Pharmaceutical composition for preventing or treating muscle diseases

Abstract

To provide use of an elder berry extract for preventing or treating muscle diseases.SOLUTION: Provided is a pharmaceutical composition for preventing or treating a muscle disease, comprising, as an active ingredient, a monosaccharide and amino acid-bound compound or a pharmaceutically acceptable salt thereof, the monosaccharide and amino acid-bound compound being a compound represented by the following formula and the like, and the muscle disease being a disease caused by muscle loss.SELECTED DRAWING: Figure 17

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition for preventing or treating a muscle disease. [Background technology]

[0002] Muscle is the most abundant tissue in the human body, and maintaining an adequate muscle mass is necessary to maintain the body's functional capacity and prevent metabolic diseases. Muscle size is regulated by intracellular signaling processes that induce anabolism and catabolism within the muscle. When signaling reactions inducing muscle protein synthesis prevail over muscle protein breakdown, muscle protein synthesis increases, inducing muscle hypertrophy and hyperplasia.

[0003] Muscles promote calcium absorption and increase bone density. However, as the body ages, their composition changes, resulting in a redistribution of body fat and protein. After the age of 50, the rate of protein synthesis in muscle cells slows down compared to the rate of degradation, leading to a rapid decline in muscle mass and the risk of developing muscle-loss disorders.

[0004] Sarcopenia, a disease caused by muscle loss, is a condition in which muscle mass in the body is reduced to approximately 13-24% of the body mass. This indicates a decrease in muscle mass, protein content, fiber diameter, muscle strength production, and fatigue resistance. Sarcopenia can occur for a variety of reasons, including sepsis, cancer, renal failure, glucocorticoid excess, denervation, muscle disuse, and aging. Major causes include a gradual decrease in the quantity and quality of skeletal muscle due to aging and weight loss, including fat and body fat components, due to an inappropriate diet.

[0005] Sarcopenia results from an imbalance between protein synthesis and degradation. When sarcopenia develops, physical activity significantly decreases, reducing quality of life and even leading to injuries in daily life. Furthermore, excessive exercise can lead to muscle fatigue and damage, which can cause pain and temporary loss of mobility.

[0006] In this regard, Korean Patent Publication No. 10-2022-0113912 relates to a composition for preventing, improving, or treating muscle diseases, which contains castor extract as an active ingredient, and indicates that castor extract can be used to prevent, improve, or treat muscle diseases without side effects by increasing the size of muscle cells and improving exercise performance. Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a use of elderberry extract for the prevention, improvement or treatment of muscle diseases. Another object of the present invention is to provide a use of elderberry extract for muscle strengthening.

[0008] It is still another object of the present invention to provide use of a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof, for the prevention, amelioration or treatment of a muscle disease.

[0009] It is still another object of the present invention to provide a use of a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof, for muscle strengthening. [Means for solving the problem]

[0010] In order to achieve the above-mentioned objects, the present invention provides a pharmaceutical composition for preventing or treating muscle diseases, which contains elderberry extract as an active ingredient.

[0011] The present invention also provides a health functional food for preventing or improving muscle diseases, which contains elderberry extract as an active ingredient. The present invention also provides a health functional food for strengthening muscles, which contains elderberry extract as an active ingredient.

[0012] The present invention also provides a method for preventing, ameliorating, or treating a muscle disease, comprising the step of administering an elderberry extract to an individual.

[0013] The present invention also provides use of an elderberry extract for the manufacture of a medicament for the prevention, amelioration or treatment of a muscle disease.

[0014] The present invention also provides a pharmaceutical composition for preventing or treating a muscle disease, which comprises a monosaccharide and amino acid-bound compound, or a pharmaceutically acceptable salt thereof, as an active ingredient.

[0015] The present invention also provides a health functional food for preventing or improving muscle diseases, which contains, as an active ingredient, a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof.

[0016] The present invention also provides a health functional food for strengthening muscles, which contains, as an active ingredient, a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof.

[0017] The present invention also provides a method for preventing, ameliorating, or treating a muscle disease, comprising the step of administering to an individual a monosaccharide and amino acid-bound compound, or a pharmaceutically acceptable salt thereof.

