Composition for preventing or treating muscular disease, comprising oxicam-based compound

Abstract

To provide a composition for the purpose of preventing, alleviating, or treating a muscular disease.SOLUTION: The present invention relates to a composition for the purpose of preventing, alleviating, or treating a muscular disease, comprising an oxicam-based compound, wherein the composition increases muscle mass and strengthens muscles through the effects of promoting differentiation of myoblasts and increasing muscle fibers, and can thereby have treatment effects for various muscular diseases and have muscle strengthening or motor performance increasing effects, wherein the oxicam-based compound is lornoxicam or meloxicam.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a composition containing an oxicam compound for preventing, improving or treating a muscle disease. . [Background technology]

[0002] Muscles play an important role in bodily functions such as energy metabolism and athletic ability, and they are important components of aging. muscle atrophy due to nutritional imbalance or lack of exercise; muscle atrophy due to other diseases such as other cancers They can be damaged or weakened by a variety of factors, including disease and aging.

[0003] Sarcopenia, a major muscle-damaging disorder, is a condition that occurs with aging. Muscle strength decreases as skeletal muscle mass decreases The most notable feature of sarcopenia is a decrease in muscle mass and a change in the type of muscle fiber. As we age, the number of type 1 and type 2 muscle fibers decreases at similar rates. In contrast, the thickness of type 1 muscle fibers is more significantly reduced in sarcopenic patients. Such sarcopenia has been reported to cause muscle weakness and functional impairment in the elderly. (Roubenoff R., Can. J. Appl. Physiol. 26, 78-89,2001).

[0004] Muscle atrophy is caused by malnutrition and prolonged muscle use. It is induced when there is no normal balance between protein synthesis and degradation, resulting in muscle It occurs due to the breakdown of proteins.

[0005] Various treatment methods are currently being developed to fundamentally treat these muscle diseases, especially stem cell Mechanisms that promote muscle cell differentiation from cells to strengthen muscles or promote muscle regeneration These methods are not a fundamental cure for muscle diseases. Treatment is possible, so research into various therapeutic substances that make this possible is necessary. be.

[0006] The present inventors have selected compounds from various compounds that have the effect of improving muscle diseases through the above-mentioned effects. As a result of this research, the present invention has been completed. Summary of the Invention [Problem to be solved by the invention]

[0007] The present inventors have found that oxicam compounds promote myoblast differentiation and improve muscle energy metabolism. This has been found to be particularly effective in treating muscle diseases such as muscular atrophy. The invention was completed.

[0008] Therefore, an object of the present invention is to provide a method for treating a rheumatoid arthritis comprising administering to a patient a therapeutically effective amount of an oxicam compound or a pharmaceutically acceptable salt thereof. The present invention provides a pharmaceutical composition for preventing or treating muscular diseases.

[0009] Another object of the present invention is to provide a method for treating muscle disease comprising administering an oxicam compound or a nutrient-acceptable salt thereof to a patient. The present invention provides a functional health food composition for preventing or improving diseases.

[0010] Yet another object of the present invention is to provide a muscle strengthening composition containing an oxicam compound. is.

[0011] Yet another object of the present invention is to provide a composition for enhancing athletic performance, which comprises an oxicam compound. To do this.

[0012] A further object of the present invention is to provide a composition for promoting muscle stem cell differentiation, which comprises an oxicam compound. The purpose is to provide.

[0013] Yet another object of the present invention is to provide a composition for muscle regeneration containing an oxicam compound. is.

[0014] It is yet another object of the present invention to provide a composition for increasing muscle mass, which comprises an oxicam compound. And so.

[0015] Yet another object of the present invention is to provide a feed additive composition containing an oxicam compound. is. [Means for solving the problem]

[0016] To achieve the above object, the present invention provides an oxicam compound or a pharmaceutically acceptable salt thereof. The present invention provides a pharmaceutical composition for preventing or treating a muscle disease, which comprises a compound capable of inhibiting or treating a muscle disease.

[0017] In order to achieve another object of the present invention, the present invention provides an oxicam compound or its food product. The present invention provides a functional health food composition for preventing or improving muscle diseases, which contains a salt acceptable to the subject.

[0018] In order to achieve still another object of the present invention, the present invention provides a muscle strengthening agent containing an oxicam compound. A composition for use in treating a skin condition is provided.

[0019] In order to achieve yet another object of the present invention, the present invention provides a method for the treatment of athletic performance comprising administering an oxicam compound. A composition for enhancing performance is provided.

[0020] In order to achieve yet another object of the present invention, the present invention provides a muscle stem cell containing an oxicam compound. A composition for promoting cell differentiation is provided.

