkim acid in L6 muscle cells.
도 8은 L6 근육세포에서, 시킴산 처리에 따른 atrogin-1 및 MuRF1의 mRNA 발현량을 측정한 결과이다.8 is a result of measuring mRNA expression levels of atrogin-1 and MuRF1 according to treatment with sickimic acid in L6 muscle cells.
도 9는 L6 근육세포에서, 시킴산 처리에 따른 미오디(MyoD), 미오게닌(myogenin) 및 MRF4의 mRNA 발현량을 측정한 결과이다.9 is a result of measuring the mRNA expression levels of MioD, myogenin and MRF4 according to Sikkim acid treatment in L6 muscle cells.
도 10은 근위축 유도 쥐에서, 시킴산 처리에 따른 근력 향상 효과를 확인한 결과이다.10 is a result of confirming the effect of improving muscle strength according to the treatment of shikimic acid in atrophy-induced rats.
도 11은 근위축 유도 쥐에서, 시킴산 처리에 따른 전경골근 무게 증가 효과를 확인한 결과이다.11 is a result of confirming the effect of increasing the weight of the tibialis anterior muscle according to the treatment with shikimic acid in the atrophy-induced rats.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 팔각회향(Illicium verum) 추출물 또는 이의 분획물을 유효성분으로 포함하는 근육 질환 예방 및 치료용 또는 근 기능 개선용 조성물을 제공한다.The present invention provides a octagonal fennel (Illicium verum) extracts or fractions muscle disorders or for the prevention and treatment of muscle function improving composition comprising as an active ingredient thereof.
또한, 본 발명은 시킴산(shikimic acid) 또는 이의 염을 유효성분으로 포함하는 근육 질환 예방 및 치료용 또는 근 기능 개선용 조성물을 제공한다.In addition, the present invention provides a composition for preventing and treating muscle disease or improving muscle function, including shikimic acid or a salt thereof as an active ingredient.
본 발명자들은 근 기능을 개선시키는 천연물질을 개발하고자 노력한 결과, 팔각회향 추출물 또는 이의 활성성분 시킴산이 근 기능에 관여하는 유전자의 발현 및 활성을 증가시킴으로써 근육 질환을 예방 또는 치료하는 효과가 있음을 규명하였다.The present inventors tried to develop a natural substance that improves muscle function, and found that the octagonal fennel extract or its active ingredient, shikimic acid, has an effect of preventing or treating muscle diseases by increasing the expression and activity of genes involved in muscle function. Did.
본 발명의 조성물은 팔각회향 추출물을 유효성분으로 포함할 수 있다.The composition of the present invention may include an octagonal fennel extract as an active ingredient.
본 발명의 조성물에서 이용되는 '팔각회향'은 붓순나무과(Illiciaceae)에 속하며, 이의 아종, 변종을 포함함은 물론이다. 또한, 본 발명의 팔각회향은 당업계에서 자명하고 본 발명의 예방 및 치료 목적과 동일 또는 유사한 용도로 사용할 수 있는 동속생약을 포함한다.The'octagonal fennel' used in the composition of the present invention belongs to the family Illiciaceae, of course, including its subspecies and varieties. In addition, the octagonal fennel of the present invention is self-evident in the art and includes a homogenous drug that can be used for the same or similar purposes as the prevention and treatment purposes of the present invention.
본 명세서에서, '추출물'은 어떤 물질을 용매에 녹여 그 활성성분 또는 특성 성분을 분리한 것을 의미한다. 구체적으로 식물에 추출용매를 가하여 추출한 추출물 및 추출용매로 추출하여 제조한 추출물에 분획용매를 가하여 분획한 분획물을 포함한다. 따라서, 본 발명의 팔각회향 추출물은 팔각회향에 용매를 가하여 추출한 추출물과 이에 다시 분획용매를 가하여 분획한 분획물을 모두 포함하는 의미이다.In the present specification,'extract' means that a substance is dissolved in a solvent to separate its active ingredient or characteristic ingredient. Specifically, the extract includes an extract extracted by adding an extractant to a plant and a fraction fractionated by adding a fractional solvent to the extract prepared by extracting with an extractant. Accordingly, the octagonal fennel extract of the present invention is meant to include both the extract extracted by adding a solvent to the octagonal fennel and the fractions fractionated by adding a fractional solvent thereto.
상기 팔각회향 추출물은 팔각회향을 물, 탄소수 1 내지 6의 유기용매 또는 이들의 혼합물, 아세톤, 에테르, 벤젠, 클로로포름, 에틸아세테이트, 메틸렌 클로라이드, 헥산, 시클로헥산, 석유에테르, 아임계 유체 및 초임계 유체로 이루어진 군으로부터 선택되는 어느 하나 이상의 용매로 추출한 것일 수 있다.The octagonal fennel extract contains octagonal fennel water, an organic solvent having 1 to 6 carbon atoms, or a mixture thereof, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether, subcritical fluid and supercritical It may be extracted with any one or more solvents selected from the group consisting of fluids.
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 적어도 어느 하나의 용매를 포함할 수 있다.The organic solvent, alcohol having 1 to 6 carbon atoms (acecohol), acetone (acetone), ether (ether), benzene (benzene), chloroform (chloroform), ethyl acetate (ethyl acetate), methylene chloride (methylene chloride), hexane It may include at least one solvent selected from the group consisting of (hexane), cyclohexane and petroleum ether.
본 발명은, 상기 팔각회향 추출물의 추출방법 및 상기 추출방법으로부터 추출된 팔각회향 추출물을 포함할 수 있다.The present invention may include an extraction method of the octagonal fennel extract and an octagonal extract extracted from the extraction method.
또한, 본 발명의 조성물은 유효성분으로 시킴산을 포함할 수 있다.In addition, the composition of the present invention may contain sikkim acid as an active ingredient.
상기"시킴산"은 천연물로부터 추출 및 정제되거나, 합성된 화합물로서, 하기 화학식 1로 표시된다.The "shimkimic acid" is a compound extracted or purified from natural products or synthesized, and is represented by the following Chemical Formula 1.
[화학식 1][Formula 1]
Figure PCTKR2019018726-appb-I000002
Figure PCTKR2019018726-appb-I000002
상기 시킴산은 팔각회향으로부터 유래한 것일 수 있다. 이때, 상기 팔각회향 추출물에 용매를 가하여 층분리하는 방법으로 제조될 수 있다.The shikimic acid may be derived from octagonal fennel. At this time, it may be prepared by a method of layer separation by adding a solvent to the octagonal fennel extract.
본 발명의 시킴산은 동일한 효능을 갖는 범위 내에서 수화물, 유도체 등을 포함할 수 있고, 이의 용매 화합물이나 입체 이성질체 또한 포함할 수 있다.The shikimic acid of the present invention may include hydrates, derivatives, and the like within a range having the same efficacy, and may also include solvent compounds or stereoisomers thereof.
상기 시킴산의 수득방법은 특별히 한정되지 않으며, 공지된 제법을 사용하여 화학적으로 합성하거나 시판되는 것을 사용할 수 있다.The method for obtaining the shikimic acid is not particularly limited, and a chemically synthesized or commercially available product may be used using a known manufacturing method.
본 발명의 일 실시예에 따르면, 상기 팔각회향 추출물은 100 MPa 이상의 초고압 조건 하에서 팔각회향을 추출하여 수득할 수 있다.According to one embodiment of the present invention, the octagonal fennel extract can be obtained by extracting octagonal fennel under ultra high pressure conditions of 100 MPa or more.
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. 이러한 근 기능은 간이 주관하며, 근육량에 비례한다. 용어 '근 기능 개선'은 근 기능을 더 좋게 향상시키는 것을 말한다.In this specification,'muscle' refers to the tendon, muscle, tendon, and'muscle function' refers to the ability to exert power by contraction of the muscle, and the muscle exerts the greatest contractile force to overcome resistance. Includes muscle power, the ability to perform, muscle endurance, the ability to indicate how long or how many times a muscle can repeat contractions and relaxations at a given weight, and wits, the ability to exert strong power in a short period of time. This muscle function is subject to the liver and is proportional to muscle mass. The term'muscle function improvement' refers to improving muscle function better.
본 발명의 일 구현예에서는, 본 발명에 따른 유효성분인 팔각회향 추출물 또는 시킴산이 근육세포에서 mTOR의 활성을 증가시키는 것으로부터 근육 단백질의 합성을 증가시키는 능력이 우수한 것을 확인하였다.In one embodiment of the present invention, it was confirmed that the octagonal fennel extract or sickimic acid, which is an active ingredient according to the present invention, has an excellent ability to increase the synthesis of muscle protein from increasing the activity of mTOR in muscle cells.
또한, 본 발명의 다른 구현예에서는, 본 발명에 따른 유효성분인 팔각회향 추출물 또는 시킴산이 근육세포에서 atrogin-1 및 MuRF-1의 mRNA 발현을 감소시키는 것으로부터 근 단백질의 분해를 억제하는 능력이 우수한 것을 확인하였다.In addition, in another embodiment of the present invention, the ability to inhibit the degradation of muscle protein from octagonal fennel extract or sickimic acid, which is an active ingredient according to the present invention, reduces mRNA expression of atrogin-1 and MuRF-1 in muscle cells. It was confirmed that it was excellent.
또한, 본 발명의 다른 구현예에서는, 본 발명에 따른 유효성분인 팔각회향 추출물 또는 시킴산이 근육세포에서 미오디(MyoD), 미오게닌(myogenin) 및 MRF4의 mRNA 발현을 증가시키는 것으로부터 근육의 분화를 촉진시키는 능력이 우수한 것을 확인하였다.In addition, in another embodiment of the present invention, the octagonal fennel extract or sickimic acid, which is an active ingredient according to the present invention, increases muscle mRNA expression by increasing the mRNA expression of MioD, myogenin and MRF4 in muscle cells. It was confirmed that the ability to promote differentiation was excellent.
또한, 본 발명의 다른 구현예에서는, 본 발명에 따른 유효성분인 팔각회향 추출물 또는 시킴산이 근위축 유도쥐에서 전경골근 무게 증가 및 근력을 향상시키는 것으로부터 근육을 증가시키는 능력이 우수한 것을 확인하였다.In addition, in another embodiment of the present invention, it was confirmed that the octagonal fennel extract or shikimic acid, which is an active ingredient according to the present invention, has an excellent ability to increase muscle from increasing anterior muscle weight and improving muscle strength in musculoskeletal induced mice.
따라서, 이러한 측면에서 본 발명의 조성물은 근육 질환 예방 및 치료용 약학 조성물 일 수 있다.Therefore, in this aspect, the composition of the present invention may be a pharmaceutical composition for preventing and treating muscle disease.
본 발명에서, "예방"은 조성물의 투여로 발병을 억제하거나 발병을 지연시키는 모든 행위를 의미한다.In the present invention, "prophylaxis" refers to all actions that inhibit or delay the onset of the administration of the composition.
본 발명에서, "개선" 또는 "치료"는 조성물의 투여로 상기 질환의 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.In the present invention, "improvement" or "treatment" refers to all actions in which the symptoms of the disease are improved or beneficially altered by administration of the composition.
본 발명의 조성물은, 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. The composition of the present invention can be used for the prevention or treatment of muscle diseases caused by muscle wasting or degeneration. Muscle wasting and degeneration are caused by genetic factors, acquired factors, aging, and the like, and muscle wasting is characterized by gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or veterinary muscles and cardiac muscles.
