KR20240007869A - Composition for improvement, treatment or prevention of muscular disorders, or improvement of muscular functions comprising Lonicera japonica - Google Patents

Abstract

The present invention relates to a composition for preventing and treating muscle disease, or improving muscle function, containing honeysuckle as an active ingredient, such as honeysuckle powder or extract. Honeysuckle, such as honeysuckle powder or extract according to the present invention, has the effect of increasing the activity of mTOR, which is involved in muscle protein synthesis, and reducing the mRNA expression of MuRF-1 and atrogin-1, which is involved in muscle protein degradation, and increases muscle mass. and improve muscle function to prevent, improve, or treat various muscle diseases such as atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and sarcopenia. It can be usefully used.

Description

Composition for improvement, treatment or prevention of muscular disorders, or improvement of muscular functions comprising Lonicera japonica} containing honeysuckle as an active ingredient

The present invention relates to a composition for improving, treating or preventing muscle disease, or improving muscle function, and more specifically, to a composition for improving or treating muscle disease containing Lonicera japonica as an active ingredient, such as honeysuckle powder or honeysuckle extract. Or it relates to a composition for prevention or improvement of muscle function.

Muscle atrophy is caused by a gradual decrease in muscle mass and refers to muscle weakness and degeneration (Cell, 119(7): 907-910, 2004). Muscle atrophy is promoted by inactivity, oxidative stress, and chronic inflammation and weakens muscle function and exercise capacity (Clinical Nutrition, 26(5): 524-534, 2007). The most important factor determining muscle function is muscle mass, which is maintained by the balance of protein synthesis and breakdown. Muscular dystrophy occurs when protein degradation occurs more than synthesis (The International Journal of Biochemistry and Cell Biology, 37(10): 1985-1996, 2005).

Muscle size is regulated by intracellular signaling pathways that induce anabolism or catabolism within the muscle. If more signaling reactions that induce synthesis rather than decomposition of muscle protein occur, muscle protein synthesis increases, which is manifested as an increase in muscle size (hypertrophy) or an increase in the number of muscle fibers (hyperplasia) due to an increase in muscle protein (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 within muscle cells. Activation of mammalian target of rapamycin (mTOR) by PI3K/Akt signaling is recognized as a central growth signaling factor that integrates various growth signals within cells. mTOR induces muscle protein synthesis by activating two factors that initiate mRNA translation, 4E-binding protein (4EBP1) and phosphorylated 70-kDa ribosomal S6 kinase (p70S6K), thereby contributing to increased muscle mass (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 into the nucleus, it increases the expression of atrogin-1 and MuRF1, E3 ubiquitin ligase factors involved in protein degradation (Disease Models and Mechanisms, 6: 25- 39, 2013). As their expression level increases, protein breakdown within the muscle is promoted, resulting in a decrease in muscle mass. Therefore, promoting the activity of mTOR and suppressing the expression of atrogin-1 and MuRF1 increases the amount of muscle protein and thus increases muscle mass.

Differentiation of muscle cells and muscle formation are regulated by various muscle regulatory factors. Among them, induction of myogenin expression by MyoD activity is the most important factor in the fusion of myogenic cells and is involved in the formation of myotube cells. Muscle fibers formed through this process form bundles and ultimately form muscles (Cellular and Molecular Life Sciences, 70: 4117-4130, 2013).

Golden-and-silver flower (scientific name Lonicera japonica Thunberg) has been traditionally used for diuresis, detoxification, and treatment of arthritis, bronchitis, and tumors (The Pharmaceutical Society of Korea, 43(1): 117-123, 1999 ). So far, honeysuckle has been described as antioxidant (Microbiology and Biotechnology Letters, 46(1): 18-28, 2018), antibacterial (Korean Society of Food Science and Technology, 37(4): 642-647, 2005), and antidiabetic (Journal of the Korean Oil Chemists' Society, 29(3): 412-420, 2012), anti-inflammatory (Journal of Pharmacy and Pharmacology, 60(6): 779-786, 2008), liver protection (Modern Food Science and Technology 26 (4), 351-353, 2010), etc., its physiological activity has been reported.

However, prior to the present invention, nothing was known about the prevention and treatment of muscle diseases or improvement of muscle function using honeysuckle.

Accordingly, the present inventors searched for natural substances that have excellent muscle function regulating activity and can be safely applied, and as a result, it was confirmed that honeysuckle, such as honeysuckle powder or honeysuckle extract, has activity for improving, treating or preventing muscle diseases, or improving muscle function. Thus, the present invention was completed.

Therefore, an object of the present invention is to provide a food composition for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

Another object of the present invention is to provide a pharmaceutical composition for treating or preventing muscle disease containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

Another object of the present invention is to provide a cosmetic composition for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

Another object of the present invention is to provide a feed additive for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

To achieve the above object, the present invention provides a food composition for improving or preventing muscle disease, or improving muscle function, containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

Additionally, the present invention provides a pharmaceutical composition for treating or preventing muscle disease containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

In addition, the present invention provides a cosmetic composition for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

In addition, the present invention provides a feed additive for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract.

Additionally, the present invention provides a method of treating muscle disease by administering the composition to humans or non-human animals.

The present invention also provides new uses of honeysuckle, such as honeysuckle powder or honeysuckle extract for the manufacture of medicines for treating muscle diseases.

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

Figure 1 shows the results confirming the effect of increasing mTOR activity in L6 muscle cells following treatment with hot water extract of honeysuckle flower buds.
Figure 2 shows the results confirming the effect of reducing the mRNA expression levels of atrogin-1 and MuRF1 in L6 muscle cells following treatment with hot water extract of honeysuckle flower buds.
Figure 3 shows the results confirming the effect of increasing muscle volume due to treatment with hot water and ethanol extract of honeysuckle flower buds in mice inducing muscular atrophy.

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

The present invention provides a food composition for improving or preventing muscle disease, or improving muscle function, containing honeysuckle, such as honeysuckle powder or honeysuckle extract; Pharmaceutical compositions for treating or preventing muscle diseases containing honeysuckle, such as honeysuckle powder or honeysuckle extract; or a cosmetic composition for improving or preventing muscle disease, or improving muscle function, containing honeysuckle, such as honeysuckle powder or honeysuckle extract; or feed additives for improving or preventing muscle disease, or improving muscle function, containing honeysuckle, such as honeysuckle powder or honeysuckle extract; Alternatively, it provides a method of treating muscle disease comprising applying honeysuckle, such as honeysuckle powder or honeysuckle extract, to a human or non-human mammal.

