KR102277061B1 - Pharmaceutical composition for prevention or treatment of muscular disease comprising Codonopsis lanceolata extract as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating muscle diseases containing an extract of deodeok ( Codonopsis lanceolata ) as an active ingredient. Specifically, the deodeok extract of the present invention alleviates the decrease in muscle weight and muscle fiber cross-sectional area due to muscle atrophy, and MuRF-1 (Muscle RING-finger protein-1) and atrozine-1 ( atrogin-1), and by increasing the expression of phosphorylated mammalian target of rapamycin (p-mTOR) involved in muscle protein synthesis, it can be usefully used to prevent, improve or treat muscle diseases .

Description

A pharmaceutical composition for the prevention or treatment of muscular diseases comprising deodeok extract as an active ingredient {Pharmaceutical composition for prevention or treatment of muscular disease comprising Codonopsis lanceolata extract as an active ingredient}

The present invention relates to a pharmaceutical composition for preventing or treating muscle diseases containing an extract of deodeok ( Codonopsis lanceolata ) as an active ingredient.

Skeletal muscle is a tissue that accounts for 40 to 50% of human body weight, and one skeletal muscle is composed of a number of muscle fibers and connective tissue, has a horizontal pattern, and is involved in voluntary movement. Muscles play an important role in metabolic functions such as maintenance of energy homeostasis and thermogenesis, and the size of muscle is an intracellular signaling process that induces anabolism or catabolism within the muscle. pathways).

Muscle atrophy refers to a continuous decrease in muscle mass and is caused by muscle weakness and degeneration (McKinnell IW et al. , Cell, 119(7): 907-910, 2004). Muscle atrophy is induced by reduced activity, oxidative stress, and chronic inflammation, weakening muscle function and exercise capacity (Lee CE et el. , Clinical Nutrition, 26(5): 524-534, 2007), and is more important than protein synthesis. It occurs when degradation occurs more frequently (Nader GA, The International Journal of Biochemistry and Cell Biology, 37(10): 1985-1996, 2005).

A representative factor involved in the synthesis of muscle protein is mTOR (mammalian target of rapamycin). mTOR is an important factor regulating the initiation of protein translation. The phosphorylated form (phosphorylated mTOR, p-mTOR) is an activated form, and its expression does not change significantly when muscles are continuously immobilized, but activation is It is known to relieve muscle atrophy when increased. mTOR increases muscle mass by inducing the synthesis of muscle protein by activating 4EBP1 (4E-binding protein 1) and p70S6K (phosphorylated 70-kDa ribosomal S6 kinase), which initiate mRNA translation (Bodine et el. , SC Nature cell biology , 3(11), 1014, 2001). On the other hand, representative factors involved in the degradation of muscle proteins include atrogin-1 and MuRF-1 (muscle RING-finger protein-), which are muscle specific E3 ubiquitin ligase factors. 1) is there. Atrogine-1 and MuRF-1 were significantly increased when the activity was decreased (Foletta VC et el. , Pflugers Archiv-European Journal of Physiology, 461(3), 325-335, 2011), which Proteosome-dependent proteolysis is promoted, resulting in reduced muscle mass. Therefore, promotion of mTOR activity and inhibition of atrozine-1 and MuRF-1 expression increase the amount of muscle protein, thereby increasing muscle mass. Currently, an increase in mitochondrial production, an inhibitor of muscle proteolysis, and an anti-inflammatory agent have been suggested as a treatment method for muscle atrophy, but there is no clear therapeutic agent.

On the other hand, deodeok ( Codonopsis lanceolata ) is a plant belonging to the Campanulaceae family, and is a perennial herb that is native to or cultivated in shady places in high mountains in Korea, China, and Japan. Deodeok is called by various names such as white ginseng, saginseng, haengyeop, gideok, and jichi, and has been known for its excellent pharmacological effects such as tonic, antipyretic, antitussive, expectorant, detoxification, and drainage in oriental medicine since ancient times. It has also been reported to have alcoholic liver disease improvement and anti-inflammatory activity (Hossen MJ et el. , Phytotherapy research, 30(3), 347-356, 2016). The root of deodeok contains a lot of saponin, which gives off a characteristic scent, and has the effect of stopping coughing and relieving phlegm, and is recognized as a good food for bronchitis, tonsillitis, and sore throat.

Accordingly, the present inventors, while discovering natural substances having muscle function regulating activity, the deodeok extract alleviates the decrease in muscle weight and muscle fiber cross-sectional area due to muscle atrophy, and affects the expression of factors involved in the synthesis or degradation of muscle proteins The present invention was completed by confirming that

It is an object of the present invention to provide the use of deodeok extract for treating muscle disease.

In order to achieve the above object, the present invention deodeok ( Codonopsis lanceolata ) provides a pharmaceutical composition for the prevention or treatment of muscle diseases containing the extract as an active ingredient.

In addition, the present invention provides a health functional food for preventing or improving muscle diseases containing deodeok extract as an active ingredient.

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

In addition, the present invention provides a skin external preparation for the prevention or treatment of muscle diseases containing deodeok extract as an active ingredient.

In addition, the present invention provides a feed additive or feed composition for preventing or improving muscle disease containing deodeok extract as an active ingredient.

The deodeok extract of the present invention alleviates the decrease in muscle weight and muscle fiber cross-sectional area due to muscle atrophy, and MuRF-1 (Muscle RING-finger protein-1) and atrogin-1 (atrogin-1) involved in the breakdown of muscle protein ), and by increasing the expression of phosphorylated mammalian target of rapamycin (p-mTOR) involved in muscle protein synthesis, it can be usefully used to prevent, improve or treat muscle diseases.

