Composition for improving, treating or preventing muscular disorders comprising sulfonamide-based compounds

Abstract

To provide a composition for preventing, improving, or treating muscular disorders.SOLUTION: Provided is a pharmaceutical composition for preventing or treating muscular disorders, including sulfonamide-based compounds represented by the formula [1] or pharmaceutically acceptable salts thereof, where R1 represents H, *-(C=N)-NH2, substituted / unsubstituted acetyl, C5-6 aryl or heteroaryl; and R2 represents -NH2 or a substituent represented by the formula [1-1].SELECTED DRAWING: Figure 6

Description

Technical field

[0001] TECHNICAL FIELD The present invention relates to a composition for improving, treating or preventing muscle diseases containing a sulfonamide compound. [Background technology]

[0002] Saropenia is a disease in which muscle mass or muscle strength decreases due to illness, aging, etc. It is estimated that muscle mass gradually decreases from the age of 40 onwards, at an rate of 8% every 10 years until the age of 70, and then an even more rapid decrease of 15% every 10 years. known to occur.

[0003] Numerous follow-up studies have revealed that the physiological changes that occur in the elderly are diverse, and that muscle mass and bone density generally decrease simultaneously with increasing age. Age-related sarcopenia not only directly induces a decrease in muscle strength and increases the risk of death due to the decrease and impairment of various bodily functions, but also causes problems such as decreased metabolism and weakened immune system, and high blood pressure. It can also lead to an increased prevalence of metabolic diseases such as diabetes, arthritis, obesity, cancer, etc. In particular, age-related sarcopenia occurs in 40% of elderly people aged 80 years or older, and is a disease that is expected to increase the social and economic burden in an aging society.

[0004] PHF20 (PHD finger protein 20) is a protein also known as GLEA2 (glioma-expressed antigen 2). Recently, it has been revealed that PHF20 acts as a transcription factor to regulate muscle damage and sarcopenia by regulating the lower target protein YY1. It was reported that during muscle cell differentiation using myoblasts (C2C12), the expression of PHF20 increases and then decreases, and that this phenomenon is regulated together with the expression of YY1, a protein that inhibits muscle differentiation.

[0005] On the other hand, there are three main methods for treating sarcopenia. The first is exercise. Exercise has been reported to increase the protein synthesis capacity of skeletal muscle in the short term, increasing muscle strength and mobility in the elderly. However, it is unsuitable for long-term treatment methods. The second is drug treatment, which can include the use of testosterone or anabolic steroids, which induce virilization in women and prostate symptoms in men. ) and other side effects. Other approved regimens include DHEA (dehydroepiandrosterone) and growth hormone, but studies have reported that they are possible treatments at sites containing SARMs (Selective Androgen Receptor Modulators). Additionally, although dietary therapy is known as a treatment, nutritional assessments indicate that there is malnutrition and that modern eating habits are inadequate to maintain adequate total body mass. However, the reality is that there is no fundamental treatment or improvement agent for sarcopenia. [Summary of the invention] [Problem to be solved by the invention]

[0006] In order to solve the above-mentioned problems, the inventors of the present invention utilized a screening system constructed using PHF20 / YY1 to detect muscle differentiation in myoblasts under conditions where overexpression of PHF20 inhibits muscle differentiation. We screened for substances that could restore or alleviate inhibition of muscle differentiation. The present invention was completed by confirming that the selected compounds can be used to suppress muscle loss and promote muscle differentiation. [Means to solve the problem]

[0007] As a means to achieve the above object, the present invention provides a preventive or therapeutic pharmaceutical agent containing a sulfonamide compound represented by the following formula 1 or a salt thereof, which can be useful for treating, alleviating, alleviating, or preventing a disease. or food compositions; quasi-drug compositions; feed compositions; or compositions for feed additives; The purpose is to

[0008] As one specific example, an object of the present invention is to provide a pharmaceutical composition for preventing or treating muscle diseases, which contains a sulfonamide compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof.

[0009] As a specific example, an object of the present invention is to provide a food composition for preventing or improving muscle diseases, which contains a sulfonamide compound represented by the following formula 1 or a foodically acceptable salt thereof.

[0010] As a specific example, an object of the present invention is to provide a feed composition for preventing or improving muscle diseases, which contains a sulfonamide compound represented by the following formula 1 or a salt thereof. [ka] In Formula 1 above, R 1 is hydrogen, *-(C=N)-NH 2 , acetyl, or C 5-6 aryl or heteroaryl, and the R 1 is unsubstituted or C 1-3 Alkyl, C 1-3 is substituted with at least one substituent selected from alkoxy and phenyl, and the R 2 is -NH 2 Or a compound represented by the following chemical formula 1-1. [ka]

[0011] In the present invention, the term "prevention" refers to all actions of suppressing the onset of muscle diseases, or suppressing or delaying the deepening of muscle diseases by administering a composition.

[0012] In the present invention, the term "treatment," "improvement," "alleviation," or "alleviation" refers to any act in which the symptoms of a muscle disease are ameliorated or beneficially altered by administration of a composition.

[0013] In the present invention, "able to serve" means to play a role that helps or plays a role in suppressing or delaying the symptoms of a muscle disease or ameliorating the symptoms of a muscle disease by administering the composition of the present invention. It means that you can. It can be used as an adjuvant to enhance the effect of medicines for treatment, and has meaning as an adjunct as a health functional food or a functional food.

[0014] In this specification, TIFF2023155212000004.tif14170 means the position to be concatenated.

[0015] The terminology used in the detailed description and examples of the invention is used for purposes of explanation only and is not to be construed as limiting. A singular expression includes a plural expression unless the context clearly dictates otherwise. As used herein, terms such as "comprising" or "having" are intended to specify the presence of features, numbers, steps, acts, components, parts, or combinations thereof that are described in the specification. It should be understood that this does not exclude in advance the existence or possibility of addition of one or more other features, figures, steps, acts, components, parts or combinations thereof.

[0016] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the examples pertain. have. Terms as defined in commonly used dictionaries should be construed to have meanings consistent with the meanings they have in the context of the relevant art, and ideally unless explicitly defined in this application. or not be interpreted in an overly formal sense.

[0017] Furthermore, in the description with reference to the accompanying drawings, the same components will be given the same reference numerals regardless of the drawing numbers, and duplicate explanations thereof will be omitted. In describing the embodiments, if it is determined that detailed explanations of related known techniques may unnecessarily cloud the gist of the embodiments, the detailed explanations will be omitted.

Effect of the invention

[0018] The composition for preventing, improving or treating muscle diseases containing the sulfonamide compound or its salt of the present invention can prevent inhibition of myoblast differentiation by regulating the expression of PHF20 and YY1. This can prevent or alleviate muscle loss or promote muscle regeneration to improve muscle function and muscle mass, and is a therapeutic agent for preventing, improving or treating muscle diseases, improving muscle function or muscle mass. , food or feed, etc. [Brief explanation of drawings]

[0019]

Figure 1

[0020] In the following, the invention will be explained in more detail. The present invention relates to the use of a sulfonamide compound represented by the following formula 1 or a salt thereof for the prevention, improvement, or treatment of muscle diseases; or provide a therapeutic composition: [ka] In Formula 1 above, R 1 is hydrogen, *-(C=N)-NH 2 , acetyl, or C 5-6 aryl or heteroaryl, and the R 1 is unsubstituted or C 1-3 Alkyl, C 1-3 is substituted with at least one substituent selected from alkoxy and phenyl, and the R 2 is -NH 2 Or a compound represented by the following chemical formula 1-1. [ka]

[0021] In one embodiment, the hetero element of the heteroaryl may be one or more selected from S, N, and O, and the hetero element may include one or more, and one or more hetero elements may be elements may be included.

