WO2020218720A1 - Composition pour la prévention ou le traitement de troubles musculaires ou l'amélioration de la fonction musculaire, contenant un extrait de leonurus japonicus ou de la léonurine - Google Patents

Composition pour la prévention ou le traitement de troubles musculaires ou l'amélioration de la fonction musculaire, contenant un extrait de leonurus japonicus ou de la léonurine Download PDF

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WO2020218720A1
WO2020218720A1 PCT/KR2020/001523 KR2020001523W WO2020218720A1 WO 2020218720 A1 WO2020218720 A1 WO 2020218720A1 KR 2020001523 W KR2020001523 W KR 2020001523W WO 2020218720 A1 WO2020218720 A1 WO 2020218720A1
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muscle
motherwort
extract
leonurine
preventing
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PCT/KR2020/001523
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English (en)
Korean (ko)
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황재관
김창희
유지희
최나현
이지연
정영호
박예지
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(주)에프앤디넷
연세대학교 산학협력단
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Publication of WO2020218720A1 publication Critical patent/WO2020218720A1/fr

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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)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/533—Leonurus (motherwort)
    • 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 diseases or improving muscle function containing motherwort extract or leonurin.
  • 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 impairs 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 between protein synthesis and degradation. Muscular dystrophy occurs when protein degradation occurs more than synthetically (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 occurring within the muscle. When more signaling reactions that induce synthesis occur rather than degradation of muscle protein, muscle protein synthesis is increased, which is indicated by an increase in muscle size (hypertrophy) or an increase in the number of muscle fibers (hyperplasia) according to the increase in muscle protein (The Korea Journal). of Sports Science, 20(3): 1551-1561, 2011).
  • PI3K phosphatidylinositol-3 kinase
  • mTOR mammalian target of rapamycin
  • mTOR contributes to muscle mass increase by inducing muscle protein synthesis by activating two factors that initiate mRNA translation, 4E-binding protein (4EBP1) and phosphorylated 70-kDa ribosomal S6 kinase (p70S6K) (The Korea Journal of Sports Science, 20(3): 1551-1561, 2011; The International Journal of Biochemistry and Cell Biology, 43(9): 1267-1276, 2011).
  • 4EBP1 4E-binding protein
  • p70S6K phosphorylated 70-kDa ribosomal S6 kinase
  • the present inventors have completed the present invention by finding a natural substance that has excellent muscle function modulating activity and can be safely applied, confirming that motherwort extract or leonurin has an activity to prevent or treat muscle disease or improve muscle function.
  • an object of the present invention is to provide a pharmaceutical composition for preventing or treating muscle diseases comprising motherwort extract or leonurine of Formula 1 as an active ingredient.
  • Another object of the present invention is to provide a food composition for preventing or improving muscle diseases comprising motherwort extract or leonurine of the following formula (1) as an active ingredient.
  • Another object of the present invention is to provide a cosmetic composition for preventing or improving muscle diseases comprising motherwort extract or leonurine of Formula 1 as an active ingredient.
  • Another object of the present invention is to provide the use of motherwort extract or leonurine represented by the following formula (1) for the manufacture of a drug for preventing or treating muscle diseases.
  • Another object of the present invention is to provide a method for treating muscle disease, comprising administering to a patient with muscle disease a pharmaceutical composition comprising a motherwort extract or leonurine represented by the following formula (1) as an active ingredient.
  • the present invention provides a pharmaceutical composition for preventing or treating muscle diseases comprising motherwort extract or leonurine of Formula 1 as an active ingredient.
  • the present invention provides a food composition for preventing or improving muscle diseases comprising motherwort extract or leonurine of the following formula (1) as an active ingredient.
  • the present invention provides a cosmetic composition for preventing or improving muscle diseases comprising motherwort extract or leonurine of Formula 1 as an active ingredient.
  • the present invention provides the use of motherwort extract or leonurine represented by the following formula (1) for the manufacture of a drug for preventing or treating muscle diseases.
