A composition for the treatment of cancer cachexia containing diflunisal or a pharmaceutically acceptable salt thereof.

Abstract

The present invention relates to a composition for the improvement and treatment of muscle diseases and cachexia, wherein diflunisal has the effect of promoting the differentiation and proliferation of myogenic cells, and further has the effect of improving and treating cachexia induced by chronic diseases such as muscle diseases or cancer through effects such as increased muscle mass and recovery of muscle strength.

Description

[Technical Field]

[0001] The present invention relates to a composition for the improvement and treatment of cachexia, particularly cancer cachexia, comprising the bisteroid analgesic diflunisal or a pharmaceutically acceptable salt thereof. [Background technology]

[0002] Cachexia is a condition characterized by persistent muscle loss due to various diseases, including cancer, heart failure, and chronic illnesses. Cancer cachexia, in particular, is a complex metabolic syndrome associated with cancer. It refers to a state of systemic malnutrition in cancer patients where, even with normal dietary intake, the body's utilization of nutrients is limited, or abnormal metabolism leads to irreversible loss of body mass (weight, muscle, etc.). It is typically defined as a loss of 5% or more of body weight due to muscle and fat loss within six months of the onset of cancer. While it does not occur in all cancer patients, it has been reported that approximately 50-60% of patients with gastrointestinal, pancreatic, lung, and colorectal cancer exhibit cancer cachexia.

[0003] The exact cause of cancer cachexia is not fully understood, but inflammatory cytokines such as IL-6, TNF-α, and CRP1 (C-reactive protein 1) are known to play a central role in muscle and fat loss. There is no specific treatment for cancer cachexia; however, treatment can indirectly increase food intake by improving pain and gastrointestinal disorders, or by using appetite stimulants. Recently, drugs such as the COX-2 inhibitor celecoxib and the TNF-α inhibitor thalidomide have been used, but celecoxib has been reported to have side effects such as anemia, gastric ulcers, allergies, heart attacks, and strokes, while thalidomide has been reported to have side effects such as depression, heart failure, dyspnea, vomiting, rash, hypertension, and birth defects during pregnancy. Moreover, there is a lack of clear evidence of therapeutic efficacy.

[0004] The inventors of this invention completed the present invention after conducting research on a drug that has therapeutic effects on cachexia, particularly cancer cachexia, has few side effects, and can be safely administered to patients. [Overview of the Initiative] [Problems that the invention aims to solve]

[0005] The inventors of this invention screened various drugs, particularly NSAIDs, for use in treating muscle diseases, especially cachexia. They confirmed effects related to muscle diseases and cachexia, such as increased body weight and muscle mass, and improved muscle strength. As a result, they found that diflunisal was excellent in improving and treating the symptoms associated with the aforementioned muscle diseases or cachexia, thus completing the present invention.

[0006] Therefore, the object of the present invention is to provide a composition for the prevention or treatment of muscle disease comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0007] Another object of the present invention is to provide a pharmaceutical composition for the improvement or treatment of cachexia comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0008] A further object of the present invention is to provide a muscle-building or muscle-strengthening composition comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0009] A further object of the present invention is to provide an anticancer adjuvant composition comprising diflunisal or a pharmaceutically acceptable salt thereof. [Means for solving the problem]

[0010] To achieve the aforementioned objectives, the present invention provides a composition for the prevention or treatment of muscle disease comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0011] To achieve another objective of the present invention, the present invention provides a pharmaceutical composition for the improvement or treatment of cachexia comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0012] To achieve another objective of the present invention, the present invention provides a muscle-building or muscle-strengthening composition comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0013] To achieve yet another object of the present invention, the present invention provides an anticancer adjuvant composition comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0014] The present invention will be described in detail below.

[0015] In one aspect of the present invention, the present invention relates to a composition for the prevention or treatment of muscle diseases comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0016] The aforementioned diflunisal is a salicylate derivative, an NSAID (non-steroidal anti-inflammatory drug) class drug, and is widely used as an analgesic, particularly for musculoskeletal disorders. It is the compound shown in formula (1) below.