[0022] In one embodiment, the aryl or heteroaryl is from the group consisting of pyrrole, pyrazole, triazole, oxazole, furan, isoxazole, isothiazole, imidazole, diazole, thiadiazole, phenyl, pyridine, pyrimidine, triazine, oxazine, and thiazine. It may be selected.

[0023] In one embodiment, the aryl or heteroaryl may be thiazole, diazole, thiadiazole, phenyl, pyridine or pyrimidine.

[0024] In one embodiment, the R 1 The substituent of C 1-3 Alkyl, C 1-3 It may be at least one substituent selected from alkoxy and phenyl. In a preferred embodiment, the R 1 The substituent may be at least one selected from the group consisting of methyl, ethyl, methoxy, ethoxy and phenyl. Said R 1 may contain one substituent, and may contain one or more substituents that are the same or different from each other.

[0025] In one embodiment, the compound of formula 1 is Sulfasalazine, Sulfamethazine, Sulfathiazole, Sulfapyridine, Sulfaphenazole, Sulfameter. , Sulfamethizole, Sulfaguanidine, Sulfacetamide sodium, Sulfadoxin, Sulfadimethoxine, Sulfanilamide or Sulfadiazine. It's fine.

[0026] In one embodiment, the salt of the compound of the present invention includes any salt type commonly accepted in the art of the present invention, and may be, for example, a sodium salt.

[0027] As a specific example, the sulfasalazine may be a compound represented by the following formula 2. [ka]

[0028] As a specific example, the sulfamethazine may be a compound represented by the following formula 3. [ka]

[0029] As one specific example, the sulfathiazole may be a compound represented by the following formula 4. [ka]

[0030] As one specific example, the sulfapyridine may be a compound represented by the following formula 5. [ka]

[0031] As a specific example, the sulfaphenazole may be a compound represented by the following formula 6. [ka]

[0032] As a specific example, the sulfameter may be a compound represented by the following formula 7. [ka]

[0033] As a specific example, the sulfamethizole may be a compound represented by the following formula 8. [ka]

[0034] As one specific example, the sulfaguanidine may be a compound represented by the following formula 9. [ka]

[0035] As a specific example, the sulfacetamide may exist in the form of a sodium salt, and more specifically may be a compound represented by the following formula 10. [ka]

[0036] As a specific example, the sulfadoxin may be a compound represented by the following formula 11. [ka]

[0037] As one specific example, the sulfadimethoxine may be a compound represented by the following formula 12. [ka]

[0038] As one specific example, the sulfanilamide may be a compound represented by the following formula 13. [ka]

[0039] As a specific example, the sulfadiazine may be a compound represented by the following formula 14. [ka]

[0040] In the present invention, muscle disease refers to a disease or condition that induces decreased muscle function, decreased muscle mass, wasted muscle, or muscle degeneration or deepens such symptoms.

[0041] As used herein, the term "muscle" refers comprehensively to tendons and muscles, and "muscle function" refers to the ability of a muscle to exert force by contracting, and the term "muscle" refers to the ability of a muscle to exert force by contracting. Muscular strength, which is the ability to exert force, and muscular endurance, which is the ability of a muscle to contract and relax under a given weight for how long or how many times, and to exert strong force in a short period of time. Including explosive power, which is an ability.

[0042] As used herein, the term "improving muscle function" refers to increasing muscle mass to better improve muscle function, promoting regeneration of damaged muscles, or improving regenerative power.

[0043] As used herein, "promoting muscle regeneration" means shortening the regeneration and recovery time of damaged muscles, activating them to increase muscle mass, or reducing the degree to which muscle mass decreases.

[0044] The muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc. Muscle wasting is characterized by gradual loss of muscle mass, weakening and degeneration of muscles, especially skeletal or voluntary muscles and cardiac muscles. However, it is not limited to the type of muscle.

[0045] In one embodiment, the muscle disease includes sarcopenia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, cachexia, and myasthenia. It may be any one selected from the group consisting of:

[0046] The present invention relates to a pharmaceutical composition for preventing or treating muscle diseases containing a sulfonamide compound represented by the following formula 1 or a salt thereof; a food composition for preventing or improving muscle diseases; a feed composition for preventing or improving muscle diseases. I will provide a: [ka] In Formula 1 above, R 1 is hydrogen, *-(C=N)-NH 2 , acetyl, or C 5-6 is an aryl or heteroaryl of R 1 is unsubstituted or C 1-3 Alkyl, C 1-3 is substituted with at least one substituent selected from alkoxy and phenyl, and the R 2 is -NH 2 Or a compound represented by the following chemical formula 1-1. [ka]

[0047] In one embodiment, the hetero element of the heteroaryl may be one or more selected from S, N, and O, and the hetero element may include one or more, and one or more hetero elements may be elements may be included.

[0048] In one embodiment, the aryl or heteroaryl is from the group consisting of pyrrole, pyrazole, triazole, oxazole, furan, isoxazole, isothiazole, imidazole, diazole, thiadiazole, phenyl, pyridine, pyrimidine, triazine, oxazine, and thiazine. It may be selected.

[0049] In one embodiment, the aryl or heteroaryl may be thiazole, diazole, thiadiazole, phenyl, pyridine or pyrimidine.

[0050] In one embodiment, the R 1 The substituent of C 1-3 Alkyl, C 1-3 It may be at least one substituent selected from alkoxy and phenyl. In a preferred embodiment, the R 1 The substituent may be at least one selected from the group consisting of methyl, ethyl, methoxy, ethoxy and phenyl. Said R 1 may contain one substituent, and may contain one or more substituents that are the same or different from each other.

[0051] In one embodiment, the compound of formula 1 is Sulfasalazine, Sulfamethazine, Sulfathiazole, Sulfapyridine, Sulfaphenazole, Sulfameter. , Sulfamethizole, Sulfaguanidine, Sulfacetamide sodium, Sulfadoxin, Sulfadimethoxine, Sulfanilamide or Sulfadiazine. It's fine.

[0052] In one embodiment, the salt of the compound of the present invention includes any salt type commonly accepted in the art of the present invention, and may be, for example, a sodium salt.

[0053] As a specific example, the compound of compound 1 of the present invention inhibits muscle loss in muscle tissue by inhibiting myoblast formation by acting on PHD20 / YY1. It plays a role in promoting muscle formation.

[0054] As an example, sulfamethazine of the present invention acts on PHD20 / YY1 and inhibits myoblast formation, thereby suppressing muscle damage and promoting regeneration. We confirmed in vivo that the balance ability and grip strength recovery of Rota Rod mice were superior to those of muscle-damaged mice.

[0055] When the composition for preventing or treating muscle diseases of the present invention is a pharmaceutical composition, it can be used to prevent or treat muscle diseases caused by muscle wasting or degeneration. Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc. Muscle wasting is characterized by gradual loss of muscle mass, weakening and degeneration of muscles, especially skeletal or voluntary muscles and cardiac muscles. do. Examples of associated diseases include sarcopenia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, cachexia, and myasthenia. Examples include, but are not limited to, symptoms. Pharmaceutical compositions of the invention can include a pharmaceutically acceptable carrier.

[0056] The pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration. Carriers for oral administration can include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Carriers for parenteral administration can also include water, suitable oils, saline, aqueous glucose, glycols, and the like. It may also further contain stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol.

[0057] The pharmaceutical composition of the present invention can be administered to mammals including humans by any method. For example, it can be administered orally or parenterally, and parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, and transdermal administration. Administration may be subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal.

[0058] The pharmaceutical composition of the present invention can be formulated into a preparation for oral or parenteral administration by the administration routes as described above. When formulated, one or more buffering agents (e.g. saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g. EDTA or glutathione), fillers, fillers, binders, They may be formulated using adjuvants (eg aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.

[0059] Solid preparations for oral administration include tablets, pills, powders, granules, solutions, gels, syrups, slurries, suspensions, capsules, etc. The pharmaceutical composition of the invention may contain at least one excipient, such as starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose ( lactose), dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, or gelatin. For example, tablets or sugar-coated tablets can be obtained by combining the active ingredient with solid excipients, grinding it, adding suitable auxiliaries, and processing it into a granular mixture.