  • the present invention provides a method for treating muscle diseases comprising administering to a patient with muscle disease a pharmaceutical composition comprising a motherwort extract or leonurine represented by the following Formula 1 as an active ingredient.
  • Motherwort extract or leonurin according to the present invention has excellent effects in increasing the activity of mTOR involved in muscle protein synthesis and inhibiting the mRNA expression of MuRF1 and atrogin-1 involved in muscle protein degradation.
  • the present invention is a natural product, it can be safely used without side effects, and thus can be used as pharmaceuticals, foods or cosmetics.
  • the present invention uses motherwort extract or leonurin for preventing or treating muscle diseases, or for improving muscle function;
  • it provides a method for preventing or treating muscle disease, or improving muscle function, comprising applying motherwort extract or leonurin to mammals, including humans:
  • motherwort' refers to the drying of the above-ground part of Leonurus japonicus belonging to the family Lamiaceae (Labiatae) motherwort genus (Leonurus ssp.).
  • motherwort extract means an extract obtained by extracting motherwort.
  • the preparation method of the motherwort extract can be obtained by extracting from the motherwort with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and a subcritical or supercritical fluid.
  • 'leonurine' includes a compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof.
  • the leonurin of the present invention is a representative bioactive component present in motherwort, and can be obtained by separating and purifying from motherwort, or synthesized or purchased.
  • muscle refers to tendons and muscles generically.
  • Muscle function means the ability to exert power by contraction of muscles, and muscle function is proportional to muscle mass.
  • the term "improving muscle function" in the present specification refers to improving muscle function better by increasing muscle mass.
  • the present invention provides a pharmaceutical composition for preventing or treating muscle disease, a food composition for preventing or improving muscle function, or a cosmetic composition for improving muscle function, containing motherwort extract or leonurin as an active ingredient.
  • leonurine can be obtained by isolation and purification from motherwort extract.
  • the motherwort extract may be an ethanol extract, hot water extract, hexane extract, ethyl acetate extract, ultra-high pressure extract, subcritical extract, and supercritical extract.
  • the motherwort extract can be obtained by extracting the motherwort with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, subcritical and supercritical. It can also be obtained by extracting the motherwort under ultra-high pressure conditions of 100 MPa or more. If necessary, it can be prepared by additionally including filtration and concentration steps according to methods known in the art.
  • the organic solvent having 1 to 6 carbon atoms is an alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, It may be at least one selected from the group consisting of methylene chloride, hexane, cyclohexane, and petroleum ether.
  • the extraction method may be applied to motherwort and used as a composition for improving muscle function.
  • the composition for preventing or treating muscle diseases of the present invention is a pharmaceutical composition
  • it can be used for preventing or treating muscle diseases caused by muscle wasting or degeneration.
  • Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by a gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or voluntary muscles and heart muscles.
  • Examples of related diseases include atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia, and sarcopenia.
  • the composition of the present invention has an effect of increasing muscle mass, and the type of muscle is not limited.
  • the pharmaceutical composition of the present invention may contain a pharmaceutically acceptable salt of leonurine.
  • pharmaceutically acceptable refers to a physiologically acceptable and when administered to a human, usually does not cause an allergic reaction or a similar reaction, and the salt is a pharmaceutically acceptable free acid (free Acid addition salts formed by acid) are preferred.
  • the pharmaceutically acceptable salt of leonurine 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, aminooxyacetic acid, benzenesulfonic acid , p-toluenesulfonic acid or methanesulfonic acid.
  • the organic acid may be, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluor
  • 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 be preferably in the form of hydrochloride or acetate, and more preferably in the form of hydrochloride.
  • the acid addition salts mentioned above are a) directly mixing rheonurine and acid, b) dissolving and mixing it in a solvent or aqueous solvent, or c) placing rheonurine in an acid in a solvent or a submerged solvent and mixing them. It is prepared by a general salt preparation method.