[0021] In order to achieve yet another object of the present invention, the present invention provides a method for producing a muscle mass enhancing drug comprising an oxicam compound. An augmentation composition is provided.

[0022] In order to achieve still another object of the present invention, the present invention provides a feed additive containing an oxicam compound. An additive composition is provided. [Effects of the Invention]

[0023] The present invention relates to a composition containing an oxicam compound for the prevention, amelioration or treatment of a muscle disease. The composition increases muscle mass through the effects of promoting myoblast differentiation and muscle fiber regeneration. It strengthens muscles and has therapeutic effects on various muscle diseases, so it is Alternatively, it may be used as a functional health food composition. [Brief explanation of the drawings]

[0024] [Figure 1] FIG. 1 shows the effect of oxicam compounds on myoblast differentiation, comparing the relative expression levels of Myo and eMHC when myoblasts were treated with each compound. [Figure 2] FIG. 2 shows the results showing the degree of differentiation of myoblasts (C2C12) depending on the administration concentration of meloxicam. [Figure 3] FIG. 3 shows the results of confirming the expression level of MHC, a myoblast differentiation marker, depending on the administration concentration of meloxicam. [Figures 4a-4b. 5a-5b] Figures 4a and 4b show the results of confirming the expression level of MHC, a myoblast differentiation marker, following administration of meloxicam in human muscle stem cells from 17 years old, and Figures 5a and 5b show the results of confirming the expression level of MHC, a myoblast differentiation marker, in human muscle stem cells from 66 years old. [Figure 6a-6b] FIG. 6a shows the change in body weight after administration of meloxicam, and FIG. 6b shows the muscle mass increasing effect confirmed through the change in weight of hind leg muscles (TA, EDL, SOL, GAS) after administration of meloxicam. [Figure 7a-7b]FIG. 7a shows the change in body weight due to meloxicam administration, and FIG. 7b shows the change in weight of organs other than skeletal muscle. [Figure 8a-8b] Figures 8a and 8b show the muscle regeneration and muscle fiber size confirmed by H&E staining when meloxicam was administered to a mouse model with CTX-induced muscle damage on days 4, 7, and 21, respectively. [Figures 9a-9c] Figure 9a shows the relative expression levels of muscle fiber types when meloxicam was administered to a mouse model with muscle damage induced by CTX. Figure 9b shows the size of the muscle fiber cross-sectional area, and Figure 9c shows the area of ​​the expression type (MHC II type). [Figure 10] Figure 10 shows the cross-sectional area (Fig. 10a) and relative proportion (Fig. 10b) of muscle fibers in which GPDH, a glycolytic enzyme, was activated when meloxicam was administered to a mouse model of muscle damage induced by CTX. [Figure 11] FIG. 11 shows the results of grip strength test and motor performance test when meloxicam was administered to a mouse model in which muscle damage was induced by CTX. [Figure 12] FIG. 12 shows the results of comparing changes in body weight and blood glucose (FIG. 12a) and changes in hind leg muscle weight (FIG. 12b) when meloxicam was administered to 28-month-old aged mice. [Figure 13] FIG. 13 shows the results of comparing the relative expression levels of muscle fiber types when 28-month-old aged mice were administered meloxicam. [Figure 14] FIG. 14 shows the results of comparing the changes in motor performance in 28-month-old aged mice administered with meloxicam through the rotarod test. [Figure 15] Figure 15 shows the results of comparing changes in body weight and grip strength (Figure 15a) and changes in motor performance (Figure 15b) when meloxicam was administered to 14-month-old middle-aged mice. [Figure 16]Figure 16 compares the changes in body weight (Figure 16a) and intake (Figure 16b) when meloxicam was administered to 4-month-old young mice, and Figure 17 compares the changes in grip strength and motor performance (Figure 17a) and muscle mass relative to body weight (Figure 17b). DETAILED DESCRIPTION OF THE INVENTION

[0025] The present invention relates to a method for preventing or treating muscle diseases comprising an oxicam compound or a pharmaceutically acceptable salt thereof. relates to therapeutic pharmaceutical compositions.

[0026] The present invention relates to a method for preventing or treating muscle diseases comprising an oxicam compound or a nutrient-acceptable salt thereof. The present invention relates to a functional health food composition for improving or preventing the condition.

[0027] The present invention relates to a muscle strengthening composition containing an oxicam compound.

[0028] The present invention relates to a composition for enhancing exercise performance that contains an oxicam compound.

[0029] The present invention relates to a composition for promoting muscle stem cell differentiation, which comprises an oxicam compound.

[0030] The present invention relates to a composition for muscle regeneration containing an oxicam compound.

[0031] The present invention relates to a composition for increasing muscle mass, which comprises an oxicam compound.