1 is a diagram showing the muscle weight per unit body weight of a mouse gastrocnemius (Cont: untreated control group, IM: muscle atrophy induction group, HMB370: 370 mg/kg HMB (beta-hydroxy beta-methlybutyrate) group , CL200: 200 mg/kg of deodeok ( Codonopsis lanceolata ) extract group).
2 is a diagram showing the muscle weight per unit body weight of the quadriceps of a mouse (Cont: untreated control group, IM: muscle atrophy induction group, HMB370: 370 mg/kg HMB administration group, CL200: 200 mg/kg deodeok extract group).
3 is a diagram showing the size of mouse muscle fibers (Cont: untreated control group, IM: muscle atrophy induction group, HMB370: 370 mg/kg HMB administration group, CL200: 200 mg/kg deodeok extract group).
4 is a diagram illustrating the quantification of the mouse muscle fiber cross-sectional area (Cont: untreated control group, IM: muscle atrophy induction group, HMB370: 370 mg/kg HMB-administered group, CL200: 200 mg/kg deodeok extract group).
5 is a diagram showing the expression of MuRF-1 gene in the calf muscle of a mouse (Cont: untreated control group, IM: muscle atrophy induction group, HMB370: 370 mg/kg HMB administration group, CL200: 200 mg/kg deodeok extract administration group).
6 is a diagram showing the expression of atrozin-1 gene in the calf muscle of a mouse (Cont: untreated control group, IM: muscle atrophy induction group, HMB370: 370 mg/kg of HMB administration group, CL200: 200 mg/kg of deodeok extract group).
7 is a diagram showing the expression of p-mTOR, MuRF-1 or atrozin-1 protein in the calf muscle of a mouse (Cont: untreated control group, IM: muscle atrophy induction group, HMB370: 370 mg/kg HMB administration group , CL200: 200 mg/kg deodeok extract group).
8 is a diagram showing the cytotoxicity confirmation experiment results of deodeok extract in the source cells. Data from three independent experiments are expressed as mean±SD. ^{* indicates a significant difference from the control group in *} p <0.05, ^** p < 0.01, and ^*** p < 0.001 by unpaired t-test. FIG. 9 shows deodeok extract treatment in the C2C12 myoblast muscle atrophy induction model. It is a diagram showing the change in the diameter of the cell muscle fiber according to the
10 is a graph in which the diameter change of FIG. 9 is digitized. Data from three independent experiments are expressed as mean±SD. # indicates a significant difference from the normal group in ^### p < 0.001 by unpaired t-test. * indicates a significant difference from the control group in ^** p <0.01, ^*** p <0.001 by unpaired t-test.
11 is a diagram showing the expression of atrozin-1 gene in the C2C12 myoblast muscle atrophy induction model (normal group: untreated control group, induction group: muscle atrophy induction group, 2.5: 2.5 μg/mL deodeok extract group, 5: 5 μg/mL deodeok extract group, 10: 10 μg/mL deodeok extract group). Data from three independent experiments are expressed as mean±SD. # indicates a significant difference from the normal group in ^### p < 0.001 by unpaired t-test. * indicates a significant difference from the control group in ^* p <0.05, ^** p < 0.01 by unpaired t-test.
12 is a diagram showing the expression of MuRF-1 gene in the C2C12 myoblast muscle atrophy induction model (normal group: untreated control group, induction group: muscle atrophy induction group, 2.5: 2.5 μg/mL deodeok extract group, 5: 5 μg/mL deodeok extract group, 10: 10 μg/mL deodeok extract group). Data from three independent experiments are expressed as mean±SD. # indicates a significant difference from the normal group in ^### p < 0.001 by unpaired t-test. * indicates a significant difference from the control group at ^** p < 0.01 by unpaired t-test.
13 is a diagram showing the expression of MuRF-1 or atrozin-1 protein in the C2C12 myoblast induction model (N: normal group, C: muscle atrophy induction group, 2.5: 2.5 μg/mL deodeok extract group) , 5: 5 μg/mL deodeok extract group, 10: 10 μg/mL deodeok extract group)
14 is a graph showing quantification of relative expression reflecting the results of FIG. 11 . Data from three independent experiments are expressed as mean±SD. # indicates a significant difference from the normal group in ^## p < 0.01 and ^### p < 0.001 by unpaired t-test. * indicates a significant difference from the control group in ^* p <0.05, ^** p < 0.01, ^*** p < 0.001 by unpaired t-test.
15 is a diagram showing the expression of p-mTOR or mTOR protein in the C2C12 myoblast muscle atrophy induction model (N: normal group, C: muscle atrophy induction group, 2.5: 2.5 μg/mL deodeok extract group, 5: 5 μg/mL deodeok extract group, 10: 10 μg/mL deodeok extract group)
16 is a graph showing quantification of relative expression reflecting the results of FIG. 15 . Data from three independent experiments are expressed as mean±SD. * indicates a significant difference from the control group in ^** p <0.01, ^*** p <0.001 by unpaired t-test.
17 is a graph showing the protein expression level ratio of p-mTOR and mTOR. Data from three independent experiments are expressed as mean±SD. * indicates a significant difference from the control group in ^* p <0.05, ^** p < 0.01 by unpaired t-test.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating muscle diseases containing an extract of deodeok ( Codonopsis lanceolata ) as an active ingredient.

The deodeok may include both raw deodeok, dried deodeok, and the like, and specifically may be dried deodeok. In addition, the deodeok may include whole deodeok, finely chopped deodeok, powder of deodeok, and the like, and specifically may be powder of deodeok. In addition, the dried deodeok may be dried by air drying, reduced pressure drying, vacuum drying, boiling drying, spray drying, freeze drying or hot air drying, and specifically may be dried by air drying.

The deodeok extract can be prepared by a manufacturing method comprising the following steps:

1) preparing an extract by adding an extraction solvent to deodeok;

2) filtering the extract of step 1); and

3) drying the filtered filtrate of step 2) after concentrating under reduced pressure.

In the manufacturing method, deodeok may include all parts such as leaves, stems, roots, and fruits of deodeok, and specifically, may be roots.

In addition, the extraction solvent may be water, alcohol, or a mixture thereof. The alcohol may be a C ₁ to C ₂ lower alcohol, and specifically, the alcohol may be ethanol or methanol. In one embodiment of the present invention, the extraction solvent may be water. The extraction solvent may be added in an amount of 1 to 15 mL per 1 g of the weight of the deodeok used for extraction, specifically, it may be added in an amount of 10 mL. The extraction temperature may be 30 to 100 ℃, specifically, may be 60-85 ℃.