  • gaba salt gabapentin salt, pregabalin salt, nicotinic acid salt, adipate salt, hemimalonic acid salt, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt or And aspartate.
  • composition for preventing or treating muscle diseases of the present invention may further include a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration.
  • Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like.
  • the carrier for parenteral administration may include water, suitable oil, saline, aqueous glucose and glycol.
  • it may further comprise a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives are benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol.
  • Other pharmaceutically acceptable carriers may be referred to as those described in the following literature (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).
  • the pharmaceutical composition of the present invention can be administered to mammals including humans by any method.
  • it can be administered orally or parenterally, and parenteral administration methods are 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 route of administration as described above.
  • one or more buffers e.g., saline or PBS
  • antioxidants e.g., bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g., aluminum hydroxide). Side
  • suspending agents e.g., thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.
  • Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions or capsules, and the like, and such solid preparations are at least one excipient in the pharmaceutical composition of the present invention, for example , Starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose , Methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethyl-cellulose or gelatin may be mixed and prepared.
  • tablets or sugar tablets can be obtained by blending the active ingredient with a solid excipient, pulverizing it, adding a suitable auxiliary, and processing into a granule mixture.
  • Liquid preparations for oral use include suspensions, liquid solutions, emulsions, or syrups, but may include various excipients, such as wetting agents, sweetening agents, fragrances, or preservatives, in addition to water or liquid paraffin, which are simple diluents commonly used. .
  • cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and an anti-coagulant, a lubricant, a wetting agent, a fragrance, an emulsifier and a preservative may be additionally included.
  • the pharmaceutical composition of the present invention may be formulated according to a method known in the art in the form of an injection, a transdermal administration, and a nasal inhalation agent together with a suitable parenteral carrier.
  • a suitable parenteral carrier In the case of such injections, they must be sterilized and protected from contamination by microorganisms such as bacteria and fungi.
  • suitable carriers for injections include, but are not limited to, water, ethanol, polyol (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), a mixture thereof and/or a solvent or dispersion medium containing vegetable oil. I can.
  • suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine 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.
  • various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like may be additionally included.
  • the injection may further include an isotonic agent such as sugar or sodium chloride in most cases.
  • transdermal administration means that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin by topically administering the pharmaceutical composition to the skin.
  • the compounds used according to the invention can be prepared using a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas, pressurized pack or It can be conveniently delivered from a nebulizer in the form of an aerosol spray.
  • a pressurized aerosol the dosage unit can be determined by providing a valve that delivers a metered amount.
  • gelatin capsules and cartridges for use in an inhaler or insufflator can be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a formula generally known for all pharmaceutical chemistry.
  • the pharmaceutical composition for preventing or treating muscle diseases of the present invention can provide a desirable muscle disease prevention and treatment effect when it contains motherwort extract or leonurin in an effective amount.
  • the term'effective amount' refers to an amount that exhibits a higher response compared to the negative control group, and preferably refers to an amount sufficient to improve muscle function.
  • the pharmaceutical composition of the present invention may contain 0.01 to 99.99% of motherwort extract or leonurin, and the balance may be occupied by a pharmaceutically acceptable carrier.
  • the effective amount of motherwort extract or leonurin contained in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.
  • the total effective amount of the pharmaceutical composition of the present invention may be administered to a patient in a single dose, and may be administered by a fractionated treatment protocol that is administered for a long time in multiple doses.
  • the pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease.
  • motherwort extract or leonurin is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per 1 kg of body weight per day, and when administered orally, motherwort extract or leonurin Based on 1 kg of body weight per day, preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per day, so that it may be administered in divided doses 1 to several times.
  • the dose of motherwort extract or leonurin is effective administration to patients in consideration of various factors such as age, weight, health status, sex, disease severity, diet and excretion rate, as well as the route and number of treatments of the pharmaceutical composition. Since the amount is determined, considering these points, those of ordinary skill in the art will be able to determine an appropriate effective dosage according to the specific use of the motherwort extract or leonurin 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 exhibits the effects of the present invention.
