KR102124986B1 - Composition for prevention or treatment of muscular disorder or improvement of muscular functions comprising Leonurus japonicus extract or leonurine - Google Patents

Abstract

The present invention relates to a novel use of an extract of Leonurus japonicus or leonurine for preventing or treating muscular diseases, or for improving muscular functions, and particularly, to a composition for preventing or treating muscular diseases, or for improving muscular functions, including an extract of Leonurus japonicus or leonurine. The composition has effects of increasing activity of mTOR participating in muscular protein synthesis and inhibiting expression of MuRF1 and atrogin-1 participating in muscular protein decomposition, increases muscle mass, and improves muscular functions. Therefore, the composition can be used advantageously for preventing, treating or alleviating various muscular diseases, such as atony, muscular atrophy, muscular dystrophy, muscular degeneration, myasthenia, cachexia and sarcopenia.

Description

Composition for prevention or treatment of muscular disorder or improvement of muscular functions comprising Leonurus japonicus extract or leonurine

The present invention relates to a composition for preventing and treating muscle disease 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 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 protein degradation occurs more than synthesis (The International Journal of Biochemistry and Cell Biology, 37(10): 1985-1996, 2005).

Muscle size is regulated by intracellular signaling pathways that induce anabolic or catabolism in the muscles. When there are more signal transduction reactions that induce synthesis rather than degradation of muscle protein, muscle protein synthesis increases, which results in an increase in muscle size (hypertrophy, muscle hypertrophy) or an increase in muscle fiber number (hyperplasia) with muscle protein increase (The Korea Journal) of Sports Science, 20(3): 1551-1561, 2011).

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). On the other hand, when the transcription factor forhead box (FoxO) moves from the cytoplasm to the nucleus, the expression of the E3 ubiquitin ligase factors atrogin-1 and MuRF1, which are involved in proteolysis, 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 MuRF1 expression increases muscle protein and increases muscle mass.

Muscle cell differentiation and muscle formation are regulated by a variety of muscle regulatory factors. Among them, induction of myogenin expression by the activity of MyoD is the most important factor for 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).

Chinese motherwort belonging to the family Lamitae uses the outpost as a medicinal plant (Journal of Physiology & Pathology in Korean Medicine, 30(2): 81-87, 2016). Motherwort has anti-obesity (Nutrients, 10(1): 20, 2018), antioxidant (Progress in Nutrition, 20: 46-58, 2018), anti-cancer activity (Journal of Ethnopharmacology, 122: 234-239, 2014) Is reported. In addition, anti-oxidation (Stroke, 41: 2661-2668, 2010), anti-inflammatory (Inflammation, 38(1): 79-88, 2015), anti-arteriosclerosis (Cellular) for leonurine, a representative active ingredient in motherwort Physiology and Biochemistry, 43(4): 1703-1717, 2017).

However, prior to the present invention, it was not known about the prevention and treatment of muscle disease or improvement of muscle function of motherwort extract or Leonurin.

Accordingly, the present inventors have completed the present invention by confirming that as a result of exploring natural substances that have excellent muscle function control activity and can be safely applied, the motherwort extract or leonurin has activity to prevent or treat muscle disease, or improve muscle function.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating muscle disease comprising motherwort extract or Leonurin of Formula 1 as an active ingredient.

[Formula 1]

Another object of the present invention is to provide a food composition for preventing or improving muscle disease comprising motherwort extract or Leonurin of Formula 1 below as an active ingredient.

[Formula 1]

Another object of the present invention is to provide a cosmetic composition for preventing or improving muscle disease comprising motherwort extract or Leonurin of Formula 1 as an active ingredient.

[Formula 1]

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating muscle disease comprising motherwort extract or Leonurin of Formula 1 as an active ingredient.

[Formula 1]

In addition, the present invention provides a food composition for preventing or improving muscle disease comprising motherwort extract or Leonurin of Formula 1 as an active ingredient.

[Formula 1]

In addition, the present invention provides a cosmetic composition for preventing or improving muscle disease comprising motherwort extract or Leonurin of Formula 1 as an active ingredient.

[Formula 1]

Motherwort extract or Leonurin according to the present invention has an excellent effect in suppressing mRNA expression of MuRF1 and atrogin-1 involved in muscle protein degradation and increasing the activity of mTOR involved in muscle protein synthesis. In addition, by increasing muscle mass and improving muscle function, it is possible to prevent, treat, or improve muscle function caused by various diseases and muscle loss. Since the present invention is a natural product, it can be safely used without side effects, and thus can be used as a medicine, food, or cosmetic.