[0018] Furthermore, the present invention provides a use of the monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for the prevention, amelioration or treatment of a muscle disease. [Effects of the Invention]

[0019] The elderberry extract or monosaccharide and amino acid-bound compound according to the present invention has the effects of inhibiting muscle cell death, suppressing the expression of inflammatory cytokines, promoting testosterone secretion, suppressing the expression of muscle atrophy markers and myostatin, and restoring muscle in animal models of sarcopenia, making it useful for the treatment of muscle diseases. [Brief explanation of the drawings]

[0020] [Figure 1] FIG. 1 shows the results of microscopic observation of the morphology of myoblasts before (A) and after (B) differentiation into myotubes in one embodiment of the present invention. [Figure 2] FIG. 1 shows the results of microscopic observation of the morphology of myotubes before (A) and after (B) dexamethasone treatment in one embodiment of the present invention. [Figure 3] 1 is a graph showing the results of confirming the muscle cell death inhibitory effect of an elderberry hot water extract (A), an elderberry 50% ethanol extract (B), or an elderberry 100% ethanol extract (C) in one embodiment of the present invention. [Figure 4] 1 shows graphs showing the results of confirming the muscle cell death inhibitory effect of FL (A), FV (B), or FI (C) in one embodiment of the present invention. [Figure 5] 1 is a graph showing the results of confirming the muscle cell death inhibitory effect of various types of amino acids (leucine, valine, arginine, tyrosine, methionine, or phenylalanine) in one embodiment of the present invention. [Figure 6] 1 is a graph showing the results of confirming the muscle cell death inhibitory effect of various types of amino acids (isoleucine, tryptophan, glycine, lysine, or threonine) in one embodiment of the present invention. [Figure 7] 1 is a graph showing the results of confirming the inhibitory effect of an elderberry hot water extract (A), an elderberry 50% ethanol extract (B), or an elderberry 100% ethanol extract (C) on TNFα expression in one embodiment of the present invention. [Figure 8]1 is a graph showing the results of confirming the inhibitory effect of FL (A), FV (B), or FI (C) on TNFα expression in one embodiment of the present invention. [Figure 9] 1 is a graph showing the results of confirming the testosterone secretion promoting effect of elderberry hot water extract (A) or FL (B) in one embodiment of the present invention. [Figure 10] 1 is a graph showing the results of confirming the inhibitory effect of an elderberry hot water extract (A), an elderberry 50% ethanol extract (B), or an elderberry 100% ethanol extract (C) on the expression of the MuRF-1 gene in one embodiment of the present invention. [Figure 11] 1 is a graph showing the results of confirming the inhibitory effect of an elderberry hot water extract (A), an elderberry 50% ethanol extract (B), or an elderberry 100% ethanol extract (C) on the expression of the atrogin-1 gene in one embodiment of the present invention. [Figure 12] This is a graph showing the results of confirming the inhibitory effect of elderberry hot water extract (A), elderberry 50% ethanol extract (B), or elderberry 100% ethanol extract (C) on myostatin gene expression in one embodiment of the present invention. [Figure 13] 1 shows graphs showing the results of confirming the inhibitory effect of FL (A), FV (B), or FI (C) on the expression of the MuRF-1 gene in one embodiment of the present invention. [Figure 14] 1 is a graph showing the results of confirming the inhibitory effect of FL(A), FV(V), or FI(C) on the expression of the atrogin-1 gene in one embodiment of the present invention. [Figure 15] In one embodiment of the present invention, this is a graph showing the results of confirming the effect of FL (A), FV (B), or FI (C) on suppressing the expression of the myostatin gene. [Figure 16] FIG. 1 is a schematic diagram showing an animal experimental design using an animal model of sarcopenia in one embodiment of the present invention. [Figure 17]1 is a graph showing the results of examining changes in body weight in an animal model of sarcopenia in response to an elderberry hot water extract (A) or FL (B) in one embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0021] The present invention will be described in detail below. The present invention provides a pharmaceutical composition for preventing or treating muscle diseases, which contains elderberry extract as an active ingredient.

[0022] In this specification, "elderberry" refers to a black-purple berry fruit that usually ripens in the fall and is therefore sometimes called black elder. In North America, elderberry extract is also called sambucol. Elderberries are known to be rich in vitamins A, B, and C, as well as anthocyanins, and have been reported to be effective in preventing colds and strengthening the immune system.

[0023] The elderberry extract can be produced by a production method including the following steps: 1) adding an extracting solvent to elderberry to prepare an extract; 2) filtering the extract of step 1); and 3) A step of concentrating the filtrate obtained in step 2) under reduced pressure and then drying it.

[0024] The extraction solvent can be water, alcohol, or a mixture thereof. The alcohol is a C1-C2 lower alcohol, specifically, ethanol, methanol, or spirits. The extraction solvent may be added in an amount of 1 to 100 times, 1 to 70 times, 1 to 50 times, 1 to 30 times, or 1 to 15 times the weight of elderberry. When alcohol is used as the extraction solvent, the alcohol may be 10 to 100%, 20 to 100%, 30 to 100%, or 40 to 100% alcohol.