  • the pharmaceutical composition for preventing or treating muscle diseases of the present invention may be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, or methods using a biological response modifier.
  • the pharmaceutical composition for preventing or treating muscle diseases of the present invention may also be provided in the form of an external preparation comprising motherwort extract or leonurin as an active ingredient.
  • the pharmaceutical composition for preventing or treating muscle diseases of the present invention is used as an external preparation for skin, additionally fatty substances, organic solvents, solubilizing agents, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents ), fragrance, surfactant, water, ionic emulsifier, nonionic emulsifier, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic activator, lipophilic activator Or it may contain adjuvants commonly used in the field of dermatology such as any other ingredients commonly used in skin external preparations such as lipid vesicles. In addition, the above ingredients may be introduced in amounts generally used in the field of dermatology.
  • the pharmaceutical composition for preventing or treating muscle diseases of the present invention is provided as an external preparation for skin, it may be a formulation such as an ointment, patch, gel, cream, or spray, but is not limited thereto.
  • the composition for preventing muscle disease or improving muscle function of the present invention may also be a food composition.
  • a food composition for preventing muscle disease or improving muscle function of the present invention it may be used for preventing or improving muscle disease caused by muscle wasting or degeneration.
  • Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by a gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or voluntary muscles and heart muscles.
  • related diseases include atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia, and sarcopenia.
  • the composition of the present invention has an effect of increasing muscle mass, and the type of muscle is not limited.
  • the food composition of the present invention includes all forms such as functional food, nutritional supplement, health food, food additives and feed, and contains humans or animals including livestock. It is targeted for 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.
  • General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruit, canned food, jam, marmalade, etc.), fish, meat and processed foods thereof (e.g. ham, sausage) Corn beef), bread and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, sweets, dairy products (e.g.
  • motherwort extract itself can be prepared in the form of tea, juice, and drink, and liquefied, granulated, encapsulated, and powdered so that it can be consumed (health drink). have.
  • motherwort extract or leonurin in the form of a food additive, it may be prepared and used in the form of a powder or a concentrate. In addition, it may be prepared in the form of a composition by mixing motherwort extract or leonurin with a known active ingredient known to have an effect of preventing muscle disease and improving muscle function.
  • the health drink composition may contain various flavors or natural carbohydrates as an additional component, such as a conventional beverage.
  • the natural carbohydrates described above include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, and erythritol.
  • Sweeteners include natural sweeteners such as taumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can be used.
  • the proportion of the natural carbohydrate is generally about 0.01%, preferably about 0.02% per 100 mL of the composition of the present invention.
  • Motherwort extract or leonurin may be contained as an active ingredient in a food composition for preventing muscle disease and improving muscle function, and the amount is not particularly limited to an amount effective to achieve an action for preventing muscle disease and improving muscle function, but It is preferably 0.01 to 100% by weight based on the total weight of the composition.
  • the food composition of the present invention may be prepared by mixing motherwort extract or leonurin with other active ingredients known to be effective in preventing muscle disease and improving muscle function.
  • the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, It may contain glycerin, alcohol or a carbonating agent.
  • the health food of the present invention may contain flesh for the manufacture of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients may be used independently or in combination. The ratio of these additives is not very important, but it is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
  • the composition for improving muscle function of the present invention may also be a cosmetic composition.
  • the cosmetic composition of the present invention contains motherwort extract or leonurin as an active ingredient, and basic cosmetic composition (face wash such as lotion, cream, essence, cleansing foam and cleansing water, pack, body oil) with dermatologically acceptable excipients, It can be prepared in the form of a color cosmetic composition (foundation, lipstick, mascara, makeup base), hair product composition (shampoo, conditioner, hair conditioner, hair gel), and soap.