[0032] The present invention relates to a feed additive composition containing an oxicam compound.

[0033] The present invention will be described in detail below.

[0034] In one aspect of the present invention, the present invention provides the oxicam compounds and their pharmaceutically acceptable salts. The oxicam compound (oxicam-based compound) of the present invention may contain a non-steroidal anti-inflammatory drug. Compounds that are widely used as anti-inflammatory agents, more specifically lornoxicam, Piroxicam, tenoxicam, ampiroxicam, droxicam, meloxicam, clono It may include texicam, tenoxicam and piroxicam, more preferably meloxicam. cam (chemical 1), piroxicam (chemical 2), tenoxicam (chemical 3) or lornoxicam (chemical 4) may be used, but is not limited to, the oxicam compound. It is represented by the following chemical formula.

[0035] [ka]

[0036] [ka]

[0037] [ka]

[0038] [ka]

[0039] In one embodiment of the present invention, the oxicam compounds of the present invention are useful for treating aging, muscle decline, muscle wasting, and other conditions. , and has the effect of preventing or treating muscle diseases caused by muscle degeneration or muscle damage. "Muscle disease" refers to muscle damage or loss caused by aging or disease, resulting in weakened muscles. This means having a condition that can be caused by genetic predisposition, high blood pressure, impaired glucose tolerance, diabetes, obesity, or lipid Age-related diseases such as dyscrasias, atherosclerosis or cardiovascular disease, cancer, autoimmune disorders Epidemics, infectious diseases, AIDS, chronic inflammatory diseases, arthritis, malnutrition, kidney disease, chronic obstructive pulmonary disease Obstructive pulmonary disease, emphysema, rickets, chronic lower spinal pain, peripheral nerve injury, central nerve injury and chemical Loss of movement due to chronic illness, fracture, trauma, or long-term bed rest This can be caused by a variety of factors, including ageing and dementia.

[0040] The muscle disorders include, but are not limited to, atony, muscle atrophy, Muscular atrophy, muscle degeneration, muscle stiffness, amyotrophic lateral sclerosis, Myasthenia, cachexia and senile sarcopenia At least one muscle disease selected from the group consisting of: Muscle-related diseases caused by muscle atrophy, more specifically, senile muscular atrophy or cancer-induced muscular atrophy (muscle atrophy) cular atrophy), muscle degeneration, muscle rigidity, amyotrophic lateral sclerosis, myasthenia, Cachexia, sarcopenia and muscle loss That is, in one embodiment, the muscle disease may include atony. , muscular atrophy, muscle degeneration, muscle stiffness, amyotrophic lateral sclerosis Sclerosis, myasthenia, cachexia and sarcopenia ) means a disease selected from the group comprising

[0041] In the present invention, the preventive, therapeutic or ameliorative effects on muscle diseases are achieved by promoting myoblast differentiation. In one embodiment of the present invention, the oxicam compound may be It was confirmed that the differentiation of myoblasts was promoted. It refers to the process by which myoblasts fuse to form multinucleated myotubes. However, cells at the differentiation stage that form myotubes express markers such as Pax7-, MyoD+, and MyoG. The cells in the early stage of differentiation that form the myotubes may be differentiated using myosin D (M Myogenic transcription factors such as yo D The expression of myosin G (MyoG) increases during the metaphase. In the late stage, when the myosin heavy chain (MHC) In one embodiment of the present invention, meloxicam, an oxicam compound, is administered to myoblasts. It was confirmed that when cells were treated with the agent, the expression of MHC and myogenin increased.

[0042] The oxicam compounds of the present invention can improve athletic performance. " refers to the ability to exercise using muscle strength, and muscle strength includes muscle mass, muscle endurance, and acidity. This is enhanced by increasing metabolic muscle mass, improving muscle recovery and intramuscular energy balance, The oxicam compound of the present invention is also enhanced by the reduction of fatigue-causing substances in the muscles. Through the effects on the various muscles listed above, it has the effect of improving the ability to perform exercise using muscles. In relation to the above-mentioned effects, in one embodiment of the present invention, the group administered with meloxicam showed It was confirmed that actual muscle strength and athletic performance increased in .

[0043] That is, the oxicam compounds of the present invention are effective in increasing muscle mass, improving muscle strength, and increasing muscle recovery. It has the effect of improving muscle function, and is useful for the prevention, treatment and treatment of muscle-related diseases. It can have an improving effect.