본 발명에서 근육 소모 또는 퇴화로 인한 근육 질환은 이에 제한되는 것은 아니나, 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근무력증, 악액질(cachexia) 및 근육감소증(sarcopenia)을 포함하며, 이러한 질환으로 인해 근육의 자연적 회복이 어려운 다양한 질환의 치료 또는 예방에 사용될 수 있다. 본 발명의 조성물은 근육량 증대 효과가 있으며, 근육은 그 종류를 제한하지 않는다.In the present invention, muscle diseases due to muscle wasting or degeneration are not limited thereto, but atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and muscular dystrophy ( sarcopenia), and these diseases can be used to treat or prevent various diseases in which natural recovery of muscles is difficult. The composition of the present invention has an effect of increasing muscle mass, and the muscle does not limit its type.
본 발명의 약학 조성물은 시킴산 또는 이의 약제학적으로 허용 가능한 염을 포함할 수 있다. 본 명세서에서 용어 "약학적으로 허용 가능한"이란 생리학적으로 허용되고 인간에게 투여될 때, 통상적으로 알레르기 반응 또는 이와 유사한 반응을 일으키지 않는 것을 말하며, 상기 염으로는 약제학적으로 허용 가능한 유리산(free acid)에 의하여 형성된 산 부가염이 바람직하다. The pharmaceutical composition of the present invention may include shikimic acid or a pharmaceutically acceptable salt thereof. As used herein, the term "pharmaceutically acceptable" means physiologically acceptable and does not generally cause an allergic reaction or similar reaction when administered to a human being, and the salt may include a pharmaceutically acceptable free acid (free). acid) is preferred.
상기 약제학적으로 허용 가능한 염은 유기산 또는 무기산을 이용하여 형성된 산 부가염일 수 있으며, 상기 유기산은 예를 들면 포름산, 아세트산, 프로피온산, 락트산, 부티르산, 이소부티르산, 트리플루오로아세트산, 말산, 말레산, 말론산, 푸마르산, 숙신산, 숙신산 모노아미드, 글루탐산, 타르타르산, 옥살산, 시트르산, 글리콜산, 글루쿠론산, 아스코르브산, 벤조산, 프탈산, 살리실산, 안트라닐산, 디클로로아세트산, 아미노옥시 아세트산, 벤젠술폰산, p-톨루엔술폰산 또는 메탄술폰산을 포함한다. 무기산은 예를 들면 염산, 브롬산, 황산, 인산, 질산, 탄산 또는 붕산을 포함한다. 산 부가염은 바람직하게는 염산염 또는 아세트산염 형태일 수 있으며, 보다 바람직하게는 염산염 형태일 수 있다.The pharmaceutically acceptable salt may be an acid addition salt formed using an organic acid or an inorganic acid, and the organic acid may be, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, Malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid, aminooxy acetic acid, benzenesulfonic acid, p- Toluenesulfonic acid or methanesulfonic acid. Inorganic acids include, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid or boric acid. The acid addition salt may preferably be in the form of a hydrochloride or acetate, more preferably in the form of a hydrochloride.
상기 언급된 산 부가염은 a) 시킴산을 직접 혼합하거나, b) 이를 용매 또는 함수 용매 중에 용해시키고 혼합시키거나, 또는 c) 시킴산을 용매 또는 수하 용매 중의 산에 위치시키고 이들을 혼합하는 일반적인 염 제조방법으로 제조된다.The above-mentioned acid addition salts are a) general salts that directly mix sickimic acid, or b) dissolve and mix it in a solvent or hydrous solvent, or c) place skimic acid in an acid in a solvent or a water-based solvent and mix them. It is manufactured by the manufacturing method.
위와는 별도로 추가적으로 염이 가능한 형태는 가바염, 가바펜틴염, 프레가발린염, 니코틴산염, 아디페이트염, 헤미말론산염, 시스테인염, 아세틸시스테인염, 메티오닌염, 아르기닌염, 라이신염, 오르니틴염 또는 아스파르트산염 등이 있다. Apart from the above, possible salt forms include gaba salt, gabapentin salt, pregabalin salt, nicotinate salt, adipate salt, hemimalonate salt, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt or And aspartates.
또한, 본 발명의 근육 질환 예방 및 치료용 약학 조성물은 약학적으로 허용 가능한 담체를 더 포함할 수 있다.In addition, the pharmaceutical composition for preventing and treating muscle diseases of the present invention may further 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).Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. In addition, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose and glycols, and the like. In addition, stabilizers and preservatives may be further included. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-parabens and chlorobutanol. As other pharmaceutically acceptable carriers, reference may be made to those described in the following documents (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 can be administered by any method to mammals, including humans. For example, it can be administered orally or parenterally, and the parenteral administration method is not limited thereto, but intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal , Intranasal, intestinal, 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 (e.g. saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g. EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g. aluminum hydroxide) Side), suspension, thickener wetting agent, disintegrant or surfactant, diluent or excipient.
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, etc. These solid preparations include at least one excipient in the pharmaceutical composition of the present invention. , 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 hydroxypropyl methyl-cellulose or gelatin, and the like. For example, tablets or tablets of sugar can be obtained by mixing the active ingredient with a solid excipient and then grinding it and adding a suitable adjuvant to the granular 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 preparations for oral use include suspensions, intravenous solutions, emulsions or syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances or preservatives, in addition to water or liquid paraffin, a simple diluent commonly used. .
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, if necessary, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and may further include an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, and a preservative. .
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 포함하는 용매 또는 분산매질일 수 있다. 보다 바람직하게는, 적합한 담체로는 행크스 용액, 링거 용액, 트리에탄올 아민이 함유된 phosphate buffered saline (PBS) 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 compositions of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations and nasal inhalants, along with suitable parenteral carriers. The injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media including 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, phosphate buffered saline (PBS) containing triethanol amine or sterile water for injection, isotonic solutions such as 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 paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, the injection may further include isotonic agents such as sugars or sodium chloride in most cases.
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 dosage forms, ointments, creams, lotions, gels, external solutions, pasta agents, linen agents, aerosols, and the like are included. In the above,'transdermal administration' means that the pharmaceutical composition is topically administered to the skin to deliver an effective amount of the active ingredient contained in the pharmaceutical composition into 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.For inhalation dosages, the compounds used in accordance with the present invention may be pressurized packs or, using suitable propellants, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered in the form of an aerosol spray from a nebulizer. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges used in inhalers or insufflators can be formulated to contain a powder mixture of a 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 prescription generally known to all pharmaceutical chemistries.
본 발명의 근육 질환 예방 및 치료용 약학 조성물은 팔각회향 추출물 또는 시킴산을 유효량으로 포함 할 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 팔각회향 추출물 또는 시킴산이 0.01 내지 99.99% 포함될 수 있으며, 잔량은 약학적으로 허용 가능한 담체가 차지할 수 있다. 본 발명의 약학 조성물에 포함되는 팔각회향 추출물 또는 시킴산의 유효량은 조성물이 제품화되는 형태 등에 따라 달라질 것이다.The pharmaceutical composition for preventing and treating muscle diseases of the present invention can provide a desirable muscle disease prevention and treatment effect when an octagonal fennel extract or shikimic acid is included in an effective amount. In the present specification, the term'effective amount' refers to an amount that exhibits a higher response than a negative control, and preferably refers to an amount sufficient to improve muscle function. The pharmaceutical composition of the present invention may include octagonal fennel extract or shikimic acid in an amount of 0.01 to 99.99%, and the remaining amount may be occupied by a pharmaceutically acceptable carrier. The effective amount of the octagonal fennel extract or shikimic acid included 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 팔각회향 추출물 또는 시킴산을 기준으로 하루에 체중 1 kg당 바람직하게 0.01 내지 50 mg, 더 바람직하게는 0.1 내지 30 mg의 양으로 투여되도록, 그리고 경구 투여시는 팔각회향 추출물 또는 시킴산을 기준으로 하루에 체중 1 kg당 바람직하게 0.01 내지 100 mg, 더 바람직하게는 0.01 내지 10 mg의 양으로 투여되도록 1 내지 수회에 나누어 투여할 수 있다. 그러나 상기 팔각회향 추출물 또는 시킴산의 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 팔각회향 추출물 또는 시킴산을 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 can be administered to a patient in a single dose, and can be administered by a fractionated treatment protocol that is administered for a long time in multiple doses. The pharmaceutical composition of the present invention can vary the content of the active ingredient according to the degree of 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 kg of body weight per day based on the octagonal extract or shikimic acid, and when administered orally, the octagonal extract or It may be administered in divided portions of 1 to several times to be administered in an amount of preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per kg of body weight per day, based on sikimic acid. However, the dose of the octagonal fennel extract or shikimic acid is effective for the patient considering various factors such as the patient's age, weight, health status, sex, disease severity, diet and excretion rate, as well as the administration route and number of treatments of the pharmaceutical composition. Since the dosage is determined, a person having ordinary skill in the art in consideration of this point will be able to determine the appropriate effective dosage according to the specific use of the octagonal extract or sickimic acid for the prevention and treatment of muscle diseases. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, route of administration and method of administration as long as it shows the effects of the present invention.
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 diseases of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormonal therapy, chemotherapy, or biological response modifiers.
본 발명의 근육 질환 예방 및 치료용 또는 근 기능 개선용 조성물은 팔각회향 추출물 또는 시킴산을 단독으로 함유할 수 있다. 추가적인 성분을 포함하게 되면 본 발명의 조성물의 근 기능 개선 효과가 더욱 증진될 수 있을 것이다. 상기 성분 추가 시에는 복합 사용에 따른 피부 안전성, 제형화의 용이성, 유효성분들의 안정성을 고려할 수 있다.The composition for preventing and treating muscle diseases or improving muscle function of the present invention may contain octagonal fennel extract or shikimic acid alone. The inclusion of additional components may further enhance the muscle function improving effect of the composition of the present invention. When adding the above components, skin safety, ease of formulation, and stability of active ingredients according to the combined use may be considered.
다른 측면에서, 본 발명의 약학 조성물은 팔각회향 추출물 또는 시킴산을 유효성분으로 포함하는 외용제의 제형으로 제공할 수 있다.In another aspect, the pharmaceutical composition of the present invention may be provided as a formulation of an external preparation containing octagonal fennel extract or shikimic acid 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 preventing and treating muscle diseases of the present invention is used as an external preparation for skin, fat substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents ), fragrance, surfactant, water, ionic emulsifier, nonionic emulsifier, filler, metal ion blocker, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic active agent, lipophilic active agent Or it may contain an adjuvant commonly used in the field of skin science, such as any other ingredients commonly used in external preparations for skin such as lipid vesicles. In addition, the ingredients may be introduced in an amount commonly used in the field of skin science.
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 diseases of the present invention is provided as an external preparation for skin, it is not limited thereto, and may be a formulation such as ointment, patch, gel, cream or spray.
다른 측면에서, 본 발명의 조성물은 근육 질환 예방 또는 근 기능 개선용식품 조성물로 제조될 수 있다. 본 발명의 근육 질환 예방 및 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 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근무력증, 악액질(cachexia) 및 근육감소증(sarcopenia) 등을 들 수 있다. 본 발명의 조성물은 근육량 증대 효과가 있으며, 근육은 그 종류를 제한하지 않는다.In another aspect, the composition of the present invention may be prepared as a food composition for preventing muscle disease or improving muscle function. When the food composition for preventing muscle disease and improving muscle function of the present invention is a food composition, it can be used to prevent or improve muscle disease due to muscle wasting or degeneration. Muscle wasting and degeneration are caused by genetic factors, acquired factors, aging, and the like, and muscle wasting is characterized by gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or veterinary muscles and cardiac muscles. Examples of related diseases include atony, muscle atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and sarcopenia. The composition of the present invention has an effect of increasing muscle mass, and the muscle does not limit its type.