  • the excipient is not limited thereto, but may include, for example, an emollient, a skin penetration enhancer, a colorant, a fragrance, an emulsifier, a thickener, and a solvent.
  • an emollient for example, an emollient, a skin penetration enhancer, a colorant, a fragrance, an emulsifier, a thickener, and a solvent.
  • flavoring, coloring, disinfecting agents, antioxidants, preservatives, moisturizing agents, etc. may be additionally included
  • thickeners, inorganic salts, synthetic polymer materials, etc. may be included for the purpose of improving physical properties.
  • it can be easily prepared by adding motherwort extract or leonurine to a common face wash and soap base.
  • a cream it can be prepared by adding motherwort extract or leonurine or a salt thereof to a cream base of a general oil-in-water (O/W) type.
  • Synthetic or natural materials such as proteins, minerals, vitamins, etc. for the purpose of improving physical properties, such as fragrances, chelating agents, coloring agents, antioxidants, preservatives, etc., may be additionally added.
  • the content of motherwort extract or leonurin contained in the cosmetic composition of the present invention is not limited thereto, 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 composition.
  • the content is less than 0.001% by weight, the desired anti-aging or wrinkle improvement effect cannot be expected, and when the content is more than 10% by weight, there may be difficulties in safety or formulation.
  • the dried motherwort was pulverized with a mixer, and then 10 g of the crushed motherwort 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, and then hot water extract of motherwort was obtained.
  • the dried motherwort was pulverized with a mixer, and then 10 g of the pulverized motherwort 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, and then a motherwort hexane extract was obtained.
  • the dried motherwort was pulverized with a mixer, and then 50 g of the crushed motherwort was put into a subcritical water reactor of a subcritical extraction apparatus (Biovan, Gyeonggi, Korea) together with 1 L of 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 perform extraction. After 20 minutes, the extract was transferred to a storage tank supplied with cooling water, rapidly cooled to 30° C., and then centrifuged at 3,600 rpm for 30 minutes to separate the floating residue, and only the supernatant was taken. A freeze dryer (Ilshin Lab Co. Ltd., Seoul, Korea) was used to remove all of the solvent to obtain a motherwort subcritical extract.
  • a freeze dryer Ilshin Lab Co. Ltd., Seoul, Korea
  • the dried motherwort was pulverized with a mixer, and then 1 g of the crushed motherwort was charged into a sample cartridge and extracted using a supercritical fluid extraction device (SFX 3560, Isco Inc., Lincoln, NE, USA).
  • a supercritical fluid extraction device SFX 3560, Isco Inc., Lincoln, NE, USA.
  • the extraction pressure was 300 bar
  • the extraction temperature was 50 °C
  • the flow rate of supercritical carbon dioxide was 60 mL/min
  • the extraction time was 2 hours.
  • the pressure of the extraction device was lowered to release the supercritical fluid state to obtain a motherwort supercritical extract.
  • Example 2 Effect of motherwort extract on increasing mTOR activity, a key biomarker for muscle protein synthesis
  • mTOR protein When mTOR protein is phosphorylated and activated, it is known that 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 muscle production induction activity of motherwort, the activity of mTOR was confirmed using the mTOR Sandwich ELISA kit (Cell Signaling Technology, Beverly, MA, USA).
  • L6 myoblasts ATCC; Manassas, VA, USA
  • DMEM Dulbecco's modified Eagle's media
  • FBS fetal bovine serum
  • HS horse serum
  • the motherwort extract prepared in Example 1 was dissolved in DMEM (Hyclone) at 10 ⁇ g/mL, respectively, and then treated on 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 by Bradford (Bio-Rad Laboratories Inc., Hercules, CA, USA) method, and then the cell lysate was dispensed into microwells and cultured at 37° C. for 2 hours. After incubation, after washing a total of 4 times with a washing buffer, a detection antibody was treated and incubated for 1 hour at 37°C. After washing a total of 4 times with a washing buffer, the secondary antibody was removed.