[0017] [ka]

[0018] Diflunisal is a competitive COX-I and COX-II inhibitor (with high affinity for COX-I) that acts on a mechanism that inhibits the conversion of arachidonic acid in the body into prostaglandin precursors, which cause inflammation and fever. It is used as an anti-inflammatory and analgesic agent.

[0019] In the present invention, diflunisal has preventive, ameliorative, or therapeutic effects on muscle diseases exhibiting symptoms such as decreased muscle function, muscle loss, muscle atrophy, muscle wasting, or muscle degeneration. The term "muscle disease" refers to a state in which muscle strength is weakened due to damage or loss caused by aging or disease, and this can be caused by various factors, including genetic predisposition; age-related diseases such as hypertension, impaired glucose tolerance, diabetes, obesity, dyslipidemia, atherosclerosis, or cardiovascular disease; chronic diseases such as cancer, autoimmune diseases, infectious diseases, AIDS, chronic inflammatory diseases, arthritis, dystrophy, kidney disease, chronic obstructive pulmonary disease, emphysema, rickets, chronic lower spinal pain, peripheral nerve injury, central neuron injury, and chemical injury; loss of exercise due to fractures, trauma, or prolonged bed rest; and aging.

[0020] The "muscle disease" of the present invention is not limited to, but may be one or more muscle diseases selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myoricosis, amyotrophic axonal sclerosis, myasthenia gravis, cachexia, and senile sarcopenia. Specifically, it may be a disease such as senile muscular atrophy, muscle disease due to cancer and chronic disease, or muscle atrophy due to muscle disuse. More specifically, it may include senile muscular atrophy or cancer-induced muscular atrophy, muscular dystrophy, muscle degeneration, myoricosis, amyotrophic axonal sclerosis, myasthenia gravis, cachexia, senile sarcopenia, and muscle loss. In particular, in the present invention, the muscle disease may be one that is induced by cancer or a chronic disease.

[0021] Diflunisal of the present invention has an improving or therapeutic effect on cachexia, especially among muscle diseases. The "cachexia" is a severe systemic wasting syndrome that appears at the end stage of diseases such as cancer, tuberculosis, diabetes, AIDS, etc., and is particularly common in patients with digestive tract cancer and lung cancer. Loss of appetite, weight loss accompanied by muscle and fat loss, and decline in physical strength are the main symptoms, and the weight decreases even with normal food intake, leading to a state of weight loss. Cachexia can reduce the therapeutic effect of the disease and may shorten the patient's life expectancy.

[0022] In the present invention, the cachexia to be improved or treated may be cancer cachexia induced by cancer.

[0023] In the present invention, the cancer that may cause cachexia is not limited to its type, and may include melanoma, leukemia, lymphoma, myeloma, myelodysplastic syndrome, breast cancer, head and neck cancer, esophageal cancer, gastric cancer, colorectal cancer (= colon cancer), rectal cancer, anal cancer, hepatocellular carcinoma, cholangiocarcinoma, gallbladder cancer, pancreatic cancer, lung cancer (non-small cell lung cancer, small cell lung cancer), thymic cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, ovarian cancer, cervical cancer, sarcoma, gastrointestinal stromal tumor, cancer of unknown primary site, mesothelioma, neuroendocrine tumor, skin cancer, hematological cancer, etc., and more preferably, it may be cancer of digestive organs such as gastric cancer, colorectal cancer, rectal cancer, liver cancer, gallbladder cancer, pancreatic cancer, or lung cancer.

[0024] The pharmaceutical composition according to the present invention may improve and treat one or more symptoms caused by cachexia selected from the group consisting of loss of appetite, weight loss, increased fatigue, muscle weakness, decreased motility, muscle loss, fat loss, and hematotoxicity, and preferably may improve and treat weight loss, muscle loss, muscle weakness, or decreased motility.

[0025] In relation to the aforementioned effects, in one embodiment of the present invention, when diflunisal was administered to a cancer cachexia induction model, it was found to have effects on weight gain, muscle mass and strength enhancement, and long-term weight loss of the spleen, liver, heart, etc., was improved compared to the control group in the cachexia induction model. This indicates that diflunisal can be used to improve and treat cancer cachexia.