1 is a result confirming the effect of increasing the activity of mTOR according to the motherwort extract treatment in L6 muscle cells.
2 is a result confirming the effect of increasing the protein expression level of p-mTOR, p-p70S6K, p-4EBP-1 according to the treatment of motherwort hydrothermal extract in L6 muscle cells.
Figure 3 is a result confirming the effect of reducing the mRNA expression of atrogin-1 and MuRF1 according to the motherwort hydrothermal extract treatment in L6 muscle cells.
4 is a result confirming the effect of reducing the mRNA expression level of atrogin-1 and MuRF1 according to the treatment of motherwort ethanol extract in L6 muscle cells.
5 is a result confirming the effect of increasing the mRNA expression and expression of MyoD and myogenin according to the motherwort hydrothermal extract treatment in L6 muscle cells.
Figure 6 is a result confirming the effect of increasing the protein expression level of p-mTOR, p-p70S6K, p-4EBP-1 according to Leonurin treatment in L6 muscle cells.
7 is a result confirming the effect of reducing the mRNA expression level of atrogin-1 and MuRF1 according to the treatment of Leonurin in L6 muscle cells.
8 is a result confirming the effect of increasing the mRNA expression and expression of MyoD and myogenin according to the treatment of motherwort hydrothermal extract in L6 muscle cells.
9 is a result of confirming the effect of improving muscle strength according to the motherwort hydrothermal extract and leonurin treatment in muscle atrophy-induced mice.
10 is a result confirming the effect of increasing the weight of the tibialis anterior muscle according to the motherwort hydrothermal extract and leonurin treatment in the atrophy-induced rat.

Hereinafter, the configuration of the present invention will be described in detail.

The present invention is a motherwort extract or Leonurin for preventing or treating muscle diseases, or for improving muscle function; A composition for preventing or treating muscle disease comprising motherwort extract or leonurin, or for improving muscle function; Or providing a method for preventing or treating muscle disease, or improving muscle function, comprising applying motherwort extract or leonurin to a mammal, including a human:

In the present specification,'motherwort' is a dry part of the ground of Leonurus japonicus belonging to the family Lamitae (Leonurus ssp.).

In the present specification, the term'motherwort extract' means an extract obtained by extracting motherwort. The method for preparing motherwort extract can be obtained by extracting from 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.

'Leonurin' in the present specification includes a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

[Formula 1]

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 and purchased.

In this specification,'muscular' refers to tendons and muscles in general.'Muscular function' refers to the ability to exert power by contraction of muscle, and muscle function is proportional to muscle mass. As used herein, the term'improve muscle function' refers to increasing muscle mass to improve muscle function better.

The present invention provides a pharmaceutical composition for preventing or treating muscle disease containing motherwort extract or leonurin as an active ingredient, a food composition for preventing muscle disease or improving muscle function, or a cosmetic composition for improving muscle function.

In one embodiment, leonurin can be obtained by isolation and purification from motherwort extract.

In one embodiment, the motherwort extract may be ethanol extract, hot water extract, hexane extract, ethyl acetate extract, ultra high pressure extract, subcritical extract, supercritical extract.

In one embodiment, motherwort extract can be obtained by extracting 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. Motherwort can also be obtained by extraction under ultra high pressure conditions of 100 MPa or more. If necessary, it may be prepared by additionally including a filtration and concentration step according to methods known in the art.

In one embodiment, the organic solvent having 1 to 6 carbon atoms is an alcohol having 1 to 6 carbon atoms, acetone (acetone), ether (ether), benzene (benzene), chloroform (chloroform), ethyl acetate (ethyl acetate), It may be one or more selected from the group consisting of methylene chloride, hexane, cyclohexane, and petroleum ether.

The extraction method may be applied to motherwort to be used as a composition for improving muscle function.

When the composition for preventing or treating muscle disease of the present invention is a pharmaceutical composition, it may be used for the prevention or treatment of 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, muscle 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 pharmaceutical composition of the present invention may include a pharmaceutically acceptable salt of leonurin. As used herein, the term “pharmaceutically acceptable” refers to a physiologically acceptable drug that, when administered to a human, typically does not cause an allergic reaction or similar reaction, and the salt is a pharmaceutically acceptable free acid (free). acid) is preferred.

The pharmaceutically acceptable salt of the Leonurin 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.

The above-mentioned acid addition salts are either a) direct mixing of leonurin and an acid, or b) dissolving and mixing it in a solvent or water-containing solvent, or c) positioning of leonurin in an acid in a solvent or a water-soluble solvent and mixing them. It is prepared by the general salt preparation method.

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.

In addition, the pharmaceutical 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. 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).

The pharmaceutical composition of the present invention can be administered to mammals, including humans, by any method. 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.

