Composition for treating cancer cachexia comprising diflunisal or pharmaceutically acceptable salt thereof

Abstract

The present invention relates to a composition for improving and treating muscle diseases and cachexia, comprising diflunisal having an effect of promoting differentiation and proliferation of muscle-derived cells, further, it is possible to improve and treat cachexia induced by chronic diseases such as muscle diseases or cancers by means of muscle mass increase, muscle strength recovery, and the like.

Description

Technical Field

[0001] The present invention relates to a composition comprising diflunisal or a pharmaceutically acceptable salt thereof as a non-steroidal analgesic for improving and treating cachexia, especially cancer cachexia. Background Art

[0002] Cachexia is a condition that causes persistent muscle loss in various diseases, including cancer, renal failure, and chronic illnesses. Cancer cachexia, in particular, is a complex metabolic syndrome that can occur with cancer. Even when cancer patients are provided with nutrients through a normal diet, their utilization is limited or their metabolism is abnormal, preventing them from being restored. Cachexia is a state of systemic malnutrition that results in a loss of body mass (weight, muscle, etc.). It is generally defined as a loss of more than 5% of body weight due to muscle and fat loss within six months of cancer onset. While not common to all cancer patients, cancer cachexia has been reported in approximately 50-60% of patients with gastrointestinal, pancreatic, lung, and colorectal cancers.

[0003] The cause of cancer cachexia has not yet been precisely determined, but it is known that inflammatory cytokines such as interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), and C-reactive protein 1 (CRP1) play a key role in the loss of muscle and fat. There is no specific treatment for cancer cachexia, but it can be indirectly improved by improving pain or gastrointestinal disorders, or by using appetite enhancers. Recently, drugs such as celecoxib, a COX-2 inhibitor, and thalidomide, a TNF-α inhibitor, have also been used. However, celecoxib has been reported to have side effects such as anemia, gastric ulcers, allergies, heart failure, and stroke, while thalidomide has been reported to have side effects such as depression, heart failure, dyspnea, vomiting, rash, hypertension, and a high incidence of fetal malformations during pregnancy. Furthermore, there is a lack of clear evidence regarding the effectiveness of these treatments.

[0004] The present inventors have completed the present invention by researching a drug that has a therapeutic effect on cachexia, particularly cancer cachexia, and can be safely administered to patients with minimal side effects. Summary of the Invention

[0005] Technical issues

[0006] The present inventors have screened various drugs for use in the treatment of muscle diseases, particularly cachexia, particularly among nonsteroidal anti-inflammatory drugs (NSAIDs). The inventors have examined their effects on muscle diseases and cachexia, such as increases in body weight and muscle mass, and improvements in muscle strength. As a result, they have discovered that diflunisal is particularly effective in ameliorating and treating symptoms associated with these muscle diseases or cachexia, leading to the completion of the present invention.

[0007] Therefore, an object of the present invention is to provide a composition for preventing or treating muscle diseases, comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0008] Another object of the present invention is to provide a pharmaceutical composition comprising diflunisal or a pharmaceutically acceptable salt thereof for improving or treating cachexia.

[0009] Another object of the present invention is to provide a method comprising diflunisal or a pharmaceutically acceptable salt thereof. A composition for muscle strengthening or muscle strength enhancement.

[0010] Another object of the present invention is to provide an anticancer adjuvant composition comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0011] Technical Solution

[0012] To achieve the above objectives, the present invention provides a composition comprising diflunisal or a pharmaceutically acceptable salt thereof for preventing or treating muscle diseases.

[0013] In order to achieve another object of the present invention, the present invention provides a pharmaceutical composition for improving or treating cachexia, comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0014] In order to achieve another object of the present invention, the present invention provides a composition for muscle strengthening or muscle strength enhancement comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0015] To achieve another object of the present invention, the present invention provides an anticancer adjuvant composition comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0016] Hereinafter, the present invention will be described in detail.