[0060] In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, internal solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water or liquid paraffin, various excipients may be used, such as Humectants, sweetening agents, flavoring agents or preservatives, etc. may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid, sodium alginate, etc. may be added as a disintegrant depending on the case, and an anti-aggregating agent, a lubricant, a wetting agent, a fragrance, an emulsifier, a preservative, etc. may be further included.

[0061] As one embodiment, when administered parenterally, the pharmaceutical compositions of the present invention may be in the form of injections, transdermal administrations and nasal inhalants, together with suitable parenteral carriers, as known in the art. The method can be formulated into a dosage form. In the case of injections, they must be sterilized and must be protected from contamination by microorganisms such as bacteria and fungi. For injectables, examples of suitable carriers include, but are not limited to, water, ethanol, polyols (such as glycerol, propylene glycol and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils. It may be a containing solvent or dispersion medium. More preferably, suitable carriers include Hank's solution, Ringer's solution, PBS (phosphate buffered saline) containing triethanolamine or sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. Isotonic solutions and the like can be used. In order to protect the injection from microbial contamination, it may further contain various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In addition, in most cases, the injection may further include an isotonic agent such as sugar or sodium chloride.

[0062] As a specific example, in the case of transdermal preparations, forms such as ointments, creams, lotions, gels, external liquids, pastes, liniments, air rolls, etc. are included. In the above, "transdermal administration" means that a pharmaceutical composition is locally administered to the skin so that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin.

[0063] As one example, for inhalation administration, the compounds used according to the invention may be administered using a suitable propellant, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered in the form of an aerosol spray from a pressurized pack or atomizer. In the case of pressurized aerosols, dosage units can be determined by providing a valve to transmit a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators can be formulated to contain a powder mixture of the compound and a suitable powder base, such as lactose or starch.

[0064] When the pharmaceutical composition for preventing or treating muscle diseases of the present invention contains the compound of formula 1 in an effective amount, it can provide a preferable effect for preventing or treating muscle diseases.

[0065] In one specific example, a method for treating muscle diseases is provided, which comprises the step of administering a therapeutically effective amount of the compound represented by formula 1 or a pharmaceutically acceptable salt thereof to a subject.

[0066] Preferably, the treatment method may further include the step of identifying a subject in need of prevention or treatment of the muscle disease before the administration step.

[0067] As used herein, the term "therapeutically effective amount" or "effective amount" refers to an amount that exhibits a greater response than a negative control group, preferably an amount sufficient to improve muscle function. The pharmaceutical composition of the present invention may contain 0.01 to 99.99% of the compound of formula 1 or a pharmaceutically acceptable salt thereof, and the remaining amount may be occupied by a pharmaceutically acceptable carrier. The effective amount of the compound of formula 1 or a pharmaceutically acceptable salt thereof contained in the pharmaceutical composition of the present invention may vary depending on the form in which the composition is commercialized.

[0068] The total effective amount of the pharmaceutical composition of the present invention can be administered to a subject in a single dose, or in a fractionated treatment protocol administered over an extended period of time in multiple doses. can be administered by. The content of the active ingredient in the pharmaceutical composition of the present invention may vary depending on the severity of the disease. The administration volume is determined by considering various factors such as the administration route of the pharmaceutical composition and the number of treatments, as well as the age, weight, health condition, sex, severity of disease, diet, and excretion rate of the subject. An effective dosage for the body can be determined. Considering these points, a person of ordinary skill in the art would understand that the compound of formula 1 or its pharmaceutically acceptable salt is suitable for specific uses for the prevention or treatment of muscle diseases. Effective doses can be determined. The pharmaceutical composition according to the present invention is not particularly limited in its dosage form, administration route, and administration method as long as it exhibits the effects of the present invention.

[0069] The "subject" may refer to a human or non-human mammal such as a primate, mouse, dog, cat, horse, or cow, but is not limited thereto.

[0070] The pharmaceutical composition for preventing or treating muscle diseases of the present invention can also be provided in the form of an external preparation containing the compound of formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0071] When the pharmaceutical composition of the present invention is used as an external preparation for skin, it may further include fatty substances, organic solvents, solubilizing agents, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, foaming agents, etc. foaming agents, fragrances, surfactants, water, ionic emulsifiers, non-ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils , containing adjuvants commonly used in the dermatological field, such as dyes, pigments, hydrophilic active agents, lipophilic active agents or lipid vesicles, and any other ingredients commonly used in topical skin preparations. I can do it. Also, said ingredients may be introduced in amounts commonly used in the dermatological field.

[0072] When the pharmaceutical composition of the present invention is provided as an external preparation for the skin, it may be in the form of, but not limited to, an ointment, patch, gel, cream, or spray.

[0073] In another embodiment, the composition for preventing or improving muscle diseases of the present invention may be a food composition for preventing or improving muscle diseases containing the compound of formula 1 or a food-logically acceptable salt thereof.

[0074] When the composition for preventing or treating muscle diseases of the present invention is a food composition, it can be used to prevent or treat muscle diseases caused by muscle wasting or degeneration. Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc. Muscle wasting is characterized by gradual loss of muscle mass, weakening and degeneration of muscles, especially skeletal or voluntary muscles and cardiac muscles. do. Examples of associated diseases include sarcopenia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, cachexia, and myasthenia. Examples include, but are not limited to, symptoms.

[0075] In one embodiment, the food composition includes all forms of functional food, nutritional supplement, health food, food additives, feed, etc. This means that animals such as humans or livestock are eaten. Food compositions of the above type can be prepared in various forms by conventional methods known in the art.

[0076] Food compositions of the above type can be prepared in various forms by conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic drinks), fruits and their processed foods (e.g. canned fruit, bottled fruit, jam, marmalade, etc.), fish, meat and their processed foods (e.g. ham). , sausage corned beef, etc.), breads and noodles (e.g. udon, soba, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, candy, dairy products (e.g. butter, cheese, etc.), edible vegetable oils, Manufactured by adding the compound of chemical formula 1 of the present invention or its food-logically acceptable salt to margarine, vegetable protein, retort food, frozen food, various seasonings (e.g. miso, soy sauce, sauce, etc.) can. In addition, nutritional supplements can be produced by adding the compound of formula 1 of the present invention or a food-wise acceptable salt thereof to capsules, tablets, pills, etc., but are not limited thereto. In addition, functional health foods include, but are not limited to, the compound of chemical formula 1 of the present invention or its food-acceptable salts, which are produced and consumed in the form of tea, juice, and drinks (health drinks). It can be liquefied, granulated, encapsulated, or powdered and ingested.

[0077] Furthermore, in order to use the compound of Chemical Formula 1 of the present invention or a food-wise acceptable salt thereof in the form of a food additive, it can be prepared and used in the form of a powder or a concentrated liquid. In addition, a composition can be prepared by mixing the compound of formula 1 of the present invention or a foodically acceptable salt thereof with a known active ingredient known to have the effect of preventing or improving muscle diseases.

[0078] When the composition for preventing or improving muscle diseases of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients like a normal drink. I can do it. The natural carbohydrates mentioned above may be monosaccharides such as glucose, fructose; disaccharides such as maltose, sucrose; polysaccharides such as dextrin, cyclodextrin; sugar alcohols such as xylitol, sorbitol, erythritol, etc. As the sweetener, natural sweeteners such as thaumatin and stevia extract; synthetic sweeteners such as saccharin and aspartame can be used. The proportion of said natural carbohydrates is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the invention.