[0026] In another aspect of the present invention, diflunisal can be used as a composition having muscle-enhancing, muscle-strengthening, or athletic performance-enhancing effects.

[0027] In one embodiment of the present invention, it was confirmed that diflunisal has the effect of promoting the proliferation and differentiation of muscle stem cells. In the embodiment, diflunisal increased the expression of Pax7, a proliferation marker for muscle stem cells, and increased the expression levels of MHC and Myogenin, which are differentiation markers for muscle stem cells. Furthermore, it was confirmed that the diameter of muscle fibers increased when diflunisal was administered. In other words, diflunisal can promote the proliferation and differentiation of muscle stem cells, and as a result, it can have the effect of improving muscle function, such as increased muscle mass, improved muscle strength, enhanced muscle recovery ability, and reduced muscle fatigue. The above effects were confirmed not only in a disease-induced model (cancer cachexia) but also in normal muscle.

[0028] Furthermore, diflunisal can improve athletic performance. "Athletic performance" refers to the ability to perform exercises using muscle strength, and in this case, muscle strength can be improved by increasing muscle mass, muscle endurance, oxidative muscle mass, muscle recovery ability, and intramuscular energy balance, and can also be enhanced by reducing fatigue substances in the muscles. In one embodiment of the present invention, the effect of improving muscle endurance and increasing muscle mass in mice was confirmed, and it was found that athletic performance can be improved by this.

[0029] The compositions of the present invention can be used in a variety of applications, including pharmaceuticals, health functional foods, functional foods, animal feed, and cell culture medium compositions. By promoting the proliferation and differentiation of muscle stem cells and improving muscle mass and muscle endurance, they may have preventive, ameliorative, or therapeutic effects on muscle cell rings and cachexia.

[0030] As used herein, the term "prevention" means all actions that can suppress or delay the onset of a disease by administering the pharmaceutical composition according to the present invention.

[0031] As used herein, the term "improvement" means all actions that can alleviate or delay the onset of symptoms induced by a disease by administering the pharmaceutical composition according to the present invention.

[0032] As used herein, the term "treatment" means all actions by which the administration of the pharmaceutical composition according to the present invention improves or benefits a symptom.

[0033] The pharmaceutical composition of the present invention can be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, and sterile injection solutions, by conventional methods, and may further contain carriers or excipients necessary for such formulation. In addition to the active ingredient, pharmaceutically acceptable carriers, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, magnesium stearate, and mineral oil. When formulation, it is prepared using commonly used fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients.

[0034] For example, solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and such solid preparations are prepared by mixing the extract or compound with at least one excipient (e.g., men, starch, calcium carbonate, sucrose or lactose, gelatin, etc.). In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as humectants, sweeteners, fragrances, and preservatives can be used.

[0035] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Examples of suppository bases include witepsol, macrogol, tween 61, cocoa butter, lauric acid butter, and glycerogenous gelatin.

[0036] The pharmaceutical composition of the present invention can be administered orally or parenterally (intravenous injection, subcutaneously, intraperitoneally, or topically) by a desired method, and the dosage depends on the patient's condition and weight, the severity of the disease and the form of the drug, the route of administration and the time, and a person skilled in the art can select an appropriate form.

[0037] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means a reasonable amount applicable to medical treatment and sufficient to treat a disease, the criteria of which are determined according to the patient's disease, severity, drug activity, sensitivity to the drug, administration time, route of administration and excretion rate, duration of treatment, concomitant components and other factors. The pharmaceutical composition of the present invention can be administered individually or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. Considering all of the above factors, the dosage can be determined at a level that minimizes side effects, which is a level that can be easily determined by those skilled in the art. Specifically, the dosage of the pharmaceutical composition may vary depending on the patient's age, weight, severity, sex, etc., and generally, an amount of 0.001 to 150 mg, preferably 0.01 to 100 mg per kg of body weight can be administered daily or every other day, 1 to 3 times a day. However, this is illustrative, and the dosage can be set as needed.