In this specification, 'Honeysuckle' refers to the flower buds, leaves, branches, or mixtures of Lonicera japonica belonging to the Caprifoliaceae family, and is a powder or extract of Honeysuckle.

In this specification, 'pulverized honeysuckle' refers to the dried flower buds, leaves, or branches of honeysuckle that can be used by mammals, including humans, according to a method that can be easily performed by a person skilled in the art to which the present invention pertains. It can be prepared and used in a form that can be easily ingested, and the active ingredient is easily released from the dry powder in the intestine after ingestion, and as a result, can be easily absorbed into the mammalian body. The shape or size of the dried pulverized particles for this purpose is not limited, but the more the surface of the particles is maximized, the easier it is to release the active ingredients described above, so it is preferable to prepare them in the form of fine powder as much as possible. In one embodiment of the present invention, dried honeysuckle flower buds, leaves, or branches were pulverized with a mixer to prepare a pulverized product.

In this specification, 'Honeysuckle extract' is obtained by extracting the flower buds, leaves, or branches of Honeysuckle using a suitable solvent, and is an extract, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, or a crude product thereof, or Includes all forms of purified products. The method for preparing the honeysuckle extract may be obtained by extracting the honeysuckle extract with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and an ultra-high pressure, subcritical or supercritical fluid, but is not limited to this.

The honeysuckle extract of the present invention can be prepared using conventional extraction methods in the industry, such as heat extraction, cold needle extraction, ultrasonic extraction, filtration, and reflux extraction, and honeysuckle can be purchased and used commercially, collected from nature, or Cultivated ones can be used.

The honeysuckle extract according to the present invention can be separated according to a conventional method known in the art for producing extracts from natural products, that is, using a conventional solvent under conventional temperature and pressure conditions.

The term 'fraction' used in this specification refers to a result obtained by a fractionation method that separates a specific component or group from a mixture containing various components. In the present invention, it refers to the result obtained by a fractionation method for separating specific components or specific groups from the honeysuckle extract prepared as described above.

In order to obtain the honeysuckle fraction according to the present invention, conventional fractionation solvents known in the art, such as water, polar solvents such as anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms such as ethanol and methanol, and hexane, butanol, and ethyl Non-polar solvents such as acetate, chloroform, dichloromethane, or mixed solvents thereof may be used, but are not limited thereto.

The honeysuckle fraction of the present invention may also include one obtained by additionally applying a purification process. For example, fractions obtained by passing the honeysuckle extract according to the present invention through an ultrafiltration membrane with a certain molecular weight cut-off value, various chromatographic methods (separation according to size, charge, hydrophobicity or affinity) Fractions obtained through various purification methods, such as separation by (produced for), are also included in the honeysuckle extract of the present invention.

In this specification, 'muscle' comprehensively refers to tendons, muscles, and tendons, and 'muscle function' refers to the ability to exert force by contracting muscles, and the ability of muscles to exert maximum contractile force to overcome resistance. These include muscular strength, which is the ability to exercise, muscular endurance, which is the ability to express how long or how many times a muscle can repeat contraction and relaxation with a given weight, and quickness, which is the ability to exert strong force in a short period of time. The term 'improvement of muscle function' in this specification refers to improving muscle function better by increasing muscle mass.

The present invention relates to a pharmaceutical composition for preventing and treating muscle diseases or improving muscle function, comprising honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract; Food compositions for preventing and improving muscle disease or improving muscle function; Provided is a cosmetic composition for preventing and improving muscle disease or improving muscle function.

In one embodiment, the honeysuckle extract may be a hot water extract, an ethanol extract, an ethyl acetate extract, a hexane extract, an ultra-high pressure extract, a subcritical extract, or a supercritical extract, but is not limited thereto.

In one embodiment, the honeysuckle extract can be obtained by extracting the flower buds, leaves, and branches of the honeysuckle with one or more solvents selected from the group consisting of water, organic solvents having 1 to 6 carbon atoms, and subcritical and supercritical fluids. Gold and silver coins can also be obtained by extraction 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.

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

In a specific embodiment of the present invention, the present inventors repeatedly extracted the dried honeysuckle powder at room temperature using hot water, ethanol, ethyl acetate, and hexane as solvents, or ultra-high pressure extraction, subcritical fluid extraction, and supercritical fluid extraction. A honeysuckle extract was prepared using this method.

The prepared hot water extract of honeysuckle flower buds was treated with L6 muscle cells to confirm its activity within the muscle cells. As a result, it was confirmed that the protein expression of p-mTOR, which is involved in muscle protein synthesis, was significantly increased (Figure 1). . In addition, it was confirmed that ethanol extract, ethyl acetate extract, hexane extract, ultra-high pressure extract, subcritical extract, and supercritical extract of honeysuckle flower buds significantly increased the protein expression of p-mTOR in L6 muscle cells (Table 1). . Additionally, it was confirmed that the hot water extract and ethanol extract of honeysuckle leaves and stems significantly increased the protein expression of p-mTOR in L6 muscle cells (Table 2).

As a result of treating L6 muscle cells with the hot water extract of honeysuckle flower buds and confirming its activity within muscle cells, it was confirmed that it suppresses the mRNA expression of MuRF-1 and atrogin-1, which are involved in muscle protein degradation (Figure 2). In addition, it was confirmed that ethanol extract, ethyl acetate extract, hexane extract, ultra-high pressure extract, subcritical extract, and supercritical extract of honeysuckle flower buds also significantly reduced the mRNA expression of MuRF-1 and atrogin-1 in L6 muscle cells. (Table 3). Additionally, it was confirmed that hot water extract and ethanol extract of honeysuckle leaves and stems also significantly reduced the mRNA expression of MuRF-1 and atrogin-1 in L6 muscle cells (Table 4).

As a result of treatment with hot water extract and ethanol extract of honeysuckle flower buds using a mouse animal model of immobilization using a skin stapler, muscle volume increased compared to the muscle atrophy group (Figure 3). In addition, as a result of treatment with pulverized honeysuckle flower buds and hot water and ethanol extracts of honeysuckle flower buds in the same immobilized mouse animal model, muscle strength and muscle weight were significantly increased compared to the muscular atrophy group (Table 5, Table 6).

When the composition for preventing and 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 occur due to genetic factors, acquired factors, aging, etc. Muscle wasting is characterized by gradual loss of muscle mass and weakness and degeneration of muscles, especially skeletal or voluntary muscles and cardiac muscles. Examples of diseases related to this include atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and sarcopenia. The composition of the present invention has the effect of increasing muscle mass, and the type of muscle is not limited.