The extraction method may be shaking extraction, Soxhlet extraction, or reflux extraction. At this time, the extraction time may be 1 to 10 hours, or 3 to 6 hours, specifically 4-5 hours. The extraction may be 1 to 5 times, specifically 3 times.

On the other hand, the vacuum concentration in step 3) may use a vacuum vacuum concentrator or a vacuum rotary evaporator. In addition, the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, in particular, may be freeze drying.

The muscle disease refers to a disease caused by muscle wasting or degeneration, and sarcopenia, dystonia (atony), muscular atrophy (muscular atrophy), muscular dystrophy (muscular dystrophy), muscle degeneration, myotonic dystrophy, It may include any one or more selected from the group consisting of amyotrophic lateral sclerosis, myasthenia and cachexia. The muscle wasting is characterized by a gradual loss of muscle mass and weakening and degeneration of voluntary muscles such as skeletal muscles or involuntary muscles such as cardiac muscles. Muscle wasting and degeneration is caused by genetic factors, acquired factors, aging, and the like.

The pharmaceutical composition of the present invention may alleviate a decrease in muscle weight and a cross-sectional area of a muscle fiber due to muscle atrophy, and increase the expression of a gene or protein of MuRF-1 or atrozin-1, which is a factor involved in the degradation of muscle protein It may be to relieve the muscle protein, or it may be to increase the expression of p-mTOR protein, a factor involved in the synthesis of muscle protein.

In a specific embodiment of the present invention, the present inventors prepared a deodeok extract, and oral administration of the deodeok extract to a mouse induced muscle atrophy alleviates the decrease in muscle weight and muscle fiber cross-sectional area (see FIGS. 1 to 4), The increase in the expression of the gene or protein of MuRF-1 or atrozin-1 involved in the degradation of muscle protein is alleviated (see FIGS. 5 to 7), and the expression of the p-mTOR protein involved in the synthesis of muscle protein is increased (see FIG. 7) was confirmed. In addition, in the myoblast muscle atrophy induction model, when the deodeok extract was treated, the size of myofibrils increased (see FIGS. 9 to 10), and the expression of factors involved in the synthesis or degradation of muscle proteins was changed (FIG. 11 to 17) were confirmed. Therefore, the deodeok extract can be usefully used for the prevention or treatment of muscle diseases.

The pharmaceutical composition according to the present invention may contain 10 to 95% by weight of the active ingredient, deodeok extract, based on the total weight of the composition. In addition, the pharmaceutical composition of the present invention may further include at least one active ingredient exhibiting the same or similar function in addition to the active ingredient.

The pharmaceutical composition of the present invention may include a carrier, a diluent, an excipient or a mixture thereof commonly used in biological agents. Any pharmaceutically acceptable carrier may be used as long as it is suitable for delivering the composition in vivo. Specifically, the carrier is Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, Ringer's solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or mixtures thereof. In addition, conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed.

When formulating the composition, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents or excipients such as surfactants may be added.

The composition of the present invention may be formulated as an oral preparation or a parenteral preparation. Oral formulations may include solid formulations and liquid formulations. The solid preparation may be a tablet, pill, powder, granule, capsule, or troche, and the solid preparation may be prepared by adding at least one excipient to the composition. The excipient may be starch, calcium carbonate, sucrose, lactose, gelatin, or a mixture thereof. In addition, the solid formulation may contain a lubricant, examples of which include magnesium stearate, talc, and the like. Meanwhile, the liquid formulation may be a suspension, an internal solution, an emulsion, or a syrup. In this case, the liquid formulation may include excipients such as wetting agents, sweetening agents, fragrances, and preservatives.

The parenteral formulation may include injections, suppositories, powders for respiratory inhalation, aerosols for sprays, powders and creams, and the like. The injection may include a sterile aqueous solution, a non-aqueous solution, a suspension solution, an emulsion, and the like. In this case, as the non-aqueous solvent or suspension solution, vegetable oils such as propylene glycol, polyethylene glycol, and olive oil, or injectable esters such as ethyl oleate may be used.

The composition of the present invention may be administered orally or parenterally according to a desired method. Parenteral administration may include intraperitoneal, intrarectal, subcutaneous, intravenous, intramuscular or intrathoracic injection.

The composition may be administered in a pharmaceutically effective amount. This may vary depending on the type of disease, severity, drug activity, patient's sensitivity to the drug, administration time, administration route, treatment period, drugs used simultaneously, and the like. However, for a desirable effect, the amount of the active ingredient included in the pharmaceutical composition according to the present invention may be 0.0001 to 500 mg/kg, specifically 0.001 to 300 mg/kg. The administration may be once to several times a day.

The composition of the present invention may be administered alone or in combination with other therapeutic agents. When administered in combination, the administration may be sequential or simultaneous.

In addition, the present invention provides a health functional food for preventing or improving muscle diseases containing deodeok extract as an active ingredient.

Deodeok or deodeok extract according to the present invention is as described above. In an embodiment of the present invention, the extract may be a root extract of deodeok, and may be extracted using water as a solvent.

The muscle disease may include any one or more selected from the group consisting of sarcopenia, dystonia, muscular atrophy, muscular dystrophy, muscle degeneration, muscle stiffness, amyotrophic axonal sclerosis, myasthenia gravis and cachexia.

The health functional food of the present invention may alleviate a decrease in muscle weight and muscle fiber cross-sectional area due to muscle atrophy, and increase the expression of a gene or protein of MuRF-1 or atrozin-1, which is a factor involved in the degradation of muscle protein It may be to relieve the muscle protein, or it may be to increase the expression of p-mTOR protein, a factor involved in the synthesis of muscle protein.