[0038] Furthermore, the composition of the present invention may be a food or a functional food for health, and in particular, the term "functional food for health" means a food manufactured and processed using raw materials or components that have a useful function for the human body, in accordance with Act No. 6727 on Functional Foods for Health, and "functional" means being taken for the purpose of regulating nutrients with respect to the structure and function of the human body, or obtaining a useful effect for health purposes such as physiological effects.

[0039] The food or health functional food of the present invention can be manufactured and processed as a pharmaceutical administration form such as powder, granules, tablets, capsules, pills, suspensions, emulsions, or syrups, or as a health functional food such as tea bags, infused tea, beverages, candies, jellies, or gums, for the purpose of preventing and improving muscle diseases.

[0040] The food or health functional food composition of the present invention can be used as a food additive and can be manufactured as a product alone or in combination with other components. It may also contain nutrients, vitamins, electrolytes, flavorings, colorings and enhancers, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages. These components can be used alone or in combination, and in appropriate amounts.

[0041] In yet another aspect of the present invention, the present invention relates to a composition for adjunct to anticancer treatment comprising diflunisal. The adjunct to anticancer treatment composition is used adjunct to cancer treatment using known anticancer agents and can be administered simultaneously or sequentially with the anticancer agent as an adjunct to anticancer treatment. [Effects of the Invention]

[0042] The present invention relates to a composition containing diflunisal or a pharmaceutically acceptable salt thereof that has the effect of improving and treating cachexia, particularly cancer cachexia, and the composition is used to improve and treat various symptoms associated with cachexia, such as weight gain, as well as increase muscle mass and muscle endurance and increase exercise performance by promoting the proliferation and differentiation of myobiocytes. [Brief explanation of the drawing]

[0043] [Figure 1] This figure shows the results of a cytotoxicity evaluation (MTT assay) of diflunisal using C2C12 cells, which are muscle stem cells. [Figure 2] This is the result of in vitro confirmation of the muscle cell differentiation-enhancing effect of diflunisal, and is based on MHC fluorescence staining. [Figure 3] This is a result of in vitro confirmation of the muscle cell differentiation-enhancing effect of diflunisal, and the results were obtained by quantitatively comparing the diameter of myotubes. [Figure 4]This document presents the results of in vitro confirmation of the muscle cell differentiation-enhancing effect of diflunisal, along with immunoblot analysis results for MHC and myogenin. [Figure 5a] This study confirmed the stem cell proliferation-enhancing effect of diflunisal administration, and presents the results of BrdU staining and quantification of BrdU+ (BrdU-positive) cells for muscle stem cells (C2C12). [Figure 5b] This study confirmed the stem cell proliferation-enhancing effect of diflunisal administration, and presents the results of BrdU staining and quantification of BrdU+ (BrdU-positive) cells for muscle stem cells (C2C12). [Figure 6] This study confirmed the stem cell proliferation-enhancing effect of diflunisal administration, and the results are from qRT-PCR analysis of Pax7 expression, a stem cell marker. [Figure 7] In mice with muscle damage induced by CTX, the effect of diflunisal administration on promoting muscle stem cell proliferation was confirmed in the tibialis anterior muscle. Figure 8 shows the BrdU stained image and the quantitative results of BrdU+ (benign BrdU) cells. [Figure 8] This study confirmed the effect of diflunisal administration on promoting muscle stem cell proliferation in the tibialis anterior muscle (TA) of mice in which muscle damage was induced with CTX, and the results represent the quantification of BrdU+ (benign BrdU) cells. [Figure 9a] In a model in which muscle atrophy was induced by dexamethasone administration, the effect of administering diflunisal to improve muscle atrophy was confirmed. Figure 9a shows the results of MHC immunostaining, and Figure 9b shows the results of immunostaining analysis for atrogin-1 expression. [Figure 9b] In a model in which muscle atrophy was induced by dexamethasone administration, the effect of administering diflunisal to improve muscle atrophy was confirmed. Figure 9a shows the results of MHC immunostaining, and Figure 9b shows the results of immunostaining analysis for atrogin-1 expression. [Figure 10] This study confirmed the effect of diflunisal on improving muscle atrophy in a model in which dexamethasone administration induced muscle atrophy, and the results are based on qRT-PCR analysis of MuRF1 expression. [Figure 11]This study confirmed the in vitro improvement effect of diflunisal administration on cancer cell culture medium (CM)-induced cachexia, and also confirmed the concentration-dependent myotubular regeneration and improvement effect of diflunisal administration in a cancer cachexia induction model using MHC immunohistochemistry. [Figure 12] This report confirms the in vitro improvement effect of diflunisal administration on cachexia induced by cancer cell culture medium (CM), and the results were quantified by measuring the diameter of the myotubes. [Figure 13] This diagram shows the induction of cancer cachexia in vivo and the confirmation of its effect upon administration of flunisal, and is a schematic representation of the method used for the aforementioned in vivo experiment. [Figure 14] This study involved inducing cancer cachexia in vivo and confirming the effects of diflunisal administration. The results show measurements of body weight, tumor weight, and body weight excluding tumors after diflunisal administration. [Figure 15] This report examines the effects of diflunisal administration on motor function (grip strength) in cancer cachexia induced in vivo. [Figure 16] This shows the results of atrogin-1 and MuRF1 mRNA expression analysis (qRT-PCR) regarding the effects of diflunisal administration on cancer cachexia induced in vivo. [Figure 17] This shows the results of muscle mass measurements for each part of the hind leg, as part of the effects of diflunisal administration on cancer cachexia induced in vivo. [Figure 18] This study examined the ameliorative effects of diflunisal on cancer cachexia induced in vivo, specifically focusing on changes in inguinal white fat (IWAT), visceral fat (EWAT), liver, heart, and spleen weight. [Modes for carrying out the invention]