The pharmaceutical composition for preventing and treating muscle diseases or improving muscle function of the present invention may include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Carriers for parenteral administration may also include water, suitable oils, saline solutions, aqueous glucose and glycols, and the like. Additionally, stabilizers and preservatives may be additionally included. Suitable stabilizers include antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. As for other pharmaceutically acceptable carriers, those described in the following literature may be referred to (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 to this, but is intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, and intraperitoneal. , may be administered intranasally, enterally, topically, sublingually, or rectally.

The pharmaceutical composition of the present invention can be formulated into a formulation for oral administration or parenteral administration according to the administration route described above. When formulated, one or more buffers (e.g. saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g. EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g. aluminum hydroxide) side), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.

Solid preparations for oral administration include tablets, pills, powders, granules, solutions, gels, syrups, slurries, suspensions or capsules, and such solid preparations may contain at least one excipient, for example, 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 hydroxypropylmethyl-cellulose or gelatin can be mixed. For example, tablets or sugar tablets can be obtained by combining the active ingredient with a solid excipient, grinding it, adding suitable auxiliaries and processing it into a granule mixture.

In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, or syrups. In addition to the commonly used simple diluents such as water or liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, or preservatives may be included. .

In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, and preservatives may be additionally included. .

When administered parenterally, the pharmaceutical composition of the present invention can be formulated with a suitable parenteral carrier in the form of injections, transdermal administration, and nasal inhalation according to methods known in the art. The above injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), mixtures thereof, and/or vegetable oils. You can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. etc. can be used. In order to protect the injection from microbial contamination, it may additionally contain various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. Additionally, in most cases, the injection may additionally contain an isotonic agent such as sugar or sodium chloride.

In the case of transdermal administration, forms such as ointments, creams, lotions, gels, external solutions, paste preparations, linear preparations, and aerol preparations are included. In the above, 'transdermal administration' means administering a pharmaceutical composition topically to the skin so that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin.

For inhalation administration, the compounds used according to the invention may be packaged in pressurized packs or using a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered in the form of an aerosol spray from a nebulizer. For pressurized aerosols, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators 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 text generally known in all pharmaceutical chemistry.

The pharmaceutical composition for preventing and treating muscle disease, or improving muscle function, of the present invention can provide desirable effects for preventing and treating muscle disease or improving muscle function when it contains an effective amount of honeysuckle extract. In this specification, the term 'effective amount' refers to an amount that produces a greater response than the negative control, 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 honeysuckle extract, and the remaining amount may be comprised of a pharmaceutically acceptable carrier. The effective amount of honeysuckle extract included in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.

The total effective amount of the pharmaceutical composition of the present invention can be administered to a patient in a single dose, or can be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease. When administered parenterally, it is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per kg of body weight per day based on the honeysuckle extract, and when administered orally, the weight is administered per day based on the honeysuckle extract. It can be administered in one to several divided doses, preferably in an amount of 0.01 to 100 mg per kg, more preferably 0.01 to 10 mg per kg. However, the effective dose for the patient is determined by considering various factors such as the route of administration and number of treatments of the pharmaceutical composition, as well as the patient's age, weight, health status, gender, severity of the disease, diet, and excretion rate. Therefore, taking this into account, a person with ordinary knowledge in the art will be able to determine an appropriate effective dosage of the honeysuckle extract according to a specific use for preventing and treating muscle diseases. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route, and administration method as long as it exhibits the effects of the present invention.

The pharmaceutical composition for preventing and treating muscle diseases or improving muscle function of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemical therapy, or biological response modifiers.

The pharmaceutical composition for preventing and treating muscle diseases or improving muscle function of the present invention can also be provided in the form of an external preparation containing a honeysuckle extract as an active ingredient.

When the pharmaceutical composition for preventing and treating muscle disease or improving muscle function according to the present invention is used as an external skin agent, fatty substances, organic solvents, solubilizers, thickening and gelling agents, softeners, antioxidants, suspending agents, and stabilizers are added. Topicals, foaming agents, fragrances, surfactants, water, ionic emulsifiers, non-ionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, It may contain adjuvants commonly used in the field of dermatology, such as hydrophilic active agents, lipophilic active agents, or any other ingredients commonly used in topical skin preparations, such as lipid vesicles. Additionally, the ingredients may be introduced in amounts commonly used in the field of dermatology.

When the pharmaceutical composition for preventing and treating muscle disease or improving muscle function of the present invention is provided as an external skin preparation, it is not limited thereto, but may be in the form of an ointment, patch, gel, cream, or spray.

The composition for preventing and improving muscle disease or improving muscle function of the present invention may also be a food composition. When the food composition of the present invention is used to prevent and improve muscle disease or improve muscle function, it can be used to prevent or improve muscle disease caused by muscle wasting or degeneration. Muscle wasting and degeneration occur due to genetic factors, acquired factors, aging, etc. Muscle wasting is characterized by gradual loss of muscle mass and weakness and degeneration of muscles, especially skeletal or voluntary muscles and cardiac muscles. Examples of diseases related to this include atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and sarcopenia. The composition of the present invention has the 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 is used for use in humans or animals including livestock. It is intended for consumption. 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 their processed foods (e.g. canned fruit, bottled foods, jam, mamalade, etc.), fish, meat and their processed foods (e.g. ham, sausages, etc.) corned beef, etc.), bread and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , vegetable proteins, retort foods, frozen foods, and various seasonings (e.g., soybean paste, soy sauce, sauce, etc.) can be manufactured by adding honeysuckle powder or honeysuckle extract. In addition, nutritional supplements are not limited to this, but can be prepared by adding honeysuckle powder or honeysuckle extract to capsules, tablets, pills, etc. In addition, health functional foods are not limited to this, but for example, honeysuckle powder or honeysuckle extract itself can be manufactured in the form of tea, juice, and drinks and liquefied, granulated, encapsulated, and powdered for drinking (health drinks). It can be consumed. Additionally, in order to use honeysuckle powder or honeysuckle extract in the form of a food additive, it can be prepared and used in the form of powder or concentrate. Additionally, it can be prepared in the form of a composition by mixing honeysuckle powder or honeysuckle extract with known active ingredients that are known to be effective in preventing and improving muscle disease or improving muscle function.

When the composition for preventing and improving muscle disease or improving muscle function of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients like ordinary drinks. . The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; polysaccharides such as dextrins and cyclodextrins; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as thaumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention.