In a specific embodiment of the present invention, the present inventors prepared a deodeok extract, and oral administration of the deodeok extract to a mouse induced muscle atrophy alleviates the decrease in muscle weight and muscle fiber cross-sectional area (see FIGS. 1 to 4), It was confirmed that the expression of factors involved in the synthesis or degradation of muscle protein was changed (see FIGS. 5 to 7 ). In addition, in the myoblast muscle atrophy induction model, when the deodeok extract was treated, the size of myofibrils increased (see FIGS. 9 to 10), and the expression of factors involved in the synthesis or degradation of muscle proteins was changed (FIG. 11 to 17) were confirmed. Therefore, the deodeok extract can be usefully used for the prevention or improvement of muscle diseases.

As used herein, the term "health functional food" refers to a food manufactured using raw materials or ingredients having a useful function or nutrients that are easily deficient in daily meals, and refers to food that helps maintain human health, but is not limited thereto. and is used in the meaning of including all health foods in the ordinary sense.

The form and type of health functional food is not particularly limited. Specifically, the health functional food may be in the form of tablets, capsules, powders, granules, liquids and pills. The health functional food may include various flavoring agents, sweetening agents, or natural carbohydrates as additional ingredients. The sweetener may be a natural or synthetic sweetener, and examples of the natural sweetener include thaumatin, stevia extract, and the like. Meanwhile, examples of synthetic sweeteners include saccharin, aspartame, and the like. In addition, the natural carbohydrates may be monosaccharides, disaccharides, polysaccharides, oligosaccharides and sugar alcohols.

The health functional food of the present invention includes, in addition to the above-described additional ingredients, nutrients, vitamins, electrolytes, flavoring agents, colorants, pextan and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives , glycerin, alcohol, and the like may be further included. These components may be used independently or in combination. The proportion of the additive may be selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The deodeok extract of the present invention can be added to food as it is or used with other foods or food ingredients. In this case, the content of the added active ingredient may be determined according to the purpose. In general, the content in the health functional food may be 0.01 to 90 parts by weight of the total weight of the food. However, in the case of long-term ingestion for health and hygiene purposes or for health control, the amount may be less than the above range, and if there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

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

Deodeok or deodeok extract according to the present invention is as described above. In an embodiment of the present invention, the extract may be a root extract of deodeok, and may be extracted using water as a solvent.

The muscle disease may include any one or more selected from the group consisting of sarcopenia, dystonia, muscular atrophy, muscular dystrophy, muscle degeneration, muscle stiffness, amyotrophic axonal sclerosis, myasthenia gravis and cachexia.

The cosmetic composition of the present invention may alleviate a decrease in muscle weight and a cross-sectional area of a muscle fiber due to muscle atrophy, and increase the expression of a gene or protein of MuRF-1 or atrozin-1, which is a factor involved in the degradation of muscle protein. It may be to relieve, or it may be to increase the expression of p-mTOR protein, a factor involved in the synthesis of muscle protein.

In a specific embodiment of the present invention, the present inventors prepared a deodeok extract, and oral administration of the deodeok extract to a mouse induced muscle atrophy alleviates the decrease in muscle weight and muscle fiber cross-sectional area (see FIGS. 1 to 4), It was confirmed that the expression of factors involved in the synthesis or degradation of muscle protein was changed (see FIGS. 5 to 7 ). In addition, in the myoblast muscle atrophy induction model, when the deodeok extract was treated, the size of myofibrils increased (see FIGS. 9 to 10), and the expression of factors involved in the synthesis or degradation of muscle proteins was changed (FIG. 11 to 17) were confirmed. Therefore, the deodeok extract can be usefully used for the prevention or improvement of muscle diseases.

The cosmetic composition of the present invention may contain the deodeok extract in an amount of 0.1 to 50% by weight.

In addition, the cosmetic composition of the present invention may be formulated in a conventionally prepared cosmetic formulation. The cosmetic composition may be formulated as a solution, suspension, emulsion, paste, gel, lotion, cream, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray. Specifically, it may be a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or mixtures thereof. In addition, when the formulation of the cosmetic composition is powder or spray, it may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof. In particular, in the case of a spray, chlorofluorohydrocarbon, propane/butane or dimethyl ether may be further included.

When the formulation of the cosmetic composition of the present invention is a solution or emulsion, it may include a solvent, a solvating agent, an emulsifying agent, or a mixture thereof as a carrier. Examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, sorbitan fatty acid ester, and the like.

When the formulation of the cosmetic composition of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester; microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tragacanth or mixtures thereof. In addition, when the formulation of the cosmetic composition is surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ethersulfide, alklamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerol fatty acid ester, or mixtures thereof.

In addition to the carrier component, the cosmetic composition of the present invention may include an antioxidant, a stabilizer, a solubilizer, a moisturizer, a pigment, a fragrance, a sunscreen, a color developer, a surfactant, or a mixture thereof as an adjuvant. The adjuvant may be used as long as it is a material commonly used in the preparation of a cosmetic composition.

In addition, the present invention provides a skin external preparation for the prevention or treatment of muscle diseases containing deodeok extract as an active ingredient.

Deodeok or deodeok extract according to the present invention is as described above. In an embodiment of the present invention, the extract may be a root extract of deodeok, and may be extracted using water as a solvent.

The muscle disease may include any one or more selected from the group consisting of sarcopenia, dystonia, muscular atrophy, muscular dystrophy, muscle degeneration, muscle stiffness, amyotrophic axonal sclerosis, myasthenia gravis and cachexia.

The external preparation for skin of the present invention may alleviate a decrease in muscle weight and a cross-sectional area of a muscle fiber due to muscle atrophy, and increase the expression of a gene or protein of MuRF-1 or atrozin-1, which is a factor involved in the degradation of muscle protein. It may be to relieve, or it may be to increase the expression of p-mTOR protein, a factor involved in the synthesis of muscle protein.