  • DMEM Hyclone
  • L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at 1 ⁇ 10 5 cells/mL.
  • DMEM Hyclone
  • FBS Hyclone
  • the medium in the well was removed and the cells were treated with DMEM (Hyclone) containing 2% HS (Hyclone) to induce myotube differentiation. Differentiation was carried out for a total of 6 days by replacing with a new medium once every 2 days.
  • DMEM Hyclone
  • TNF- ⁇ tumor necrosis factor alpha
  • the obtained cell lysate was centrifuged at 13,000 rpm for 10 minutes to obtain a supernatant.
  • the protein concentration in the supernatant was quantified by Bradford (Bio-Rad Laboratories Inc.), and then the protein was heated for 5 minutes and developed on a 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gel to separate the protein.
  • SDS-PAGE 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis
  • ⁇ -tubulin antibody Cell Signaling Technology
  • BSA bovine serum albumin
  • HRP Horseradish peroxidase conjugated secondary antibody
  • BSA horseradish peroxidase conjugated secondary antibody
  • the protein band was colored using ECL Western Blot detection reagent (GE Healthcare, Piscataway, NJ, USA), and the colored protein band was confirmed using the G;BOX EF imaging system (Syngene, Cambridge, UK). The results are shown in FIG. 2.
  • L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at 1 ⁇ 10 5 cells/mL.
  • DMEM Hyclone
  • FBS Hyclone
  • the medium in the well was removed and the cells were treated with DMEM (Hyclone) containing 2% HS (Hyclone) to induce myotube differentiation. Differentiation was carried out for a total of 6 days by replacing with a new medium once every 2 days.
  • Example 1-1 After differentiation, the motherwort hot-water extract prepared in Example 1-1 was dissolved in DMEM (Hyclone) containing 50 ng/mL TNF- ⁇ (PeproTech) at a concentration of 40 and 80 ⁇ g/mL in DMEM, and then in cells. Treated and incubated for 12 hours. Total RNA was isolated using a TRIzol reagent (Takara, Otsu, Japan). The isolated total RNA was quantified using NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, MA, USA.
  • DMEM Hyclone
  • TNF- ⁇ PeproTech
  • 16 ⁇ L of quantified RNA was prepared at 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, Foster City, CA, USA). It was synthesized with cDNA under conditions. Of the 16 ⁇ L of the generated cDNA, 1 ⁇ L of cDNA, the following specific primers (Bioneer, Daejeon, Korea) and PCR premix (ELPIS-Biotech) were used for 30 seconds at 95 °C, 30 seconds at 60 °C, 45 seconds at 72 °C. PCR was performed by repeating 35 times.
  • Reverse primer 5'-AGCCTGGAAGATGTCGTTGG-3' (SEQ ID NO: 2)
  • Reverse primer 5'-CGTCTTCGTGTTCCTTGCAC-3' (SEQ ID NO: 4)
  • Reverse primer 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6)
  • the amplified cDNA as a result of PCR was separated by electrophoresis with 1.5% agarose gel, and the cDNA band was confirmed using a G;BOX EF imaging system (Syngene, Cambridge, UK).
  • Example 4 The experiment was carried out in the same manner as in Example 4, except that the ethanol extract of motherwort prepared in Example 1-3 was treated with the cells at concentrations of 10 and 20 ⁇ g/mL.
  • the ethanol extract of motherwort of the present invention has excellent ability to inhibit the expression of biomarkers involved in the decomposition of muscle proteins in muscle cells.
  • L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at 1 ⁇ 10 5 cells/mL.
  • DMEM Hyclone
  • FBS Hyclone
  • Example 1-1 The hot water extract of motherwort prepared in was dissolved at a concentration of 40 and 80 ⁇ g/mL, and then treated on the cells to induce myotube differentiation. Differentiation was carried out for a total of 6 days by replacing with a new medium once every 2 days.