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), a suspending agent, a thickening agent wetting agent, a disintegrating agent or a surfactant, a diluent or an excipient.

Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions or capsules, etc. These solid preparations include, for example, 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 blending the active ingredient with a solid excipient and then grinding it and adding a suitable adjuvant to the granule mixture.

In addition to simple excipients, lubricants such as magnesium styrene 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. .

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, lubricant, wetting agent, fragrance, emulsifier, and preservative. .

When administered parenterally, the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations, and nasal inhalants together with suitable parenteral carriers. 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 be. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with 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, various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, etc. may be further included. In addition, the injection may additionally contain isotonic agents such as sugars or sodium chloride in most cases.

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.

In the case of inhalation dosages, the compounds used according to the invention may be used in the form of pressurized packs, using suitable propellants, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. 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.

The pharmaceutical composition for preventing or treating muscle diseases of the present invention may provide a desirable muscle disease prevention and treatment effect when the motherwort extract or leonurin 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 motherwort extract or leonurin may be included in the pharmaceutical composition of the present invention 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 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 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 parenterally administered, 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 motherwort extract or leonurin, and motherwort extract or leonurin when administered orally. It may be divided into 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 1 kg of body weight per day. However, the dose of motherwort extract or leonurine is effective for the patient taking into consideration various factors such as the patient's age, weight, health status, sex, disease severity, diet and excretion rate, as well as the route and frequency of administration of the pharmaceutical composition. Since the amount is determined, those who have 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 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 shows 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 methods using surgery, radiation therapy, hormone therapy, chemotherapy or biological response modifiers.

The pharmaceutical composition for preventing or treating muscle diseases of the present invention may also be provided as a formulation of an external preparation containing motherwort extract or leonurin as an active ingredient.

When the pharmaceutical composition for preventing or 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.

When the pharmaceutical composition for preventing or treating muscle diseases of the present invention is provided as an external preparation for skin, the present invention is not limited thereto, and may be a formulation such as ointment, patch, gel, cream or spray.

The composition for preventing muscle disease or improving muscle function of the present invention may also be a food composition. When the food composition for preventing muscle disease or improving muscle function of the present invention is a food composition, it may be used for preventing or improving 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, muscle 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. Drinks (including alcoholic beverages), fruits and processed foods thereof (e.g. canned fruits, canned 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 vegetable oil, margarine , Vegetable protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding motherwort extract. In addition, it is not limited to the nutritional supplement, but may be prepared by adding motherwort extract or leonurin to capsules, tablets, pills, etc. In addition, the health functional food is not limited to this, for example, the motherwort extract itself can be consumed by liquefaction, granulation, encapsulation, and powdering to prepare and drink (health drink) in the form of tea, juice, and drink. have. In addition, in order to use motherwort extract or leonurin in the form of food additives, it can be prepared and used in powder or concentrate form. In addition, motherwort extract or Leonurin can be prepared in the form of a composition by mixing with known active ingredients known to have muscle disease prevention and muscle function improvement effects.

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 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; 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 can be used. The ratio of the above natural carbohydrate is generally about 0.01 g per 100 mL of the composition of the present invention, preferably about 0.02 g.

The motherwort extract or Leonurin 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 may be prepared by mixing with motherwort extract or leonurin together with other active ingredients known to be effective in the composition for preventing muscle disease and improving muscle function.

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.

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 a basic cosmetic composition (face wash, cream, essence, cleansing foam and cleansing water-like cleansing agent, pack, and vodioyl), along with dermatologically acceptable excipients, It can be prepared in the form of color cosmetic composition (foundation, lipstick, mascara, makeup base), hair product composition (shampoo, conditioner, hair conditioner, hair gel) and soap.

The excipients include, but are not limited to, for example, skin emollients, skin penetration enhancers, colorants, fragrances, emulsifiers, thickeners and solvents. In addition, it may further include flavoring agents, coloring agents, disinfecting agents, antioxidants, preservatives and moisturizing agents, and may include thickeners, inorganic salts, synthetic polymer materials, etc. for the purpose of improving physical properties. 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 motherwort extract or leonurin to a conventional cleanser and soap base. When preparing a cream, it can be prepared by adding motherwort extract or Leonurin 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.

The content of motherwort extract or leonurin 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 relative to the total weight of the total composition. If the content is less than 0.001% by weight, the desired anti-aging or anti-wrinkle effect cannot be expected, and if it is more than 10% by weight, there may be difficulty in safety or preparation of the formulation.

Hereinafter, the present invention will be described in more detail through examples.

However, the following examples are only to illustrate the present invention, the content of the present invention is not limited to the following examples.