In a specific embodiment of the present invention, the present inventors prepared a deodeok extract, and oral administration of the deodeok extract to a mouse induced muscle atrophy alleviates the decrease in muscle weight and muscle fiber cross-sectional area (see FIGS. 1 to 4), It was confirmed that the expression of factors involved in the synthesis or degradation of muscle protein was changed (see FIGS. 5 to 7 ). In addition, in the myoblast muscle atrophy induction model, when the deodeok extract was treated, the size of myofibrils increased (see FIGS. 9 to 10), and the expression of factors involved in the synthesis or degradation of muscle proteins was changed (FIG. 11 to 17) were confirmed. Therefore, the deodeok extract can be usefully used for the prevention or treatment of muscle diseases.

The external preparation for skin of the present invention may include a pharmaceutically acceptable carrier and excipient. The carrier and excipient may include preservatives, stabilizers, wetting agents, emulsification accelerators and buffers, and the like. Specifically, the excipient may be lactose, dextrin, starch, mannitol, sorbitol, glucose, saccharose, microcrystalline cellulose, gelatin, polyvinylpyrrolidone, or a mixture thereof. The external preparation for skin may be appropriately prepared according to a method well known in the art. The external preparation for skin may be prepared in the form of powder, gel, ointment, cream, liquid and aerosol.

In addition, the present invention provides a feed additive or feed composition for preventing or improving muscle disease containing deodeok extract as an active ingredient.

As used herein, "feed" means any natural or artificial diet, meal, etc., or a component of the meal, intended for or suitable for being eaten, ingested, and digested by an animal. The feed can be classified in various ways according to nutritional value, main ingredient, distribution, moisture content, compounding state and processing type, etc., and the feed includes roughage, rich feed, supplementary feed, protein feed, starch feed, fat feed or fiber feed. It can be used, but is not limited thereto.

The term "feed additive" as used herein refers to a substance added to feed for the purpose of various effects, such as nutrient supplementation and weight loss prevention, enhancement of digestibility of fiber in feed, improvement of oil quality, prevention of reproductive disorders and improvement of fertility rate, prevention of high temperature stress in summer do. The feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act, and mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, and trace minerals such as zinc, copper, cobalt, selenium, and kerotene. , vitamin E, vitamins A, D, E, nicotinic acid, vitamins such as vitamin B complex, protective amino acids such as methionine and lyic acid, protective fatty acids such as fatty acid calcium salt, probiotics (lactic acid bacteria), yeast culture, mold Live bacteria such as fermented products, yeast agents, and the like may be further included.

Deodeok or deodeok extract according to the present invention is as described above. In an embodiment of the present invention, the extract may be a root extract of deodeok, and may be extracted using water as a solvent.

The muscle disease may include any one or more selected from the group consisting of sarcopenia, dystonia, muscular atrophy, muscular dystrophy, muscle degeneration, muscle stiffness, amyotrophic axonal sclerosis, myasthenia gravis and cachexia.

The feed additive or composition for feed of the present invention may be to alleviate a decrease in muscle weight and muscle fiber cross-sectional area due to muscle atrophy, and a gene or protein of MuRF-1 or atrozin-1, which is a factor involved in the degradation of muscle protein. It may be to alleviate the increase in expression, or it may be to increase the expression of p-mTOR protein, which is a factor involved in the synthesis of muscle protein.

In a specific embodiment of the present invention, the present inventors prepared a deodeok extract, and oral administration of the deodeok extract to a mouse induced muscle atrophy alleviates the decrease in muscle weight and muscle fiber cross-sectional area (see FIGS. 1 to 4), It was confirmed that the expression of factors involved in the synthesis or degradation of muscle protein was changed (see FIGS. 5 to 7 ). In addition, in the myoblast muscle atrophy induction model, when the deodeok extract was treated, the size of myofibrils increased (see FIGS. 9 to 10), and the expression of factors involved in the synthesis or degradation of muscle proteins was changed (FIG. 11 to 17) were confirmed. Therefore, the deodeok extract can be usefully used for the prevention or improvement of muscle diseases.

Since the feed additive of the present invention has the effect of preventing or improving muscle disease, it is possible to prevent muscle disease by continuously ingesting it to poultry, livestock, and the like, and to improve muscle disease that has already occurred.

In addition, the feed additive may additionally contain an acceptable carrier such as poultry and livestock. A known carrier, stabilizer, etc. may be added to the feed additive, and if necessary, various nutrients such as vitamins, amino acids, minerals, antioxidants, antibiotics, antibacterial agents and other additives may be added. , granules, pellets, suspensions, and the like.

When the feed additive is supplied, it may be supplied alone or mixed with feed for poultry and livestock.

Since the composition for feed of the present invention has the effect of preventing or improving muscle disease, it is possible to prevent muscle disease by continuously ingesting it to poultry, livestock, and the like, and to improve muscle disease that has already occurred.

In addition, the composition for feed can be prepared by adding deodeok extract in an appropriate effective concentration range according to various feed preparation methods known in the art.

The composition for feed is not particularly limited as long as it is an individual for the purpose of preventing or improving muscle disease, and is applicable to any individual. For example, non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cattle, sheep, pigs, goats, etc., birds, or fish can be applied to any subject.

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Example 1. Preparation of deodeok extract 1

Deodeok extract was prepared by purchasing deodeok root of good quality and not deteriorated. Specifically, the purchased deodeok was air-dried for 4 days, pulverized, and the dried powder was circulated and dehydrated with water. 1000 mL of distilled water was added to 100 g of deodeok powder, and the extract was extracted three times at 85° C. for 4 hours at room temperature. After cooling to , filtration and concentration to prepare a deodeok extract. The prepared extract was prepared by freeze-drying to make a powder.

Example 2. Preparation of deodeok extract 2

Deodeok extracts were prepared by purchasing deodeok roots of good quality and unaltered (sandodeok, a luxury brand in Jonamsan, Yongmunsan, Yangpyeong, Gyeonggi-do). Specifically, the purchased deodeok was air-dried for 4 days, then pulverized, and the dried powder was cycle-washed and dehydrated with water. 1000 mL of distilled water was added to 100 g of deodeok powder, shaken at 60° C. for 5 hours, and extracted three times. After cooling the extract to room temperature, it was filtered and concentrated to prepare a deodeok extract. The prepared extract was prepared by freeze-drying to make a powder.