[0079] The compound represented by formula 1 of the present invention or a foodically acceptable salt thereof may be included as an active ingredient in a food composition for preventing or improving muscle diseases, and the amount thereof may be The amount is not particularly limited to an amount effective to achieve the effect, but is preferably from 0.01 to 100% by weight based on the total weight of the entire composition. The food composition of the present invention is produced by mixing the compound of formula 1 or a foodically acceptable salt thereof with other active ingredients known to be effective in compositions for preventing or improving muscle diseases. obtain.

[0080] In addition to the above, the health food of the present invention also contains various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, It can contain pH regulators, stabilizers, preservatives, glycerin, alcohol or carbonating agents, etc. In addition, the health food of the present invention can contain fruit pulp for the production of natural fruit juice, fruit juice drink, or vegetable drink. Such components can be used individually or in admixture. Although the proportion of such additives is not particularly important, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

[0081] In another embodiment, the composition for preventing or improving muscle diseases of the present invention may be a feed composition for preventing or improving muscle diseases or a feed additive composition containing the compound of formula 1 or a salt thereof.

[0082] In the present invention, the term "feed" means any natural or artificial diet, meal, etc. or a component of said meal intended for or suitable for eating, ingesting, and digesting by an animal, and which includes the Feed containing the disease prevention or ameliorating composition as an active ingredient can be manufactured into various forms of feed known in the art, and preferably includes concentrate feed, forage, and / or special feed. It is not limited to this.

[0083] In the present invention, the term "feed additive" refers to various effects such as nutrient supplementation and prevention of weight loss, enhancement of digestibility of fiber in feed, improvement of milk quality, prevention of reproductive disorders and improvement of conception rate, and prevention of summer high temperature stress. Includes substances added to feed for feeding purposes. The feed additive of the present invention falls under the category of supplementary feed under the Feed Management Act, and contains mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and composite minerals, as well as trace amounts of zinc, copper, cobalt, selenium, etc. Mineral preparations, carotene, vitamins A, D, E, nicotinic acid, vitamin B complex, etc., protected amino acids such as methionine, lysine, protected fatty acids such as fatty acid calcium salts, probiotics ( Live bacteria such as lactic acid bacteria (lactic acid bacteria), yeast cultures, and mold ferments, yeast agents, and the like may further be included.

[0084] Concentrate feed includes seeds and nuts including grains such as wheat, oats, and corn; byproducts obtained by refining grains such as rice bran, bran, and wheat bran; beans; canola; Oil cakes are by-products obtained by extracting oil from sesame, flaxseed, coconut, etc.; starch cakes are the residue left after starch is removed from sweet potatoes, potatoes, etc.; residual starch grains, which are the main components; fishmeal, fish cakes. , fish soluble, which is concentrated fresh liquid obtained from fish, meat meal, blood meal, feather meal, skim milk powder, cheese from milk, and the leftover liquid when making casein from skim milk. Examples include, but are not limited to, animal feed such as dried whey, yeast, chlorella, and seaweed.

[0085] Forage, a silo is filled with raw grass feed such as wild grass, grass, grass clippings, feed turnips, feed beets, root vegetables such as rutabaga, which is a type of turnip, fresh grass, green clippings, grains, etc. Examples include, but are not limited to, silage, which is stored feed that has been fermented with lactic acid, wild grass, hay obtained by cutting and drying grass, straw from seed crops, and leaves from leguminous plants. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea and its derivative diureido isobutane, supplementing ingredients that tend to be insufficient when only natural feed ingredients are mixed, or improving the storability of the feed. There are feed additives and dietary supplements, which are substances that are added in small amounts to compounded feed to increase the feed, but are not limited thereto.

[0086] The feed additive for preventing or improving muscle diseases according to the present invention is prepared by adding the compound of formula 1 of the present invention or a salt thereof in an appropriate effective concentration range using various feed manufacturing methods known in the art. Manufacturable.

[0087] To avoid redundant descriptions, unless otherwise specified, the definitions and explanations for each component of the present invention described in Chemical Formula 1 and the pharmaceutical composition section apply mutatis mutandis to the food composition and feed composition.

[0088] In the following, embodiments will be described in detail with reference to the accompanying drawings. However, since various changes may be made to the embodiments, the scope of the patent application is not restricted or limited by these embodiments. It is to be understood that all modifications, equivalents, and alternatives to the embodiments are within the scope of this invention.

[0089] Example 1. HTS (High Throughput Screening) system construction 1-1.Cell line production In order to discover substances that control sarcopenia, we constructed an HTS (High Throughput Screening) system using cell lines that can easily monitor the differentiation state of muscle cells.

[0090] Specifically, mouse normal myoblast C2C12 cell line was obtained from ATCC (Manassas, VA, USA). C2C12 cells were passaged every 2-3 days in 100 mm dishes containing DMEM supplemented with 10% fetal bovine serum, 2 mM glutamine, 100 units / ml penicillin and 100 ug / ml streptomycin (Life Technologies, Grand Island, NY, USA). Cultured. The cells were incubated with 5% CO in a humidified state. 2 , and grown by culturing at 37°C. While used in experiments, the cells were harvested at the end of treatment for further analysis.

[0091] C2C12 myoblasts, which are stable cells in which PHF20 is overexpressed, were transformed with a plasmid containing both the YY1 promoter and GFP, which suppresses muscle differentiation, to form a stable cell line. ) was created. By quantifying and measuring the extent to which the GFP signal is suppressed, the degree of muscle differentiation was utilized as a quantitative measurement index (Figure 1).

[0092] 1-2. Confirmation of PHF20 expression by selection marker concentration Quantitative measurement system for muscle differentiation In order to construct an HTS system that can be used as a measurement index, stable cell lines were treated with hygromycin, a selection marker for GFP expression, at different concentrations. ) were selected.

[0093] Specifically, C2C12 cell lines transformed with PHF20 and YY1-promoter-GFP plasmids were treated with 1 mg / ml of neomycin, 2 ug / ml of puromycin, and 150 ug / ml of hygromycin (1 set). , 50ug / ml (2Set) or 250ug / ml (3Set) to select only completely transformed cell lines. The culture medium was replaced once every two days, and neomycin, puromycin, and hygromycin were treated at different concentrations for about 1 month.

[0094] In addition, three different types of C2C12-PHF20 / YY1-promoter-GFP cell lines (150ug / ml (1Set), 50ug / ml (2Set), or 250ug / ml (3Set)) were collected in a 96-well white cell line. 0.5×10 on plate 5 After culturing for 24 hours and treating the cells at 70% confluence with doxycycline at 0, 50ng / ml, 250ng / ml, 500ng / ml, or 1000ng / ml, the cells were cultured for 24 hours. Cultured for hours. Cells were collected to confirm GFP fluorescence depending on the concentration of doxycycline, and fluorescence was confirmed using GloMax Microplate readers Explorer (Promega, Madison, Wisconsin, USA). After setting the Extension value to 435-488, measurements were taken to quantify the degree of GFP emission.

[0095] In addition, for Western blot analysis, C2C12 cells were left on ice, washed twice with cold PBS, and treated with cell lysis buffer (50 mmol / L Tris-HCl, pH 7.5, 1% (v / v) Nonidet PBS. -40, 250mmol / L NaCl, 0.1mmol / L phenyl methyl sulfonyl fluoride, 0.1mmol / L sodium vanadate, 20mmol / L β-glycerol phosphate, 2mmol / L DTT, 1mmol / L Leupeptin and 10mmol / L PNPP) at 4°C. , and allowed to dissolve for 30 minutes. The cell lysate was then centrifuged at 16,000 xg for 20 minutes at 4°C. The supernatant was collected and used for SDS-PAGE, and the protein content was evaluated by bovine serum albumin protein analysis. Proteins were mixed with sample buffer containing β-mercaptoethanol and heated at 100 °C for 2 min. 40 μg of each cell lysate was fractionated by SDS-PAGE on a 10% polyacrylamide gel and transferred to a nitrocellulose membrane. After blocking for 1 hour at room temperature with 5% skim milk dissolved in TBS (tri-buffered saline) containing 0.02% Tween 20, the mixture was reacted with primary antibody (1:1000 dilution) overnight at 4°C. Actin (1:5000 dilution) was used as a volume control group. After incubation with primary antibody, incubate with goat anti-mouse or anti-rabbit HRP (horse radish peroxidase)-conjugated antibody diluted 1:2000 in TBS / Tween-20 containing 5% skim milk for 1 hour at room temperature. Blots were washed 4 times with TBS / Tween-20 before reacting for 2 hours. After washing with TBS / Tween-20, the blot was used for antigen detection using an enhanced chemiluminescence system. Proteins were visualized with an ECL-chemiluminescence kit (GE Healthcare, Life Sciences).