[Example 1] Preparation of the extract above the motherwort

<1-1> Preparation of Ikmocho hot water extract

After crushing the dried motherwort with a mixer, 10 g of the ground 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 to obtain a motherwort hot water extract.

<1-2> Preparation of 50% ethanol extract of motherwort

After crushing the dried motherwort with a mixer, 10 g of the ground motherwort sample was added to 100 mL of 50% ethanol mixed with ethanol and water in 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 50% ethanol extract of motherwort.

<1-3> Preparation of ethanol extract of motherwort

After crushing the dried motherwort with a mixer, 10 g of the ground motherwort 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 motherwort ethanol extract.

<1-4> Preparation of motherwort ethyl acetate extract

After crushing the dried motherwort with a mixer, 10 g of the ground motherwort 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 component to obtain the motherwort ethyl acetate extract.

<1-5> Preparation of motherwort hexane extract

After crushing the dried motherwort with a mixer, 10 g of the ground 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 to obtain the motherwort hexane extract.

<1-6> Preparation of subcritical extract of motherwort

After crushing the dried motherwort with a mixer, 50 g of the ground motherwort 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 to which cooling water was supplied, 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. The motherwort subcritical extract was obtained by removing all of the solvent using a freeze dryer (Ilshin Lab Co. Ltd., Seoul, Korea).

<1-7> Preparation of supercritical extract of motherwort

The dried motherwort was ground with a mixer, and then 1 g of the ground motherwort 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 is completed, the pressure of the extraction device is lowered to release the supercritical fluid state to obtain a motherwort supercritical extract.

[Example 2] Increased effect of mTOR activity of core biomarker of muscle protein synthesis of motherwort extract

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, to confirm the activity of inducing muscle growth of motherwort, mTOR activity was confirmed using mTOR Sandwich ELISA kit (Cell Signaling Technology, Beverly, MA, USA).

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, the motherwort extract prepared in Example 1 was dissolved in DMEM (Hyclone) at 10 μg/mL, and then treated in 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 the method of Bradford (Bio-Rad Laboratories Inc., Hercules, CA, USA), and then the cell lysate was dispensed into a microwell and incubated at 37°C for 2 hours. After cultivation, after washing 4 times with washing buffer, the detection antibody was treated and incubated for 1 hour at 37°C. After washing 4 times with washing buffer solution, the secondary antibody was washed. Put in and incubate for 30 minutes at 37 ℃. Finally, after washing 4 times with washing buffer solution, TMB substrate was added to each well, incubated at 37°C for 10 minutes, and stop solution was added to stop TMB reaction. After 2 minutes, absorbance was measured at a wavelength of 450 nm to measure mTOR level in the root canal cells treated with motherwort hot water extract and motherwort ethanol extract. The results are shown in FIG. 1.

As a result, as shown in FIG. 1, it was confirmed that mTOR activity was significantly ( ^** p <0.01) increased in L6 muscle cells as the motherwort extract was treated. This means that the motherwort extract of the present invention has an excellent ability to increase muscle production in muscle cells.

[Example 3] The effect of increasing the expression level of biomarkers of muscle protein synthesis of motherwort hot water extract

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 carried out for a total of 6 days by replacing with a new medium once every 2 days. After differentiation, 50 ng/mL tumor necrosis factor alpha (TNF-α; PeproTech, Rocky Hills, NJ, USA) in DMEM (Hyclone) containing motherwort hydrothermal extract prepared in Example 1-1 above was added to 40 DMEM. After dissolving at a concentration of 80 μg/mL, the cells were treated and cultured for 12 hours. After incubation, cells were lysed with NP-40 buffer solution (ELPIS-Biotech, Daejeon, Korea) containing protease inhibitor cocktail (Sigma-Aldrich St. Louis, MO, USA) to obtain cell lysates. 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. The isolated protein was transferred to a nitrocellulose membrane. Then, dilute p-mTOR, mTOR, p-p70S6K, p70S6K, p-4EBP1, 4EBP1, or α-tubulin antibody (Cell Signaling Technology) in 2.5% bovine serum albumin (BSA) at a ratio of 1:1000, respectively. The protein transferred to the cellulose membrane was reacted 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). A secondary antibody (Bethyl Laboratories, Inc., Montgomery, TA, USA) conjugated with horseradish peroxidase (HRP), which recognizes the primary antibody, was diluted in 2.5% BSA to 1:5000 to room temperature with a nitrocellulose membrane for 2 hours. Was reacted, and washed three times for 10 minutes using TBST. The protein band was developed using an ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ, USA), and the protein band developed using a G;BOX EF imaging system (Syngene, Cambridge, UK) was identified. The results are shown in FIG. 2.