본 발명의 식품 조성물은 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health 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 of functional foods, nutritional supplements, health foods, food additives and feeds, and includes animals including humans or livestock. Subject to eating. 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 팔각회향 추출물을 첨가하여 제조할 수 있다. 또한, 영양보조제로는 이에 한정되지 않지만 캡슐, 타블렛, 환 등에 팔각회향 추출물을 첨가하여 제조할 수 있다. 또한, 건강기능식품으로는 이에 한정되지 않지만 예를 들면, 팔각회향 추출물 자체를 차, 쥬스 및 드링크의 형태로 제조하여 음용(건강음료)할 수 있도록 액상화, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다. 또한, 팔각회향 추출물을 식품 첨가제의 형태로 사용하기 위해서는 분말 또는 농축액 형태로 제조하여 사용할 수 있다. 또한, 팔각회향 추출물과 근육 질환 예방 및 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. Beverages (including alcoholic beverages), fruits and processed foods thereof (e.g. canned fruits, bottled foods, jams, marmalades, etc.), fish, meat and processed foods (e.g. ham, sausages) Corn beef, etc.), breads and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), juice, various drinks, cookies, syrup, dairy products (e.g. butter, cheese, etc.), edible plant maintenance, margarine , Plant protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding octagonal extract. In addition, it is not limited to the nutritional supplement, but may be prepared by adding octagonal fennel extract to capsules, tablets, pills, and the like. In addition, the health functional food is not limited to this, for example, the octagonal fennel extract itself is prepared in the form of tea, juice, and drink to be consumed by liquefaction, granulation, encapsulation, and powdering for drinking (health drink). Can. In addition, in order to use the octagonal fennel extract as a food additive, it can be prepared and used in the form of a powder or a concentrate. In addition, the octagonal fennel extract can be prepared in the form of a composition by mixing with a known active ingredient known to have an effect of preventing muscle disease and improving muscle function.
본 발명의 근육 질환 예방 및 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; 말토스, 슈크로스와 같은 디사카라이드; 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드; 자일리톨, 소르비톨, 에리트리톨 등의 당알콜일 수 있다. 감미제는 타우마틴, 스테비아 추출물과 같은 천연 감미제; 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 mL 당 일반적으로 약 0.01 ~0.04 g, 바람직하게는 약 0.02 ~0.03 g이다.When the composition for preventing muscle disease and 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, etc., as additional ingredients, as in conventional beverages. The above-described natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin and cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.
팔각회향 추출물 또는 시킴산은 근육 질환 예방 및 근 기능 개선용 식품 조성물의 유효성분으로 함유될 수 있는데, 그 양은 근육 질환 예방 및 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 팔각회향 추출물과 함께 근육 질환 예방 및 근 기능 개선용 조성물에 효과가 있는 것으로 알려진 다른 활성 성분과 함께 혼합하여 제조될 수 있다.The octagonal fennel extract or shikimic acid may be contained as an active ingredient in a food composition for preventing muscle disease and improving muscle function, but the amount is not particularly limited to an amount effective to achieve an effect for preventing muscle disease and improving muscle function, It is preferred that it is 0.01 to 100% by weight relative to the total weight of the composition. The food composition of the present invention can be prepared by mixing with an octagonal fennel extract along with other active ingredients known to be effective in the composition for preventing muscle disease and improving muscle function.
상기 외에 본 발명의 건강식품은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산, 펙트산의 염, 알긴산, 알긴산의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올 또는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강식품은 천연 과일주스, 과일주스 음료, 또는 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01 ~ 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acids, alginic acids, salts of alginic acids, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, Glycerin, alcohol or carbonic acid. In addition, the health food of the present invention may contain natural fruit juice, fruit juice beverage, or flesh for the production of vegetable beverages. These ingredients may be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
본 발명의 식품 조성물은 근육 질환 예방 또는 근 기능 개선을 목적으로 사료첨가제 또는 사료 조성물에 첨가할 수 있다.The food composition of the present invention may be added to a feed additive or feed composition for the purpose of preventing muscle disease or improving muscle function.
본 발명에서 용어, "사료첨가제"는 영양소 보충 및 체중감소 예방, 사료 내 섬유소의 소화 이용성 증진, 유질 개선, 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.In the present invention, the term, "feed additive" is a substance added for the purpose of various effects, such as supplementing nutrients and preventing weight loss, improving digestibility of fiber in feed, improving oil quality, preventing reproductive disorders and improving fertility rates, and preventing high-temperature stress in summer. It includes. The feed additive of the present invention corresponds to supplementary feeds under the feed management method, and mineral preparations such as sodium hydrogen carbonate, bentonite, magnesium oxide, and complex minerals, mineral preparations such as zinc, copper, cobalt, and selenium, and keratin , Vitamin A, D, E, nicotinic acid, vitamins such as vitamin B complex, protective amino acids such as methionine and lysine, protective fatty acids such as fatty acid calcium salts, probiotics (lactic acid bacteria), yeast culture, mold fermentation, etc. Probiotics, yeasts, etc. 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 골 형성 촉진용 조성물을 유효성분으로 포함하는 사료는 당업계에 공지된 다양한 형태의 사료로 제조가능하며, 바람직하게는 농후 사료, 조사료 및/또는 특수사료가 포함될 수 있으나, 이로 한정되지 않는다.The term "feed" in the present invention, any natural or artificial diet for the animal to eat, eat, digest or suitable for this, or a component of the meal, or the composition of the meal, bone formation promoting composition according to the present invention Feed containing as an active ingredient can be prepared in various forms of feed known in the art, and preferably may include a rich feed, forage and/or special feed, 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, 혈분, 우모분, 탈지분유, 우유에서 치즈, 탈지유에서 카제인을 제조할 때의 잔액인 훼이(whey)를 건조한 건조훼이 등의 동물질사료, 효모, 클로렐라, 해조류가 있으나 이에 제한되지 않는다.Thick feed contains seeds, wheat, oats, corn and other grains, and by-products obtained by refining and harvesting grains such as rice bran, wheat bran, barley bran, etc., rice, soybean, fluid, sesame, flaxseed, coco palm, etc. Fish-soluble (fish fish), which is a concentrate of fresh liquid obtained from fish, fish meal, fish residues, fish residues, etc. soluble), meat powder, blood powder, milk powder, skim milk powder, cheese from milk, whey, which is the balance when manufacturing casein from skim milk, and animal feed materials such as dry whey, yeast, chlorella, and algae, but are not limited thereto. Does not.
조사료에는 야초, 목초, 풋베기 등의 생초 사료, 사료용 순무, 사료용 비트, 순무의 일종인 루터베어거 등의 뿌리채소류, 생초, 풋베기작물, 곡실 등을 사일로에 채워 놓고 젖산발효시킨 저장사료인 사일리지(silage), 야초, 목초를 베어 건조시킨 건초, 종축용 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.Forage, forages such as wild grasses, grasses, and foot cuttings, feed turnips, feed beets, root vegetables such as Luther Bearer, a kind of turnip, raw grass, green grass crops, grain rooms, etc., are filled with silos and stored as lactic acid fermentation. Silage, silage, hay that has been cut with grasses, straw for breeding crops, and leaves of legumes, but is not limited thereto. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivatives such as diureid isobutane, and supplements with ingredients that are difficult to lack when only natural feed raw materials are blended, or added to feeds to increase feed storage. There are, but are not limited to, feed additives and dietary supplements that are added in trace amounts.
본 발명에 따른 상기 사료 첨가제는 당업계에 공지된 다양한 사료 제조방법에 따라 적절한 유효 농도 범위에서 팔각회향 추출물 또는 시킴산을 첨가하여 제조 가능하다.The feed additive according to the present invention can be prepared by adding octagonal fennel extract or sickimic acid in an appropriate effective concentration range according to various methods for preparing feeds known in the art.
본 발명에 따른 사료 첨가제는 근 기능 개선을 목적으로 하는 개체이면 제한 없이 적용가능하다. 예를 들면, 원숭이, 개, 고양이, 토끼, 모르모트, 랫트, 마우스, 소, 양, 돼지, 염소 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 object aimed at improving muscle function. For example, it can be applied to any individual such as non-human animals such as monkeys, dogs, cats, rabbits, mormots, rats, mice, cows, sheep, pigs, goats, birds and fish.
또 다른 측면에서, 본 발명의 근 기능 개선용 조성물은 또한 화장료 조성물일 수 있다. 본 발명의 화장료 조성물은 팔각회향 추출물 또는 시킴산을 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.In another aspect, the composition for improving muscle function of the present invention may also be a cosmetic composition. The cosmetic composition of the present invention contains an octagonal fennel extract or shikimic acid as an active ingredient, and a basic cosmetic composition (a face wash, cream, essence, cleansing foam and cleansing water-like cleanser, pack, and vodioyl) along with dermatologically acceptable excipients. , Color cosmetics composition (foundation, lipstick, mascara, makeup base), hair product composition (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 팔각회향 추출물을 첨가하여 용이하게 제조할 수 있다. 크림을 제조하는 경우에는 일반적인 수중유적형(O/W)의 크림베이스에 팔각회향 추출물 또는 이의 염을 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 excipients include, but are not limited to, for example, skin emollients, skin penetration enhancers, colorants, fragrances, emulsifiers, thickening agents and solvents. In addition, fragrances, pigments, fungicides, antioxidants, preservatives and moisturizers may be further included, and for the purpose of improving physical properties, may include thickeners, inorganic salts, synthetic polymer materials, and the like. For example, in the case of preparing the cleanser and soap with the cosmetic composition of the present invention, it can be easily prepared by adding an octagonal fennel extract to a conventional cleanser and soap base. When preparing a cream, it can be prepared by adding an octagonal fennel extract or a salt thereof to a cream base of a general oil-in-water type (O/W). Synthetic or natural materials such as proteins, minerals, vitamins, etc. for the purpose of improving physical properties may be additionally added to fragrances, chelating agents, pigments, antioxidants, preservatives, and the like.
본 발명의 화장료 조성물에 함유되는 팔각회향 추출물 또는 시킴산의 함량은 이에 한정되지 않지만 전체 조성물 총중량에 대하여 0.001 내지 10 중량%인 것이 바람직하고, 0.01 내지 5중량%인 것이 더욱 바람직하다. The content of the octagonal fennel extract or shikimic acid contained in the cosmetic composition of the present invention is not limited to this, but is preferably 0.001 to 10% by weight, and more preferably 0.01 to 5% by weight based on the total weight of the total composition.
본 발명의 또 다른 실시예에 따르면, 팔각회향 추출물 또는 상기 화학식 1로 표시되는 시킴산을 유효성분으로 함유하는 조성물을 개체에 투여 또는 복용시키는 단계를 포함하는 근육 질환 예방 또는 치료방법이 제공될 수 있다.According to another embodiment of the present invention, a method for preventing or treating muscle disease comprising the step of administering or taking a composition containing an octagonal fennel extract or a composition containing shikimic acid represented by Formula 1 as an active ingredient in an individual may be provided. have.