Honeysuckle powder or honeysuckle extract may be contained as an active ingredient in a food composition for preventing and improving muscle disease or improving muscle function, and the amount is effective to achieve the action for preventing and improving muscle disease or improving muscle function. The amount is not particularly limited, but is preferably 0.01 to 100% by weight based on the total weight of the entire composition. The food composition of the present invention can be prepared by mixing honeysuckle powder or honeysuckle extract with other active ingredients known to be effective in compositions for preventing and improving muscle disease or improving muscle function.

In addition to the above, the health food of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, It may contain glycerin, alcohol, or carbonating agent. In addition, the health food of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients can be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The composition for preventing and improving muscle disease or improving muscle function of the present invention may also be a cosmetic composition. The cosmetic composition of the present invention contains honeysuckle powder or honeysuckle extract as an active ingredient and is used as a basic cosmetic composition (toilet water, cream, essence, cleansing agent such as cleansing foam and cleansing water, pack, body oil) along with dermatologically acceptable excipients. , color cosmetic compositions (foundation, lipstick, mascara, makeup base), hair product compositions (shampoo, rinse, hair conditioner, hair gel), and soap.

The excipients are not limited thereto, but may include, for example, skin emollients, skin penetration enhancers, colorants, fragrances, emulsifiers, thickeners, and solvents. In addition, it may additionally contain fragrances, pigments, disinfectants, antioxidants, preservatives, and moisturizers, and may include thickeners, inorganic salts, synthetic polymer materials, etc. for the purpose of improving physical properties. For example, when preparing a face wash or soap using the cosmetic composition of the present invention, it can be easily prepared by adding honeysuckle powder or honeysuckle extract to a regular face wash or soap base. When manufacturing cream, it can be produced by adding honeysuckle, honeysuckle powder, or honeysuckle extract to a general oil-in-water (O/W) cream base. Here, synthetic or natural materials such as proteins, minerals, and vitamins can be added for the purpose of improving physical properties, such as fragrances, chelating agents, pigments, antioxidants, and preservatives.

The content of honeysuckle powder or honeysuckle extract contained in the cosmetic composition of the present invention is not limited thereto, but is preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, based on the total weight of the entire 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 exceeds 10% by weight, there may be difficulties in safety or formulation manufacturing.

The composition of the present invention can be added to a feed additive or a feed composition containing the same for the purpose of preventing or improving muscle disease.

In the present invention, the term 'feed additive' refers to a substance added to feed for the purpose of various effects such as supplementing nutrients and preventing weight loss, improving digestibility of fiber in feed, improving milk quality, preventing reproductive disorders and improving conception rate, and preventing high temperature stress in the summer. Includes. The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act, and includes mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, mineral preparations such as trace minerals such as zinc, copper, cobalt, and selenium, and kerotene. , vitamins such as vitamins A, D, E, nicotinic acid, and vitamin B complex, protective amino acids such as methionine and lysine, protective fatty acids such as fatty acid calcium salts, probiotics (lactic acid bacteria), yeast cultures, and live bacteria such as mold fermentation products, Yeast agents, etc. may be additionally included.

In the present invention, the term 'feed' refers to any natural or artificial diet, meal, etc., or an ingredient of the meal, for or suitable for eating, ingestion, and digestion by animals, and to prevent or improve muscle disease according to the present invention. Feed containing the composition as an active ingredient can be manufactured from various types of feed known in the art, and may preferably include concentrated feed, roughage and/or special feed, but is not limited thereto.

Concentrated feed includes seeds and fruits, including grains such as wheat, oats, and corn; bran, which includes rice bran, bran, and barley bran, which are by-products obtained from refining grains; soybeans; oil, sesame seeds, linseed, and coco oil; It is a product made by concentrating residues such as residual starch, which is the main component of the starch residue remaining after removing the starch from the by-products such as seed jelly, sweet potatoes, and potatoes, fishmeal, fish residue, and fresh liquid obtained from fish. Fish soluble, meat meal, blood meal, feather meal, skim milk powder, animal feed such as dried whey, which is the residue from the production of cheese from milk and casein from skim milk, yeast, These include, but are not limited to, chlorella and seaweed.

Forage includes raw grass feed such as wild grass, grass, and green cuttings, turnips for feed, beets for feed, root vegetables such as rutterbearger, a type of turnip, raw grass, green cuttings, and grains, etc., in silos. These include, but are not limited to, silage, which is stored feed that has been filled and fermented with lactic acid, field grass, hay made by cutting and drying grass, straw from breeding crops, and leaves from legumes. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea and its derivative diureide isobutane, and mixed feeds to supplement ingredients that are likely to be lacking when mixing only natural feed ingredients, or to increase the storability of the feed. Substances added in trace amounts include feed additives and dietary supplements, but are not limited to these.

The feed additive for preventing or improving muscle disease according to the present invention can be manufactured by adding honeysuckle powder or honeysuckle extract in an appropriate effective concentration range according to various feed manufacturing methods known in the art.

The feed additive according to the present invention can be applied without limitation to any object aimed at preventing or improving muscle disease. For example, it can be applied to any entity, including non-human animals such as cows, horses, pigs, goats, sheep, dogs, cats, rabbits, birds, and fish.

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

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

[Example 1] Preparation of honeysuckle flower bud extract

<1-1> Preparation of honeysuckle flower bud hot water extract

The dried honeysuckle flower buds were pulverized with a mixer, and then 10 g of the crushed honeysuckle flower bud 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 in a vacuum rotary concentrator to remove solvent components, thereby obtaining a hot water extract of honeysuckle flower buds.

<1-2> Preparation of ethanol extract of honeysuckle flower buds

The dried honeysuckle flower buds were pulverized with a mixer, and then 10 g of the crushed honeysuckle flower bud 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 in a vacuum rotary concentrator to remove solvent components, thereby obtaining an ethanol extract of honeysuckle flower buds.

<1-3> Preparation of ethyl acetate extract from honeysuckle flower buds

The dried honeysuckle flower buds were pulverized with a mixer, and then 10 g of the crushed honeysuckle flower bud 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 in a vacuum rotary concentrator to remove solvent components, and then an ethyl acetate extract of honeysuckle flower buds was obtained.