[0044] [Best mode for carrying out the invention] The following examples will be given in detail to illustrate this specification. However, the examples described herein can be modified in various different forms, and the scope of this specification should not be construed as being limited to the examples described below. The examples herein are provided to give a more complete explanation of this specification to a person of average knowledge of the art.

[0045] Example 1. Culture and differentiation induction of myofigrous cell lines C2C 12 This is a myobiogenic cell line obtained from C3H mice and is widely used in muscle cell differentiation studies. The cells were cultured in a cell culture medium (GM), and muscle cell differentiation was induced in a differentiation medium (DM). The cell culture medium used was DMEM supplemented with 15% fetal bovine serum (FBS), and the differentiation medium used was DMEM containing 2% horse serum.

[0046] To induce muscle cell differentiation, cells were dispensed into cell culture medium and cultured. When the cell density reached approximately 80-90%, the medium was replaced with differentiation medium, and differentiation was induced for approximately 2-3 days.

[0047] Experimental Example 1. Evaluation of cytotoxicity of diflunisal Cytotoxicity was evaluated using the MTT assay with diflunisal (CAS No. 22494-42-4, W06). C2C 12 Myofibrillators were treated with diflunisal at concentrations of 0, 1, 10, 100, 1,000, and 10,000 nM for 18 hours, and cell viability was confirmed by MTT analysis. Myofibrillators (1.0 × 10) were placed on cell culture medium. 4 Cells (per well) were reacted with 5 mg / mL MTT for 4 hours, and the eluted reaction product was then measured by absorbance at 570 nm. As a result, the cells showed almost the same viability as the control group up to a diflunisal concentration of ~10,000 nM used for MTT analysis (Figure 1).

[0048] Experimental Example 2. Confirmation of the stem cell differentiation-enhancing effect of diflunisal. To analyze the effects of diflunisal on muscle stem cell differentiation, MHC immunofluorescence staining was performed (Figure 2). C2C 12 Cell lines were treated with DMSO (control group) and diflunisal (control group) at concentrations of 10, 100, 1,000, and 10,000 nM, respectively. Myotube differentiation was then induced, and the degree of myotube formation was compared and analyzed by immunofluorescence staining using MHC antibodies on day 3 of differentiation induction. The results showed that myotube differentiation was enhanced in a concentration-dependent manner in diflunisal-treated cells compared to the control group. Furthermore, quantification of myotube diameter also confirmed a concentration-dependent increase in diflunisal treatment (Figure 3).