  • Reverse primer 5'-GGTGTAACAACCATACCCCACT-3' (SEQ ID NO: 8)
  • Reverse primer 5'-GGCCACTCACTGTCTCTCAAA-3' (SEQ ID NO: 10)
  • Reverse primer 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6)
  • the amplified cDNA as a result of PCR was separated by electrophoresis with 1.5% agarose gel, and the cDNA band was confirmed using a G;BOX EF imaging system (Syngene).
  • Example 3 The experiment was carried out in the same manner as in Example 3, except that the cells were treated with Leonurine at 20 and 40 ⁇ M.
  • the experiment was carried out in the same manner as in Example 4, except that the cells were treated with Leonurine at 20 and 40 ⁇ M.
  • Example 6 The experiment was carried out in the same manner as in Example 6, except that the cells were treated with Leonurine at 20 and 40 ⁇ M.
  • mice 7-week-old male mice (C57BL/6J; DBL, Korea) were purchased and tested. All animals were reared at the Yonsei Laboratory Animal Reaserch Center (YLARC, Seoul, Korea), and the environment of the breeding room was maintained at a temperature of 23 ⁇ 2°C and a relative humidity of 55 ⁇ 10%. Before the start of the experiment, a total of 20 mice were randomly divided into 5 mice per group. After acclimating for 1 week, anesthesia was induced by intraperitoneal injection of 325 mg/kg of tribromoethanol (Sigma-Aldrich).
  • the right hindlimb gastrocnemius muscle and right sole of the rats in the muscle atrophy group and the sample administration group were treated with a stapler using a skin stapler (Unidus, Chungcheongbuk-do, Korea). It was injured and the right hind limb was immobilized and kept in this condition for a week.
  • the stapler core fixed on the calf muscle and the sole of the foot was removed, and for another week, the hot water extract of motherwort prepared in Example 1-1 was 150 mg/kg or 300 mg/kg and the concentration of Leonurine 30 mg/kg Was administered orally every day.
  • the normal group and the muscular atrophy group were orally administered with saline instead of the sample.
  • mice After the oral administration period was over, the muscle strength of the mice was measured using a muscle strength meter (Panlab, Barcelona, Spain). The tail of the rat was pulled with a constant force until the rat released the youngest of the strength tester, and a total of 5 consecutive tests were performed per animal.
  • muscle strength was significantly reduced in the muscular atrophy group compared to the normal group ( ## p ⁇ 0.01), but the muscle strength was significantly decreased by treatment with hot water extract of motherwort and leonurin ( ** p ⁇ 0.01). ).
  • the hot water extract of motherwort and leonurin of the present invention have excellent effects of increasing muscle strength reduced due to muscle atrophy.
  • the experimental animal was intraperitoneally injected with 325 mg/kg of tribromomethanol (Sigma-Aldrich) and sacrificed through cardiac blood collection after anesthesia. After confirming that the heartbeat had stopped, the uninjured tibialis anterior muscle was removed from the right hind limb and the weight was measured.
  • tribromomethanol Sigma-Aldrich
  • the weight of the tibialis anterior muscle in the muscular atrophy group was significantly reduced ( ## p ⁇ 0.01) compared to the normal group, but the weight was significant ( * p ⁇ 0.05, ** p ⁇ 0.01).
  • the hot water extract of motherwort and leonurin of the present invention have excellent effects of increasing the weight of the reduced muscle due to muscle atrophy.
  • Example 10-1 After inducing muscle atrophy in the same manner as in Example 10-1, the ethanol extract of motherwort prepared in Example 1-3 was orally administered daily at a concentration of 200 mg/kg.
  • the motherwort ethanol extract-administered group had muscle strength and muscle weight of 23.91% ( ** p ⁇ 0.01) compared to the muscle atrophy group, respectively. ) And 17.28% ( ** p ⁇ 0.01). This means that the ethanol extract of motherwort of this onset is excellent in increasing muscle strength and muscle weight, which is reduced due to muscle atrophy.