<1-4> Preparation of honeysuckle flower bud hexane extract

The dried honeysuckle flower buds were pulverized with a mixer, and then 10 g of the crushed honeysuckle flower bud sample was added to 100 mL of hexane and extracted at 40°C for 3 hours. The extracted sample was filtered under reduced pressure using Whatman No. 1 filter paper, and the filtered extract was concentrated using a vacuum rotary concentrator to remove solvent components, thereby obtaining a hexane extract of honeysuckle flower buds.

<1-5> Preparation of ultra-high pressure extract of honeysuckle flower buds

After pulverizing the dried honeysuckle flower buds with a mixer, place 1 g of the crushed honeysuckle flower buds and 76 mL of 18% ethanol in a polyethylene pack, seal it, and use an ultra-high pressure extraction device (Frescal MFP-7000; Mitsubishi Heavy Industries). and extracted. Ultra-high pressure extraction conditions were extraction pressure of 320 MPa and extraction time of 5 min. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was concentrated in a vacuum rotary concentrator to remove solvent components, thereby obtaining an ultra-high pressure extract of honeysuckle flower buds.

<1-6> Preparation of honeysuckle flower bud subcritical extract

The dried honeysuckle flower buds were pulverized with a mixer, and then 50 g of the crushed honeysuckle flower buds were placed in a subcritical water reactor of a subcritical extraction device (Biovan, Gyeonggi, Korea) along 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 and rapidly cooled to 30°C. Then, it was centrifuged at 3,600 rpm for 30 minutes to separate the floating residue, and only the supernatant was collected. A subcritical extract of honeysuckle flower buds was obtained by removing all solvents using a freeze dryer (Ilshin Lab Co. Ltd., Seoul, Korea).

<1-7> Preparation of honeysuckle flower bud supercritical extract

The dried honeysuckle flower buds were pulverized with a mixer, and then 10 g of the crushed honeysuckle flower buds were filled into a sample cartridge and extracted using a supercritical fluid extraction device (SFX 3560, Isco Inc., Lincoln, NE, USA). The supercritical extraction conditions were extraction pressure of 300 bar, extraction temperature of 50°C, supercritical carbon dioxide flow rate of 60 mL/min, and extraction time of 2 hours. Once the supercritical fluid extraction was completed, the pressure of the extraction device was lowered to release the supercritical fluid state to obtain a supercritical extract of honeysuckle flower buds.

[Example 2] Preparation of honeysuckle leaf extract

<2-1> Preparation of honeysuckle leaf hot water extract

The dried honeysuckle leaves were pulverized with a mixer, and then 10 g of the crushed honeysuckle leaf sample was added to 100 mL of water and extracted at 80°C for 3 hours. The extracted sample was filtered under reduced pressure using Whatman No. 1 filter paper, and the filtered extract was concentrated in a vacuum rotary concentrator to remove solvent components, thereby obtaining a hot water extract of honeysuckle leaves.

<2-2> Preparation of honeysuckle leaf ethanol extract

The dried honeysuckle leaves were pulverized with a mixer, and then 10 g of the crushed honeysuckle leaf 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 using Whatman No. 1 filter paper, and the filtered extract was concentrated in a vacuum rotary concentrator to remove solvent components, thereby obtaining an ethanol extract of honeysuckle leaves.

[Example 3] Preparation of honeysuckle branch extract

<3-1> Preparation of honeysuckle branch thermal water extract

The dried honeysuckle branches were pulverized with a mixer, and then 10 g of the crushed honeysuckle branch 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 in a vacuum rotary concentrator to remove solvent components, thereby obtaining a hot water extract of Honeysuckle branch.

<3-2> Preparation of ethanol extract of honeysuckle branches

The dried honeysuckle branches were pulverized with a mixer, and then 10 g of the crushed honeysuckle branch 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 using Whatman No. 1 filter paper, and the filtered extract was concentrated in a vacuum rotary concentrator to remove solvent components, thereby obtaining an ethanol extract of honeysuckle branches.

[Example 4] Effect of hot water extract of honeysuckle flower buds on promoting muscle protein synthesis

It is known that when mTOR protein is phosphorylated and activated, it can induce the activation of proteins involved in muscle protein synthesis and muscle mass increase in the PI3K/Akt signaling pathway within muscle cells. Therefore, in order to confirm the muscle creation-inducing activity of the hot water extract of honeysuckle flower buds, the activity of mTOR was confirmed using the mTOR Sandwich ELISA kit (Cell Signaling Technology, Beverly, MA, USA).

L6 myoblasts (ATCC; Manassas, VA, USA) were seeded in 6-well plates at 1 × 10 with Dulbecco's modified Eagle's media (DMEM; Hyclone) containing 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA). After seeding, the cells were cultured for 24 hours to reach 10 ⁵ cells/well. After culture, the medium in the well was removed, replaced with DMEM (Hyclone) containing 2% horse serum (HS; Hyclone), and cultured for an additional 6 days to differentiate L6 cells into myotube cells. Next, the hot water extract of the honeysuckle flower bud of Example 1-1 was dissolved in DMEM (Hyclone) at a concentration of 10 μg/mL and 20 μg/mL, then treated with cells and cultured for 12 hours. After culturing, cells were lysed by treatment with cell lysis buffer. The protein in the obtained cell lysate was quantified at a concentration of 1 mg/mL using Bradford (Bio-Rad Laboratories Inc., Hercules, CA, USA). 50 μL of cell lysate was dispensed into each microwell with anti-mTOR antibody attached and left at 37°C for 2 hours. After washing a total of 4 times with washing buffer, treatment with detection antibody and leaving at 37°C for 1 hour, washing again with washing buffer a total of 4 times, horseradish peroxidase ) conjugated secondary antibody was added and left at 37°C for 30 minutes. Finally, after washing a total of four times with the washing buffer solution, TMB substrate was added to each well, left at 37°C for 10 minutes, and stop buffer solution was added to stop the TMB reaction. After 2 minutes, the absorbance was measured at a wavelength of 450 nm to measure the level of mTOR in the root canal cells treated with the hot water extract of honeysuckle flower buds. The experiment was repeated a total of three times, and each measured value was calculated as a percentage (%) of the control group and expressed as mean ± standard deviation. Differences between groups were confirmed through Duncan multi-range analysis using one-way analysis of variance using the SPSS25.0 statistical package (SPSS Inc., Chicago, IL, USA). At this time, if the p value was less than 5%, it was indicated as statistically significant.

As a result, as shown in Figure 1, it was confirmed that the activity of mTOR increased significantly ( ^** p < 0.01) when treated with the hot water extract of honeysuckle flower buds compared to the control group that was not treated with the extract. This means that the hot water extract of honeysuckle flower buds of the present invention has an excellent ability to promote muscle protein synthesis in muscle cells.