[0044] The oxicam compounds of the present invention have potential for preventing muscle diseases through muscle regeneration in injured mice. In one embodiment of the present invention, meloxicam may have a preventive, therapeutic or ameliorative effect. The effect of administering the drug to a mouse model of muscle injury (CTX-injury) was confirmed. As a result, it was confirmed that muscle mass increased compared to the control group, and when meloxicam was administered, When observing the degree of muscle and muscle fiber regeneration, it was confirmed that there was a finer increase. In addition, when oxicam compounds such as meloxicam were administered, the type 2 muscle fibers (MHC The rate of type II was significantly increased compared to the control group. Muscle tissue in mice damaged or weakened by aging, as well as muscles damaged by It promotes muscle stem cell differentiation, increases muscle fiber size, and increases muscle mass. It can have the effect of improving strength and athletic performance.

[0045] In addition, the oxicam compounds of the present invention are effective in regenerating muscles damaged by muscle diseases or aging. In addition, it also improves muscle strength and function through muscle regeneration and muscle mass increase effects in normal muscles. In one embodiment of the present invention, young mice (14 months, 4 months) muscle stem cell differentiation and muscle mass increase effects were confirmed, and Meloxicam was also effective in the muscles of young people. It has been confirmed that long-term administration of this drug has the effect of increasing muscle strength and improving exercise performance. did.

[0046] The composition of the present invention is useful for pharmaceuticals, health functional foods, functional foods, animal feeds, cell culture media, etc. It may be used for various purposes such as compositions, promoting myoblast differentiation or intramuscular myogenesis. It promotes mitochondrial activity and has preventive, ameliorative or therapeutic effects on muscle diseases.

[0047] As used herein, the term "prevention" refers to the prevention of a disease by administering the pharmaceutical composition according to the present invention. It refers to any action that can prevent or delay the onset of muscle disease.

[0048] As used herein, the term "treatment" refers to the administration of the pharmaceutical composition according to the present invention. Any action that improves or favorably alters symptoms.

[0049] The pharmaceutical composition of the present invention can be prepared by a conventional method into powders, granules, tablets, capsules, Oral dosage forms such as suspensions, emulsions, syrups, aerosols, topical preparations, suppositories and sterilized It may be formulated into an injection form for use, and a carrier or excipient necessary for the formulation may be added. A pharmaceutically acceptable carrier that may be further included in the active ingredient, Excipients and diluents include lactose, dextrose, sucrose, sorbitol, Mannitol, xylitol, erythritol, maltitol, starch, acacia gum , alginate, gelatin, calcium phosphate, calcium silicate, cellulose Methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate Dibenzoate, Propyl Hydroxybenzoate, Magnesium Stearate and Minerals When formulating, commonly used fillers, extenders, binders, wetting agents, etc. The formulation is prepared using a diluent or excipient such as a disintegrant, a surfactant, or the like.

[0050] For example, solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc. Such solid formulations may contain the extract or compound in combination with at least one excipient, at least Tomomen, starch, calcium carbonate , sucrose or lactose, gelatin, etc. In addition to simple excipients, magnesium stearate and talc are also used. Lubricants are also used. Liquid preparations for oral administration include suspensions, liquid contents, emulsions, and syrups. In addition to the commonly used simple diluents water and liquid paraffin, there are various Excipients such as wetting agents, sweetening agents, flavoring agents, preservatives, and the like may also be included.

[0051] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspension solvents include propylene glycol e glycol), polyethylene glycol, vegetable oils such as olive oil, ethyl oleate Injectable esters such as benzophenone may also be used. Witepsol, Macrogol, Tween 61, Cocoa butter, Lauri Fat, glycerol gelatin, etc. may also be used.

[0052] The pharmaceutical composition of the present invention can be administered orally or parenterally (intravenously, subcutaneously) according to the desired method. The dosage may be determined depending on the condition and weight of the patient, the severity of the disease, and the amount of the drug administered. The dosage form may vary depending on the drug form, route of administration, and time, and an appropriate dosage form may be selected by a person skilled in the art. good.

[0053] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. "Practically effective amount" means a reasonable amount applicable to medical treatment, sufficient to treat a disease. The criteria for this are based on the patient's disease, severity, drug activity, drug sensitivity, It depends on the administration time, route of administration and excretion rate, duration of treatment, concomitant ingredients, and other factors. The pharmaceutical composition of the present invention may be used in combination with an individual therapeutic agent or other therapeutic agents. The above elements may be administered sequentially or simultaneously with conventional therapeutic agents. Taking all these factors into consideration, the dosage may be determined at a level that minimizes side effects. This can be easily determined by one skilled in the art. The dosage varies depending on the patient's age, weight, severity, and sex, and is generally 0.001 to 150 mg per 1000 mg, preferably 0.01 to 50 mg, more preferably 0. A dose of 0.1 to 5 mg may be administered daily or every other day, 1 to 3 times a day. The dosage is variable and can be adjusted as needed.