[0096] As shown in Figures 2a and 2b, the results of Example 1-1 using a fluorescence microscope and Western blot analysis on cell lines confirmed that the strength of the invention increased in a doxycycline dose-dependent manner. . As a result of a comparative analysis of the luminescence level under each condition, it was found that the condition of treating hygromycin at 250ug / ml was the most efficient, and the experiment was fixed under the condition of treating doxycycline at 250ng / ml. It progressed.

[0097] 1-3. Confirmation of effectiveness by doxycycline concentration and treatment time The C2C12-PHF20 / YY1-promoter-GFP cell line was treated with doxycycline at 50 ng / ml or 250 ng / ml, and after 24 hours, the degree of green light emission was confirmed using a fluorescence microscope.

[0098] In addition, the C2C12-PHF20 / YY1-promoter-GFP cell line was treated with 250 ng / ml of doxycycline for 12, 24, 36, and 48 hours, respectively, using GloMax Microplate readers Explorer (Promega, Madison, Wisconsin, USA). The degree of green light emission was confirmed.

[0099] As a result, as shown in FIG. 3, it was confirmed that under the condition of a fixed doxycycline concentration (250 ng / ml), the degree of luminescence increased as the reaction time increased.

[0100] Example 2: Candidate substance screening 2-1.Candidate substance screening In order to screen substances using the HTS system constructed in Example 1, we purchased an FDA-approved drug library (APExBIO, Discovery Probe) and confirmed the degree of GFP inhibition for a total of 1670 compounds.

[0101] Specifically, 0.5 × 10 of the C2C12-PHF20 / YY1-promoter-GFP cell line according to Example 1 was placed in a 96-well plate. 5 The cells were cultured for 24 hours and 70% confluent cells were treated with doxycycline at 250 ng / ml. After 24 hours, each compound was treated at a final concentration of 10 uM. After 24 hours of compound treatment, the degree of green luminescence was confirmed using GloMax Microplate Readers Explorer (Promega, Madison, Wisconsin, USA).

[0102] Intensity of fluorescence: 4 x 10 C2C12-PHF20 / YY1-promoter-GFP cells in a 6-well culture plate. 5 and cultured for 24 hours. Thereafter, 70% confluent cells were treated with doxycycline at 250 ng / ml for 24 hours, and then treated with sulfamethazine, ciclopirox, or IOX1 at a final concentration of 10 uM for 24 hours. Thereafter, the degree of green luminescence was confirmed using GloMax Microplate Readers Explorer (Promega, Madison, Wisconsin, USA).

[0103] As a result, 13 compounds with the effect of reducing green fluorescence were found: Sulfasalazine, Sulfamethazine, Sulfathiazole, Sulfapyridine, Sulfaphenazole, Sulfameter, Sulfamethizole, Sulfaguanidine, Sulfacetamide sodium, Sulfadoxin, Sulfadimethoxine, Sulfanilamide and Sulfadiazine ( Sulfadiazine) was derived as a candidate substance (Figure 4).

[0104] 2-2. Quantitative evaluation of ability to inhibit myoblast differentiation The effectiveness of sulfamethazine, one of the candidate substances, was confirmed. Sulfamethazine was purchased from Sigma (catalog No. S8876), dissolved in DMSO at 10 mM, and used for subsequent cell experiments.

[0105] In order to quantitatively measure the ability of sulfamethazine to inhibit myoblast formation by concentration, the HTS system constructed in Example 1 was treated with 1 nM, 10 nM, 100 nM, 1 μM, or 10 μM of sulfamethazine, respectively. green fluorescence was measured.

[0106] As a result, as shown in Figure 6, it was confirmed that the higher the concentration of sulfamethazine treated, the weaker the fluorescence intensity was. This is because sulfamethazine acts on PHF20 to inhibit myoblast formation. It means having the ability to suppress the expression of , and means that the suppressing ability improves in a concentration-dependent manner.

[0107] In addition, the IC of sulfamethazine was determined based on the green fluorescence intensity obtained in the experiment described above. 50 The value was confirmed to be 0.591uM (Figure 6).

[0108] In addition, C2C12 muscle cell line was treated with each compound at different concentrations for 24 hours, and protein expression (PHF20, YY1.MyoD1) was confirmed through Western blotting. It was confirmed that the protein expression of PHF20 and YY1 decreased and the expression of MyoD increased in the groups treated with each compound (Figure 7).

[0109] 2-3.MF20 (Anti-Myosin) staining Place sterile coverslips in a 12-well plate, 1 x 10 per well. 5 C2C12 / Tet-On PHF20 cells were plated and then grown at 37°C. Cells were treated with doxycycline for 24 hours. After 24 hours, cells were differentiated with DM (2% HS) for 5 days, and on the 4th day of differentiation, each sulfonamide compound was treated at different concentrations. Thereafter, on day 5 of differentiation, the cells were washed twice with PBS at 37°C, reacted with 4% paraformaldehyde for 1 hour, and fixed on a coverslip. Cells were then washed twice with PBS. Cells were reacted with Triton x-100 in PBS for 30 minutes before blocking. Coverslips were blocked with 1% BSA for 1 hour at room temperature with shaking. Anti-Myosin (MF20) antibody was added to 1% BSA (1:200) and reacted overnight at 4°C. The coverslips were then washed three times each with PBS, and Alexa fluor 568 secondary antibody (1:1000) in 1% BSA was added and allowed to react for 1 hour with mixing in the dark at room temperature. Thereafter, the coverslips were washed three times with PBS, mounted on slides using mounting medium containing DAPI VECTASHIELD (St.Louis, USA), and images of the stained cells were taken using a Zeiss. This is shown in Figures 8a to 8c.

[0110] Specifically, Figure 8a shows that the C2C12-PHF20 inducible cell line was treated with doxycyclin to differentiate into myotubes from a group in which PHF20 was overexpressed and a control group not treated with doxycyclin. , muscle function and development on day 5 of differentiation after drug (sulfamethazine, sulfathiazole, sulfadiazine, sulfadoxine, sulfadimethoxine, sodium sulfacetamide, sulfaphenazole, sulfameter) treatment on day 4 of differentiation. This is an image of cell immunofluorescence staining to confirm the differentiation ability of muscle cells after staining with MF20 (heavy chain of myosin II), which is involved in muscle cells.

[0111] Figure 8a shows cellular immunofluorescence to confirm the differentiation potential of muscle cells when treated with sulfamethazine, sulfathiazole, sulfadiazine, sulfadoxine, sulfadimethoxine, sodium sulfacetamide, sulfaphenazole, and sulfameter. Figure 8b is a cell immunofluorescence staining image to confirm the differentiation ability of muscle cells when treated with sulfamethazine, sulfapyridine, sulfanilamide, and sulfamethizole; This is an image of cell immunofluorescence staining to confirm the differentiation ability of muscle cells when treated with different concentrations.

[0112] Example 3: Confirmation of effectiveness in elderly mouse model 3-1. Elderly mouse model construction and treadmill adaptation experiment Thirty-two 48-week-old C57BL / 6J male mice were obtained from KBSI and kept at a constant room temperature and a 12-h night / day cycle for 2 weeks before the experiment, and the mice were fed a standard rodent diet and had free access to water. I decided to ingest it.