또한, 팔각회향 추출물 또는 상기 화학식 1로 표시되는 시킴산을 유효성분으로 함유하는 조성물의 근육 질환 예방 또는 치료 용도가 제공될 수 있다.In addition, the use of the octagonal fennel extract or the composition containing shikimic acid represented by the formula (1) as an active ingredient may be provided for the prevention or treatment of muscle disease.
이하 본 발명을 실시예를 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 merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.
[실시예 1] 팔각회향 추출물의 제조[Example 1] Preparation of octagonal fennel extract
<1-1> 팔각회향 열수 추출물의 제조<1-1> Preparation of octagonal fennel hot water extract
건조된 팔각회향을 믹서로 분쇄한 다음, 분쇄한 팔각회향 시료 10 g을 물 100 mL에 넣고 3시간 80℃에서 추출하였다. 추출된 시료는 와트만(Whatman) 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 팔각회향 열수 추출물을 얻었다.After crushing the dried octagonal fennel, 10 g of the crushed octagonal 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 the solvent component to obtain an octagonal fennel hot water extract.
<1-2> 팔각회향 50% 에탄올 추출물의 제조<1-2> Preparation of 50% Ethanol Extract of Octagonal Fennel
건조된 팔각회향을 믹서로 분쇄한 다음, 분쇄한 팔각회향 시료 10 g을 에탄올과 물을 중량비 50:50으로 혼합한 50% 에탄올 100 mL에 넣고 3시간 40℃에서 추출하였다. 추출된 시료는 와트만 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 팔각회향 50% 에탄올 추출물을 얻었다.After crushing the dried octagonal fennel, 10 g of the crushed octagonal sample was added to 100 mL of 50% ethanol mixed with ethanol and water at a weight ratio of 50:50 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 the solvent component to obtain an octagonal 50% ethanol extract.
<1-3> 팔각회향 에탄올 추출물의 제조<1-3> Preparation of octagonal ethanol extract
건조된 팔각회향을 믹서로 분쇄한 다음, 분쇄한 팔각회향 시료 10 g을 100% 에탄올 100 mL에 넣고 3시간 40℃에서 추출하였다. 추출된 시료는 와트만 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 팔각회향 에탄올 추출물을 얻었다.After crushing the dried octagonal fennel, 10 g of the crushed octagonal 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 the solvent component to obtain an octagonal ethanol extract.
<1-4> 팔각회향 에틸아세테이트 추출물의 제조<1-4> Preparation of octagonal ethyl acetate extract
건조된 팔각회향을 믹서로 분쇄한 다음, 분쇄한 팔각회향 시료 10 g을 에틸아세테이트 100 mL에 넣고 3시간 40℃에서 추출하였다. 추출된 시료는 와트만 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 팔각회향 에틸아세테이트 추출물을 얻었다.After crushing the dried octagonal fennel, 10 g of the crushed octagonal 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 the solvent components to obtain an octagonal ethyl acetate extract.
<1-5> 팔각회향 헥산 추출물의 제조<1-5> Preparation of octagonal hexane extract
건조된 팔각회향을 믹서로 분쇄한 다음, 분쇄한 팔각회향 시료 10 g을 헥산 100 mL에 넣고 3시간 40℃에서 추출하였다. 추출된 시료는 와트만 1번 여과지로 감압여과하고, 여과된 추출액을 진공 회전 농축기로 농축하여 용매성분을 제거한 후 팔각회향 헥산 추출물을 얻었다.After crushing the dried octagonal fennel, 10 g of the crushed octagonal 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 the solvent component to obtain an octagonal hexane extract.
<1-6> 팔각회향 아임계 추출물의 제조<1-6> Preparation of subcritical extract of octagonal fennel
건조된 팔각회향을 믹서로 분쇄한 다음, 분쇄한 팔각회향 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 팔각회향 아임계 추출물을 얻었다.After crushing the dried octagonal fennel, 50 g of the crushed octagonal fennel was placed in a subcritical water reactor of a subcritical extraction apparatus (Biovan, Gyeonggi, Korea) together 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 to extract. After 20 minutes, the extract was transferred to a storage tank supplied with cooling water, rapidly cooled to 30°C, and centrifuged at 3,600 rpm for 30 minutes to separate the floating residue, and only the supernatant was taken. Octagonal fennel subcritical extract was obtained by removing all solvents using a freeze dryer (Ilshin Lab Co. Ltd., Seoul, Korea).
<1-7> 팔각회향 초임계 추출물의 제조<1-7> Preparation of supercritical extract of octagonal fennel
건조된 팔각회향을 믹서로 분쇄한 다음, 분쇄한 팔각회향 1 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 팔각회향 초임계 추출물을 얻었다.After crushing the dried octagonal fennel, 1 g of the crushed octagonal fennel was charged into a sample cartridge and extracted using a supercritical fluid extraction device (SFX 3560, Isco Inc., Lincoln, NE, USA). For the supercritical extraction conditions, the extraction pressure was 300 bar, the extraction temperature was 50°C, the supercritical carbon dioxide flow rate was 60 mL/min, and the extraction time was 2 hours. When the supercritical fluid extraction was completed, the pressure of the extraction device was lowered to release the supercritical fluid state to obtain an octagonal supercritical extract.
[실시예 2] 팔각회향 추출물의 mTOR 활성 증가 효과[Example 2] Effect of increasing mTOR activity of octagonal fennel 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, 팔각회향의 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 muscle mass increase in the PI3K/Akt signaling pathway in muscle cells. Thus, in order to confirm the activity of octagonal muscle production, mTOR activity was confirmed using an 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 Х 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. 그런 다음, 상기 실시예 1에서 제조한 팔각회향 추출물을 각각 10 μg/mL씩 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 팔각회향 추출물을 처리한 근관 세포 내 mTOR 수준을 측정하였다. 그 결과를 도 1에 나타내었다.L6 myoblasts (ATCC; Manassas, VA, USA) 1 Х in a 6-well plate with Dulbecco's modified Eagle's media (DMEM; Hyclone) with 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) After seeding to be 10 5 cells/well, the cells were cultured for 24 hours. After incubation, the medium in the well was removed and exchanged with DMEM (Hyclone) containing 2% horse serum (HS; Hyclone) for 6 days to incubate L6 cells to differentiate into myotubes. Then, each of the octagonal fennel extract prepared in Example 1 was dissolved in DMEM (Hyclone) at 10 μg/mL, and then treated with cells and cultured for 12 hours. After incubation, cells were lysed by treatment with a cell lysis buffer. The protein in the obtained cell lysate was quantified at a concentration of 1 mg/mL using Bradford (Bio-Rad Laboratories Inc., Hercules, CA, USA). Cell lysate was dispensed in 50 μL of each microwell attached with anti-mTOR antibody and left at 37°C for 2 hours. After washing a total of 4 times with a washing buffer, treated with a detection antibody, and allowed to stand for 1 hour at 37°C. After washing a total of 4 times with a washing buffer solution, horseradish peroxidase ) Conjugated secondary antibody was added and left at 37°C for 30 minutes. Finally, after washing four times with washing buffer solution, TMB substrate was placed in each well and left at 37° C. for 10 minutes, and a stop solution was added to stop the reaction of TMB. After 2 minutes, the absorbance was measured at a wavelength of 450 nm to measure the mTOR level in the root canal cells treated with the octagonal extract. The results are shown in FIG. 1.
그 결과, 도 1에 나타난 바와 같이 팔각회향 추출물을 처리함에 따라 L6 근육세포에서 mTOR의 활성이 유의적(** p < 0.01)으로 증가한 것을 확인할 수 있었다. As a result, as shown in FIG. 1, it was confirmed that the activity of mTOR in L6 muscle cells was significantly increased ( ** p <0.01) as the octagonal fennel extract was treated.
이는 본 발명의 팔각회향 추출물이 근육세포 내에서 근 단백질 합성을 촉진시키는 능력이 우수하다는 것을 의미한다.This means that the octagonal fennel extract of the present invention has an excellent ability to promote muscle protein synthesis in muscle cells.
[실시예 3] 팔각회향 열수 추출물, 50% 에탄올 추출물, 에탄올 추출물의 mTOR 단백질 발현량 증가 효과[Example 3] Octagonal fennel hot water extract, 50% ethanol extract, the effect of increasing the amount of mTOR protein expression of ethanol extract
근육세포인 L6 myoblast (ATCC)를 10% FBS (Hyclone)가 함유된 DMEM (Hyclone)와 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)이 함유된 DMEM (Hyclone)에 상기 실시예 1-1에서 제조한 팔각회향 열수 추출물(10 μg/mL), 실시예 1-2에서 제조한 팔각회향 50% 에탄올 추출물(10 μg/mL), 실시예 1-3에서 제조한 팔각회향 에탄올 추출물(10 μg/mL)을 DMEM (Hyclone)에 녹인 후, 세포에 처리하여 myotube 분화를 유도하였다. 이 때, 시료 대신 0.01% DMSO를 처리한 군을 대조군으로 하였다. 이 과정을 2일간 6일 동안 진행하여 분화시킨 후, proteinase inhibitor cocktail (Sigma-Adrich, St. Louis, MO, USA)이 포함된 NP-40 완충용액(ELPIS-Biotech, Daejeon, Korea)으로 용해시켰다. 완충용액에 용해된 세포를 1.5 mL 튜브(tube)로 옮겨 13,000 rpm으로 10분간 원심 분리하여 상등액만을 취하였다. 상등액을 브래드포드(Bradford, Bio-Rad Laboratories Inc.)를 이용하여 정량하였다. 정량된 단백질을 5분간 끓인 후 10% SDS-PAGE로 전기영동하여 분리하였으며, 분리된 단백질들을 니트로셀룰로스 막으로 전달하였다. p-mTOR 1차 항체(Cell signaling technology)를 2.5% bovine serum albumin (BSA)에 1:1000의 비율로 희석하여 니트로셀룰로스 막에 전달된 단백질과 20시간 동안 상온에서 반응시켰다. 1차 항체를 반응시킨 다음 Tris-buffer Saline Tween20 (TBST)을 이용하여 니트로셀룰로스 막을 10분간 3회 세척하였다. 세척 후, 1차 항체를 인지하는 horseradish peroxidase가 접합된 anti-rabbit 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. Protein band는 ECL western blotting detection reagents (Amersham, Tokyo, Japan)를 사용하여 발색하였으며, G;BOX EF imaging system (Syngene, Cambridge, UK)을 이용하여 발색된 protein band를 확인하였다. 그 결과를 도 2에 나타내었다.The muscle cell, L6 myoblast (ATCC), was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) to 1 Х 10 5 cells/mL. When the cell density was about 80-85%, the medium in the well was removed and octagonal hot water extract (10 μg/) prepared in Example 1-1 in DMEM (Hyclone) containing 2% HS (Hyclone) mL), octagonal fennel ethanol extract (10 μg/mL) prepared in Example 1-2, and octagonal ethanol extract (10 μg/mL) prepared in Example 1-3 was dissolved in DMEM (Hyclone). , Cells were treated to induce myotube differentiation. At this time, the group treated with 0.01% DMSO instead of the sample was used as a control. After this process was performed for 6 days for 2 days and differentiated, it was dissolved with an NP-40 buffer solution (ELPIS-Biotech, Daejeon, Korea) containing a proteinase inhibitor cocktail (Sigma-Adrich, St. Louis, MO, USA). . The cells lysed in the buffer solution were transferred to a 1.5 mL tube and centrifuged at 13,000 rpm for 10 minutes to take only the supernatant. The supernatant was quantified using Bradford, Bio-Rad Laboratories Inc. The quantified protein was boiled for 5 minutes and then separated by electrophoresis with 10% SDS-PAGE, and the separated proteins were transferred to a nitrocellulose membrane. The p-mTOR primary antibody (Cell signaling technology) was diluted in 2.5% bovine serum albumin (BSA) at a ratio of 1:1000 and reacted with the protein delivered to the nitrocellulose membrane at room temperature for 20 hours. After reacting the primary antibody, the nitrocellulose membrane was washed three times for 10 minutes using Tris-buffer Saline Tween20 (TBST). After washing, the anti-rabbit secondary antibody (Bethyl Laboratories, Inc., Montgomery, TA, USA) conjugated with horseradish peroxidase recognizing the primary antibody was diluted to 1:5000 in 2.5% BSA with nitrocellulose membrane 2 The mixture was reacted at room temperature for an hour, and washed three times with 10 minutes using TBST. Protein bands were developed using ECL western blotting detection reagents (Amersham, Tokyo, Japan), and protein bands developed using a G;BOX EF imaging system (Syngene, Cambridge, UK) were identified. The results are shown in FIG. 2.