As a result, as shown in FIG. 2, the protein expression levels of p-mTOR, p-p70S6K and p-4EBP-1 involved in muscle protein synthesis by TNF-α were significantly decreased ( ^## p <0.01). On the other hand, it was confirmed that the protein expression levels of p-mTOR, p-p70S6K and p-4EBP-1 increased significantly ( ^** p <0.01) as the motherwort hydrothermal extract was treated. This means that the motherwort hydrothermal extract of the present invention is excellent in the ability to increase the expression of biomarkers involved in muscle protein synthesis in muscle cells.

[Example 4] Inhibitory effect of the expression of biomarkers on muscle proteolysis of motherwort hot water extract

, 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 30초, 72 ℃에서 45초를 35번 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 carried out for a total of 6 days by replacing with a new medium once every 2 days. After differentiation, the motherwort hydrothermal extract prepared in Example 1-1 was dissolved in DMEM at a concentration of 40 and 80 μg/mL in DMEM (Hyclone) containing 50 ng/mL TNF-α (PeproTech), and then in cells. Treated and incubated for 12 hours. Total RNA was isolated using 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) 42

, 55 minutes and 70 minutes

, Was synthesized with cDNA at 15 minutes. 16 μL of the generated cDNA, 1 μL of cDNA, the following specific primers (Bioneer, Daejeon, Korea) and PCR premix (ELPIS-Biotech) were performed at 95°C for 30 seconds, 60°C for 30 seconds, and 72°C for 45 seconds. PCR was repeated 35 times.

MuRF1

Forward primer: 5'-CCGGACGGAAATGCTATGGA-3' (SEQ ID NO: 1)

Reverse primer: 5'-AGCCTGGAAGATGTCGTTGG-3' (SEQ ID NO: 2)

Atrogin-1

Forward primer: 5'-ATGTCTGGAGGTCGTTTCCG-3' (SEQ ID NO: 3)

Reverse primer: 5'-CGTCTTCGTGTTCCTTGCAC-3' (SEQ ID NO: 4)

β-Actin

Forward primer: 5'-TGACAGGATGCAGAAGGAGATTAC-3' (SEQ ID NO: 5)

Reverse primer: 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6)

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, Cambridge, UK).

As a result, as shown in Figure 3, while the mRNA expression level of atrogin-1 and MuRF1 involved in muscle protein degradation by TNF-α increased significantly ( ^## p <0.01), as the motherwort hydrothermal extract was treated, It was confirmed that the mRNA expression levels of atrogin-1 and MuRF1 were significantly decreased ( ^* p <0.05, ^** p <0.01). This means that the motherwort hydrothermal extract of the present invention is excellent in inhibiting the expression of biomarkers involved in muscle protein degradation in muscle cells.

[Example 5] Inhibitory effect of the expression of biomarkers on muscle protein degradation of motherwort ethanol extract

Experiments were conducted in the same manner as in Example 4, except that the motherwort ethanol extract prepared in Example 1-3 was treated to cells at concentrations of 10 and 20 μg/mL.

As a result, as shown in FIG. 4, mRNA expression levels of atrogin-1 and MuRF1 involved in muscle protein degradation by TNF-α increased, whereas mRNA expression levels of atrogin-1 and MuRF1 were increased by processing motherwort ethanol extract. The decrease was confirmed. This means that the motherwort ethanol extract of the present invention is excellent in inhibiting the expression of biomarkers involved in muscle protein degradation in muscle cells.

[Example 6] Effect of increasing the expression level of muscle differentiation biomarkers of motherwort hot water extract

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). The motherwort hydrothermal extract prepared in was dissolved at concentrations of 40 and 80 μg/mL, and then treated with 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. At this time, a 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). 16 μL of the generated cDNA, 1 μL of cDNA, the following specific primers (Bioneer), and PCR premix (ELPIS-Biotech) were repeated 35 times at 95°C for 30 seconds, 60°C for 30 seconds, and 72°C for 45 seconds. PCR was performed.

MyoD

Forward primer: 5'-TGCCTCGGAGATAATACAGCC-3' (SEQ ID NO: 7)

Reverse primer: 5'-GGTGTAACAACCATACCCCACT-3' (SEQ ID NO: 8)

Myogenin

Forward primer: 5'-AGAGAGCCCCCTTGTTAATGC-3' (SEQ ID NO: 9)

Reverse primer: 5'-GGCCACTCACTGTCTCTCAAA-3' (SEQ ID NO: 10)

β-Actin:

Forward primer: 5'-TGACAGGATGCAGAAGGAGATTAC-3' (SEQ ID NO: 5)

Reverse primer: 5'-TAAAACGCAGCTCAGTAACAGTC-3' (SEQ ID NO: 6)

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).