[0025] The extraction method may be shaking extraction, cold maceration extraction, reflux extraction, or ultrasonic extraction. The extraction time may be 1 to 20 hours, 2 to 20 hours, 1 to 10 hours, 2 to 10 hours, 1 to 5 hours, or 2 to 5 hours. The extraction may be repeated one or more times.

[0026] In the above step 3), a vacuum vacuum concentrator or a vacuum rotary evaporator can be used for the vacuum concentration. The drying can be vacuum drying, vacuum drying, boiling drying, spray drying, or freeze drying, and specifically, freeze drying.

[0027] The muscle disease may be a disease caused by muscle loss. In another aspect, the muscle disease may be a progressive disease including loss of walking ability due to gradual loss of muscle strength, weakening of respiratory muscles, weakening of cardiac function, etc. The muscle disease may be a congenital disease or an acquired disease. For example, the muscle loss disease may be myatony, muscular atrophy, muscular dystrophy, muscle degeneration, myotonia, muscular dystrophy, amyotrophic lateral sclerosis, myasthenia, cachexia, or sarcopenia.

[0028] The pharmaceutical composition of the present invention may contain 10 to 95 wt% of the active ingredient, elderberry extract, based on the total weight of the composition. In addition, the pharmaceutical composition of the present invention may further contain one or more active ingredients exhibiting the same or similar functions in addition to the active ingredient.

[0029] The pharmaceutical compositions of the present invention may contain carriers, diluents, excipients, or mixtures thereof commonly used in biological preparations. Pharmaceutically acceptable carriers can be used without limitation as long as they are suitable for delivering the composition into the body. Specific examples of such carriers include compounds listed in the Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, infusion solutions, dextrose solutions, maltodextrin solutions, glycerol, ethanol, or mixtures thereof. Conventional additives such as antioxidants, buffers, and bacteriostatic agents may also be added as needed.

[0030] When the above composition is formulated, commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. may be added.

[0031] The compositions of the present invention can be formulated into oral or parenteral formulations. Oral formulations include solid and liquid formulations. Examples of solid formulations include tablets, pills, powders, granules, capsules, and lozenges. These solid formulations can be prepared by adding at least one or more excipients to the composition. The excipients can be starch, calcium carbonate, sucrose, lactose, gelatin, or mixtures thereof. The solid formulations can also contain lubricants, such as magnesium stearate and talc. The liquid formulations can also include suspensions, oral solutions, emulsions, or syrups. The liquid formulations can also contain excipients such as wetting agents, sweeteners, flavoring agents, and preservatives.

[0032] The parenteral preparations include injections, suppositories, powders for respiratory inhalation, aerosols for sprays, powders, and creams. The injections may be sterilized aqueous solutions, non-aqueous solvents, suspension injections, emulsions, etc. In addition, non-aqueous solvents or suspension injections may be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, or injectable esters such as ethyl oleate.

[0033] The present invention also provides a health functional food for preventing or improving muscle diseases, which contains elderberry extract as an active ingredient. The elderberry extract contained in the health functional food according to the present invention can have the characteristics described above. The muscle disease can also have the characteristics described above.

[0034] The elderberry extract of the present invention may be added to food directly or together with other foods or food ingredients. In this case, the content of the active ingredient to be added is determined depending on the purpose, but it may generally be 0.01 to 90 parts by weight based on the total weight of the functional health food.

[0035] The form and type of the health functional food are not particularly limited. Specifically, the health functional food may be in the form of tablets, capsules, powders, granules, liquids, or pills. The health functional food may contain various flavorings, sweeteners, or natural carbohydrates as additional ingredients. The sweeteners may be natural or synthetic sweeteners. Examples of natural sweeteners include thaumatin and stevia extract. Examples of synthetic sweeteners include saccharin and aspartame. Examples of natural carbohydrates include monosaccharides, disaccharides, polysaccharides, oligosaccharides, and sugar alcohols.

[0036] In addition to the above-mentioned additional ingredients, the health functional food of the present invention may further contain nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, etc. These ingredients may be used alone or in combination. The proportion of the above additives may be in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

[0037] The present invention also provides a health functional food for strengthening muscles, which contains elderberry extract as an active ingredient. The elderberry extract contained in the health functional food according to the present invention can have the characteristics described above. The health functional food can also have the characteristics described above.

[0038] The present invention also provides a method for preventing, ameliorating, or treating a muscle disease, comprising the step of administering an elderberry extract to an individual. The elderberry extract used in the method according to the invention may have the characteristics as described above. The muscle disorder may also have the characteristics as described above.

[0039] The individual may be a mammal, and in particular a human. The administration method can be oral or parenteral, depending on the desired method. Parenteral administration includes intraperitoneal, rectal, subcutaneous, intravenous, intramuscular, or thoracic injection modes.