[0054] The composition of the present invention may be a food or a functional health food, and in particular, "Functional Health Foods" are foods that have beneficial functions for the human body based on Act No. 6727 on Functional Health Foods. The term "functional" refers to food that has been manufactured and processed using raw materials or ingredients that have the following properties: Useful for health applications such as regulating nutrients for structure and function or physiological effects It means to take something for the purpose of getting an effect.

[0055] The food or health functional food of the present invention is a powder formulation for the purpose of preventing and improving muscle diseases. Pharmaceutical administration of granules, tablets, capsules, pills, suspensions, emulsions, syrups, etc. Healthy functional foods in the form of tea bags, infused tea, beverages, candy, jelly, gum, etc. It can be manufactured and processed as a product.

[0056] The food or functional health food composition of the present invention may be used as a food additive, It may be formulated alone or in combination with other ingredients. It may also contain nutrients, vitamins, lysates, flavourings, colourings and enhancers, pectinic acid and its salts, alginic acid and its salts, organic Acid, protective colloid thickener, pH adjuster, stabilizer, preservative, glycerin, alcohol, charcoal Carbonation agents used in acidic drinks may also be included. The above ingredients may be used alone or in combination. They may be used individually or in combination in appropriate amounts.

[0057] In one embodiment, the present invention relates to a muscle strengthening composition containing an oxicam compound. do.

[0058] The aforementioned "muscle strengthening" refers to the strengthening of physical performance, strengthening of maximum endurance, increasing muscle mass, and muscle recovery. Strengthening of the body's natural circulation, reducing muscle fatigue, improving energy balance, or a combination of these effects. The composition increases muscle mass through its ability to differentiate myoblasts into muscle cells, resulting in overall It can increase muscle mass, strengthening maximal endurance and thereby enhancing physical performance. This can reduce muscle fatigue. It also helps to replace muscle cells quickly, which can help with muscle damage. The muscle strengthening composition of the present invention can be used in a food composition or a food additive. The food composition may be produced in the form of a food product, particularly in the form of a health food composition. Therefore, the muscle strengthening composition of the present invention is effective in reducing muscle loss due to aging. It can be used not only for small amounts but also as a supplement for muscle building and muscle strengthening for the general public. stomach.

[0059] In another embodiment, the present invention provides a feed or feed additive comprising an oxicam compound. The present invention relates to a composition for use.

[0060] In the present invention, "feed" refers to organic or inorganic nutrients necessary to sustain the life of an animal. The feed refers to a substance that provides the energy and nutrients required by animals such as livestock. It contains nutrients such as protein, lipids, vitamins, and minerals, and is found in grains, roots, fruits, and food processing by-products. Plant feed such as algae, fiber, oils and fats, starch, yeast, and grain by-products Proteins, minerals, fats and oils, minerals, fats and oils, animal proteins such as single-cell proteins Examples include, but are not limited to, feed.

[0061] In the present invention, the term "feed additive" refers to a feed additive that is used to improve the productivity and health of animals. It means a substance added to food, including but not limited to growth promotion, disease prevention, etc. Amino acid preparations, vitamin preparations, enzyme preparations, flavoring agents, silicate preparations, buffering agents, extractants, etc. It may further contain oligosaccharides.

[0062] MODE FOR CARRYING OUT THE INVENTION

[0063] In the following, the present invention will be described in detail with reference to examples in order to specifically explain the present invention. The embodiments described herein may be modified in various other forms and the scope of the present disclosure is set forth below. The examples herein should not be construed as limiting the scope of the invention. These and other related drawings are provided to enable those skilled in the art to more completely understand the present invention.

[0064] Example 1: Isolation and culture of muscle cells C2Cl2 is a myoblast cell line derived from live mice of the C3H strain and has been widely used in muscle cell differentiation studies. The C2C12 cells are cultured in both a general cell culture medium and a differentiation medium. The normal cell culture medium (GM, growth media) was used for 10 DMEM supplemented with 100% fetal bovine serum was used. The differentiation medium (DM) was 2% DMEM containing human serum was used.

[0065] In addition, the rectus abdominis (Rectus Abdominis) of 17- and 66-year-old Caucasian men was Human Skeletal Muscle Stem Cells (skMDC) isolated from human muscle cells scle Cells(Standard Donors),Cat NO:SK-11 11) was purchased from MyoSite and used.

[0066] Example 2: Creation of a mouse model of muscle injury (CTX-injury) The experimental animals used were 30 C57BL / 6 male mice aged 5 months. Five mice with similar blood pressure were assigned to each group, and the control group was treated with meloxicam. The mice in the experimental group were given 0.02 mg of meloxicam. The mice were orally administered with 20 μM cardiotoxin (cardiotoxin) at a dose of 1 / kg for 7 days. 50 μl of otoxin (CTX) was administered to the tibialis anterior (TA) muscle. Muscle damage was induced by direct injection into the meat, followed by oral administration of meloxicam (0.02 mg / kg) for 21 days. Administered.