그 결과, 도 2에 나타낸 바와 같이 팔각회향 열수 추출물, 50% 에탄올 추출물, 에탄올 추출물의 처리에 의해 L6 근육세포에서 p-mTOR의 발현량이 증가한 것을 확인할 수 있었다.As a result, as shown in FIG. 2, it was confirmed that the expression level of p-mTOR in L6 muscle cells increased by treatment of octagonal fennel hot water extract, 50% ethanol extract, and ethanol extract.
이는 본 발명의 팔각회향 열수 추출물, 50% 에탄올 추출물, 에탄올 추출물이 근육세포 내에서 근 단백질 합성을 촉진시키는 능력이 우수하다는 것을 의미한다.This means that the octagonal fennel hot water extract, 50% ethanol extract, and ethanol extract of the present invention have excellent ability to promote muscle protein synthesis in muscle cells.
[실시예 4] 팔각회향 열수, 50% 에탄올 추출물, 에탄올 추출물의 atrogin-1과 MuRF-1의 mRNA 발현량 감소 효과[Example 4] Octagonal fennel hot water, 50% ethanol extract, effect of reducing mRNA expression levels of atrogin-1 and MuRF-1 of ethanol extract
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 팔각회향 열수 추출물(10 μg/mL), 실시예 1-2에서 제조한 팔각회향 50% 에탄올 추출물(10 μg/mL), 실시예 1-3에서 제조한 팔각회향 에탄올 추출물(10 μg/mL)을 DMEM (Hyclone)에 녹인 후, 세포에 처리하였다. 6시간 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. The muscle cell L6 myoblast (ATCC) was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) to 1 Х 10 5 cells/mL. When the cell density was about 80-85%, the medium in the well was removed and DMEM (Hyclone) containing 2% HS (Hyclone) was treated to the cells to induce myotube differentiation. Differentiation was performed every 6 days for a total of 6 days by replacing with a new medium. After differentiation, octagonal hot water extract (10 μg/) prepared in Example 1-1 in DMEM (Hyclone) containing 50 ng/mL tumor necrosis factor alpha (TNF-α; PeproTech, Rocky Hills, NJ, USA) mL), octagonal fennel ethanol extract (10 μg/mL) prepared in Example 1-2, and octagonal ethanol extract (10 μg/mL) prepared in Example 1-3 was dissolved in DMEM (Hyclone). , Cells were treated. After 6 hours, total RNA was isolated using a TRIzol reagent (Takara, Otsu, Japan). Total RNA isolated was quantified using a nanodrop (NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, MA, USA). Quantitative 16 μL of RNA using Reverse Transcriptase Premix (ELPIS-Biotech) and 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 with cDNA under conditions. 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 at 95°C for 30 seconds, 60°C for 1 minute, and 72°C for 1 minute. PCR was repeated 30 times.
Atrogin-1Atrogin-1
Forward primer: 5'-GTTACTGCAACAAGGAGAATCTGTT-3'(서열번호 1)Forward primer: 5'-GTTACTGCAACAAGGAGAATCTGTT-3' (SEQ ID NO: 1)
Reverse primer: 5'-CCGTATGAGTCTTATGTTTTGCTGG-3'(서열번호 2)Reverse primer: 5'-CCGTATGAGTCTTATGTTTTGCTGG-3' (SEQ ID NO: 2)
MuRF-1MuRF-1
Forward primer: 5'-CCGGACGGAAATGCTATGGA-3'(서열번호 3)Forward primer: 5'-CCGGACGGAAATGCTATGGA-3' (SEQ ID NO: 3)
Reverse primer: 5'-AGCCTGGAAGATGTCGTTGG-3'(서열번호 4)Reverse primer: 5'-AGCCTGGAAGATGTCGTTGG-3' (SEQ ID NO: 4)
*?*-Actin:*?*-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. 그 결과를 도 3에 나타내었다.As a result of PCR, the amplified cDNA was separated by electrophoresis with a 1.5% agarose gel, and the cDNA band was confirmed using a G;BOX EF imaging system (Syngene). The results are shown in FIG. 3.
그 결과, 도 3에 나타낸 바와 같이 TNF-α를 처리함에 따라 증가한 atrogin-1 및 MuRF-1의 mRNA 발현이 팔각회향 열수 추출물, 50% 에탄올 추출물, 에탄올 추출물의 처리에 의해 감소함을 알 수 있었다. As a result, as shown in FIG. 3, it was found that the mRNA expression of atrogin-1 and MuRF-1 increased with TNF-α treatment was decreased by treatment with octagonal hot water extract, 50% ethanol extract, and ethanol extract. .
이는 본 발명의 팔각회향 열수 추출물, 50% 에탄올 추출물, 에탄올 추출물이 근육세포 내에서 근 단백질 분해를 억제하는 능력이 우수하다는 것을 의미한다.This means that the octagonal fennel hot water extract, 50% ethanol extract, and ethanol extract of the present invention have excellent ability to inhibit muscle protein degradation in muscle cells.
[실시예 5] 팔각회향 열수 추출물의 MyoD, myogenin 및 MRF4의 mRNA 발현량 증가 효과[Example 5] Effect of increasing mRNA expression of MyoD, myogenin and MRF4 of octagonal hot water extract
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 50 ng/mL TNF-α (PeproTech)와 2% HS (Hyclone)가 함유된 DMEM (Hyclone)에 상기 실시예 1-1에서 제조한 팔각회향 열수 추출물을 녹인 후, 세포에 처리하여 myotube 분화를 유도하였다. 2일에 한 번씩 새로운 배지로 교체하여 총 6일 동안 분화를 진행하였다. 이 때, TNF-α 가 함유되지 않고 2% HS (Hyclone)가 함유된 DMEM (Hyclone)를 세포한 군을 대조군으로 하였다. TRIzol시약(Takara)을 사용하여 총 RNA를 분리하였다. 분리한 총 RNA는 나노드랍(NanoDrop 1000; Thermo Fisher Scientific Inc.)을 이용하여 정량하였다. 정량된 16 μL의 RNA를 Reverse Transcriptase Premix (ELPIS-Biotech)와 PCR 기계(Gene Amp PCR System 2700; Applied Biosystems)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. The muscle cell L6 myoblast (ATCC) was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) to 1 Х 10 5 cells/mL. When the cell density was about 80-85%, the medium in the well was removed, and Example 1-1 was added to DMEM (Hyclone) containing 50 ng/mL TNF-α (PeproTech) and 2% HS (Hyclone). After dissolving the octagonal fennel hot water extract prepared in the cells, the cells were treated to induce myotube differentiation. Differentiation was performed every 6 days for a total of 6 days by replacing with a new medium. At this time, the group containing DMEM (Hyclone) containing 2% HS (Hyclone) without TNF-α was used as a control. Total RNA was isolated using TRIzol reagent (Takara). Total RNA isolated was quantified using a nanodrop (NanoDrop 1000; Thermo Fisher Scientific Inc.). Quantitative 16 μL of RNA was synthesized by cDNA under conditions of 42°C, 55 minutes and 70°C, 15 minutes using Reverse Transcriptase Premix (ELPIS-Biotech) and PCR machine (Gene Amp PCR System 2700; Applied Biosystems). 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 at 95°C for 30 seconds, 60°C for 1 minute, and 72°C for 1 minute. PCR was repeated 30 times.
MyoDMyoD
Forward primer: 5'-TGCCTCGGAGATAATACAGCC-3'(서열번호 7)Forward primer: 5'-TGCCTCGGAGATAATACAGCC-3' (SEQ ID NO: 7)
Reverse primer: 5'-GGTGTAACAACCATACCCCACT-3'(서열번호 8)Reverse primer: 5'-GGTGTAACAACCATACCCCACT-3' (SEQ ID NO: 8)
MyogeninMyogenin
Forward primer: 5'-AGAGAGCCCCCTTGTTAATGC-3'Forward primer: 5'-AGAGAGCCCCCTTGTTAATGC-3'
Reverse primer: 5'-GGCCACTCACTGTCTCTCAAA-3'Reverse primer: 5'-GGCCACTCACTGTCTCTCAAA-3'
MRF4MRF4
Forward primer: 5'-CTTGAGGGTGCGGATTTCCT-3'Forward primer: 5'-CTTGAGGGTGCGGATTTCCT-3'
Reverse primer: 5'-GGGAGTTTGCGTTCCTCTGA-3'Reverse primer: 5'-GGGAGTTTGCGTTCCTCTGA-3'
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. 그 결과를 도 4에 나타내었다.As a result of PCR, the amplified cDNA was separated by electrophoresis with a 1.5% agarose gel, and the cDNA band was confirmed using a G;BOX EF imaging system (Syngene). The results are shown in FIG. 4.
그 결과, 도 4에 나타낸 바와 같이 TNF-α를 처리함에 따라 유의적(## p < 0.01)으로 감소한 MyoD, myogenin 및 MRF4의 mRNA 발현이 팔각회향 열수 추출물의 처리에 의해 유의적(* p < 0.05, ** p < 0.01)으로 증가함을 알 수 있었다. As a result, as shown in FIG. 4, mRNA expression of MyoD, myogenin and MRF4 decreased significantly ( ## p <0.01) as TNF-α was treated, which was significant by treatment of octagonal hot water extract ( * p < 0.05, ** p <0.01).
이는 본 발명의 팔각회향 열수 추출물이 근육의 분화를 촉진시키는 능력이 우수하다는 것을 의미한다.This means that the octagonal hot water extract of the present invention has an excellent ability to promote muscle differentiation.