[Example 5] Effect of extract of honeysuckle flower buds on promoting muscle protein synthesis

mTOR activity was measured in the same manner as in Example 1, except that 20 μg/mL of the honeysuckle flower bud extracts of Examples 1-2 to 1-7 were each treated.

As a result, as shown in Table 1, it was confirmed that treatment with the honeysuckle flower bud extract significantly ( ^** p < 0.01) increased the activity of mTOR compared to the control group that was not treated with the extract. This means that the honeysuckle flower bud extract of the present invention has an excellent ability to promote muscle protein synthesis within muscle cells.

experimental group Relative mTOR activity (%) Example 1-2 135.8±3.9** Example 1-3 128.4±2.6** Example 1-4 132.0±4.5** Examples 1-5 133.5±3.1** Example 1-6 125.3±4.8** Example 1-7 136.1±2.8**

[Example 6] Effect of extracts of honeysuckle leaves and branches on promoting muscle protein synthesis

mTOR in the same manner as in Example 1, except that the honeysuckle leaf extract of Examples 2-1 to 2-2 and the honeysuckle branch extract of Examples 3-1 to 3-2 were each treated with 20 μg/mL. Activity was measured.

As a result, as shown in Table 2, it was confirmed that the activity of mTOR increased significantly ( ^** p < 0.01) when treated with honeysuckle leaf and branch extracts compared to the control group not treated with the extract. This means that the honeysuckle leaf and branch extract of the present invention has an excellent ability to promote muscle protein synthesis in muscle cells.

experimental group Relative mTOR activity (%) Example 2-1 124.5±3.1** Example 2-2 129.1±4.4** Example 3-1 118.5±2.0** Example 3-2 122.1±3.6**

[Example 7] Inhibitory effect of muscle protein degradation of hot water extract of honeysuckle flower buds

L6 myoblast (ATCC), a muscle cell, was placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone) at 1 × 10 ⁵ cell/mL. When the cell density reached about 80-85%, the medium in the well was removed and the cells were treated with DMEM (Hyclone) containing 2% HS (Hyclone) to induce myotube differentiation. Differentiation was carried out for a total of 6 days, with new medium replaced every 2 days. After differentiation, 10 μg/mL of the hot water extract of the honeysuckle flower bud of Example 1-1 was added to DMEM (Hyclone) containing 50 ng/mL tumor necrosis factor alpha (TNF-α; PeproTech, Rocky Hills, NJ, USA). , were dissolved in DMEM (Hyclone) at concentrations of 20 μg/mL and 40 μg/mL, and then treated with cells. After 12 hours, total RNA was isolated using TRIzol reagent (Takara, Otsu, Japan). The isolated total RNA was quantified using NanoDrop 1000 (Thermo Fisher Scientific Inc., Waltham, MA, USA). 16 μL of quantified RNA was incubated at 42°C for 55 minutes and at 70°C for 15 minutes using Reverse Transcriptase Premix (ELPIS-Biotech) and a PCR machine (Gene Amp PCR System 2700; Applied Biosystems, Foster City, CA, USA). cDNA was synthesized under these conditions. Among 16 μL of generated cDNA, 1 μL of cDNA, the following specific primers (Bioneer, Daejeon, Korea), and PCR premix (ELPIS-Biotech) were used for 30 seconds at 95°C, 1 minute at 60°C, and 1 minute at 72°C. PCR was repeated 30 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'-GTTACTGCAACAAGGAGAATCTGTT-3' (SEQ ID NO: 3)

Reverse primer: 5'-CCGTATGAGTCTTATGTTTTGCTGG-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 using a 1.5% agarose gel, and the cDNA band was confirmed using the G;BOX EF imaging system (Syngene).

As a result, as shown in Figure 2, it was found that the mRNA expression of MuRF1 and atrogin-1, which increased with treatment with TNF-α, was decreased by hot water extract of honeysuckle flower buds. This means that the hot water extract of honeysuckle flower buds of the present invention has an excellent ability to inhibit muscle protein degradation within muscle cells.

[Example 8] Inhibitory effect of muscle protein degradation of honeysuckle flower bud extract

The same method as Example 7 was repeated a total of three times, except that 20 μg/mL of the honeysuckle flower bud extract of Examples 1-2 to 1-7 was treated each. Next, the band density of MuRF1 and atrogin-1 mRNA was measured using the Image J program (National Institute of Health, Bethesda, MD, USA), and then each measured value was calculated as a percentage (%) relative to the control group and averaged ± Expressed as standard deviation.

As a result, as shown in Table 3, the mRNA expression of MuRF1 and atrogin-1 increased significantly ( ^## p < 0.01) with treatment with TNF-α, but both significantly ( ^{* *} p < 0.01). This means that the honeysuckle flower bud extract of the present invention has an excellent ability to inhibit muscle protein degradation within muscle cells.

experimental group Relative MuRF1
Expression amount (%) Relative Atrogin-1
Expression amount (%) TNF-α 292.6±36.9** 317.0±35.7** Example 1-2 135.7±40.5** 154.3±22.6** Example 1-3 140.3±45.9** 170.9±33.2** Example 1-4 129.6±33.0** 137.1±43.2** Examples 1-5 143.6±38.1** 148.0±29.0** Example 1-6 172.4±32.5** 162.5±31.8** Example 1-7 138.5±43.2** 142.5±31.2*

[Example 9] Inhibitory effect of root protein decomposition of honeysuckle leaf and branch extract

Except that the honeysuckle leaf extract of Examples 2-1 to 2-2 and the honeysuckle branch extract of Examples 3-1 to 3-2 were each treated at 20 μg/mL, a total of The procedure was repeated 3 times. Next, the band density of MuRF1 and atrogin-1 mRNA was measured using the Image J program (National Institute of Health, Bethesda, MD, USA), and then each measured value was calculated as a percentage (%) relative to the control group and averaged ± Expressed as standard deviation.