[0017] In one embodiment of the present invention, the present invention relates to a composition for preventing muscular dystrophy comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0018] Diflunisal, a salicylic acid derivative, belongs to the nonsteroidal anti-inflammatory drug (NSAID) series of drugs and is widely used as an analgesic for musculoskeletal diseases. It is a compound represented by the following chemical formula 1.

[0019] Chemical formula 1

[0020]

[0021] Diflunisal is a competitive COX-I and COX-II inhibitor (with high affinity for COX-I). It plays a role in the mechanism of inhibiting the conversion of arachidonic acid in the body from prostaglandins that induce inflammation and inducing inflammation, and is used as an anti-inflammatory agent and analgesic.

[0022] In the present invention, diflunisal has the effect of preventing, ameliorating, or treating muscle diseases characterized by symptoms such as decreased muscle function, muscle loss, muscle atrophy, muscle wasting, or muscle degeneration. The term "muscle disease" refers to a state of damaged or weakened muscle due to aging or disease, and is caused by a variety of factors, including: genetic factors; 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, acquired immunodeficiency syndrome (AIDS), chronic inflammatory diseases, arthritis, malnutrition, kidney disease, chronic obstructive pulmonary disease, emphysema, rickets, chronic low back pain, peripheral nerve damage, central nervous system damage, and chemical damage; loss of motor function due to fractures, trauma, or prolonged bed rest; and aging.

[0023] The "muscle disease" of the present invention may be one or more muscle diseases selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myotonia, amyotrophic lateral sclerosis, myasthenia, cachexia, and sarcopenia, but is not limited thereto. Specifically, it may be diseases such as senile amyotrophy, muscle diseases caused by cancer and chronic diseases, or muscle atrophy caused by muscle disuse. More specifically, it may include senile amyotrophy or muscle atrophy caused by cancer, muscular dystrophy, muscle degeneration, myotonia, amyotrophic lateral sclerosis, myasthenia, cachexia, sarcopenia, and muscle loss. In particular, the muscle disease in the present invention may be a muscle disease induced by cancer or a chronic disease.

[0024] Among muscle diseases, diflunisal of the present invention is particularly effective in ameliorating or treating cachexia. Cachexia is a severe, debilitating syndrome that can occur with diseases such as cancer, tuberculosis, diabetes, and acquired facial disfigurement syndrome. It is particularly common in patients with gastrointestinal cancers and lung cancer. Key symptoms include decreased appetite, weight loss associated with muscle and fat loss, and decreased strength, meaning weight loss occurs even with a normal diet. Cachexia can reduce the effectiveness of treatment and shorten a patient's life expectancy.

[0025] In the present invention, the cachexia to be improved or treated may be cancer cachexia induced by cancer.

[0026] In the present invention, the types of cancers that can induce cachexia are not limited and can 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, stem cell liver cancer, bile duct cancer, 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, blood cancer, etc. More preferably, it can be digestive organ cancer such as gastric cancer, colorectal cancer, rectal cancer, liver cancer, gallbladder cancer, pancreatic cancer or lung cancer.

[0027] The pharmaceutical composition of the present invention can improve and treat symptoms caused by one or more cachexia selected from the group consisting of decreased appetite, weight loss, increased fatigue, decreased muscle strength, decreased exercise capacity, muscle loss, fat loss and hematopoietic toxicity. Preferably, it can improve and treat weight loss, muscle loss, decreased muscle strength or decreased exercise capacity.

[0028] Regarding these effects, in one embodiment of the present invention, administration of diflunisal to a cancer cachexia-induced model resulted in increased body weight, muscle mass, and muscle strength. Furthermore, in the cachexia-induced model, it was confirmed that decreased weights of organs such as the spleen, liver, and heart were improved compared to the untreated group. This indicates that diflunisal can be used to improve and treat cancer cachexia.

[0029] In another embodiment of the present invention, the diflunisal can be used as a composition having the effects of enhancing muscle, strengthening muscle strength, or improving exercise performance.