[실시예 6] 동물모델에서 팔각회향 열수 추출물의 근 기능 및 근육량 증가 효과[Example 6] Effect of muscle function and muscle mass increase of octagonal hot water extract in animal models
<6-1> 동물의 사육 및 근위축 유도<6-1> Animal breeding and muscle atrophy
실험동물로 생후 8주된 수컷쥐(C57BL/6J; Narabiotech, Seoul, Korea)를 구입하여 실험을 진행하였다. 모든 동물의 사육은 연세실험동물연구센터(Yonsei Laboratory Animal Reaserch Center; YLARC, Seoul, Korea)에서 진행되었으며, 사육실 환경은 온도 23 ± 2℃, 상대습도 55 ± 10%로 유지시켰다. 7일간 적응시킨 후, 300 mg/kg 트리브로모메탄올(tribromoethanol, Sigma-Aldrich)을 복강 주사하여 마취를 유도하였다. 마취 후, 근위축군과 시료 투여군에 속한 쥐의 오른쪽 뒷다리(hindlimb) 장딴지근육(gastrocnemius muscle)과 오른쪽 발바닥을 피부 스테이플러(skin stapler)(Unidus, Chungcheongbuk-do, Korea)를 사용하여 스테이플러 심으로 근육을 손상시키고 오른쪽 뒷다리가 움직이지 못하게 하였으며 이 상태를 일주일 간 유지시켰다. 일주일 후, 장딴지근과 발바닥에 고정되어 있던 스테이플러 심을 제거하고 다시 일주일간 상기 실시예 1-1에서 제조한 팔각회향 열수 추출물을 100 mg/kg 또는 200 mg/kg의 농도로 매일 경구투여하였다. 이 때, 정상군과 근위축군은 팔각회향 추출물 대신에 식염수로 경구투여를 실시하였다.As an experimental animal, an 8-week-old male rat (C57BL/6J; Narabiotech, Seoul, Korea) was purchased and tested. Breeding of all animals was conducted at the Yonsei Laboratory Animal Reaserch Center (YLARC, Seoul, Korea), and the breeding room environment was maintained at 23±2℃ and relative humidity 55±10%. After 7 days of adaptation, anesthesia was induced by intraperitoneal injection of 300 mg/kg tribromoethanol (Sigma-Aldrich). After anesthesia, the right hindlimb gastrocnemius muscle and the right sole of the rats belonging to the muscle atrophy group and the sample administration group were muscled with a stapler core using a skin stapler (Unidus, Chungcheongbuk-do, Korea). Damage was made and the right hind leg was immobilized and this condition was maintained for a week. One week later, the stapler core fixed to the calf muscle and sole was removed, and the octagonal fennel hot water extract prepared in Example 1-1 was orally administered daily for a week at a concentration of 100 mg/kg or 200 mg/kg. At this time, the normal group and the atrophic group were administered orally with saline instead of the octagonal fennel extract.
<6-2> 팔각회향 열수 추출물의 근력 향상 효과 확인<6-2> Confirm the effect of octagonal fennel hot water extract to improve muscle strength
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, the muscle strength of the rats was measured using a muscle strength meter (Panlab, Barcelona, Spain). The rat's tail was pulled with constant force until the rat placed the youngest of the muscular strength tester, and a total of 5 consecutive tests were performed per head.
그 결과, 도 5에 나타낸 바와 같이 정상군에 비해 근위축군에서 근력이 유의적(## p < 0.01)으로 감소하였으나, 팔각회향 열수 추출물을 처리함에 따라 근력이 유의적(** p < 0.01)으로 증가한 것을 확인하였다. As a result, as shown in FIG. 5, the muscle strength was significantly decreased ( ## p <0.01) in the atrophy group compared to the normal group, but the muscle strength was significant as the octagonal hot water extract was treated ( ** p <0.01). It was confirmed that the increase.
이는 본 발명의 팔각회향 열수 추출물이 근위축으로 인해 감소한 근력을 증가시키는 효과가 우수하다는 것을 의미한다.This means that the octagonal fennel hot water extract of the present invention has an excellent effect of increasing the decreased muscle strength due to muscular atrophy.
<6-3> 팔각회향 열수 추출물의 근육 무게 증가 효과<6-3> Effects of octagonal fennel hot water extract to increase muscle weight
근력 측정이 끝난 후, 실험동물에 300 mg/kg 트리브로모메탄올(Sigma-Aldrich)을 복강 주사하여 마취 후 심채혈을 통해 희생을 하였다. 심장박동이 멈춘 것을 확인한 뒤, 오른쪽 뒷다리에서 전경골근(tibialis anterior muscle)을 적출하여 무게를 측정하였다.After the measurement of muscle strength, 300 mg/kg tribromomethanol (Sigma-Aldrich) was injected intraperitoneally into the experimental animal and then sacrificed through cardiac blood after anesthesia. After confirming that the heartbeat had stopped, weight was measured by extracting the tibialis anterior muscle from the right hind leg.
그 결과, 도 6에 나타낸 바와 같이 정상군에 비해 근위축군의 전경골근 무게가 유의적(## p < 0.01)으로 감소하였으나, 팔각회향 열수 추출물을 처리함에 전경골근 무게가 유의적(* p < 0.05, ** p < 0.01)으로 증가한 것을 확인하였다. As a result, as shown in FIG. 6, the weight of the forearm muscle of the atrophic group decreased significantly ( ## p <0.01) compared to the normal group, but the weight of the forearm muscle was significantly ( * p < 0.05, ** p <0.01).
이는 본 발명의 팔각회향 열수 추출물이 근위축으로 인해 감소한 근육의 무게를 증가시키는 효과가 우수하다는 것을 의미한다.This means that the octagonal hot water extract of the present invention has an excellent effect of increasing the weight of muscles reduced due to muscular atrophy.
[실시예 7] 동물모델에서 팔각회향 에탄올 추출물의 근 기능 및 근육량 증가 효과[Example 7] Effect of muscle function and muscle mass increase of octagonal ethanol extract in animal model
상기 실시예 6-1과 동일한 방법으로 근위축을 유도한 다음 상기 실시예 1-3에서 제조한 팔각회향 에탄올 추출물을 200 mg/kg의 농도로 매일 경구투여하였다. 다음, 상기 실시예 6-2와 실시예 6-3과 동일한 방법으로 근력과 근육 무게를 측정한 결과, 팔각회향 에탄올 추출물 투여군은 근위축군에 비하여 근력과 근육 무게가 각각 14.38%(** p < 0.01)와 18.66%(** p < 0.01) 증가하였다. 이는 본 발병의 팔각회향 에탄올 추출물이 근위축으로 인해 감소한 근력과 근육의 무게를 증가시키는 효과가 우수하다는 것을 의미한다.The muscle atrophy was induced in the same manner as in Example 6-1, and then the octagonal ethanol extract prepared in Example 1-3 was orally administered daily at a concentration of 200 mg/kg. Next, as a result of measuring muscle strength and muscle weight in the same manner as in Example 6-2 and Example 6-3, the octagonal ethanol extract-administered group had 14.38% ( ** p < 0.01) and 18.66% ( ** p <0.01). This means that the octagonal ethanol extract of the present disease is excellent in reducing muscle strength and muscle weight due to muscular atrophy.
[실시예 8] 동물모델에서 팔각회향 초임계 추출물의 근 기능 및 근육량 증가 효과[Example 8] Effect of muscle function and muscle mass increase of octagonal supercritical extract in animal model
상기 실시예 6-1과 동일한 방법으로 근위축을 유도한 다음 상기 실시예 1-7에서 제조한 팔각회향 초임계 추출물을 200 mg/kg의 농도로 매일 경구투여하였다. 다음, 상기 실시예 6-2와 실시예 6-3과 동일한 방법으로 근력과 근육 무게를 측정한 결과, 팔각회향 초임계 추출물 투여군은 근위축군에 비하여 근력과 근육 무게가 각각 18.70%(** p < 0.01)와 22.95%(** p < 0.01) 증가하였다. 이는 본 발병의 팔각회향 초임계 추출물이 근위축으로 인해 감소한 근력과 근육의 무게를 증가시키는 효과가 우수하다는 것을 의미한다.The muscle atrophy was induced in the same manner as in Example 6-1, and then the octagonal supercritical extract prepared in Example 1-7 was orally administered daily at a concentration of 200 mg/kg. Next, as a result of measuring muscle strength and muscle weight in the same manner as in Example 6-2 and Example 6-3, the octagonal supercritical extract-administered group had a muscle strength and a muscle weight of 18.70% ( ** p , respectively) compared to the muscular atrophy group. <0.01) and 22.95% ( ** p <0.01). This means that the octagonal supercritical extract of the present onset is excellent in reducing muscle strength due to muscular atrophy and increasing muscle weight.
[실시예 9] 시킴산의 mTOR 활성 증가 효과[Example 9] Increase of mTOR activity of shikimic acid
시킴산을 5 μM 및 10 μM의 농도로 세포에 처리하였다는 점을 제외하고, 상기 실시예 2와 동일한 방법으로 실험을 진행하였다. Experiments were conducted in the same manner as in Example 2, except that the cells were treated with concentrations of sickimic acid at 5 μM and 10 μM.
그 결과, 도 7에 나타낸 바와 같이 시킴산을 처리함에 따라 L6 근육세포에서 mTOR의 활성이 유의적(** p < 0.01)으로 증가한 것을 확인할 수 있었다.As a result, as shown in FIG. 7, it was confirmed that the activity of mTOR in L6 muscle cells increased significantly ( ** p <0.01) as treated with shikimic acid.
이는 본 발명의 시킴산이 근육세포 내에서 근 단백질 합성을 촉진시키는 능력이 우수하다는 것을 의미한다.This means that the shikimic acid of the present invention has an excellent ability to promote muscle protein synthesis in muscle cells.
[실시예 10] 시킴산의 atrogin-1과 MuRF-1의 mRNA 발현량 감소 효과[Example 10] Sikkim acid atrogin-1 and MuRF-1 mRNA expression reduction effect
시킴산을 20 μM 및 40 μM의 농도로 세포에 처리하였다는 점을 제외하고, 상기 실시예 4와 동일한 방법으로 실험을 진행하였다.Experiments were conducted in the same manner as in Example 4, except that the cells were treated with concentrations of sickimic acid at a concentration of 20 μM and 40 μM.
그 결과, 그 결과, 도 8에 나타낸 바와 같이 TNF-α를 처리함에 따라 유의적(## p < 0.01)으로 증가한 atrogin-1 및 MuRF-1의 mRNA 발현이 시킴산의 처리에 의해 유의적(** p < 0.01)으로 감소함을 알 수 있었다. As a result, as a result, as shown in FIG. 8, the mRNA expression of atrogin-1 and MuRF-1 increased significantly ( ## p <0.01) with TNF-α treatment, which was significant by treatment with sickimic acid ( ** p <0.01).
이는 본 발명의 시킴산이 근육세포 내에서 근 단백질 분해를 억제하는 능력이 우수하다는 것을 의미한다.This means that the shikimic acid of the present invention is excellent in inhibiting muscle protein degradation in muscle cells.
[실시예 11] 시킴산의 MyoD, myogenin 및 MRF4의 mRNA 발현량 증가 효과[Example 11] The effect of increasing the mRNA expression level of MyoD, myogenin and MRF4 of Shikimic acid
시킴산을 20 μM 및 40 μM의 농도로 세포에 처리하였다는 점을 제외하고, 상기 실시예 5와 동일한 방법으로 실험을 진행하였다.Experiments were conducted in the same manner as in Example 5, except that the cells were treated with concentrations of sickimic acid at a concentration of 20 μM and 40 μM.
그 결과, 도 9에 나타낸 바와 같이 TNF-α를 처리함에 따라 유의적(## p < 0.01)으로 감소한 MyoD, myogenin 및 MRF4의 mRNA 발현이 시킴산의 처리에 의해 유의적(* p < 0.05, ** p < 0.01)으로 증가함을 알 수 있었다. As a result, as shown in Fig. 9, the mRNA expression of MyoD, myogenin and MRF4 decreased significantly ( ## p <0.01) with TNF-α treatment was significant ( * p <0.05, ** p <0.01).