[0040] The above-mentioned administration can be performed in a pharmaceutically effective amount. This varies depending on the type and severity of the disease, the activity of the drug, the patient's sensitivity to the drug, the administration time, the administration route, the treatment period, and other factors. However, to achieve the desired effect, the amount of the active ingredient contained in the above-mentioned administration can be 0.0001 to 1,000 mg / kg, specifically 0.001 to 500 mg / kg. The above-mentioned administration can be performed once or several times a day.

[0041] The above administrations may be administered alone or in combination with other therapeutic agents. When administered in combination, the administration may be sequential or simultaneous. The present invention also provides use of an elderberry extract for the manufacture of a medicament for the prevention, amelioration or treatment of a muscle disease.

[0042] The elderberry extract used in the manufacture of the medicament according to the present invention may have the characteristics as described above. The muscle disease may also have the characteristics as described above. The present invention also provides a pharmaceutical composition for preventing or treating a muscle disease, which comprises a monosaccharide and amino acid-bound compound, or a pharmaceutically acceptable salt thereof, as an active ingredient.

[0043] As used herein, "monosaccharide" refers to the smallest unit of carbohydrates, also known as simple sugar. The monosaccharide is generally a colorless, water-soluble crystalline solid, and examples thereof include glucose, fructose, and galactose. In one embodiment of the present invention, the monosaccharide may be fructose. The "fructose" is a monosaccharide containing six carbon atoms and a ketose having a ketone group, and is represented by the formula C6H 12 It is represented by the chemical formula O6. Fructose is an intermediate in the process of glucose breakdown and glycogen synthesis in the sugar metabolism of living organisms.

[0044] Furthermore, in this specification, the term "amino acid" refers to a compound that is a basic building block of proteins that make up living organisms and contains both a chemically basic amino group (-NH2) and an acidic carboxyl group (-COOH). Generally, proteins are decomposed into various amino acids when hydrolyzed with acid or enzymes such as pepsin or trypsin. The amino acids thus produced are absorbed into the body and rearranged to synthesize proteins that make up the body. The amino acids are classified into non-essential amino acids and essential amino acids.

[0045] The amino acids include any amino acids known in the art, and specifically, the amino acids may be branched-chain amino acids (BCAAs). Branched-chain amino acids (BCAAs) refer to amino acids having a branched chain with a central carbon atom bonded to three or more carbon atoms and an aliphatic side chain. Specific examples of BCAAs include leucine, valine, and isoleucine.

[0046] The monosaccharide and amino acid-bound compound according to the present invention is a compound obtained by binding a monosaccharide and an amino acid having the above-mentioned characteristics by a method well known in the art. As an example, the monosaccharide and amino acid-bound compound may be a compound in the form of fructose bound to BCAA. Specifically, the monosaccharide and amino acid-bound compound is a compound in the form of fructose bound to leucine, valine, or isoleucine, and more specifically, the monosaccharide and amino acid-bound compound may be a compound represented by the following [Chemical Formula 1] to [Chemical Formula 3].

[0047] [C1] JPEG2025134854000002.jpg51166

[0048] [Case 2] JPEG2025134854000003.jpg48166

[0049] [C3] JPEG2025134854000004.jpg51166

[0050] Furthermore, the present invention can include not only the monosaccharide and amino acid-linked compounds described above, but also pharmaceutically acceptable salts thereof.

[0051] Here, "pharmaceutically acceptable salt" means a salt that is suitable for use in contact with the tissues of humans and lower animals without inducing excessive toxicity, irritation, allergic reactions, etc., within the scope of pure medical judgment. The above-mentioned pharmaceutically acceptable salts are well known in the art and are described in detail, for example, in the literature (SM Berge et al., J. Pharmaceutical Sciences, 66, 1, 1977). The salts can be prepared in the same reaction system during the final isolation and purification of the compound of the present invention, or can be prepared separately by reacting with an inorganic or organic base. Specific examples of base addition salts include alkali salts and alkaline earth metal salts such as ammonium, lithium, sodium, potassium, magnesium, and calcium salts; salts with organic bases; salts with primary, secondary, and tertiary aliphatic and aromatic amines, such as methylamine, ethylamine, propylamine, isopropylamine, the four butylamine isomers, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline, isoquinoline, benzathine, N-methyl-D-glucamine, 2-amino-2-(hydroxymethyl)-1,3-propanediol, and hydrabamine; and salts with amino acids such as arginine and lysine.

[0052] The present invention also includes hydrates or solvates of the monosaccharide and amino acid-bound compounds, and derivatives thereof. The solvent for the solvates is not particularly limited, and any solvent known in the art can be used without limitation.