[0030] In one embodiment of the present invention, diflunisal was confirmed to promote the proliferation and differentiation of muscle stem cells. In the embodiment, diflunisal increased the expression of Pax7, a muscle stem cell proliferation marker, and increased the expression of myosin heavy chain (MHC) and myogenin, muscle stem cell differentiation markers. Furthermore, an increase in muscle fiber diameter was observed following diflunisal administration. In other words, diflunisal promotes the proliferation and differentiation of muscle stem cells, resulting in effects that improve muscle function, such as increased muscle mass, improved muscle strength, enhanced muscle resilience, and reduced muscle fatigue. Similar effects were observed not only in models of induced disease (cancer cachexia), but also in normal muscle.

[0031] Furthermore, diflunisal can improve motor performance. "Motor performance" refers to the ability to perform movements using muscle strength. In this context, muscle strength can be improved through muscle stamina, muscle endurance, increased oxidative muscle mass, muscle recovery, and intramuscular energy balance. It can also be enhanced by reducing fatigue substances within the muscle. In one embodiment of the present invention, improved muscle endurance and increased muscle mass were confirmed in mice, indicating that this can improve motor performance.

[0032] The composition of the present invention can be used for various purposes such as medicines, health functional foods, functional foods, animal feed, cell culture fluid compositions, etc., and can have the effect of preventing, improving or treating muscle diseases and cachexia based on the effects of promoting the proliferation and differentiation of muscle stem cells, increasing muscle mass, and improving muscle endurance.

[0033] In this specification, the term "prevention" refers to all actions to suppress a disease or delay its onset by administering the pharmaceutical composition of the present invention.

[0034] In this specification, the term "improvement" refers to all actions to alleviate symptoms induced by a disease or delay the onset of the disease by administering the pharmaceutical composition of the present invention.

[0035] In this specification, the term "treatment" refers to all actions to improve or cure symptoms by administering the pharmaceutical composition of the present invention.

[0036] The pharmaceutical composition of the present invention can be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, sprays according to common methods, and the form of external-use agent, suppository and sterile injection solution is used, and can also include carrier or excipient required for the preparation. Pharmaceutically acceptable carriers, excipients and diluents that can also be included in the active ingredient include lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate and mineral oil. During preparation, diluents or excipients such as commonly used fillers, extenders, bonding agents, wetting agents, disintegrants, surfactants can be used to prepare.

[0037] For example, solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, and the like. These solid preparations contain at least one excipient in the extract or compound and can be prepared by mixing starch, calcium carbonate, sucrose or lactose, gelatin, and the like. Furthermore, in addition to simple excipients, lubricants such as magnesium stearate are also used. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, and the like. In addition to water and liquid paraffin, which are commonly used as simple diluents, they can also contain a variety of excipients, such as wetting agents, sweeteners, fragrances, and preservatives.

[0038] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, oils, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, polyethylene glycol, Tween 61, cocoa butter, tartar resin, and glycerinated gelatin.

[0039] The pharmaceutical composition of the present invention can be administered orally or parenterally (intravenously, subcutaneously, intraperitoneally or by lesion application) according to the target method. The dosage varies depending on the patient's condition and weight, the severity of the disease, the form of the drug, the route of administration and the time of administration. The appropriate form can be selected by relevant practitioners.

[0040] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, a "pharmaceutically effective amount" is a reasonable amount applicable in medical treatment, which refers to a sufficient amount to treat the disease. Its standard can be determined based on the patient's disease, the severity of the disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the metabolic ratio, the treatment period, the ingredients used in combination, and other matters. The pharmaceutical composition of the present invention can be administered as a single therapeutic agent, or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with existing therapeutic agents. The dosage can be determined at the minimum level without side effects after considering all the above factors, which can be easily determined by relevant practitioners. Specifically, the dosage of the pharmaceutical composition varies according to the patient's age, weight, severity of the disease, sex, etc., and is generally administered in an amount of 0.001 mg to 150 mg per 1 kg of body weight, preferably, in an amount of 0.01 mg to 100 mg per day or every other day, once to three times a day. However, this is only an example, and the dosage can be set differently as needed.

[0041] Furthermore, the composition of the present invention may be a video or a health functional food. In particular, the "health functional food" refers to a food prepared and processed using raw materials or ingredients that have effective functionality for the human body in accordance with Law No. 6727 on Health Functional Foods, and "functional" means that it is taken for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological effects on the structure and function of the human body.