[0113] Treadmill adaptation was performed randomly for 3 days at Grade 0, speed (cm / sec) 15, 10 minutes, electrical stimulation 02AM (when electrical stimulation occurs 4 or more times in a row, it is defined as weakness), and depending on the treadmill results. Mice with similar abilities were distributed into each group and divided into 4 groups of 8 mice per group.

[0114] 3-2. Confirmation of treadmill exercise power depending on sulfasalazine dosage in elderly mouse model In order to confirm the treadmill exercise power depending on the dose of sulfasalazine, the mice divided in the above 3-1 were treated with group 1 with PBS (Hyclone Dulbecco's Phaosphate Buffer saline Cat.SH30028.02) and group 2 with sulfasalazine (Sigma, Cat. Nr S0883.CAS Number 599-79-1) 5mpk, Group 3 sulfasalazine 50mpk, Group 4 sulfasalazine 500mpk were orally administered at 10:30 every day for 28 days. To reduce errors in treadmill exercise performance, starve for 3 hours before treadmill measurement, and then run the treadmill four times at 7-day intervals (0 days, 7 days, 14 days, 28 days) according to Table 1 below. did.

[0115]

table 1

[0116] The mice's weights were measured for a total of 28 days to see if there were any changes. As shown in Figure 9, no differences in body weight between groups were observed during the entire experimental period.

[0117] As a result of checking the treadmill exercise force, as shown in Figure 10, it was confirmed that the higher the dose of sulfasalazine, the more the treadmill exercise force of the mice increased ((*)p≦0.05 vs G1, (**) p≦0.01 vs G1, (***)p≦0.001 vs G1).

[0118] 3-3. Confirmation of recovery of rotarod balance ability and grip strength by administration of sulfasalazine in elderly mouse model

[0119] In order to confirm the rotarod balancing ability and grip strength recovery effect by sulfasalazine dosage, 50-week-old C57BL / 6J male mice were administered PBS Group 1, sulfasalazine 5 mg / kg Group 2, sulfasalazine 50 mg / kg Group 3, and sulfasalazine 500 mg / kg. The animals were divided into Group 4 and orally administered at 10 am every day for 28 days, and their body weights were measured. The rotarod was measured on days 0, 7, 14, 21, and 28 of the animal groups constructed in Group 2 under the conditions shown in Table 2 below, and the results are shown in Figure 11 ((*) p≦0.05 vs G1). Grip strength was measured on 2nd and 4th paws of mice on days 0, 9, 16, 23, and 28 in the same animal group as the rotarod, and the results are shown in FIGS. 12a and 12b.

[0120] [Table 2]

[0121] As shown in FIGS. 11a and 11b, it was confirmed that the recovery of the balance ability of mice using rotarod improved as the sulfasalazine dose increased and the administration day became longer. In addition, as shown in Figures 12a and 12b, in both 2paw and 4paw measurements, an improvement in grip strength was observed with an increase in the sulfasalazine dose compared to the control group, Group 1. We were able to confirm that grip strength improved as the days got longer.

[0122] Example 4: Effect confirmed in high-fat diet CTX-induced sarcopenia mouse model 4-1. High-fat diet CTX-induced sarcopenia mouse model construction and weight confirmation To establish a high-fat diet CTX-induced sarcopenia mouse model, 23-week-old C57BL / 6J male mice on 60% HFD for 19 weeks were treated with cardiotoxin (LATOXAN, Portes-les-Valence, France). A highly concentrated stock solution was prepared by dissolving 1 mg / ml in PBS. To apply to mice, dilute it again in PBS (Hyclone Dulbecco's Phaosphate Buffer saline Cat.SH30028.02) at a final concentration of 0.03 mg / ml, and use a 1 ml syringe (24G) to inject the quadriceps muscles ( A high-fat diet CTX-induced sarcopenia mouse model was established by intramuscular injection into the CTX quadriceps muscle.

[0123] Group 1: PBS was orally administered to high-fat diet mice without CTX injection; Group 2: PBS was orally administered to high-fat diet mice injected with CTX; sulfasalazine 5 mg / kg orally to high-fat diet mice injected with CTX. Group 3 was administered with sulfasalazine 50 mg / kg orally to high-fat diet mice injected with CTX, Group 5 was orally administered sulfasalazine 500 mg / kg to high-fat diet mice injected with CTX, and the group was orally administered at 10 a.m. every day. After administration for 14 days, body weights were measured, and the results are shown in FIG.

[0124] As shown in FIG. 13, no differences in body weight between groups were observed during the entire experimental period.

[0125] 4-2. Confirmation of rotarod balance ability and grip strength recovery ability of high-fat diet CTX-induced mice Measurement before injecting CTX into the sarcopenic mouse constructed in 4-1 above is set as day 0, and on days 4, 8, and 12 after injecting CTX into the muscle, perform the measurement under the conditions shown in Table 3 below. Measurements were made using a rota rod (Harvard & Panlab, LE8205), and the results are shown in FIG. In the same animal group as the rotarod, the measurement before CTX injection was taken as day 0, and on the 5th, 9th, and 13th days, grip strength was measured on the 2nd and 4th paws of the mice, and the results were reported. Shown in Figures 15a and 15b ((*)p≦0.05 vs G2).

[0126] [Table 3]

[0127] As shown in FIG. 14, it was confirmed that the recovery of the balance ability of the mice on the rotarod improved as the sulfasalazine dose increased and the day of administration became longer than in the CTX-injected group.

[0128] In addition, as shown in Figures 15a and 15b, in both 2- and 4-paw measurements, an increase in sulfasalazine dose resulted in an improvement in grip strength compared to the CTX injection group, and the longer the day of administration, the better the grip strength. I was able to confirm that.

[0129] Example 5: Effect confirmed in mouse model of sarcopenia caused by Velcro injury 5-1. Construction of sarcopenia mouse model due to Velcro injury The left leg of an 8-week-old C57BL / 6J male mouse was anesthetized with isoflurane, a 5 cm sports tape was wrapped around the entire leg from above the ankle, and then a 10 mm wide Velcro was wrapped to secure it. The condition of the Velcro was checked every day (Figure 16). Meanwhile, in the present invention, Velcro injury means that muscles are wrapped with Velcro so that they cannot move, and such Velcro causes muscle damage.

[0130] 5-2. Confirmation of treadmill exercise power according to sulfasalazine dosage in mouse model of sarcopenia caused by Velcro injury To confirm the amount of treadmill exercise depending on the dose of sulfasalazine, Group 1 group (n=5) in which PBS was orally administered to uninjured 8-week-old C57BL / 6J male mice, and Group 2 group in which PBS was orally administered to mice with Velcro injuries. (n=5), Group 3 group (n=5) in which sulfasalazine 5 mg / kg was orally administered to Velcro injured mice, Group 4 group (n=5) in which sulfasalazine 50 mg / kg was orally administered to Velcro injured mice, Velcro injured mice The animals were divided into Group 5 groups (n=5) to which 500 mg / kg of sulfasalazine was orally administered at 10 a.m. every day for 14 days, and their body weights were measured. The results are shown in FIG. After 14 days, undo the Velcro and measure the treadmill on days 0, 3, 6, 10, and 14, starve for 3 hours before measuring, warm up at 15 cm / sec for 2 minutes, and change to 25 cm / sec. After that, the speed was increased at 15 cm / sec every minute until it became weak (Table 4).

[0131] [Table 4]

[0132] The mice's weights were measured for a total of 14 days to see if there were any changes.

[0133] As shown in FIG. 17, no difference in body weight between groups was observed during the entire experimental period.