[0053] The muscle disease may be a disease caused by muscle loss. In another aspect, the muscle disease may be a progressive disease including loss of walking ability due to gradual loss of muscle strength, weakening of respiratory muscles, weakening of cardiac function, etc. The muscle disease may be a congenital disease or an acquired disease. For example, the muscle loss disease may be myasthenia, muscle atrophy, muscle dystrophy, muscle degeneration, muscle rigidity, muscular dystrophy, amyotrophic lateral sclerosis, myasthenia, cachexia, or sarcopenia.

[0054] The pharmaceutical composition of the present invention may contain 10 to 95% by weight of the active ingredient, a monosaccharide and amino acid-linked compound, or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In addition to the active ingredient, the pharmaceutical composition of the present invention may further contain one or more active ingredients having the same or similar function.

[0055] The pharmaceutical compositions of the present invention may contain carriers, diluents, excipients, or mixtures thereof commonly used in biological preparations. Pharmaceutically acceptable carriers can be used without limitation as long as they are suitable for delivering the composition into the body. Specific examples of such carriers include compounds listed in the Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, infusion solutions, dextrose solutions, maltodextrin solutions, glycerol, ethanol, or mixtures thereof. Conventional additives such as antioxidants, buffers, and bacteriostatic agents may also be added as needed.

[0056] When the above composition is formulated, commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. may be added.

[0057] The composition according to the present invention can be formulated into oral or parenteral formulations. Oral formulations include solid and liquid formulations. Examples of solid formulations include tablets, pills, powders, granules, capsules, and lozenges. These solid formulations can be prepared by adding at least one excipient to the composition. The excipient can be starch, calcium carbonate, sucrose, lactose, gelatin, or a mixture thereof. The solid formulation can also contain a lubricant, such as magnesium stearate or talc. The liquid formulation can also include a suspension, oral solution, emulsion, or syrup. The liquid formulation can also contain excipients such as wetting agents, sweeteners, flavoring agents, and preservatives.

[0058] The parenteral preparations include injections, suppositories, powders for respiratory inhalation, aerosols for sprays, powders, and creams. The injections may be sterilized aqueous solutions, non-aqueous solvents, suspension injections, emulsions, etc. In addition, non-aqueous solvents or suspension injections may be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, or injectable esters such as ethyl oleate.

[0059] The present invention also provides a health functional food for preventing or improving muscle diseases, which contains, as an active ingredient, a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof.

[0060] The monosaccharide and amino acid bond compound or a pharmaceutically acceptable salt thereof contained in the health functional food according to the present invention may have the characteristics described above. The muscle disease may also have the characteristics described above.

[0061] The monosaccharide and amino acid bond compound or its pharmaceutically acceptable salt according to the present invention may be added to a food product as is or may be used together with other foods or food ingredients. In this case, the content of the active ingredient to be added may be determined depending on the purpose, but may generally be 0.01 to 90 parts by weight based on the total weight of the functional health food.

[0062] The form and type of the health functional food are not particularly limited. Specifically, the health functional food may be in the form of tablets, capsules, powders, granules, liquids, or pills. The health functional food may contain various flavorings, sweeteners, or natural carbohydrates as additional ingredients. The sweeteners may be natural or synthetic sweeteners. Examples of natural sweeteners include thaumatin and stevia extract. Examples of synthetic sweeteners include saccharin and aspartame. Examples of natural carbohydrates include monosaccharides, disaccharides, polysaccharides, oligosaccharides, and sugar alcohols.

[0063] In addition to the above-mentioned additional ingredients, the health functional food of the present invention may further contain nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, etc. These ingredients may be used alone or in combination. The proportion of the above additives may be in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

[0064] The present invention also provides a health functional food for strengthening muscles, which contains, as an active ingredient, a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof.

[0065] The monosaccharide and amino acid bond compound or a pharmaceutically acceptable salt thereof contained in the health functional food according to the present invention may have the characteristics as described above. The health functional food may also have the characteristics as described above.

[0066] The present invention also provides a method for preventing, ameliorating, or treating a muscle disease, comprising the step of administering to an individual a monosaccharide and amino acid-bound compound, or a pharmaceutically acceptable salt thereof.

[0067] The monosaccharide and amino acid conjugate compound or a pharmaceutically acceptable salt thereof used in the method according to the present invention can have the characteristics as described above. The muscle disease can also have the characteristics as described above.

[0068] The individual may be a mammal, and in particular a human. The administration method can be oral or parenteral, depending on the desired method. Parenteral administration can be by intraperitoneal, rectal, subcutaneous, intravenous, intramuscular, or thoracic injection.