[0067] <Experimental Example 1> Confirmation of myoblast differentiation enhancement effect 1-1. Enhancement of differentiation of mouse muscle stem cells The C2Cl2 cell line of Example 1 was dispensed into cell culture medium and cultured in DMEM medium for 24 hours. After that, the differentiation medium was added with oxicam compounds, meloxicam (F03) and lornoxicam. (F03-1), piroxicam (F03-2) and tenoxicam (F03-3) , differentiation markers Myogenin and MHC (myosin heavy chain The relative expression levels of C2C12 cells induced to differentiate in Example 1 were analyzed. , lysis buffer (20mM Tris-HCl, pH8.0, 150mM NaCl, 1% Triton X, Proteinase Inhibitor) After lysis, the cell lysate samples were quantified and equal amounts of protein were analyzed by SDS-PAGE electrophoresis. After electrophoresis, the sample was transferred to a PVDF membrane. Blocking was performed with silk and TTBS (0.03% Tween 20, Tris 2.42 The differentiation markers were washed in TTBS containing 5% BSA (1 L, 9 g of NaCl, pH 7.4). The primary antibody against MHC (myosin heavy chain), which is the target of the antibody, was diluted to 1:500. After dilution, the mixture was added and incubated overnight at 4°C. The secondary antibody was diluted 1:5000 in TTBS containing lk and then incubated at room temperature. After that, ECL (Enhanced Chemiluminescent solution) Then, the membrane was exposed to an X-ray film. The protein expression level was confirmed.

[0068] As a result, as shown in Figure 1, the oxicam compound-treated group showed a significantly higher oxicam activity than the control group (DMSO). When treated with HCl, it was confirmed that the relative expression level of myoblast differentiation markers increased.

[0069] In addition, among the compounds, meloxicam was administered to C2C12 cells at 1, 10, and 100 The medium was changed every other day for 3 days. After differentiation, the cells were examined under a microscope and disrupted to detect differentiation markers. Western blot analysis was performed using Myogenin and MHC antibodies. 2. After washing the cells with 1X PBS, they were soaked in 4% paraformaldehyde. After fixation with ethylenediamine at room temperature, the cells were then cultured in permeabilization buffer. n buffer) was added and reacted at room temperature. PBST (blocking buffer), PBS containing 0.05% Tween 20 S was used to inhibit non-specific antibody binding. After adding the primary antibody against MHC, the reaction was allowed to proceed at room temperature. Blocking buffer Add secondary antibody diluted 1:5000 in blocking buffer and incubate at room temperature. After reacting with 40% glycerol in DW) After application and fixation, photographs were taken under a fluorescent microscope and the results were analyzed.

[0070] As a result, as shown in Figure 2, myoblasts treated with high concentrations of meloxicam expressed MHC(m The expression level of the yosin heavy chain was also increased as shown in Figure 3. It was confirmed that this was the case.

[0071] 1-2. Enhancement of human muscle stem cell differentiation To confirm whether meloxicam also has the effect of promoting differentiation in human muscle stem cells, To this end, an experiment was conducted.

[0072] Rectus Abdominis of 17-year-old and 66-year-old Caucasian men Human skeletal muscle stem cells (skMDC) isolated from e Cells(Standard Donors),Cat NO:SK-1111) The cells were purchased from MyoSite and used. Human muscle stem cells were dispensed into cell culture medium. After culturing for 24 hours, the differentiation medium was supplemented with meloxicam (1 μM), an oxicam compound. The cells were treated with MHC (myosin) markers for myofiber differentiation for 4 days. The degree of differentiation into muscle fibers was measured by immunostaining for mitochondrial heavy chain (MHC). The muscle fiber diameter was analyzed.

[0073] As a result, as shown in Figure 4, muscle stem cells extracted from a 17-year-old subject were able to form myotubes. It was confirmed that the diameter increased, and as can be seen in Figure 5, the sample was extracted from a 66-year-old subject. It was confirmed that muscle stem cells showed an increase in both expression level and myotube diameter.