As a result, as shown in FIG. 5, mRNA expression of MyoD and myogenin decreased significantly ( ^## p <0.01) as treated with TNF-α was significant by treatment of motherwort hydrothermal extract ( ^** p <0.01). It was found that the increase. This means that the motherwort hydrothermal extract of the present invention has an excellent ability to promote muscle differentiation.

[Example 7] The effect of increase in the amount of expression of biomarkers of muscle protein synthesis of leonurin

The experiment was conducted in the same manner as in Example 3, except that the cells were treated with 20 and 40 μM Leonurin.

As a result, as shown in FIG. 6, the protein expression levels of p-mTOR, p-p70S6K and p-4EBP-1 involved in muscle protein synthesis by TNF-α were significant ( ^# p <0.05, ^## p <0.01), while the level of protein expression of p-mTOR, p-p70S6K and p-4EBP-1 increased significantly ( ^* p <0.05, ^** p <0.01) as the treatment with Leonurin increased. there was. This means that the Leonurin of the present invention has excellent ability to increase the expression of biomarkers involved in muscle protein synthesis in muscle cells.

[Example 8] Leonurin's muscle protein degradation biomarker expression inhibitory effect

The experiment was conducted in the same manner as in Example 4, except that the cells were treated with 20 and 40 μM Leonurin.

As a result, as shown in FIG. 7, mRNA expression levels of atrogin-1 and MuRF1 involved in muscle protein degradation by TNF-α increased significantly ( ^## p <0.01), whereas atrogin as treated with Leonurin It was confirmed that mRNA expression levels of -1 and MuRF1 were significantly decreased ( ^** p <0.01). This means that the Leonurin of the present invention is excellent in inhibiting the expression of biomarkers involved in muscle protein degradation in muscle cells.

[Example 9] Leonurin's muscle differentiation biomarker expression increase effect

The experiment was conducted in the same manner as in Example 6, except that the cells were treated with 20 and 40 μM Leonurin.

As a result, as shown in FIG. 8, mRNA expression of MyoD and myogenin decreased significantly ( ^## p <0.01) as treated with TNF-α was significant by treatment of Leonurin ( ^* p <0.05, ^** p <0.01). This means that the present invention has excellent ability to promote muscle differentiation.

[Example 10] Effect of muscle function and muscle mass increase of motherwort hydrothermal extract and leonurin in animal models

<10-1> Animal breeding and muscle atrophy

As a laboratory animal, a 7-week-old male rat (C57BL/6J; DBL, Korea) was purchased and tested. The breeding of all animals was carried out at the Yonsei Laboratory Animal Reaserch Center (YLARC, Seoul, Korea), and the breeding room environment was maintained at a temperature of 23 ± 2°C and a relative humidity of 55 ± 10%. Prior to the start of the experiment, a total of 20 mice were randomly divided into 5 mice per group. After acclimatization for 1 week, anesthesia was induced by intraperitoneal injection of 325 mg/kg of tribromoethanol (Sigma-Aldrich). After anesthesia, the right hindlimb gastrocnemius muscle and the right sole of the rats in the atrophy group and the sample administration group were muscled with the 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 motherwort hydrothermal extract prepared in Example 1-1 was 150 mg/kg or 300 mg/kg and Leonurin at a concentration of 30 mg/kg for another week. Orally daily. At this time, the normal group and the muscular atrophy group were orally administered with saline instead of the sample.

<10-2> Confirm the effect of motherwort hot water extract and Leonurin to improve muscle strength

After the oral administration period, the muscle strength of the rat 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.

As a result, as shown in FIG. 9, muscle strength was significantly decreased ( ^## p <0.01) in the muscular atrophy group compared to the normal group, but the muscle strength was significant ( ^** p <0.01 as treated with motherwort hydrothermal extract and leonurin). ). This means that the motherwort hydrothermal extract and leonurin of the present invention have an excellent effect of increasing the decreased muscular strength due to muscular atrophy.

<10-3> Confirm the effect of motherwort hydrothermal extract and leonurin to increase muscle weight

After the muscle strength measurement was completed, the experimental animal was sacrificed through cardiac blood after anesthesia by intraperitoneal injection of 325 mg/kg of tribromomethanol (Sigma-Aldrich). After confirming that the heartbeat stopped, weight was measured by extracting the intact tibialis anterior muscle from the right hind leg.