[0069] The above-mentioned administration can be performed in a pharmaceutically effective amount. This varies depending on the type and severity of the disease, the activity of the drug, the patient's sensitivity to the drug, the administration time, the administration route, the treatment period, and other factors. However, to achieve the desired effect, the amount of the active ingredient contained in the above-mentioned administration can be 0.0001 to 1,000 mg / kg, specifically 0.001 to 500 mg / kg. The above-mentioned administration can be performed once or several times a day.

[0070] The above administrations may be administered alone or in combination with other therapeutic agents. When administered in combination, the administration may be sequential or simultaneous. Furthermore, the present invention provides a use of the monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for the prevention, amelioration or treatment of a muscle disease.

[0071] The elderberry extract used in the manufacture of the medicament according to the present invention may have the characteristics as described above. The muscle disease may also have the characteristics as described above. [Example]

[0072] The present invention will be described in detail below based on the following examples. However, the following examples are merely illustrative of the present invention and the present invention is not limited thereto. Anything that has substantially the same configuration as the technical idea described in the claims of the present invention and exhibits similar effects is included in the technical scope of the present invention.

[0073] Example 1. Preparation of Elderberry Hot Water Extract A hot water extract was prepared from dried elderberries in the following manner. First, dried elderberries were mixed with 10 times their weight in purified water and extracted at 80°C for 3 hours to obtain an extract. The resulting extract was filtered using a metal detector and concentrated to obtain an elderberry concentrate. The elderberry concentrate was mixed with 30% maltodextrin and freeze-dried to produce a powder of elderberry hot water extract.

[0074] Example 2. Preparation of 50% ethanol extract of elderberry A powder of 50% ethanol extract of elderberry was produced in the same manner as in Example 1 above, except that 50% ethanol was added instead of purified water and extraction was carried out for 6 hours.

[0075] Example 3. Preparation of 100% ethanol extract of elderberry A powder of 100% ethanol extract of elderberry was produced in the same manner as in Example 1 above, except that 100% ethanol was added instead of purified water and extraction was carried out for 6 hours.

[0076] Example 4. Analysis of elderberry extract components Liquid chromatography / mass spectrometry (LC / MS) was performed on the elderberry hot water extract prepared above to confirm the presence of FL (fructose-leucine) in the extract. Specifically, 200 mg of elderberry hot water extract powder was dissolved in water, followed by the addition of methanol and ultrasonic treatment for at least 60 minutes to prepare the sample. A calibration curve was prepared using 0.2 mg / L FL to determine the quantification range. Analysis was performed using the sample prepared under the above conditions, and this was repeated three times. The FL content in the elderberry hot water extract was calculated using the peak area and test solution RT in the usual manner, and the results are shown in Table 1.

[0077] [Table 1]

[0078] As shown in Table 1, the FL content in the samples was found to be 1.02% on average.

[0079] Experimental Example 1. Inhibition of muscle cell death The following method was used to confirm whether the elderberry extract and the compound containing monosaccharides and amino acids produced above inhibit the death of muscle cells.

[0080] 1-1. Preparation of myotube cells First, myoblast cell C2Cl2 cell line (ATCC, USA) was cultured in DMEM medium containing 10% FBS (fetal bovine serum) and 1% antibiotic-antimycotic at 37°C and 5% CO2. The culture medium of the cultured cells was replaced with culture medium containing 2% HS (horse serum) and further cultured for 72 hours. The cultured cells were observed under a microscope to confirm differentiation of the myoblasts into myotubes, and then treated with 10 μM dexamethasone for 24 hours. After the reaction, the cells were again observed under a microscope. Figure 1 shows the results of confirming differentiation into myotubes, and Figure 2 shows the results of microscopic observation after dexamethasone treatment.

[0081] As shown in Figure 1, myoblasts were differentiated into myotubes. Furthermore, as shown in Figure 2, it was confirmed that dexamethasone treatment caused muscle loss by reducing the thickness of myotubes.

[0082] 1-2. Elderberry extract inhibits muscle cell death The cell lines treated with dexamethasone as described above were then treated with 12.5, 25, 50, or 100 μg / ml of elderberry hot water extract, or 100, 200, 300, or 400 μg / ml of elderberry 50% ethanol extract, or 100, 200, 300, or 400 μg / ml of elderberry 100% ethanol extract. After 24 hours, the cell culture medium was removed, 5 mg / ml of MTT reagent was added per well, and the cells were further cultured for 4 hours. DMSO was then added to dissolve the precipitate, and the absorbance at 570 nm was measured using a microplate reader. Cell viability was calculated from the above measurements using the standard method, and the results are shown in Figure 3.

[0083] As shown in Figure 3, the cell viability decreased by dexamethasone was significantly restored by treatment with elderberry extract.