Experimental Example 1. Confirmation of changes in muscle weight and muscle fiber cross-sectional area according to administration of deodeok extract

It was confirmed by animal experiments whether deodeok extract improved the reduction of muscle weight and muscle fiber cross-sectional area according to muscle atrophy.

1-1. Confirmation of relief of muscle weight loss

Whether the deodeok extract relieves muscle weight loss due to muscle atrophy was confirmed by the following method.

First, 5-week-old male C57BL/6 mice were acclimatized to the animal laboratory environment for 7 days, and then divided into 5 groups according to the experimental conditions. Specifically, the experimental group was an untreated control group (normal group, Cont), muscle atrophy induction group (induction group, IM), muscle atrophy induction and 370 mg/kg of beta-hydroxy beta-methylbutylate (beta-hydroxy beta- It was divided into a positive control group (HMB administration group, HMB 370) administered with methlybutyrate, HMB), and an experimental group (deodeok extract administration group, CL 200) to which muscle atrophy was induced and 200 mg/kg of deodeok extract (Example 1) was administered. Muscle atrophy is leg immobilization by applying a fixation device made of a 1.5 mL microfuge tube, clips, and Velcro tape to one hind limb of a mouse according to 'Disease models & mechanisms, 8(9), 1059-1069, 2015'. (immobilization) was used to induce it. HMB and deodeok extract were dissolved in 0.5% carboxymethyl cellulose (CMC) and administered orally once a day for 14 days from the day of application of leg fixation. After 14 days, the gastrocnemius and quadriceps of the hind leg were excised and the weight was measured and standardized based on the weight.

As a result, as shown in FIGS. 1 and 2 , muscle atrophy was induced in the induction group, and the weight of the calf muscle and quadriceps muscle was reduced by about 40% and about 33% compared to the normal group, and the HMB administration group compared with the normal group. The weight of the calf muscle was reduced by about 24% and the quadriceps muscle by about 17%, so HMB showed about 16% of muscle atrophy protection, respectively. In the deodeok extract-treated group, the calf muscle decreased by about 27% and the quadriceps muscle compared to the normal group. The muscle weight was reduced by about 18%, so the deodeok extract showed about 13% and 15% muscle atrophy protection, respectively. From this, it can be seen that the deodeok extract relieves muscle weight loss due to muscle atrophy, and has a superior protective effect against muscle atrophy per unit dose than the positive control, HMB.

1-2. Confirmation of relief of muscle fiber cross-sectional area reduction

Whether the deodeok extract relieved the reduction of the muscle fiber cross-sectional area due to muscle atrophy was confirmed by the following method. Specifically, the calf muscle tissue extracted in Experimental Example 1-2 was fixed with 4% paraformaldehyde, and then stained with hematoxylin and eosin (H&E), and image J software (image J software). ) was used to quantify the muscle fiber cross-sectional area.

As a result, as shown in FIG. 3 , muscle atrophy was induced in the induction group, and the muscle fiber size of the calf muscle was significantly reduced compared to the normal group, and the decrease in the size of the muscle fibers was alleviated in the HMB-administered group and the deodeok extract-administered group. In addition, as shown in FIG. 4 , the cross-sectional area of muscle fibers decreased by about 50% in the induction group, by about 33% in the HMB-administered group, and by about 35% in the deodeok extract-administered group compared to the normal group, so that the HMB and deodeok extract were about 17% and 15%, respectively. % of the muscle atrophy protection. From this, it was found that the deodeok extract relieved the reduction in the cross-sectional area of muscle fibers due to muscle atrophy, and had a better protective effect on muscle atrophy per unit dose than the positive control, HMB.

Experimental Example 2. Confirmation of degradation of muscle protein or change in expression of synthetic factors according to administration of deodeok extract

It was confirmed by the following method whether the deodeok extract changes the expression of factors involved in the degradation or synthesis of muscle proteins according to muscle atrophy.

2-1. Confirmation of alleviation of increased expression of MuRF-1 (Muscle RING-finger protein-1) and atrogin-1 (atrogin-1) genes involved in muscle protein degradation

It was confirmed using qRT-PCR (quantitative real-time PCR) whether the deodeok extract relieved the increase in the expression of MuRF-1 and atrozin-1, which are involved in the degradation of muscle protein.

Specifically, mRNA was extracted from the calf muscle tissue extracted in Experimental Example 1-2 according to the manufacturer's protocol using RNA RED (Intron, Korea), and cDNA was synthesized according to the manufacturer's protocol using a cDNA synthesis kit (TaKaRa, Japan). After that, qRT-PCR was performed according to a conventionally known method using the primers of Table 1 and the ABI Step One Plus TM Real-Time PCR System (Applied Biosystems, USA).

Primer name Primer sequence (5'→3') SEQ ID NO: MuRF-1 forward AAG ACT GAG CTG AGT AAC TG One MuRF-1 reverse TAG AGG GTG TCA AAC TTC TG 2 Atrogin-1 forward AGA AAG AAA GAC ATT CAG AAC A 3 Atrogin-1 reverse GCT CCT TCG TAC TTC CTT 4

As a result, as shown in FIGS. 5 and 6, the MuRF-1 gene was expressed about 3 times in the induction group, about 1.3 times in the HMB-administered group, and about 0.8 times in the deodeok extract-administered group, compared to the normal group, and atrozine- 1 gene was expressed about 3 times in the induction group, about 1.7 times in the HMB-administered group, and about 1.1 times in the deodeok extract-administered group compared to the normal group. From this, it can be seen that the deodeok extract relieves the increase in the expression of MuRF-1 and atrozin-1 genes according to muscle atrophy, and has a better protective effect on muscle atrophy than the positive control, HMB.

2-2. Alleviation of the increase in the expression of MuRF-1 and atrozin-1 proteins involved in the degradation of muscle proteins, and confirmation of the increase in the expression of the phosphorylated mammalian target of rapamycin (p-mTOR) protein involved in the synthesis of muscle proteins

Western blot to determine whether deodeok extract relieved the increase in the expression of MuRF-1 and atrozin-1 proteins involved in muscle protein degradation and increased the expression of p-mTOR protein involved in muscle protein synthesis. was confirmed using .