[0074] <Experimental Example 2> Confirmation of muscle recovery effect in muscle injury mouse model

[0075] 2-1. Increased muscle mass in muscle-damaged mouse models Experiments to confirm muscle recovery effects using a CTX-injured mouse model After 21 days of CTX-injury, meloxicam was administered to the hind limbs (hind limbs). Tibialis anterior muscle (TA) , tibialis anterior), EDL (Extensor digitorum longus mus) cle, extensor digitorum longus), Sol (Soleus muscle, soleus muscle), Gas (Ga The weights of the strocnemius muscle and gastrocnemius muscle were measured and compared. The results were 5.8% for TA, 14.1% for EDL, 5.4% for Gas, and 14.1% for the control group, respectively. It was confirmed that muscle mass increased by 3.4% in the Sol and Sol groups (Fig. 6). This result is in contrast to the fact that there was almost no change in the weight of the melo These results show that xicam specifically increases skeletal muscle mass.

[0076] 2-2. Muscle regeneration and muscle fiber size increase in muscle injury mouse models Meloxicam enhances muscle regeneration and myofibers in a CTX-injured mouse model To confirm the effect of increasing fiber size, TA muscle isolated from the tissue of experimental animals was used. H&E staining (hematoxylin & eosin staining) was performed. 4% paraformaldehyde-fixed frozen sections The sections were stained with hematoxylin and eosin for contrast staining. eosine), fixed with mounting solution, and observed under a light microscope. The muscle recovery rate and muscle fiber size were observed on days 4, 7, and 21 after muscle injury. The increase in the amount of HCl was confirmed as shown in Figure 8.

[0077] To further confirm the type of muscle that was increased, The relative expression levels of mRNA were compared. As a result, as shown in Figure 9, the meloxicam-administered group In the 21st day after injury, the amount of Type II muscle fibers was significantly increased compared to the control group. It was confirmed that

[0078] Furthermore, a laminin antibody was used to analyze the size of myofibers in TA muscle tissue. Immunohistochemical staining was performed and observed under a fluorescence microscope. As shown in the bottom photograph of Figure 9, Cross-sectional area of ​​muscle fibers (CSA) as determined by laminin staining It was confirmed that the cross-sectional area of ​​metabolic muscle fibers expressing MHC increased. This indicates that the number of muscle fibers and the resulting muscle mass are increased. The increase can be attributed to an increase in the diameter and proportion of metabolic muscle fibers.

[0079] 2-3. Increased activity of metabolic enzymes in muscle of a mouse model of muscle injury Meloxicam was administered to CTX-injured mice 21 days after injury. In the treated group, the metabolic enzymes in muscle mitochondria (glycolytic enzymes) The effect of increasing enzyme activity was confirmed.

[0080] The enzyme GPDH (Glycerol-3-phosphate dehydrogenase) was added to TA muscle sections isolated from experimental animals. α-glycerol, a substrate for α-phosphate dehydrogenase Phosphate was added and the color change of muscle fibers was observed according to the activity of GPDH. The muscle sections were fixed in Merck aquatex and observed under a microscope. As shown in Figure 10, in the meloxicam-treated group, the size of metabolic muscle fibers increased (left). ), and it was confirmed that the proportion of muscle fibers in which GPDH was activated was higher than in the control group. These results suggest that meloxicam administration increases the activity of metabolic enzymes in the muscle. It can be seen that...

[0081] 2-4. Increased muscle strength and exercise capacity in mouse models with muscle injury After inducing CTX-injury, muscle strength and To confirm exercise capacity, experiments were conducted to evaluate muscle strength and exercise capacity. Grip strength tests were conducted to check the muscle strength of the groups. Grip strength testing was performed using a BIOSEB mouse grip strength measuring device. The mouse was placed on a wire mesh attached to an instrument panel that allowed the strength of the force to be monitored. The mouse's grip strength against the wire mesh was measured while pulling its tail. The measurement was repeated four times. The average value was divided by the body weight. In an experiment to evaluate motor skills, the Sicam group showed an increase of approximately 21.9% compared to the control group. The meloxicam group also showed an increase in exercise capacity of up to 72.3%. To confirm the motor performance of the treatment and control groups, a rotarod test was conducted. The Rotarod test was performed using a mouse The distance taken by a mouse to fall from a rotating rotor was measured using a rotarod device. The rotarod test was performed after adaptive training. The experiments were carried out at 8 rpm for 5 minutes, 10 rpm for 3 minutes, and 13 rpm for 1 minute for two days. The constant rotation test was performed at 13 rpm until the mouse fell off. The accelerated rotarod test was performed at 40 The measurements were taken at an acceleration rate of 5 rpm to 18 rpm per second. The experiment was stopped at 500 seconds and measurements were taken.

[0082] <Experimental Example 3> Confirmation of age-dependent muscle mass and athletic ability increases during long-term administration of meloxicam recognition 3-1. Changes in muscle mass, muscle type, and exercise capacity in aging mice After 4 months of meloxicam administration to 24-month-old mice, the hind limbs (h The changes in the intestinal and skeletal muscle mass were measured and compared by weight. As shown in Figure 12, an increase in muscle mass was observed in the hindlimb muscles and EDL muscles ( (bottom), which indicates that muscle mass gain occurs at a relatively high rate compared to overall weight gain (top). It is known that he was born.