[0084] 1-3. Monosaccharide and amino acid-binding compounds inhibit muscle cell death In addition to FL, which was confirmed to be present in elderberry extract, we confirmed that FV (fructose-valine) and FL (fructose-isoleucine), compounds in which the monosaccharide fructose is linked to the BCAAs valine or isoleucine, inhibited muscle cell death. Experiments were conducted under the same conditions as in Experiments 1-2 above, except that FL, FV, or FI were used at concentrations of 2.5, 5, 10, or 20 μg / ml instead of elderberry extract. Control groups included leucine, valine, isoleucine, tryptophan, arginine, tyrosine, glycine, lysine, methionine, phenylalanine, and threonine. Cell viability calculations for cells treated with FL, FV, or FL are shown in Figure 4, and cell viability calculations for cells treated with amino acids are shown in Figures 5 and 6.

[0085] As shown in Figure 4, the cell viability decreased by dexamethasone was significantly restored by FL, FV, or FI. In contrast, as shown in Figures 5 and 6, amino acid treatment did not restore cell viability.

[0086] Experimental Example 2: Inhibition of inflammatory cytokine expression The following method was used to confirm whether the elderberry extract and the compound containing monosaccharides and BCAAs prepared above inhibit the expression of inflammatory cytokines.

[0087] 2-1. Elderberry extract inhibits TNFα expression The cells treated with elderberry extract were trypsinized in the same manner as in Experimental Example 1-2 above, and the cells alone were collected. The collected cells were centrifuged at 3,000 rpm for 5 minutes, and the supernatant was removed. TM (GeneAll, Korea) and Hybrid-R TM Total RNA was extracted using the following methods: (GeneAll, Korea). The concentration of the extracted RNA was measured using the nanodrop method, and cDNA was synthesized using the RNA as a template. cDNA was synthesized using the RNA, oligo-(dT) primer, and 2x HyperScript TM The cDNA was synthesized using a mixture containing RT Master Mix (GeneAll, Korea). The synthesized cDNA was then used as a template for PCR amplification using Power SYBR TM Real-time PCR (qPCR) was performed under standard conditions using Green PCR Master Mix to confirm TNFα expression. The graph showing the results of confirming TNFα mRNA expression levels is shown in Figure 7.

[0088] As shown in FIG. 7, the expression level of TNFα, which was increased by dexamethasone, was significantly suppressed by treatment with elderberry extract.

[0089] 2-2. Monosaccharide and amino acid-binding compounds inhibit TNFα expression Cells treated with FL, FV, or FI in the same manner as in Experimental Example 1-3 above were treated with trypsin to recover the cells alone, and the TNFα expression level was confirmed under the same conditions and method as in Experimental Example 2-1. The graph of the results is shown in Figure 8.

[0090] As shown in FIG. 8, the expression level of TNFα, which was increased by dexamethasone, was significantly suppressed by treatment with FL, FV, or FI.

[0091] Therefore, the above results indicate that the expression of inflammatory cytokines, which directly induce muscle wasting, is significantly suppressed by elderberry extract, as well as monosaccharide and amino acid-linked compounds, and that these components suppress the death of muscle cells induced by inflammatory cytokines.

[0092] Experimental example 3. Promoting male hormone secretion Whether the elderberry extract and the compound containing monosaccharides and BCAA prepared above inhibit muscle loss caused by the male hormone testosterone was confirmed by ELISA analysis as follows.

[0093] First, mouse Leydig cells, the TM3 cell line (ATCC, USA), were cultured using standard methods. The cultured cells were treated with 500 μM hydrogen peroxide (HO) for 6 hours, and then 12.5, 25, 50, or 100 μg / ml of elderberry hot water extract or 1.25, 2.5, 5, or 10 μg / ml of FL was added. Next, standard ELISA was performed using the cell culture medium to confirm testosterone levels, and the results are shown in Figure 9.

[0094] As shown in FIG. 9, the testosterone concentration suppressed by hydrogen peroxide was significantly restored by the elderberry hot water extract or FL.

[0095] Experimental Example 4: Inhibition of muscle protein expression We used qPCR to confirm whether the elderberry extract and the monosaccharide and BCAA-conjugated compounds inhibited the expression of muscle proteins, specifically, muscle atrophy markers MuRF-1 (muscle RING-finger protein-1) and atrogin-1, and myostatin, a negative regulator of skeletal muscle mass.

[0096] 4-1. Elderberry extract inhibits muscle protein expression As described in Experimental Example 1-2, qPCR was performed using cells treated with elderberry hot water extract, 50% ethanol extract, or 100% ethanol extract. The results of confirming the expression levels of MuRF-1, atrogin-1, and myostatin mRNA are shown in Figures 10 to 12.