Specifically, from the calf muscle tissue extracted in Experimental Example 1-2, cOmplete ^TM protease inhibitor cocktail tablets (cOmplete ^TM protease inhibitor cocktail tablets, Roche Diagnostics, USA) 4, sucrose (sucrose) 17.115 g, 1 M Tris Proteins were lysed according to the manufacturer's protocol using a lysis buffer containing 2 mL of buffer (tris buffer, Trizma base, pH 7.4), 0.2 mL of 0.5 M EDTA (pH 8.0), and distilled water (DW). extracted. Using the Pierce ^TM BCA Protein Assay Kit (Pierce ^TM BCA Protein Assay Kit, Thermo Fisher Scientific, USA), check the protein concentration according to the manufacturer's protocol and adjust it to a certain concentration, and then add the protein at the same concentration to 7.5% sodium dodecyl sulfate- Electrophoresis on polyacrylamide gel (Sodium Dodecyl Sulfate (SDS)-polyacrylamide gel) and transfer to a polyvinylidene fluoride (PVDF) membrane through electroblotting. After blocking the membrane for 1 hour at room temperature using 5% skim milk, it was incubated overnight with a primary antibody of MuRF-1, atrozin-1, or p-mTOR protein at 4°C. , After incubation with the secondary antibody for 2 hours the next day, it was developed with a LAS3000 luminescent image analyzer (LAS3000 luminescent image analyzer, Fujifilm, Japan). Antibodies include anti-MuRF-1 antibody (Santa Cruz Biotechnology, USA), anti-atrozine-1 antibody (Santa Cruz Biotechnology, USA), anti-p-mTOR antibody (Cell Signaling Technology, USA), and HRP ( Horseradish peroxidase)-conjugated secondary antibody (Santa Cruz Biotechnology, USA) was used.

As a result, as shown in FIG. 7 , there was no difference in expression of p-mTOR protein in the induction group compared to the normal group, but the expression increased in the HMB-administered group and the deodeok extract-administered group, and MuRF-1 and atrozine-1 The expression of the protein was significantly increased in the induction group compared to the normal group, but the expression was reduced to the level of the normal group in the HMB-administered group and the deodeok extract-administered group. From this, it was found that the deodeok extract relieved the increase in the expression of MuRF-1 and atrozin-1 protein according to muscle atrophy, increased the expression of p-mTOR protein, and had a better protective effect on the muscle atrophy than the positive control, HMB. Could know.

Experimental Example 3. Confirmation of myoblast cytotoxicity of deodeok extract

In order to confirm the cytotoxicity of the deodeok extract in myoblasts, the mouse-derived myoblast (Myoblast) C2C12 cell line was purchased from the American Type Culture Collection (ATCC, USA) and an experiment was performed.

Specifically, the experimental group was a normal group, an induction group (dexamethasone (Sigma Aldrich Co.) treatment), a deodeok extract treatment group of Example 2 (5, 10, 15, 20, 30, 40, 50 μg/mL) a total of 7 By setting the concentration, the cytotoxicity of C2C12 myoblasts was confirmed. Cytotoxicity was measured using Cell counting kit-8 (Dojindo Laboratories, Japan). The results obtained in the experiment were expressed as mean ± standard error, and one-way ANOVA was performed using SPSS to verify statistical significance for each experimental group. did.

As a result, as shown in FIG. 8, it was confirmed that there was no toxicity up to 10 μg/mL. Therefore, in subsequent cell experiments, efficacy and mechanism of action studies were performed at concentrations of 2.5, 5, and 10 μg/ml.

Experimental Example 4. Confirmation of muscle atrophy improvement effect by administration of deodeok extract

In order to confirm the improvement effect of deodeok extract, a muscle atrophy model was made with C2C12, a mouse-derived myoblast, and the improvement effect of muscle atrophy was confirmed.

First, C2C12 cells (ATCC No. CRL-1772??) were cultured in DMEM medium containing 10% FBS and 1% penicillin/streptomycin at 37 ° C., 5% CO ₂ conditions, and 2% horse serum, 1% penicillin/ It was differentiated into myotubes under DMEM medium containing streptomycin composition. The differentiated myofibrils were seeded in a 6-well plate with 10 ⁵ cells per well, and when the cells grew about 80%, the cells were exchanged for a differentiation medium containing 2% horse serum and cultured for 9 days, every 2 days. It was replaced with differentiation medium. On the 9th day of differentiation, 50 μM of dexamethasone was treated for 48 hours to create a muscle atrophy model. The deodeok extract was treated simultaneously with dexamethasone at each concentration for 48 hours to confirm the inhibitory effect of muscle atrophy. The picture was taken at 30 magnification using an optical microscope and shown in FIG. 9, and 8 muscle fibers were randomly selected from the picture and their length was measured using the Image J program. The results are shown in FIG. 10 . The results obtained in the experiment were expressed as mean ± standard error, and one-way ANOVA was performed using SPSS to verify the statistical significance of each experimental group, and the difference between the experimental groups was verified for significance at the level of p < 0.05 with the Bonferroni correction. did. The experimental groups are summarized and shown in Table 2 below.

number experimental group dexamethasone Deodeok extract of Example 2 One normal group - - 2 judo group 50 μM - 3 deodeok
extract treatment group 50 μM 2.5 μg/mL 4 50 μM 5 μg/mL 5 50 μM 10 μg/mL

As a result, as shown in FIG. 9, it can be confirmed that the length of the muscle fiber is increased in a concentration-dependent manner of the deodeok extract. In addition, as shown in FIG. 10 , as a result of quantifying the diameter and length of muscle fibers, the induced group decreased by 58% compared to the normal group, and the deodeok extract was treated with 2.5, 5, and 10 μg/ml, respectively, compared to the induced group. It was confirmed that the length of the muscle fiber decreased by about 34%, 62%, and 79% was increased. From this, it can be seen that the deodeok extract of the present invention is excellent in the effect of improving muscle atrophy.