[0083] In addition, the muscle types expressed were confirmed through the relative expression levels of mRNA. As shown in Figure 13, the meloxicam-administered group showed a significant increase in the development of Myh Type II muscle fibers. In other words, meloxicam-treated aged mice showed increased muscle mass and metabolic activity. It was confirmed that the proportion of muscle fibers increased.

[0084] In addition, to confirm the changes in motor ability between the meloxicam-treated and control groups, a rotarod test was performed. As a result, as shown in Figure 14, It was confirmed that the exercise capacity of the xicam-treated group increased by more than 50% compared to the control group. The effect of increasing exercise capacity was confirmed not only in aged mice but also in 7-month-old mice. In other words, the effect of meloxicam on improving athletic performance is not limited to aging muscles, but extends to normal muscles as well. It was also confirmed that the effect was observed.

[0085] 3-2. Changes in muscle strength and motor ability in mice (4 months old) Meloxicam administration caused changes in muscle strength and exercise capacity even in relatively young mice. Experiments were conducted to measure the effects of meloxicam on the 21st day after CTX-injury induction. Grip strength test to measure muscle strength in the CAM-treated and control groups Grip strength test The strength of the grip was measured using a mouse grip strength measuring device manufactured by BIOSEB. A mouse is placed on a wire mesh attached to a meter panel that can be used for feeding. The mouse is then pulled by its tail to move towards the wire mesh. The force required to grasp the object was measured. The force was calculated by dividing the average value of four consecutive measurements by the body weight. As a result, as shown in Figure 15, in terms of muscle strength, the meloxicam group showed an improvement of approximately 21% compared to the control group. 0.9% increased in the meloxicam group in a study to assess exercise capacity. We confirmed that exercise capacity increased by up to 72.3%.

[0086] 3-3. Changes in muscle strength and motor ability in normal mice (4 months old) after long-term administration Changes in muscle strength and exercise capacity in 4-month-old normal mice after long-term administration of meloxicam First, we conducted an experiment to observe the effects of meloxicam on normal mice after administering it for two months. When comparing the weight change between the 2 groups and the control group, no significant change was observed (Fig. 16a). No changes were observed (Figure 16b). Grip test and treadmill test were performed. The treadmill test was performed by Columbus Institute of Technology. Measurements were performed using a treadmill for mice manufactured by ments. Measurements were performed at a gradient of 10%. The speed started at 8 m / min and increased by 1 m / min every 2 min until the mouse became fatigued. The results of the grip strength test confirmed that muscle strength was improved in the meloxicam group, and exercise The duration of the effect was also improved compared to the control group (Fig. 17a). It was confirmed that the muscle mass relative to body weight in the group administered methicone was greater than that in the control group. In other words, the experimental results show that meloxicam administration also significantly increased the risk of inflammatory bowel disease in normal young mice (Fig. 17b). At the same time, muscle function improves, and muscle strength and athletic performance increase.

[0087] The present invention has been described above with a focus on its preferred embodiments. A person skilled in the art would understand that the present invention can be realized without departing from the essential characteristics of the present invention. It will be understood that the disclosed embodiments may be embodied in various modified forms. The examples should be considered in an illustrative rather than a restrictive sense. and all other aspects within the scope of the claims and their equivalents, rather than the foregoing description. The differences must be construed as included in the present invention.

Claims

1. A composition for strengthening muscle strength, improving muscle function, enhancing exercise performance, promoting differentiation of muscle stem cells, regenerating muscle, or increasing muscle mass in patients with a muscle disease selected from the group consisting of hypotonia, muscle atrophy, muscle degeneration, muscle rigidity, amyotrophic lateral sclerosis, myasthenia gravis, and cachexia, or in aged or normal muscles, comprising: It contains an oxicam compound and a pharmaceutically acceptable salt thereof as an active ingredient, A composition characterized in that the oxicam compound is lornoxicam or meloxicam.

2. 2. The composition according to claim 1, characterized in that the composition is one or more selected from the group consisting of food, functional food, health functional food, pharmaceutical, feed, and feed additive.

3. Use of the composition according to claim 1 in the manufacture of a medicament for strengthening muscle strength, improving muscle function, enhancing exercise performance, promoting differentiation of muscle stem cells, regenerating muscle, or increasing muscle mass in patients with a muscle disease selected from the group consisting of hypotonia, muscle atrophy, muscle degeneration, muscle rigidity, amyotrophic lateral sclerosis, myasthenia, and cachexia, or in aged or normal muscles.

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