[0097] As shown in Figures 10 to 12, the mRNA expression levels of MuRF-1, atrogin-1, and myostatin, which were increased by dexamethasone, were significantly reduced by the elderberry extract.

[0098] 4-2. Monosaccharide and amino acid-binding compounds inhibit muscle protein expression As described in Experimental Examples 1-3, qPCR was performed as described above using cells treated with FL, FV, or FL. The results of confirming the expression levels of MuRF-1, atrogin-1, and myostatin mRNA are shown in graphs in Figures 10 to 12.

[0099] As shown in Figures 10 to 12, the expression levels of MuRF-1, atrogin-1, and myostatin mRNA, which were increased by dexamethasone, were significantly reduced by FL, FV, or FI.

[0100] Therefore, the above results demonstrate that elderberry extract or monosaccharide and amino acid-bound compounds can have a therapeutic effect on sarcopenia by suppressing the expression of muscle atrophy markers and myostatin.

[0101] Experimental Example 5: Confirmation of effectiveness in an animal model of sarcopenia The therapeutic effects of the elderberry extract and the compound to which monosaccharides and BCAAs are bound were confirmed using an animal model of sarcopenia.

[0102] Specifically, 7-week-old male C57BL / 6 mice (Samtako Bio Korea, Korea) were acclimated and then measured for body weight (g), lean body weight (g), fat mass (g), and body fat percentage (%) using dual-energy X-ray absorptiometry (DXA). Based on these measurements, the mice were divided into groups of 7, and 20 mg / kg of dexamethasone was administered intraperitoneally once daily for 14 days to induce sarcopenia (Figure 13). Additionally, while receiving dexamethasone, 300 mg / kg of elderberry hot water extract or 0.5 mg / kg of FL was orally administered once daily. Mice on day 0 of administration (normal mice) served as a control group. On days 7 and 14 after administration, the mice were photographed by DXA to measure their body weight, lean body mass, fat mass, and body fat percentage. The results are shown in Figure 14 and Tables 2 and 3. On day 14 after administration, the mice were sacrificed in the usual way, and the thymus, spleen, liver, and muscles were removed and weighed. The results are shown in Table 4.

[0103] [Table 2]

[0104] [Table 3]

[0105] [Table 4]

[0106] As shown in Figure 14 and Tables 2 and 3, dexamethasone significantly reduced the body weight, lean body mass, and tissue area of ​​the mice, but these were restored by administration of the elderberry hot water extract or FL. The weight of each tissue was also reduced by dexamethasone, but was restored to approximately the same level as the normal control group by administration of the elderberry hot water extract or FL. Therefore, the above results demonstrate that elderberry extract or a compound combining monosaccharides and amino acids is effective in treating sarcopenia.

Claims

1. A pharmaceutical composition for preventing or treating a muscular disease, comprising a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof, as an active ingredient, The monosaccharide and amino acid bonded compound is a compound represented by the following [Chemical Formula 1] to [Chemical Formula 3], The pharmaceutical composition for preventing or treating a muscle disease, wherein the muscle disease is a disease caused by muscle loss. [Chemical formula 1] [Chemical 2] [Chemical 3]

2. 2. The pharmaceutical composition for preventing or treating a muscle disease according to claim 1, wherein the disease caused by muscle loss is myasthenia, muscle atrophy, muscle dystrophy, muscle degeneration, muscle rigidity, muscular dystrophy, amyotrophic lateral sclerosis, myasthenia, cachexia, or sarcopenia.

3. A health functional food for preventing or improving muscle diseases, comprising a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof, as an active ingredient, The monosaccharide and amino acid bonded compound is a compound represented by the following [Chemical Formula 1] to [Chemical Formula 3], The muscle disease is a disease caused by muscle loss, and the health functional food is for preventing or improving the muscle disease. [Chemical formula 1] [Chemical 2] [Chemical 3]

4. A health functional food for strengthening muscles, comprising a monosaccharide and an amino acid bond compound, or a pharmaceutically acceptable salt thereof, as an active ingredient, The monosaccharide and amino acid-bound compound is a compound represented by the following [Chemical Formula 1] to [Chemical Formula 3], which is a health functional food for strengthening muscles. [Chemical formula 1] [Chemical 2] [Chemical 3]

5. Use of a monosaccharide and amino acid bond compound, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for the prevention, amelioration, or treatment of a muscular disease, comprising: The monosaccharide and amino acid bonded compound is a compound represented by the following [Chemical Formula 1] to [Chemical Formula 3], The muscle disease is a disease caused by muscle loss. [Chemical formula 1] [Chemical 2] [Chemical 3]

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