As a result, as shown in FIG. 10, the weight of the anterior apical muscle of the atrophic group decreased significantly ( ^## p <0.01) compared to the normal group, but the weight was significant when treating the motherwort hot water extract and leonurin ( ^* p < 0.05, ^** p <0.01). This means that the motherwort hydrothermal extract and leonurin of the present invention are excellent in increasing the weight of muscles reduced due to muscular atrophy.

[Example 11] Effect of muscle function and muscle mass increase of motherwort ethanol extract in animal models

The muscle atrophy was induced in the same manner as in Example 10-1, and then motherwort 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 10-2 and Example 10-3, the group treated with motherwort ethanol extract had 23.91% ( ^** p <0.01) of muscle strength and muscle weight, respectively, compared to the muscular atrophy group. ) And 17.28% ( ^** p <0.01). This means that the motherwort ethanol extract of the present disease is excellent in reducing muscle strength and muscle weight due to muscular atrophy.

[Example 12] Effect of muscle function and muscle mass increase of motherwort subcritical extract in animal model

After inducing muscular dystrophy in the same manner as in Example 10-1, the motherwort subcritical extract prepared in Example 1-6 was orally administered daily at a concentration of 200 mg/kg. Next, as a result of measuring the muscle strength and muscle weight in the same manner as in Example 10-2 and Example 10-3, the motherwort subcritical extract administration group had 24.97% of muscle strength and muscle weight, respectively, compared to the muscular atrophy group ( ^** p < 0.01) and 20.60% ( ^** p <0.01). This means that the motherwort subcritical extract of the present onset is excellent in reducing muscle strength due to muscular atrophy and increasing muscle weight.

Hereinafter, a manufacturing example of a drug, food or cosmetic product containing motherwort extract or leonurin as an active ingredient according to the present invention will be described, but the present invention is not intended to be limited thereto, but only to be specifically described. The drug, food or cosmetic composition of Preparation Examples 1 to 3 was prepared according to the following compositional components and composition ratios with motherwort extract or leonurin having excellent effect of preventing or treating muscle disease or improving muscle function according to a conventional method. .

[Production Example 1] Pharmaceutical

<1-1> powder

After mixing motherwort extract or Leonurin 50 mg, 2 g of crystalline cellulose, a powder was prepared by filling in an airtight bag according to a conventional powder manufacturing method.

<1-2> tablets

After mixing motherwort extract or Leonurin 50 mg, crystalline cellulose 400 mg, and magnesium stearate 5 mg, tablets were prepared by tableting according to a conventional tablet manufacturing method.

<1-3> Capsule

Capsules were prepared by mixing motherwort extract or 30 mg of leonurin, 100 mg of whey protein, 400 mg of crystalline cellulose, and 6 mg of magnesium stearate, and then filling into gelatin capsules according to a conventional capsule preparation method.

[Production Example 2] Food

<2-1> Manufacture of health food

Motherwort Extract or Leonurin 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, 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 phosphate 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, 24.8 mg of magnesium chloride can be prepared by mixing, and the compounding ratio may be arbitrarily modified. The above ingredients are mixed according to a conventional health food manufacturing method, then granules are prepared, and a conventional method is performed. Depending on the health food composition can be used.

<2-2> Preparation of health drinks

1000 mg of motherwort extract or Leonurin, 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 of preparing a health drink for 900 mL, followed by about 1 hour After stirring and heating at 85 for a while, the resulting solution is filtered, obtained in a sterilized 2 L container, sealed and sterilized, and then refrigerated and then used in the manufacture of a health drink composition.

<2-3> Chewing gum

Chewing gum was prepared in a conventional manner by mixing 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of fragrance, and 2% by weight of water and 0.1% by weight of motherwort extract or leonurin.

<2-4> Candy

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 motherwort extract or leonurin.

<2-5> Biscuit

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 A biscuit was prepared by a conventional method by mixing 7.27% by weight of milk with a motherwort extract or by weight of leonurin.

[Production Example 3] Cosmetics

<3-1> Nutrient Cosmetics (Milk Lotion)

The motherwort extract or leonurin was prepared according to the ratio of the nutrient longevity formulations in Table 1, and nutrient longevity was prepared according to a conventional method.

Ingredients Preparation Example 3-1
(weight%) Motherwort extract or Leonurin 2.0 Squalane 5.0 Wax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 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, pigments, flavors Proper Purified water to 100

<3-2> Flexible Cosmetics (Skin Lotion)

The motherwort extract or Leonurin was prepared according to the conventional method in accordance with the ratio of the softener composition in Table 2 below.

Ingredients Preparation Example 3-2
(weight%) Motherwort extract or Leonurin 2.0 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 PEG 12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservatives, pigments, flavors Proper Purified water to 100

<3-3> Nutrition Cream

Nutritious cream was prepared according to a conventional method using the motherwort extract or leonurin in the proportion of the nutritional cream formulation in Table 3 below.