[0134] To confirm the treadmill locomotor force of Velcro-injured mice and non-Velcro-injured mice, we compared the distance they ran on the treadmill on the first day after unwrapping the Velcro, as shown in Figure 18a. We confirmed that the Velcro-wrapped mice sustained the same Velcro injury in each group (Group 2 to Group 5) compared to the mice without (Group 1), and as shown in Figure 18b, the mice wrapped in Velcro were injured in the same manner as in the mice wrapped in Velcro (Group 1). Even when comparing the time spent running on the treadmill, we were able to confirm the same results as the distance run on the treadmill.

[0135] In addition, as shown in Figure 19a, as a result of comparing the distance run on the treadmill on the day when the Velcro was untied, on the 3rd, 6th, 10th, and 14th, compared to the comparison group where the Velcro was not wrapped, It was confirmed that the higher the dose of sulfasalazine was administered after the 6th day of administration and the longer the administration period, the more the distance run on the treadmill increased.

[0136] In addition, as shown in Figure 19b, as a result of comparing the time spent running on the treadmill on the day when the Velcro was untied, on the 3rd, 6th, 10th, and 14th, compared to the comparison group where the Velcro was not wrapped, After 6 days of administration, it was confirmed that the higher the dose of sulfasalazine and the longer the administration period, the more time spent running on the treadmill ((*)p≦0.05, (**)p≦0.01, (***) )p≦0.001).

[0137] Example 6: Effect confirmed in sarcopenia mouse model 6-1.Confirmation of muscle regeneration effect in CTX-induced sarcopenia mouse model The experimental animals were 7-week-old male and female C57BL / 6J mice weighing 20±3 g from DOOYEOL Bio (Seoul, South Korea). The mice were housed under constant room temperature and a 12 h night / day cycle for one week prior to the experiment, and the mice were fed a standard rodent diet and had free access to water.

[0138] To construct a CTX-induced sarcopenia mouse model, a highly concentrated stock solution was prepared by dissolving cardiotoxin (LATOXAN, Portes-les-Valence, France) in PBS at 1 mg / ml. Had made. For application to mice, it was diluted again in PBS to a final concentration of 0.03 mg / ml and injected intramuscularly into both thigh muscles of the mice using a 1 ml syringe (24G) to construct a sarcopenia mouse model.

[0139] Thereafter, in an experiment to confirm the effect of the sulfonamide compound of the present invention, the efficacy was confirmed using a CTX-induced sarcopenia mouse model.

[0140] 6-2. Muscle regeneration effect confirmed in CTX-induced sarcopenia mouse model 4uM sulfamethazine was orally administered to the mouse model described in 6-1 (24 hours after CTX administration), muscle tissues were collected from the mice on days 3, 6, and 10, and muscle differentiation was determined through H&E staining. I checked the extent.

[0141] Specifically, for mouse muscle tissue immunostaining, the muscle tissues of the comparison group and the sulfamethazine-treated group were fixed in 4% paraformaldehyde, washed, and then fixed in paraffin. After cutting the tissue to a thickness of about 4 mm, fixing it on a slide, it was stained using antibodies specific to muscle types.

[0142] As shown in Figure 20, as a result of confirming the degree of muscle differentiation of mouse muscle tissues through H&E staining, it was confirmed that the muscles in the control group (sulfamethazine non-administered group) were damaged by CTX. It was confirmed that muscles were regenerated in the group treated with sulfamethazine orally compared to the control group. In particular, it can be seen that muscle regeneration occurs from the 3rd day after sulfamethazine treatment, and that muscle regeneration occurs rapidly on the 6th and 10th days, and at the same time, the muscles become denser.

[0143] 6-3. Confirmation of the effects of sulfamethazine by muscle type In the same manner as in 6-2 above, 4 uM sulfamethazine was administered to mice, and muscle tissue was collected from the mice on the 3rd, 6th, and 10th day. The muscle tissue samples were tested for myosin type I (myosin type I, DSHB BA-75), myosin type IIa (myosin IIa, DSHB, BF-F3), and myosin type IIb (myosin type IIb, DSHB, SC-71), respectively. Confocal microscopy (confocal, Leica , Wetzlar, Germany).

[0144] As shown in Figure 21, an increase in the expression of green myosin IIa and red myosin IIb was observed in the sulfamethazine-treated group compared to the control group, and on day 6, the expression of blue-stained myosin I increased. Images on day 10 were observed to recover to be similar to day 0 of the normal control group. This means that recovery from muscle damage in the sulfamethazine-treated group was faster than in the CTX-induced muscle-depleted mice (sulfamethazine-untreated group).

[0145] 6-4. Confirmation of balance ability of Rota Rod by sulfamethazine dosage In order to confirm the ability to balance depending on the dose of sulfamethazine, 7-week-old male mice were divided into groups of 10 each, experimental groups were set up as shown in Table 5, and the experiment was conducted. Specifically, the rotarod was measured before CTX injection, and after CTX injection (day 0), the drug was orally administered once / day for 10 days, and on days 4, 9, and 14. The rotarod was measured on each day. The measurement conditions of the rotarod are as shown in FIG. 23.

[0146] [Table 5]

[0147] The mice's weights were measured for a total of 14 days to see if there were any changes. As shown in Figure 22, no changes in body weight were observed for each group during the entire experimental period, except for a decrease in body weight in all groups on day 10 when no food was provided.

[0148] As a result of confirming the balance ability of the rotarod, as shown in Figures 24a and 24b, the higher the dose of sulfamethazine, the more the mouse rotarod It was confirmed that the equilibrium capacity of In the group in which muscle loss was not induced ((-)PBS group), the rotarod balance ability improved to a level that was observed due to repeated exercise, but in the sulfamethazine administration group, the balance ability improved in a dose-dependent manner. improved.

[0149] 6-5. Confirmation of grip strength recovery due to sulfamethazine administration In order to confirm the recovery of grip strength due to the dose of sulfamethazine, 7-week-old male mice were divided into groups of 10 mice each, experimental groups were established as shown in Table 6, and the experiment was conducted. Specifically, grip strength was measured before CTX injection, and after CTX injection (day 0), the drug was orally administered once / day for 10 days, and on days 4, 9, and 14. Grip strength was measured on each day.

[0150] [Table 6]

[0151] The mice's weights were measured for a total of 14 days to see if there were any changes. As shown in Figure 22, no changes in body weight were observed for each group during the entire experimental period, except for a decrease in body weight in all groups on day 10 when food was not provided.

[0152] As a result of confirming the grip strength recovery effect, as shown in Figures 26a to 26d, the motor strength of the CTX-injected hind legs (2 paws) was significantly lower than that of the PBS-administered group ((+)PBS) after CTX injection. It was confirmed that the higher the dose of sulfamethazine was, the more the grip strength of the mice increased compared to the sulfamethazine group, and the results of measuring the grip strength of all four legs also showed that it increased in a sulfamethazine concentration-dependent manner. Similar to the rotarod group without muscle loss induction ((-)PBS group), which showed a repeated level of improvement in balance ability, the sulfamethazine treatment group showed a dose-dependent increase in grip strength. It was confirmed that recovery was improved.

[0153] Example 7: Single oral administration toxicity confirmation 7-1. Single oral administration toxicity confirmation Seven-week-old male and female C57BL / 6J mice weighing 20±3 g were obtained from DOOYEOL Bio (Seoul, South Korea). The mice were housed under constant room temperature and a 12 h night / day cycle for one week prior to the experiment, and the mice were fed a standard rodent diet and had free access to water. The experiment was divided into a control group and a test group (100 mg / kg, 500 mg / kg, 1000 mg / kg), and sulfamethazine was diluted with 12.5% ​​DMSO and 12.5% ​​cremophor and administered orally at 10 ml / kg. Control group: PBS (no sulfamethazine administered) Group 1: 100mg / kg (experimental group, sulfamethazine administration) Group 2: 500mg / kg (experimental group, sulfamethazine administration) Group 3: 1000mg / kg (experimental group, sulfamethazine administration)

[0154] The experimental period lasted for 2 weeks, and the general condition and presence or absence of death were observed at least once a day. Body weights were measured before administration and on days 1, 3, 7, and 14 after administration, and necropsy was performed on the 14th day.