이는 본 발명의 시킴산이 근육의 분화를 촉진시키는 능력이 우수하다는 것을 의미한다.This means that the shikimic acid of the present invention has excellent ability to promote muscle differentiation.
[실시예 12] 동물모델에서 시킴산의 근 기능 및 근육량 증가 효과[Example 12] Effect of muscle function and muscle mass increase of sikkim acid in animal model
<12-1> 동물의 사육 및 근위축 유도<12-1> Animal breeding and muscle atrophy
상기 실시예 6-1과 동일한 방법으로 동물을 사육하고 근위축을 유도하였다. 이때, 팔각회향 열수 추출물 대신 시킴산 50 mg/kg 및 100 mg/kg을 실험동물에 투여하였다.Animals were bred in the same manner as in Example 6-1, and muscle atrophy was induced. At this time, instead of the octagonal fennel hot water extract, 50 mg/kg and 100 mg/kg of shikimic acid were administered to the experimental animals.
<12-2> 시킴산의 근력 향상 효과 확인<12-2> Confirming the effect of improving the strength of Sikkim Mountain
경구 투여 기간이 끝나고, 상기 실시예 6-2과 동일한 방법으로 실험을 진행하여 시킴산의 근력 향상 효과를 확인하였다.After the oral administration period was over, the experiment was conducted in the same manner as in Example 6-2 to confirm the effect of improving the muscle strength of Sikkim.
그 결과, 도 10에 나타낸 바와 같이 정상군에 비해 근위축군에서 근력이 유의적(## p < 0.01)으로 감소하였으나, 시킴산을 처리함에 따라 근력이 유의적(** p < 0.01)으로 증가한 것을 확인하였다.As a result, as shown in FIG. 10, muscle strength was significantly decreased ( ## p <0.01) in the atrophy group compared to the normal group, but muscle strength was significantly increased ( ** p <0.01) as treated with shikimic acid. Was confirmed.
이는 본 발명의 시킴산이 근위축으로 인해 감소한 근력을 증가시키는 효과가 우수하다는 것을 의미한다.This means that the effect of increasing the muscle strength of the shikimic acid of the present invention is reduced due to muscle atrophy.
<12-3> 시킴산의 근육 무게 증가 효과 확인<12-3> Confirming the effect of shikimic acid to increase muscle weight
근력 측정이 끝난 후, 상기 실시예 6-3과 동일한 방법으로 실험을 진행하여 시킴산의 근육 무게 증가 효과를 확인하였다.After the muscle strength measurement was completed, an experiment was conducted in the same manner as in Example 6-3 to confirm the effect of increasing the muscle weight of sikkim acid.
그 결과, 도 11에 나타낸 바와 같이 정상군에 비해 근위축군의 전경골근 무게가 유의적(## p < 0.01)으로 감소하였으나, 시킴산을 처리함에 따라 전경골근 무게가 유의적(* p < 0.05, ** p < 0.01)으로 증가한 것을 확인하였다.As a result, the weight of the tibialis anterior proximal herd significantly compared to the normal group As shown in 11 (## p <0.01) Note that the tibialis anterior weight ever, as decreased by, Sikkim handle acid (* p <0.05 , ** p <0.01).
이는 본 발명의 시킴산이 근위축으로 인해 감소한 근육의 무게를 증가시키는 효과가 우수하다는 것을 의미한다.This means that the shikimic acid of the present invention has an excellent effect of increasing the weight of muscles reduced due to muscle atrophy.
이하, 본 발명의 팔각회향 추출물 및 시킴산을 유효성분으로 함유하는 의약품, 식품 또는 화장품의 제조예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다. 상기 근육 질환 예방 및 치료 또는 근 기능 개선 효과가 우수한 팔각회향 추출물을 가지고 하기와 같은 조성성분 및 조성비에 따라 제조예 1 내지 3의 의약품, 식품 또는 화장료 조성물을 통상적인 방법에 따라서 제조하였다.Hereinafter, an example of manufacturing a pharmaceutical, food or cosmetic product containing the octagonal fennel extract and shikimic acid of the present invention as an active ingredient will be described, but the present invention is not intended to be limited thereto, but only to be specifically described. The pharmaceutical, food or cosmetic compositions of Preparation Examples 1 to 3 were prepared according to the following compositional ingredients and composition ratios with the octagonal fennel extract excellent in preventing and treating muscle diseases or improving muscle function.
[제조예 1] 의약품[Production Example 1] Pharmaceutical
<1-1> 산제<1-1> powder
팔각회향 추출물 또는 시킴산 50 mg, 결정셀룰로오즈 2 g을 혼합한 후 통상의 산제 제조방법에 따라서 기밀포에 충진하여 산제를 제조하였다.After mixing octagonal fennel extract or 50 mg of shikimic acid and 2 g of crystalline cellulose, a powder was prepared by filling in an airtight fabric according to a conventional powder manufacturing method.
<1-2> 정제<1-2> tablets
팔각회향 추출물 또는 시킴산 50 mg, 결정셀룰로오즈 400 mg, 스테아린산 마그네슘 5 mg을 혼합한 후 통상의 정제 제조방법에 따라서 타정하여 정제를 제조하였다.After mixing the octagonal fennel extract or 50 mg of sikkim acid, 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> Capsule
팔각회향 추출물 및 시킴산 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 the octagonal fennel extract and 30 mg of shikimic acid, 100 mg of whey protein, 400 mg of crystalline cellulose, and 6 mg of magnesium stearate, the capsules were prepared by filling into gelatin capsules according to a conventional capsule preparation method.
[제조예 2] 식품[Production Example 2] Food
<2-1> 건강식품의 제조<2-1> Manufacture of healthy food
팔각회향 추출물 또는 시킴산 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.Octagonal fennel extract or shikimic acid 1000 mg, 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, pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium 25.3 mg, potassium phosphate 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 mg, carbonic acid It can be prepared by mixing 100 mg of calcium and 24.8 mg of magnesium chloride, and the compounding ratio may be arbitrarily modified. After mixing the above ingredients according to a conventional health food manufacturing method, granules are prepared, and then Depending on the method, it can be used to prepare a health food composition.
<2-2> 건강음료의 제조<2-2> Preparation of health drinks
팔각회향 추출물 또는 시킴산 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 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 L용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 건강음료 조성물 제조에 사용할 수 있다.Octagonal fennel extract or 1000 mg of shikimic acid, 1000 mg of citric acid, 100 g of oligosaccharides, 2 g of plum concentrate, and 1 g of taurine were added with purified water to mix all of the above ingredients according to the general method for preparing a health drink of 900 mL, followed by about 1 After stirring and heating at 85° C. for a period of time, the resulting solution may be filtered, obtained in a sterilized 2 L container, sealed and sterilized, and then refrigerated and used in the manufacture of a health drink composition.
<2-3> 츄잉껌<2-3> Chewing gum
껌 베이스 20 중량%, 설탕 76.9 중량%, 향료 1 중량% 및 물 2 중량% 와 팔각회향 추출물 또는 시킴산 0.1 중량%를 배합하여 통상의 방법으로 츄잉껌을 제조하였다.Chewing gum was prepared by a conventional method by mixing 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of fragrance and 2% by weight of water and 0.1% by weight of octagonal extract or shikimic acid.
<2-4> 캔디<2-4> Candy
설탕 60 중량%, 물엿 39.8 중량% 및 향료 0.1 중량%와 팔각회향 추출물 또는 시킴산 0.1 중량%를 배합하여 통상의 방법으로 캔디를 제조하였다.Candy was prepared by a conventional method by mixing 60% by weight of sugar, 39.8% by weight of starch syrup and 0.1% by weight of fragrance and 0.1% by weight of octagonal fennel extract or shikimic acid.
<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 팔각회향 추출물 또는 시킴산 1.0001중량%를 배합하여 통상의 방법으로 비스켓을 제조하였다.Power 1st class 25.59% by weight, Gravity 1st class 22.22% by weight, 4.80% by weight per white sugar, 0.73% by weight of salt, 0.78% by weight of glucose, 11.78% by weight of palm shortening, 1.54% by weight of ammonium, 0.17% by weight of sodium bicarbonate, 0.16% by weight of sodium bisulfite , 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, substitute powder 0.29% by weight, calcium phosphate 0.03% by weight, spray salt 0.29% by weight and spray Biscuits were prepared in a conventional manner by mixing 7.27% by weight of oil with 1.0001% by weight of octagonal fennel extract or shikimic acid.
[제조예 3] 화장품[Production Example 3] Cosmetics
<3-1> 영양화장수(밀크로션)<3-1> Nutrient Cosmetics (Milk Lotion)
팔각회향 추출물 또는 시킴산을 하기 표 1의 영양화장수 제형 비율대로 하여 통상적인 방법에 따라 영양화장수를 제조하였다.The octagonal fennel extract or sickimic acid was prepared according to the ratio of the nutrient composition of Table 1, and the nutrient composition was prepared according to a conventional method.
배합성분Ingredients 제조예3-1(중량%)Preparation Example 3-1 (% by weight)
팔각회향 추출물 또는 시킴산Octagonal fennel extract or shikimic acid 2.02.0
스쿠알란Squalane 5.05.0
밀납Wax 4.04.0
폴리솔베이트 60Polysorbate 60 1.51.5
솔비탄세스퀴올레이트Sorbitan sesquioleate 1.51.5
유동파라핀Floating paraffin 0.50.5
카프릴릭/카프릭트리글리세라이드Caprylic/Capric Triglyceride 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, pigments, flavors 적량Proper
정제수Purified water to 100to 100
<3-2> 유연화장수(스킨로션)<3-2> Flexible Cosmetics (Skin Lotion)
팔각회향 추출물 또는 시킴산을 하기 표 2의 유연화장수 제형 비율대로 하여 통상적인 방법에 따라 유연화장수를 제조하였다.The octagonal fennel extract or shikimic acid was prepared according to the ratio of the flexible cosmetic composition of Table 2, and the flexible cosmetic composition was prepared according to a conventional method.
배합성분 Ingredients 제조예3-2(중량%)Preparation Example 3-2 (% by weight)
팔각회향 추출물 또는 시킴산Octagonal fennel extract or shikimic acid 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, pigments, flavors 적량Proper
정제수Purified water to 100to 100
<3-3> 영양크림<3-3> Nutrition Cream
팔각회향 추출물 또는 시킴산을 하기 표 3의 영양크림 제형 비율대로 하여 통상적인 방법에 따라 영양크림을 제조하였다.An octagonal fennel extract or shikimic acid was prepared according to the ratio of the nutrient cream formulations in Table 3, to prepare a nutrient cream.
배합성분 Ingredients 제조예3-3(중량%)Preparation Example 3-3 (% by weight)
팔각회향 추출물 또는 시킴산Octagonal fennel extract or shikimic acid 2.02.0
폴리솔베이트 60Polysorbate 60 1.51.5
솔비탄세스퀴올레이트Sorbitan sesquioleate 0.50.5
PEG60 경화피마자유PEG60 hardened castor oil 2.02.0
유동파라핀Floating paraffin 1010
스쿠알란Squalane 5.05.0
카프릴릭/카프릭트리글리세라이드Caprylic/Capric Triglyceride 5.05.0
글리세린glycerin 5.05.0
부틸렌글리콜Butylene glycol 3.03.0
프로필렌글리콜Propylene glycol 3.03.0
트리에탄올아민Triethanolamine 0.20.2
방부제antiseptic 적량Proper
색소Coloring 적량Proper
향료Spices 적량Proper
정제수Purified water to 100to 100
<3-4> 마사지크림<3-4> Massage Cream
팔각회향 추출물 또는 시킴산을 하기 표 4의 마사지크림 제형 비율대로 하여 통상적인 방법에 따라 마사지크림을 제조하였다.Massage cream was prepared according to a conventional method using the octagonal fennel extract or shikimic acid in the proportion of the massage cream formulation shown in Table 4 below.