[0042] The food or health functional food of the present invention can be prepared and processed into pharmaceutical forms such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, or health functional foods such as tea bags, extracts, beverages, candies, jellies, and chewing gums for the purpose of preventing and improving muscle diseases.

[0043] The present invention The health functional food composition can be used as a food additive and can be commercialized alone or in combination with other ingredients. Furthermore, it can contain nutrients, vitamins, electrolytes, flavoring agents, colorants and synergists, pectin and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. These ingredients can be used alone or in combination, and can be used in appropriate amounts.

[0044] In another embodiment, the present invention relates to a composition for adjuvant anticancer therapy comprising diflunisal. The composition for adjuvant anticancer therapy can be used as an adjuvant in cancer treatment using a known anticancer agent and can be administered simultaneously or sequentially with the anticancer agent as an adjuvant anticancer agent.

[0045] Effects of the Invention

[0046] The present invention relates to a composition comprising diflunisal or a pharmaceutically acceptable salt thereof, which is effective in improving and treating cachexia, particularly cancer cachexia. The composition not only increases muscle mass, improves muscle endurance, and enhances mobility by promoting the proliferation and differentiation of myogenic cells, but also improves various symptoms associated with cachexia, such as weight gain, and the use of the composition for treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 The figure shows the results of evaluating the cytotoxicity of diflunisal (MTTassay) using C2C12 cells, which are muscle stem cells.

[0048] Figure 2 The results of MHC fluorescence staining are shown to confirm the effect of diflunisal on promoting muscle cell differentiation in vitro.

[0049] Figure 3 The results of confirming the effect of diflunisal on promoting muscle cell differentiation in vitro are shown, and the results of quantitative comparison of myotube diameters are shown.

[0050] Figure 4 The results of immunoblotting analysis on MHC and myogenin are shown to confirm the in vitro effect of diflunisal on promoting muscle cell differentiation.

[0051] Figure 5a 、 Figure 5b and Figure 6 Confirmation of the stem cell proliferation promoting effect of diflunisal administration, Figure 5a and Figure 5b For C2C as muscle stem cells 12 The results of BrdU staining and quantification of BrdU+ (BrdU positive) cells were Figure 6 Shown are the results of quantitative real-time polymerase chain reaction (qRT-PCR) analysis of Pax7 expression, a stem cell marker.

[0052] Figure 7 and Figure 8 The effect of diflunisal on promoting the proliferation of muscle stem cells was confirmed in the tibialis anterior (TA) muscle of mice with CTX-induced muscle damage. Figure 7 This is the BrdU staining image. Figure 8 This is the quantitative result of BrdU+ (BrdU positive) cells.

[0053] Figure 9a 、 Figure 9band Figure 10 The effect of diflunisal on improving muscle atrophy was confirmed in a dexamethasone-induced muscle atrophy model. Figure 9a The results of immunostaining analysis on MHC are shown. Figure 9b The results of immunostaining analysis on the expression of dystrophin-1 are shown. Figure 10 Shown are the results of qRT-PCR analysis of MuRF1 expression.

[0054] Figure 11 and Figure 12 To confirm the in vitro improvement effect of diflunisal administration on cachexia induced by cancer cell culture medium (CM), Figure 11 To confirm the concentration-dependent myotube regeneration and improvement effect of diflunisal administration in a cancer cachexia-induced model by MHC immunostaining, Figure 12 The results were quantified by measuring the diameter of the myotubes.

[0055] Figure 13 and Figure 14 The effect of diflunisal administration was confirmed after inducing cancer cachexia in vivo. Figure 13 A schematic diagram showing the method used for the in vivo experiment is shown. Figure 14 The results are the results of measuring body weight, tumor weight and changes in body weight excluding tumor after administration of diflunisal.