[0049] Furthermore, the degree of expression of differentiation markers was confirmed by immunoblotting. Diflunisal treatment was found to induce an increase in the expression of MHC and myogenin, which are muscle stem cell differentiation markers, and this was found to be at a significant level compared to the positive control group treated with ursolic acid (UA) (Figure 4).

[0050] Experimental Example 3. Confirmation of the stem cell proliferation-promoting effect of diflunisal. BrdU staining experiments were performed to analyze the effects of diflunisal on myofibrillar proliferation. C2C 12 Myobiocytes were treated with either DMSO or diflunisal (1 μM) for approximately 20 hours, then reacted with BrdU for 15 minutes, followed by immunofluorescence staining using an antibody against BrdU (Figure 5a). The results showed increased myobiocyte proliferation in the diflunisal-treated group. Quantitative comparisons also confirmed a significant increase in the proportion of BrdU-positive cells in the diflunisal-treated group compared to the control group (Figure 5b).

[0051] Furthermore, we checked whether diflunisal treatment increased the expression level of Pax7, a muscle growth marker. As a result, as shown in Figure 6, we confirmed that the relative expression level of Pax7 mRNA increased during diflunisal treatment.

[0052] Experimental Example 4. Effect of promoting stem cell proliferation in an animal model of muscle injury. Experiments were conducted to confirm whether diflunisal administration has an effect of enhancing stem cell proliferation in animal models in which actual muscle injury occurs. Male C57BL / 6 mice were orally administered diflunisal at a dose of 0.02 mg / kg for two weeks. On the second week, 10 μM CTX (cardiotoxic: CTX) was injected directly into the muscle at a dose of 2 μL per g of body weight to induce muscle injury. Subsequently, diflunisal was administered at a dose of 0.02 mg / kg for three days. Immunofluorescence staining was then performed on the TA muscle tissue of the mice using a BrdU antibody. As a result, as shown in Figure 7, increased myofibrillar proliferation was confirmed in the group treated with diflunisal. Furthermore, quantitative comparisons confirmed that the proportion of BrdU-positive cells increased by more than 2.5 times in the group treated with diflunisal compared to the control group (Figure 8).

[0053] Experimental Example 5: Effect of improving muscle atrophy To determine whether diflunisal can restore or protect muscles from DEX (dexamethasone)-induced muscle atrophy, C2C 12 Experiments were conducted using myofibril cells. C2C 12 Myotomies were differentiated in DM (differentiation medium) for 2 days, then pretreated with 100 μl of MDEX for 4 hours, treated with diflunisal (1 μM) along with DMSO in a vehicle, and cultured in DM for an additional 24 hours.

[0054] Dexamethasone-treated C2C 12 The degree of myotubular formation was compared and analyzed by performing immunofluorescence staining using MHC antibodies on myotubular cells. As shown in Figure 9, it was confirmed that MHC expression decreased when muscle atrophy was induced by dexamethasone, but recovered when diflunisal was administered. Furthermore, immunoblotting revealed that the expression level of Atrogin-1, a muscle atrophy marker, increased when dexamethasone was administered, but decreased again when diflunisal was administered (Figure 9).

[0055] In addition, in the qRT-PCR analysis results for MuRF1 expression, which is another muscle atrophy marker, it was confirmed that when muscle atrophy was induced, MuRF1 expression increased, but when diflunisal was administered, its expression was significantly decreased (Figure 10).

[0056] Experimental Example 6. Improvement effect on cachexia induced by cancer cell culture medium To confirm the improvement effect of diflunisal on cancer cachexia, cancer cachexia was induced in muscle cells using cancer cell culture medium (CM) (upper part of Figure 11).