As a result, as shown in Table 4, the mRNA expression of MuRF1 and atrogin-1 increased significantly ( ^## p < 0.01) with treatment with TNF-α, but both significantly (## p < 0.01) increased with treatment with honeysuckle leaf and branch extracts. ^** p < 0.01). This means that the honeysuckle leaf and branch extract of the present invention has an excellent ability to inhibit muscle protein degradation within muscle cells.

experimental group Relative MuRF1
Expression amount (%) Relative Atrogin-1
Expression amount (%) TNF-α 292.6±36.9** 317.0±35.7** Example 2-1 180.4±33.1** 203.8±33.8** Example 2-2 154.8±43.7** 138.9±41.6** Example 3-1 179.3±40.4** 180.4±43.1** Example 3-2 166.7±36.4** 201.2±37.0**

[Example 10] Effect of the honeysuckle bud extract on increasing muscle volume in an immobilization animal model

As experimental animals, 7-week-old male mice (C57BL/6J; DBL, Korea) were purchased and experiments were conducted. The breeding room environment for all animals was maintained at a temperature of 23 ± 2°C and a relative humidity of 55 ± 10%. Before starting the experiment, a total of 24 mice were randomly divided into 6 mice per group. After a week of adaptation, anesthesia was induced by intraperitoneal injection of 325 mg/kg tribromoethanol (Sigma-Aldrich). After anesthesia, the gastrocnemius muscle of the right hindlimb and the sole of the right foot of mice in the muscle atrophy group and the sample administration group were stapled using a skin stapler (Unidus, Chungcheongbuk-do, Korea). The right hind leg was damaged and unable to move, and this condition was maintained for a week. After a week, the stapler core fixed to the gastrocnemius muscle and sole was removed, and for another week, the hot water extract of the honeysuckle bud of Example 1-1 and the ethanol extract of the honeysuckle bud of Example 1-2 were administered at a concentration of 200 mg/kg, respectively. It was administered orally daily. At this time, the normal group and the muscle atrophy group were orally administered saline solution instead of the sample.

After the oral administration period was over, the subjects were anesthetized with isoflurane and the volume of the right hind limb muscles was measured using positron emission tomography/computed tomography/single photon emission tomography (microPET/CT/SPECT; Siemens Inveon, Knoxville, TN, USA). .

As a result, as shown in Figure 3, the muscle volume of the muscle atrophy group decreased compared to the normal group, but the muscle volume increased when treated with hot water and ethanol extract of honeysuckle flower buds. This means that the hot water and ethanol extract of honeysuckle flower buds of the present invention are excellent in increasing the volume of muscles reduced due to muscle atrophy.

[Example 11] Immobilization Effect of pulverized and extract of honeysuckle buds on improving muscle strength and increasing muscle mass in an animal model

<11-1> Confirmation of the effect of honeysuckle bud pulverization and extract on increasing muscle weight

In the same process as Example 10, 1,000 mg/kg of crushed honeysuckle buds, the hot water extract of honeysuckle buds of Example 1-1, and the ethanol extract of honeysuckle buds of Example 1-2 were administered at a concentration of 100 mg/kg every day. It was administered orally for 7 days.

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 mouse was pulled with a constant force until the mouse released the last part of the muscle strength measuring device, and a total of 5 consecutive tests were performed per mouse.

As a result, as shown in Table 5, the muscle strength was significantly ( ^## p < 0.01) decreased in the muscle atrophy group compared to the normal group, but when treated with honeysuckle bud grinds and extracts, the muscle strength was significantly (## p < 0.01) compared to the muscle atrophy group. ^* p < 0.01, ^** p < 0.01). This means that the pulverized material and extract of honeysuckle buds of the present invention are excellent in increasing muscle strength reduced due to muscle atrophy.

experimental group Muscle strength (g) Jeongsang-gun 217.5±12.7 Muscular atrophy group 160.1±9.8 ^## Honeysuckle bud powder 184.7±10.3* Example 1-1 198.6±12.1** Example 1-2 206.1±13.9**

<11-2> Confirmation of the effect of honeysuckle bud pulverization and extract on increasing muscle weight

After muscle strength measurements were completed, the experimental animals were anesthetized by intraperitoneal injection of 325 mg/kg of tribromomethanol (Sigma-Aldrich), and then sacrificed through cardiac blood sampling. After confirming that the heartbeat had stopped, the undamaged tibialis anterior muscle was removed from the right hind leg and its weight was measured.

As a result, as shown in Table 6, the weight of the tibialis anterior muscle of the muscle atrophy group was significantly ( ^## p < 0.01) reduced compared to the normal group, but when treated with honeysuckle bud grinds and extracts, the weight was lower than that of the muscle atrophy group. There was a significant increase ( ^* p < 0.01, ^** p < 0.01). This means that the pulverized material and extract of honeysuckle buds of the present invention are excellent in increasing the weight of muscles reduced due to muscle atrophy.

experimental group Muscle weight (mg) Jeongsang-gun 62.6±3.0 Muscular atrophy group 47.0±3.3 ^## Honeysuckle bud powder 52.9±2.5** Example 1-1 57.1±4.1** Example 1-2 58.7±3.9**

Hereinafter, examples of manufacturing medicines, foods, or cosmetics containing honeysuckle as an active ingredient, such as honeysuckle powder or honeysuckle extract according to the present invention, will be described. However, the present invention is not intended to be limited, but merely explained in detail. The pharmaceutical, food or cosmetic compositions of Preparation Examples 1 to 3 were prepared according to a conventional method using the honeysuckle powder or extract, which is excellent for preventing and treating muscle diseases or improving muscle function, according to the composition and ratio as follows. Manufactured.

[Production Example 1] Food

<1-1> Manufacturing of health food

1000 mg of honeysuckle powder or honeysuckle extract of the present invention, vitamin A acetate 70 ug, vitamin E 1.0 mg, vitamin B1 0.13 mg, vitamin B2 0.15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 ug, vitamin C 10 mg, biotin 10 ug, nicotinic acid amide 1.7 mg, folic acid 50 ug, calcium pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, monobasic potassium phosphate 15 mg, dicalcium phosphate 55 mg, potassium citrate 90 mg, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride. The mixing ratio may be modified arbitrarily. The above ingredients are mixed according to a typical health food manufacturing method, and then granules are prepared. , can be used to manufacture health food compositions according to conventional methods.

<1-2> Manufacturing of health drinks

Add purified water to 1000 mg of the honeysuckle extract of the present invention, 1000 mg of citric acid, 100 g of oligosaccharides, 2 g of plum concentrate, and 1 g of taurine to make a total of 900 mL. Mix the above ingredients according to a normal health drink manufacturing method, and then brew for about 1 hour. After stirring and heating at 85°C, the resulting solution is filtered, placed in a sterilized 2 L container, sealed, sterilized, stored in the refrigerator, and then used to manufacture a health drink composition.

<1-3> 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 flavor, 2% by weight of water, and 0.1% by weight of the honeysuckle extract of the present invention.