배합성분 Ingredients 제조예3-4(중량%)Preparation Example 3-4 (wt%)
팔각회향 추출물 또는 시킴산Octagonal fennel extract or shikimic acid 1.01.0
밀납Wax 10.010.0
폴리솔베이트 60Polysorbate 60 1.51.5
PEG 60 경화피마자유 PEG 60 hardened castor oil 2.02.0
솔비탄세스퀴올레이트Sorbitan sesquioleate 0.80.8
유동파라핀Floating paraffin 40.040.0
스쿠알란Squalane 5.05.0
카프릴릭/카프릭트리글리세라이드Caprylic/Capric Triglyceride 4.04.0
글리세린glycerin 5.05.0
부틸렌글리콜Butylene glycol 3.03.0
프로필렌글리콜Propylene glycol 3.03.0
트리에탄올아민Triethanolamine 0.20.2
방부제, 색소, 향료Preservatives, pigments, flavors 적량Proper
정제수Purified water to 100to 100
<3-5> 팩<3-5> Pack
팔각회향 추출물 또는 시킴산을 하기 표 5의 팩 제형 비율대로 하여 통상적인 방법에 따라 팩을 제조하였다.Packs were prepared according to a conventional method using the octagonal fennel extract or shikimic acid according to the pack formulation ratios in Table 5 below.
배합성분 Ingredients 제조예3-5(중량%)Preparation Example 3-5 (wt%)
팔각회향 추출물 또는 시킴산Octagonal fennel extract or shikimic acid 1.01.0
폴리비닐알콜Polyvinyl alcohol 13.013.0
소듐카르복시메틸셀룰로오스Sodium carboxymethylcellulose 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, pigments, flavors 적량Proper
정제수Purified water to 100to 100
<3-6> 젤<3-6> Gel
팔각회향 추출물 또는 시킴산을 하기 표 6의 젤 제형 비율대로 하여 통상적인 방법에 따라 젤을 제조하였다.The gel was prepared according to a conventional method using the octagonal fennel extract or shikimic acid according to the gel formulation ratio in Table 6 below.
배합성분Ingredients 제조예3-6(중량%)Preparation Example 3-6 (wt%)
팔각회향 추출물 또는 시킴산Octagonal fennel extract or shikimic acid 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 PEG 60 hardened castor oil 0.50.5
트리에탄올아민Triethanolamine 0.30.3
방부제, 색소, 향료Preservatives, pigments, flavors 적량Proper
정제수Purified water to 100to 100
본 발명은 팔각회향 추출물 및 시킴산을 함유하는 근육 질환 예방 및 치료용 또는 근 기능 개선용 조성물에 관한 것이다. 본 발명에 따른 팔각회향 추출물 및 시킴산은 근 단백질 합성에 관여하는 mTOR의 활성 증가 및 근 단백질 분해에 관여하는 MuRF-1과 atrogin-1의 mRNA 발현을 감소시키는 효과가 있으며, 근육량을 증가시키고 및 근기능을 향상시켜, 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근무력증, 악액질(cachexia) 및 근육감소증(sarcopenia) 등의 각종 근육 질환 예방, 치료 또는 개선에 유용하게 이용될 수 있으므로 산업상 이용가능성이 있다. The present invention relates to a composition for preventing and treating muscle disease or improving muscle function, comprising an octagonal fennel extract and shikimic acid. The octagonal fennel extract and shikimic acid according to the present invention have 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, increasing muscle mass and muscle function It is useful for preventing, treating or improving various muscle diseases such as atony, muscle atrophy, muscle dystrophy, muscle degeneration, myasthenia, cachexia, and sarcopenia. As it can be used, it has industrial applicability.
서열번호 1은 Atrogin-1의 Forward primer이며, 다음과 같다. 5'-GTTACTGCAACAAGGAGAATCTGTT-3'(서열번호 1) 서열 번호 2는, Atrogin-1의 Reverse primer이며 5'-CCGTATGAGTCTTATGTTTTGCTGG-3'(서열번호 2)와 같다.SEQ ID NO: 1 is the forward primer of Atrogin-1 , as follows. 5'-GTTACTGCAACAAGGAGAATCTGTT-3' (SEQ ID NO: 1) SEQ ID NO: 2 is the reverse primer of Atrogin-1 and is the same as 5'-CCGTATGAGTCTTATGTTTTGCTGG-3' (SEQ ID NO: 2).
서열번호 3은 MuRF-1의 Forward primer이며 5'-CCGGACGGAAATGCTATGGA-3'(서열번호 3)와 같고, 서열번호 4는 MuRF-1의 Reverse primer이며 5'-AGCCTGGAAGATGTCGTTGG-3'(서열번호 4)와 같다.SEQ ID NO: 3 is the forward primer of MuRF-1 and is the same as 5'-CCGGACGGAAATGCTATGGA-3' (SEQ ID NO: 3), and SEQ ID NO: 4 is the reverse primer of MuRF-1 and 5'-AGCCTGGAAGATGTCGTTGG-3' (SEQ ID NO: 4) same.
서열번호 5는 β-Actin의 Forward primer이며 5'-TGACAGGATGCAGAAGGAGATTAC-3'(서열번호 5)와 같고, 서열번호 6은 β-Actin의 Reverse primer이며 5'-TAAAACGCAGCTCAGTAACAGTC-3'(서열번호 6)와 같다.SEQ ID NO: 5 is a forward primer of β-Actin and is the same as 5'-TGACAGGATGCAGAAGGAGATTAC-3' (SEQ ID NO: 5), and SEQ ID NO: 6 is a reverse primer of β-Actin and 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6) same.
다음은, MyoD의 Forward primer서열이며, 5'-TGCCTCGGAGATAATACAGCC-3'와 같고, MyoD의 Reverse primer서열은 5'-GGTGTAACAACCATACCCCACT-3'과 같다.The following is MyoD Forward primer sequence, same as 5'-TGCCTCGGAGATAATACAGCC-3', and MyoD Reverse primer sequence is 5'-GGTGTAACAACCATACCCCACT-3'.
Myogenin의 Forward primer서열은 5'-AGAGAGCCCCCTTGTTAATGC-3'와 같으며, Reverse primer서열은 5'-GGCCACTCACTGTCTCTCAAA-3'와 같다. The forward primer sequence of Myogenin is 5'-AGAGAGCCCCCTTGTTAATGC-3', and the reverse primer sequence is 5'-GGCCACTCACTGTCTCTCAAA-3'.
MRF4의 Forward primer서열은 5'-CTTGAGGGTGCGGATTTCCT-3'와 같고, Reverse primer서열은 5'-GGGAGTTTGCGTTCCTCTGA-3'와 같다. The forward primer sequence of MRF4 is 5'-CTTGAGGGTGCGGATTTCCT-3', and the reverse primer sequence is 5'-GGGAGTTTGCGTTCCTCTGA-3'.

Claims (9)

  1. 팔각회향 추출물 또는 하기 화학식 1로 표시되는 시킴산을 유효성분으로 함유하는 근육 질환 예방 또는 치료용 약학 조성물:Pharmaceutical composition for preventing or treating muscle diseases containing octagonal fennel extract or shikimic acid represented by the following formula 1 as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2019018726-appb-I000003
    Figure PCTKR2019018726-appb-I000003
  2. 제 1 항에 있어서,According to claim 1,
    화학식 1로 표시되는 시킴산은 팔각회향 추출물로부터 분리된 것을 특징으로 하는 근육 질환 예방 또는 치료용 약학 조성물.Sikkim acid represented by Formula 1 is a pharmaceutical composition for preventing or treating muscle diseases, characterized in that isolated from the octagonal fennel extract.
  3. 제 1 항에 있어서, According to claim 1,
    팔각회향 추출물은 물, 탄소수 1 내지 6의 유기용매, 아임계 유체 및 초임계 유체로 이루어진 군으로부터 선택되는 하나 이상의 용매로 팔각회향을 추출하여 수득한 것을 특징으로 하는 근육 질환 예방 또는 치료용 약학 조성물.Octagonal fennel extract is a pharmaceutical composition for preventing or treating muscle diseases characterized by being obtained by extracting octagonal fennel with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, subcritical fluid and supercritical fluid. .
  4. 제 3 항에 있어서, The method of claim 3,
    상기 유기용매는, The organic solvent,
    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 적어도 어느 하나의 용매인 근육 질환 예방 또는 치료용 약학 조성물.Alcohols having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, and cyclo Pharmaceutical composition for preventing or treating muscle disease, which is at least one solvent selected from the group consisting of hexane and petroleum ether.
  5. 제 1 항 내지 제 4 항 중 어느 한 항에 있어서, The method according to any one of claims 1 to 4,
    상기 근육 질환은, 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근무력증, 악액질(cachexia) 및 근육감소증(sarcopenia)으로 이루어진 군에서 선택되는 하나 이상의 질병을 예방 또는 치료하는 것을 특징으로 하는 근육 질환 예방 또는 치료용 약학 조성물.The muscle disease prevents one or more diseases selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia and sarcopenia. Or pharmaceutical composition for the prevention or treatment of muscle disease characterized in that the treatment.
  6. 팔각회향 추출물 또는 하기 화학식 1로 표시되는 시킴산을 유효성분으로 함유하는 근육 질환 예방 또는 근 기능 개선용 식품 조성물:Food composition for preventing or improving muscle function or muscle function containing octagonal fennel extract or shikimic acid represented by the following Chemical Formula 1 as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2019018726-appb-I000004
    Figure PCTKR2019018726-appb-I000004
  7. 팔각회향 추출물 또는 하기 화학식 1로 표시되는 시킴산을 유효성분으로 함유하는 근 기능 개선용 화장료 조성물:Octagonal fennel extract or cosmetic composition for improving muscle function, containing shikimic acid represented by the following Chemical Formula 1 as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2019018726-appb-I000005
    Figure PCTKR2019018726-appb-I000005
  8. 팔각회향 추출물 또는 하기 화학식 1로 표시되는 시킴산을 유효성분으로 함유하는 조성물을 개체에 투여 또는 복용시키는 단계를 포함하는 근육 질환 예방 또는 치료방법:A method of preventing or treating muscle disease comprising the step of administering or taking an octagonal fennel extract or a composition containing shikimic acid represented by Formula 1 as an active ingredient in an individual:
    [화학식 1] [Formula 1]
    Figure PCTKR2019018726-appb-I000006
    Figure PCTKR2019018726-appb-I000006
  9. 팔각회향 추출물 또는 하기 화학식 1로 표시되는 시킴산을 유효성분으로 함유하는 조성물의 근육 질환 예방 또는 치료 용도:Prevention or treatment of muscle disease of octagonal fennel extract or a composition containing shikimic acid represented by the following formula 1 as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2019018726-appb-I000007
    Figure PCTKR2019018726-appb-I000007
PCT/KR2019/018726 2018-12-28 2019-12-30 Composition for preventing and treating muscle disease or improving muscle function, comprising illicium verum extract or shikimic acid WO2020139057A2 (en)

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