[0057] In the cancer cachexia induction model, diflunisal was treated at concentrations of 10, 100, and 1,000 nM, respectively, and the myotube regeneration and improvement effects were confirmed by MHC immunostaining. The diameter of the myotubes was measured to confirm the presence or absence of improvement. As a result, as shown in Figures 11 and 12, when cultured in CM culture medium, it was confirmed that cancer cachexia was induced and the diameter of the myotubes decreased, but when diflunisal was administered, the diameter of the myotubes increased in a concentration-dependent manner. This indicates that diflunisal has an effect of improving muscle loss and muscle contraction caused by cancer cachexia.

[0058] Experimental Example 7. Improvement effect on cancer cachexia in an animal model 7-1) Improvement effect on weight loss due to cancer cachexia Lewis lung cancer (LCC, 5×10 6 cells / mouse) was injected into C57BL / 6 mice to induce lung cancer. Two weeks after injection, diflunisal was orally administered daily at a dose of 0.02 or 0.2 mpk for 21 days (Figure 13). Thereafter, changes in body weight, changes in tumor weight, and changes in body weight excluding the tumor were measured for each mouse. As a result, as shown in Figure 14, there was no significant change in the overall body weight in all experimental groups, but in the cancer induction group, the tumor weight increased, and it was confirmed that the decrease in the change in body weight excluding the tumor was improved in the diflunisal administration group. In particular, the body weight excluding the tumor in the 0.02 mpk diflunisal administration group was significantly increased compared to the cancer induction group (Figure 14).

[0059] 7-2) Improvement of exercise capacity and muscle mass due to cancer cachexia To compare the motor skills of the aforementioned cancer cachexia-induced mouse model, grip strength tests were conducted in each experimental group. As shown in Figure 15, the diflunisal-administered group showed a concentration-dependent increase in grip strength and a significant increase in motor skills compared to the cancer-induced group.

[0060] Furthermore, immunoblotting analysis of the expression levels of Atrogin-1 and MuRF-1, markers associated with muscular dystrophy, revealed an increase in relative mRNA expression during cancer induction. However, administration of diflunisal resulted in a decrease in this expression again (Figure 16).

[0061] Furthermore, changes in muscle mass of the hindlimb muscles—tibialis anterior (TA), extensor digitorum longus (EDL), soleus (SOL), and gastrocnemius (GA)—were examined as shown in Figure 17. It was confirmed that muscle mass of all TA, EDL, SOL, and GAS decreased during cancer induction, and administration of diflunisal significantly improved muscle loss due to cancer cachexia (Figure 17). This confirms the effect of diflunisal in suppressing muscle loss due to cancer cachexia (Figure 17).

[0062] 7-3) Improvement effect on weight loss due to cancer cachexia Furthermore, in a mouse model of cancer cachexia, the effects of diflunisal on non-muscle tissues such as white fat, visceral fat, liver, heart, and spleen were compared by measuring the weight of each tissue. As shown in Figure 18, all models in which cancer was induced showed a decrease in weight, but it was confirmed that administration of diflunisal improved fat and long-term weight loss due to cancer cachexia.

[0063] The present invention has been discussed, focusing on its preferred embodiments. Those with ordinary skill in the art to which the present invention pertains will understand that the present invention can be realized in modified forms that do not depart from its essential characteristics. Therefore, the disclosed embodiments should be considered in an explanatory rather than restrictive manner. The scope of the present invention is defined in the claims, not in the foregoing description, and all differences within that scope should be construed as being included within the present invention.

[0064] [Modes for carrying out the invention] In one embodiment, the present invention relates to a pharmaceutical composition for the improvement or treatment of muscle diseases comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0065] The aforementioned muscle diseases can be selected from the group including atony, muscular atrophy, muscular dystrophy, muscle degeneration, myorticosis, amyotrophic axonal sclerosis, myasthenia gravis, muscle loss, and sarcopenia.

[0066] The aforementioned muscle disease may be induced by aging, decreased muscle function, muscle wasting, muscle degeneration, disuse, or muscle injury.

[0067] In another aspect, the present invention relates to a pharmaceutical composition for the improvement or treatment of cachexia comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0068] The aforementioned cachexia may be cancer cachexia, which is induced by cancer.