<1-4> Candy

Candy was prepared in a conventional manner by mixing 60% by weight of sugar, 39.8% by weight of starch syrup, 0.1% by weight of flavor, and 0.1% by weight of the honeysuckle extract of the present invention.

<1-5> Biscuits

Power grade 1 25.59% by weight, gravity grade 1 22.22% by weight, refined sugar 4.80% by weight, table salt 0.73% by weight, glucose 0.78% by weight, palm shortening 11.78% by weight, ammonium 1.54% by weight, sodium bicarbonate 0.17% by weight, sodium bisulfite 0.16% by weight , 1.45% by weight of rice flour, 0.0001% by weight of vitamin B, 0.04% by weight of milk flavor, 20.6998% by weight, 1.16% by weight of whole milk powder, 0.29% by weight of powdered milk, 0.03% by weight of monobasic calcium phosphate, 0.29% by weight of spray salt and spray. Biscuits were prepared in a conventional manner by mixing 7.27% by weight of oil and 0.8301% by weight of honeysuckle powder or extract of honeysuckle of the present invention.

[Preparation Example 2] Medicine

<2-1> Sanje

A powder was prepared by mixing 50 mg of the honeysuckle extract of the present invention and 2 g of crystalline cellulose and filling it into an airtight envelope according to a conventional powder production method.

<2-2> Tablets

Tablets were prepared by mixing 50 mg of the honeysuckle extract of the present invention, 400 mg of crystalline cellulose, and 5 mg of magnesium stearate and compressing them according to a conventional tablet manufacturing method.

<2-3> Capsules

Capsules were prepared by mixing 30 mg of the honeysuckle extract of the present invention, 100 mg of whey protein, 400 mg of crystalline cellulose, and 6 mg of magnesium stearate and filling them into gelatin capsules according to a conventional capsule manufacturing method.

[Production Example 3] Cosmetics

<3-1> Nourishing lotion (milk lotion)

Nutritional lotion was prepared according to a conventional method using the honeysuckle extract of the present invention according to the nutritional lotion formulation ratio shown in Table 3 below.

Ingredients Manufacturing Example 3-1
(weight%) honeysuckle extract 2.0 squalane 5.0 beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 liquid paraffin 0.5 Caprylic/Capric Triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 propylene glycol 3.0 Carboxy vinyl polymer 0.1 Triethanolamine 0.2 Preservatives, coloring, flavoring Appropriate amount Purified water to 100

<3-2> Soft lotion (skin lotion)

Soft lotion was prepared according to a conventional method using the honeysuckle extract of the present invention according to the softening lotion formulation ratio shown in Table 4 below.

Ingredients Manufacturing Example 3-2
(weight%) honeysuckle extract 2.0 glycerin 3.0 Butylene glycol 2.0 propylene glycol 2.0 Carboxy vinyl polymer 0.1 PEG 12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservatives, coloring, flavoring Appropriate amount Purified water to 100

<3-3> Nutritional cream

A nutritional cream was prepared according to a conventional method using the honeysuckle extract of the present invention according to the nutritional cream formulation ratio shown in Table 5 below.

Ingredients Manufacturing Example 3-3
(weight%) honeysuckle extract 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 PEG60 hydrogenated castor oil 2.0 liquid 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 Appropriate amount pigment Appropriate amount Spices Appropriate amount Purified water to 100

<3-4> Massage cream

Massage cream was prepared according to a conventional method by using the pulverized product or extract of honeysuckle according to the present invention according to the massage cream formulation ratio in Table 6 below.

Ingredients Manufacturing Example 3-4
(weight%) Honeysuckle powder or honeysuckle extract 1.0 beeswax 10.0 Polysorbate 60 1.5 PEG 60 hydrogenated castor oil 2.0 Sorbitan sesquioleate 0.8 liquid 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, coloring, flavoring Appropriate amount Purified water to 100

<3-5> Pack

A pack was prepared according to a conventional method using the honeysuckle extract of the present invention according to the pack formulation ratio shown in Table 7 below.

Ingredients Manufacturing Example 3-5
(weight%) honeysuckle extract 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, coloring, flavoring Appropriate amount Purified water to 100

<3-6> Gel

A gel was prepared according to a conventional method using the honeysuckle extract of the present invention according to the gel formulation ratio shown in Table 8 below.

Ingredients Manufacturing Example 3-6
(weight%) honeysuckle extract 0.5 Sodium ethylenediamine acetate 0.05 glycerin 5.0 Carboxy vinyl polymer 0.3 ethanol 5.0 PEG 60 hydrogenated castor oil 0.5 Triethanolamine 0.3 Preservatives, coloring, flavoring Appropriate amount Purified water to 100

As described above, the present invention provides a composition for improving, treating or preventing muscle disease, or improving muscle function, containing honeysuckle, such as honeysuckle powder or honeysuckle extract. More specifically, the honeysuckle of the present invention improves muscle mass and strength by increasing the activity of mTOR and reducing the mRNA expression level of MuRF1 and atrogin-1, thereby providing excellent effects for improving, treating or preventing muscle diseases, or improving muscle function. indicates. Therefore, the present invention has high industrial applicability because it can provide a composition that shows excellent effects for improving, treating or preventing muscle diseases, or improving muscle function.

Claims (8)

A food composition for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient. According to clause 1,
The honeysuckle is a crushed product of flower buds, leaves, or branches of honeysuckle,
The food composition for improving or preventing muscle disease or improving muscle function, characterized in that the honeysuckle extract is obtained by extracting flower buds, leaves, or branches of honeysuckle. According to clause 1,
The honeysuckle extract is obtained by extracting honeysuckle with one or more solvents selected from the group consisting of water, organic solvents having 1 to 6 carbon atoms, subcritical fluids, and supercritical fluids, for improving or preventing muscle diseases, or for muscle Food composition for improving function. According to claim 3,
The organic solvent includes alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, and hexane ( A food composition for improving or preventing muscle disease, or improving muscle function, characterized in that it is one or more solvents selected from the group consisting of hexane, cyclohexane, and petroleum ether. The method according to any one of claims 1 to 4,
The muscle disease is characterized in that it is at least one selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia gravis, cachexia, and sarcopenia. Food composition for improving or preventing muscle disease or improving muscle function. A pharmaceutical composition for treating or preventing muscle disease containing honeysuckle as an active ingredient. A cosmetic composition for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient. A feed additive for improving or preventing muscle disease or improving muscle function containing honeysuckle as an active ingredient.