Experimental Example 5. Confirmation of changes in expression of muscle protein-related factors in myoblast muscle atrophy induction model according to administration of deodeok extract

Deodeok extract relieves the increase in the expression of MuRF-1 and atrozin-1 proteins involved in the breakdown of muscle proteins in myoblast muscle atrophy induction model, and increases the expression of p-mTOR protein involved in muscle protein synthesis. It was confirmed using a Western blot (Western blot).

5-1. Confirmation of alleviation of increased expression of MuRF-1 (Muscle RING-finger protein-1) and atrogin-1 (atrogin-1) genes involved in muscle protein degradation

It was confirmed using qRT-PCR (quantitative real-time PCR) whether the deodeok extract relieved the increase in the expression of MuRF-1 and atrozin-1, which are involved in the degradation of muscle protein. Specifically, mRNA was extracted from the cells of Experimental Example 4 and analyzed, and the specific experimental method was performed in the same manner as in Experimental Example 2-1.

As a result, as shown in FIG. 11 , the atrozin-1 gene was increased about 3 times in the induction group compared to the normal group, and expression was about 3 times in the group treated with deodeok extract at 2.5, 5, and 10 μg/ml, respectively. It was decreased by 33.3, 37.0, and 55.2%, and as shown in FIG. 12 , the MuRF-1 gene increased about 6 times in the induction group compared to the normal group, but the group treated with deodeok extract at 2.5, 5, and 10 μg/ml, respectively. expression was decreased by 40.2, 44.3, and 50.3%. From this, it can be seen that the deodeok extract relieves the increase in the expression of MuRF-1 and atrozin-1 genes according to muscle atrophy, and has an excellent effect on inhibiting muscle degradation.

5-2. Alleviation of the increase in the expression of MuRF-1 and atrozin-1 proteins involved in the degradation of muscle proteins, and confirmation of the increase in the expression of the phosphorylated mammalian target of rapamycin (p-mTOR) protein involved in the synthesis of muscle proteins

Western blot to determine whether deodeok extract relieved the increase in the expression of MuRF-1 and atrozin-1 proteins involved in muscle protein degradation and increased the expression of p-mTOR protein involved in muscle protein synthesis. was confirmed using . Specifically, the protein was extracted from the cells of Experimental Example 4 and analyzed, and the specific experimental method was performed in the same manner as in Experimental Example 2-2. In addition, the ratio of p-mTOR to mTOR was measured because mTOR is phosphorylated to p-mTOR and activated, thereby participating in muscle synthesis.

As a result, as shown in FIGS. 13 and 14, the protein expression level of atrozin-1 was increased by about 2 times in the induction group compared to the normal group, and the group treated with deodeok extract at 2.5, 5, and 10 μg/ml, respectively. was decreased by 35.8, 45.8, and 50.3%, and the protein expression level of MuRF-1 increased by about 2 times in the induction group compared to the normal group, and 28.0, 28.0, and in the group treated with deodeok extract at 2.5, 5, and 10 μg/ml, respectively. 41.7, 57.4% decreased. From this, it can be seen that the deodeok extract of the present invention is excellent in the inhibitory effect on muscle degradation.

In addition, as shown in FIGS. 15 and 16 , as a result of quantifying p-mTOR, the normal group and the induced group showed similar expression levels and were similar, and the group treated with deodeok extract at 2.5, 5, and 10 μg/ml, respectively. It was confirmed that the expression was increased to 50.0, 66.9, and 139.7% compared to the induction group. As a result of quantifying mTOR, the normal group, the induced group, and the deodeok extract treated group showed similar expression levels. In addition, as shown in FIG. 17 , as a result of quantifying the ratio of p-mTOR to mTOR, the normal group and the induced group showed similar values, and in the group treated with deodeok extract at 2.5, 5, and 10 μg/ml, respectively, It was confirmed that 41.1, 67.7, and 126.9% were increased. From this, it can be seen that the deodeok extract of the present invention has an excellent effect of promoting muscle synthesis by increasing p-mTOR (active form) through phosphorylation of mTOR.

<110> JUNG, Woo Jae <120> Pharmaceutical composition for prevention or treatment of muscular disease comprising Codonopsis lanceolata extract as an active ingredient <130> 2019P-09-020 <150> KR 10-2018-0096863 <151> 2018-08-20 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MuRF-1 forward <400> 1 aagactgagc tgagtaactg 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MuRF-1 reverse <400> 2 tagagggtgt caaacttctg 20 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Atrogin-1 forward <400> 3 agaaagaaag acatcagaa ca 22 <210> 4 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Atrogin-1 reverse <400> 4 gctccttcgt acttcctt 18

Claims (10)

Deodeok ( Codonopsis lanceolata ) A pharmaceutical composition for preventing or treating sarcopenia, muscular atrophy, muscular dystrophy and muscle degeneration containing an extract as an active ingredient.
The pharmaceutical composition according to claim 1, wherein the extract is a leaf, stem, root, or fruit extract of deodeok.
The pharmaceutical composition according to claim 1, wherein the extract is _{extracted with water, a C 1} to C ₂ lower alcohol or a mixture thereof.
delete According to claim 1, wherein the pharmaceutical composition is MuRF-1 (Muscle RING-finger protein-1) or atrogin-1 (atrogin-1) characterized in that alleviating the increase in the expression of the gene or protein, the pharmaceutical composition.
The pharmaceutical composition of claim 1, wherein the pharmaceutical composition increases the expression of p-mTOR (phosphorylated mammalian target of rapamycin) protein.
A health functional food composition for preventing or improving sarcopenia, muscular atrophy, muscular dystrophy and muscle degeneration containing deodeok extract as an active ingredient.
delete delete A feed additive or feed composition for preventing or improving sarcopenia, muscular atrophy, muscular dystrophy and muscle degeneration containing deodeok extract as an active ingredient.

Images