[0155] In the present invention, a single oral administration toxicity test is a qualitative and quantitative examination of the toxicity that appears within a short period of time when a test substance is administered to a test animal in a single dose (including cases of divided administration within 24 hours). This refers to the test to be performed.

[0156] Specifically, Figure 27a shows a whole mouse for confirming the toxicity of a single oral administration of female and male mice, and Figure 27b shows a photograph of a male mouse before organ removal. Figure 27c shows a photograph of a female mouse before organ removal.

[0157] In addition, all organs of the head, thoracic cavity, and abdominal cavity were observed, and these are shown in FIGS. 28a to 39.

[0158] Specifically, FIG. 28a is a photograph of a liver of a male mouse dissected and toxicity confirmed, and FIG. 28b is a photograph of a liver of a female mouse dissected and toxicity confirmed. Figure 29a is a photograph of the lungs of a male mouse dissected and toxicity confirmed; Figure 29b is a photograph of the lungs of a female mouse dissected and toxicity confirmed; Figure 31 is a photograph of the brains of male and female mice dissected to confirm toxicity. Figure 31 is a photograph of the hearts of male and female mice dissected and toxicity confirmed. Figure 33 is a photograph of the stomachs of male and female mice dissected and toxicity confirmed; Figure 33 is a photograph of the pancreas of male and female mice dissected and toxicity confirmed; Figure 34 is a photograph of the spleen of male and female mice dissected to confirm toxicity, and Figure 35a is a photograph of the kidney of a male mouse dissected to confirm toxicity. 35b is a photograph of a female mouse kidney (Kideny) dissected and toxicity confirmed; Figure 36a is a male mouse small intestine (S.intestine) dissected and toxicity confirmed; , Fig. 37a is a photograph of dissecting the small intestine (S.intestine) of a female mouse and confirming toxicity, Figure 37a is a photograph of dissecting the large intestine (L.intestine) of a male mouse and confirming toxicity. , is a photograph of the large intestine (L.intestine) of a female mouse dissected and toxicity confirmed; Figure 38 is a photograph of a male mouse testis (testis) dissected and toxicity confirmed; This is a photograph showing the toxicity of a dissected mouse womb.

[0159] Figure 40 shows the results of measuring the body weights of male and female mice in each group on the 1st, 3rd, 7th, or 14th day after oral administration of PBS or sulfamethazine. The results of measuring the weight of each tissue of each male mouse are shown in FIG. 41a, and the results of measuring the weight of each tissue of female mice are shown in FIG. 41b. There was no peculiarity in the weight of the tissues in each group, and it was confirmed that the decrease in the weight of the female pancreas in the 500mpk group was due to the disappearance of some tissues during dissection.

[0160] 7-2. Histochemical evaluation of toxicity after single oral administration Mice that received a single oral administration as in Experimental Example 7-1 were dissected on the 14th day after administration, and histochemical toxicity was evaluated. The liver was pathologically observed through H&E staining of the dissected liver tissue.

[0161] As shown in Figures 42a and 42b, no toxicity was observed in the isolated livers of male and female mice, and no specific pathological findings were observed in the livers of each group. .

[0162] 7-3. Confirmation of hepatotoxicity, cardiac toxicity, and renal toxicity in blood As in Experiment 7-1, mice were given a single oral dose, and on the 14th day after administration, blood was collected from the heart of the mouse, serum was collected, and hepatotoxicity markers, cardiotoxicity markers, and nephrotoxicity markers were determined. The expression of each was confirmed. Specifically, blood samples were collected at the heart post-surgery on day 14 after administering sulfamethazine. The collected blood was left at room temperature for 1 hour to coagulate, and then a centrifuge (13,000 rpm) was operated for 20 minutes to separate serum. Blood analysis was conducted separately for hepatotoxicity, cardiotoxicity, and renal toxicity, and the analysis was carried out by DOOYEOL Biotech (Seoul, Korea).

[0163] As a result, as shown in FIGS. 43a and 43b, no specific phenomena related to hepatotoxicity, cardiotoxicity, and renal toxicity in male and female mice were observed in the sulfamethazine administration group.

Claims

1. A pharmaceutical composition for the prevention or treatment of muscle disease comprising a sulfonamide compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 In the above formula 1, R １ is hydrogen, *-(C=N)-NH ２ acetyl, or C ５－６ The aryl or heteroaryl is, and the R １ Is it a non-substitution, or C １－３ Alkyl, C １－３ Substituted with at least one substituent selected from alkoxy and phenyl, the R ２ is, -NH ２ is or, R1 is 2-pyridinyl, and R2 is as shown in formula 1-1 below: 【Chemistry 2】 It is a compound represented by [this symbol].

2. The aryl or heteroaryl includes thiazole, diazole, thiadiazole, phenyl, pyridine or pyrimidine, and the aryl or heteroaryl is unsubstituted or substituted with at least one substituent selected from C １－３ alkyl, C １－３ alkoxy, and phenyl, and is the pharmaceutical composition according to claim 1.

3. Said C １－３ Alkyl and C １－３ The pharmaceutical composition according to claim 1 or 2, wherein at least one substituent selected from among alkoxys is methyl or methoxy.

4. The pharmaceutical composition according to claim 1, wherein the aforementioned chemical formula 1 is represented by any one of the following chemical formulas 2 to 14: 【Transformation 3】 【Chemistry 4】 【Transformation 5】 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】

5. The pharmaceutical composition according to claim 1, wherein the muscle disease is one selected from the group consisting of sarcopenia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, cachexia, and myasthenia gravis.

6. Food compositions for the prevention or improvement of muscle diseases, comprising a sulfonamide compound represented by the following formula 1 or a food-grade salt thereof: 【Chemistry 16】 In the above formula 1, R １ is hydrogen, *-(C=N)-NH ２ acetyl, or C ５－６ The aryl or heteroaryl is, and the R １ Is it a non-substitution, or C １－３ Alkyl, C １－３ Substituted with at least one substituent selected from alkoxy and phenyl, the R ２ is, -NH ２ is or, R1 is 2-pyridinyl, and R2 is as shown in formula 1-1 below: 【Chemistry 17】 It is a compound represented by [this symbol].

7. The aryl or heteroaryl comprises thiazole, diazole, thiadiazole, phenyl, pyridine, or pyrimidine, and the aryl or heteroaryl is unsubstituted or C １－３ Alkyl, C １－３ The food composition according to claim 6, which is substituted with at least one substituent selected from alkoxy and phenyl.

8. Said C １－３ Alkyl and C １－３ The food composition according to claim 6 or 7, wherein at least one substituent selected from among alkoxys is methyl or methoxy.

9. A feed composition for the prevention or improvement of muscle disease containing a sulfonamide compound represented by the following formula 1 or a salt thereof: [Chemistry 18] In the above formula 1, R １ is hydrogen, *-(C=N)-NH ２ acetyl, or C ５－６ The aryl or heteroaryl is, and the R １ Is it a non-substitution, or C １－３ Alkyl, C １－３ Substituted with at least one substituent selected from alkoxy and phenyl, the R ２ is, -NH ２ is or, R1 is 2-pyridinyl, and R2 is as shown in formula 1-1 below: 【Chemistry 19】 It is a compound represented by [this symbol].

10. The aryl or heteroaryl comprises thiazole, diazole, thiadiazole, phenyl, pyridine, or pyrimidine, and the aryl or heteroaryl is unsubstituted or C １－３ Alkyl, C １－３ The feed composition according to claim 9, which is substituted with at least one substituent selected from alkoxy and phenyl.

11. Said C １－３ Alkyl and C １－３ The feed composition according to claim 9 or 10, wherein at least one substituent selected from among alkoxys is methyl or methoxy.

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