[0069] Furthermore, the composition improves and treats one or more symptoms of cachexia selected from the group consisting of loss of appetite, weight loss, increased fatigue, decreased muscle strength, decreased mobility, muscle loss, fat loss, and hematopoietic toxicity.

[0070] In yet another aspect of the present invention, the present invention relates to a muscle-building or muscle-strengthening composition comprising diflunisal or an acceptable salt thereof.

[0071] The aforementioned composition may have an effect of promoting the proliferation or differentiation of muscle stem cells.

[0072] Furthermore, the composition may be selected from the group consisting of pharmaceutical compositions, health functional food compositions, functional food compositions, or animal feed compositions.

[0073] In yet another aspect of the present invention, the present invention relates to an adjunct to anticancer treatment comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0074] The above composition can be administered in combination with one or more anticancer agents.

[0075] In yet another aspect of the present invention, the present invention relates to the use of diflunisal or an acceptable salt thereof for the treatment of muscle diseases.

[0076] The aforementioned muscle diseases are selected from the group including cachexia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, myorticosis, amyotrophic axonal sclerosis, myasthenia gravis, muscle loss, and sarcopenia.

[0077] Furthermore, the aforementioned muscle diseases are induced by aging, decreased muscle function, muscle wasting, muscle degeneration, disuse, or muscle injury.

[0078] In yet another aspect of the present invention, the present invention relates to a method for treating cachexia, comprising the step of administering an effective amount of diflunisal or a pharmaceutically acceptable salt thereof to an individual.

[0079] Furthermore, in the above method, diflunisal or a pharmaceutically acceptable salt thereof is administered to the individual together with one or more anticancer agents.

Claims

1. A pharmaceutical composition for the improvement or treatment of muscle diseases, comprising diflunisal or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition for improving or treating a muscle disease according to claim 1, wherein the muscle disease is selected from the group including atony, muscular atrophy, muscular dystrophy, muscle degeneration, myorticosis, amyotrophic axonal sclerosis, myasthenia gravis, muscle loss, and sarcopenia.

3. The composition according to claim 1, characterized in that the muscle disease is induced by aging, decreased muscle function, muscle wasting, muscle degeneration, disuse, or muscle injury.

4. A pharmaceutical composition for the improvement or treatment of cachexia, comprising diflunisal or a pharmaceutically acceptable salt thereof.

5. The pharmaceutical composition for improving or treating cachexia according to claim 4, characterized in that the cachexia is cancer cachexia induced by cancer.

6. The composition according to claim 4, wherein the composition improves and treats one or more symptoms of cachexia selected from the group consisting of loss of appetite, weight loss, increased fatigue, decreased muscle strength, decreased mobility, muscle loss, fat loss, and hematopoietic toxicity.

7. A composition for muscle building or strengthening, comprising diflunisal or an acceptable salt thereof.

8. The composition according to claim 7, wherein the composition has an effect of promoting the proliferation or differentiation of muscle stem cells.

9. The composition according to claim 7, wherein the composition is one or more selected from the group consisting of pharmaceutical compositions, health functional food compositions, functional food compositions, or animal feed compositions.

10. A composition for adjunct use in anticancer treatment, comprising diflunisal or a pharmaceutically acceptable salt thereof.

11. The composition according to claim 10, characterized in that the composition is administered in combination with one or more anticancer agents.

12. Use of diflunisal or its acceptable salts for the treatment of muscle diseases.

13. The use according to claim 12, wherein the muscle disease is selected from the group including cachexia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, myorticosis, amyotrophic axonal sclerosis, myasthenia gravis, muscle loss, and sarcopenia.

14. The use according to claim 12, wherein the muscle disease is induced by aging, decreased muscle function, muscle wasting, muscle degeneration, disuse, or muscle injury.

15. A method for treating cachexia, comprising the step of administering an effective amount of diflunisal or a pharmaceutically acceptable salt thereof to an individual.

16. The method for treating cachexia according to claim 15, wherein the diflunisal or a pharmaceutically acceptable salt thereof is administered to the individual together with one or more anticancer agents.

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