Ingredients Preparation Example 3-3
(weight%) Motherwort extract or Leonurin 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 PEG60 hardened castor oil 2.0 Floating paraffin 10 Squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 antiseptic Proper Coloring Proper Spices Proper Purified water to 100

<3-4> Massage Cream

Massage cream was prepared according to a conventional method using the motherwort extract or leonurin in the proportion of the massage cream formulation shown in Table 4 below.

Ingredients Preparation Example 3-4
(weight%) Motherwort extract or Leonurin 1.0 Wax 10.0 Polysorbate 60 1.5 PEG 60 hardened castor oil 2.0 Sorbitan sesquioleate 0.8 Floating paraffin 40.0 Squalane 5.0 Caprylic/Capric Triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, pigments, flavors Proper Purified water to 100

<3-5> Pack

Packs were prepared according to a conventional method using the motherwort extract or leonurin as the pack formulation ratios in Table 5 below.

Ingredients Preparation Example 3-5
(weight%) Motherwort extract or Leonurin 1.0 Polyvinyl alcohol 13.0 Sodium carboxymethylcellulose 0.2 glycerin 5.0 Allantoin 0.1 ethanol 6.0 PEG 12 nonylphenyl ether 0.3 Polysorbate 60 0.3 Preservatives, pigments, flavors Proper Purified water to 100

<3-6> Gel

Gels were prepared according to a conventional method using the motherwort extract or leonurin according to the gel formulation ratios in Table 6 below.

Ingredients Preparation Example 3-6
(weight%) Motherwort extract or Leonurin 0.5 Sodium ethylenediamine acetate 0.05 glycerin 5.0 Carboxyvinyl polymer 0.3 ethanol 5.0 PEG 60 hardened castor oil 0.5 Triethanolamine 0.3 Preservatives, pigments, flavors Proper Purified water to 100

<110> FND NET CO.,LTD. Industry-Academic Cooperation Foundation, Yonsei University <120> Composition for prevention or treatment of muscular disorder or improvement of muscular functions comprising Leonurus japonicus extract or leonurine <130> HPC-9171 <160> 10 <170> KoPatentIn 3.0 <210> 1 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for Atrogin-1 <400> 1 gttactgcaa caaggagaat ctgtt 25 <210> 2 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for Atrogin-1 <400> 2 ccgtatgagt cttatgtttt gctgg 25 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for MuRF1 <400> 3 ccggacggaa atgctatgga 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for MuRF1 <400> 4 agcctggaag atgtcgttgg 20 <210> 5 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for beta Actin <400> 5 tgacaggatg cagaaggaga ttac 24 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for beta Actin <400> 6 taaaacgcag ctcagtaaca gtc 23 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for MyoD <400> 7 tgcctcggag ataatacagc c 21 <210> 8 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for MyoD <400> 8 ggtgtaacaa ccatacccca ct 22 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for Myogenin <400> 9 agagagcccc cttgttaatg c 21 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for Myogenin <400> 10 ggccactcac tgtctctcaa a 21

Claims (8)

Pharmaceutical composition for preventing or treating muscle disease containing motherwort extract or Leonurin represented by Formula 1 as an active ingredient:
[Formula 1]

.
According to claim 1,
Leonurin represented by the formula (1) is a pharmaceutical composition for preventing or treating muscle disease, characterized in that isolated from motherwort extract.
According to claim 1,
The motherwort extract is a pharmaceutical composition for the prevention or treatment of muscle disease, characterized in that obtained by extracting the upper part of motherwort.
According to claim 1,
The motherwort extract is a pharmaceutical composition for preventing or treating muscle diseases characterized by being obtained by extracting motherwort with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid and a supercritical fluid.
The method of claim 4,
The organic solvent has 1 to 6 carbon atoms (alcohol), acetone (acetone), ether (ether), benzene (benzene), chloroform (chloroform), ethyl acetate (ethyl acetate), methylene chloride (methylene chloride), hexane ( hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) pharmaceutical composition for preventing or treating muscle diseases, characterized in that at least one solvent selected from the group consisting of.
The method according to any one of claims 1 to 4,
The muscle disease is at least one selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and sarcopenia. Pharmaceutical composition for preventing or treating muscle disease.
Food composition for preventing muscle disease or improving muscle function comprising motherwort extract or Leonurin represented by Formula 1 as an active ingredient:
[Formula 1]

.
A cosmetic composition for improving muscle function containing motherwort extract or Leonurin represented by the following Chemical Formula 1 as an active ingredient:
[Formula 1]

.

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