  • the motherwort subcritical extract prepared in Example 1-6 was orally administered daily at a concentration of 200 mg/kg.
  • the motherwort subcritical extract-administered group had muscle strength and muscle weight of 24.97%, respectively, compared to the muscle atrophy group ( ** p ⁇ 0.01) and 20.60% ( ** p ⁇ 0.01). This means that the motherwort subcritical extract of this onset has an excellent effect of increasing the muscle strength and muscle weight reduced due to muscle atrophy.
  • compositions of Preparation Examples 1 to 3 were prepared according to the following composition and composition ratio with motherwort extract or leonurine having excellent muscle disease prevention or treatment, or muscle function improvement effect according to a conventional method. .
  • the powder was prepared by filling it in an airtight cloth according to a conventional powder preparation method.
  • tablets After mixing motherwort extract or 50 mg of leonurine, 400 mg of crystalline cellulose, and 5 mg of magnesium stearate, tablets were prepared by tableting according to a conventional tablet preparation method.
  • the capsules were prepared by filling them into gelatin capsules according to a conventional capsule preparation method.
  • 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, Nicotinamide 1.7 mg, folic acid 50 ug, calcium pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, dicalcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate It can be prepared by mixing 100 mg and 24.8 mg of magnesium chloride, and the mixing ratio may be arbitrarily modified. After mixing the above ingredients according to a conventional health food manufacturing method, granules are prepared, and the usual method It can be used in the manufacture of health food composition according to.
  • a chewing gum was prepared by a conventional method by combining 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of flavor, 2% by weight of water, and 0.1% by weight of motherwort extract or leonurin.
  • Nutrient cosmetic water was prepared according to a conventional method using motherwort extract or leonurine in the ratio of the nutrient cosmetic water formulation shown in Table 1 below.
  • Example 3-1 (% by weight) Motherwort Extract or Leonurine 2.0 Squalane 5.0 Beeswax 4.0 Polysorbate 60 1.5 Sorbitansquioleate 1.5 Floating paraffin 0.5 Caprylic/Capric Triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservatives, colors, flavors Appropriate amount Purified water to 100
  • Softening lotion (skin lotion) Softening lotion was prepared according to a conventional method by using motherwort extract or leonurine in the ratio of the softening lotion formulation shown in Table 2 below.
  • a massage cream was prepared according to a conventional method by using the motherwort extract or leonurin in the ratio of the massage cream formulation shown in Table 4 below.
  • a pack was prepared according to a conventional method by using the pack motherwort extract or leonurine in the ratio of the pack formulation shown in Table 5 below.

Abstract

La présente invention concerne une nouvelle utilisation d'un extrait de Leonurus japonicus et de la léonurine, utilisé pour prévenir ou traiter des troubles musculaires ou pour améliorer la fonction musculaire, et concerne une composition qui permet de prévenir ou de traiter des troubles musculaires ou d'améliorer la fonction musculaire contenant l'extrait de Leonurus japonicus ou la léonurine. La composition selon la présente invention augmente l'activité de mTOR, lequel est impliqué dans la synthèse des protéines musculaires, a pour effet d'inhiber l'expression de MuRF1 et de l'atrogine-1, qui sont impliqués dans la dégradation des protéines musculaires, augmente la masse musculaire et améliore la fonction musculaire, et peut de ce fait être utilisable pour la prévention, le traitement ou l'amélioration de diverses maladies musculaires telles que l'atonie, l'atrophie musculaire, la dystrophie musculaire, la dégénérescence musculaire, la myasthénie, la cachexie et la sarcopénie.
PCT/KR2020/001523 2019-04-22 2020-01-31 Composition pour la prévention ou le traitement de troubles musculaires ou l'amélioration de la fonction musculaire, contenant un extrait de leonurus japonicus ou de la léonurine WO2020218720A1 (fr)

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