[0056] Figures 15 to 17 The effect of diflunisal administration on cancer cachexia induced in vivo is shown, and the results of examining exercise capacity (grip strength) are shown. Figure 15 ), dystrophin-1 and MuRF1 mRNA expression analysis (qRT-PCR) results ( Figure 16 ) and the results of measuring the muscle mass of each part of the hind legs ( Figure 17 ).

[0057] Figure 18 The improvement effect of diflunisal on cancer cachexia induced in vivo was shown, and changes in the weights of inguinal white fat (IWAT), visceral fat (EWAT), liver, heart, and spleen were confirmed. DETAILED DESCRIPTION

[0058] The following describes this specification in detail through the use of examples. However, the examples herein can be modified into various other forms, and the scope of this specification should not be construed as limited to the examples described in detail below. The examples herein are provided to more fully illustrate this specification to those skilled in the art.

[0059] Example 1. Culture and differentiation induction of myogenic cell lines

[0060] C2C l2 This is a myogenic cell line derived from C3H mice and is widely used in muscle cell differentiation research. The cells are cultured in Growth Media (GM) and induced to differentiate into muscle cells in Differentiation Media (DM). The cell culture medium is Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 15% fetal bovine serum (FBS), while the differentiation medium is supplemented with 2% horse serum.

[0061] To induce muscle cell differentiation, cells are cultured by dispensing into a cell culture medium. When the cell density reaches approximately 80% to 90%, the culture medium is replaced with a differentiation medium and differentiation is induced for approximately 2 to 3 days.

[0062] Experimental Example 1. Evaluation of the cytotoxicity of diflunisal

[0063] The cytotoxicity of diflunisal (CAS No. 22494-42-4, W06) was assessed by MTT assay. 12 Myogenic cells were treated with diflunisal at concentrations of 0 nM, 1 nM, 10 nM, 100 nM, 1000 nM, and 10000 nM, and then the cell viability was determined by MTT analysis. 4 Cells (cells / 96 wells) were reacted with 5 mg / mL MTT for 4 hours, and then the absorbance of the dissolved reactant was measured at a wavelength of 570 nm. As a result, the cell viability of diflunisal used in the MTT assay was similar to that of the control group even when treated with a concentration of 10,000 nM. Figure 1 ).

[0064] Experimental Example 2: Confirmation of the Stem Cell Differentiation-Promoting Effect of Diflunisal

[0065] In order to analyze the effect of diflunisal on the differentiation of muscle stem cells, MHC immunofluorescence staining was performed ( Figure 2). To C2C 12 The cell lines were treated with 10nM, 100nM, 1000nM, and 10,000nM of dimethyl sulfoxide (DMSO) and diflunisal, respectively, to induce muscle cell differentiation. On the third day of differentiation induction, immunofluorescence staining using an MHC antibody was performed to compare and analyze the degree of myotube formation. The results showed that muscle cell differentiation was promoted in cells treated with diflunisal in a concentration-dependent manner compared to the control group. Furthermore, quantification of myotube diameter also confirmed an increase in diflunisal-dependent treatment concentration. Figure 3 ).

[0066] At the same time, the expression level of differentiation markers was confirmed by immunoblotting. It was confirmed that diflunisal treatment induced an increase in the expression of MHC and myogenin, which are muscle stem cell differentiation markers. This was also a significant level compared to the case of ursolic acid (UA) treatment as a positive control group ( Figure 4 ).

[0067] Experimental Example 3: Confirmation of the Stem Cell Proliferation-Promoting Effect of Diflunisal

[0068] In order to analyze the effect of diflunisal on the proliferation of myogenic cells, BrdU staining experiments were performed. 12 Myogenic cells were treated with dimethyl sulfoxide or diflunisal (1 μM) as a control group for 20 hours, and then reacted with BrdU for 15 minutes. Immunofluorescence staining was performed using an antibody against BrdU. Figure 5a As a result, it was confirmed that the proliferation of myogenic cells was increased in the diflunisal-treated group. In addition, in a quantitative comparison, it was confirmed that the proportion of BrdU-positive cells was significantly increased when diflunisal was treated compared to the control group ( Figure 5b ).

[0069] Furthermore, we examined whether diflunisal treatment increased the expression of Pax7, a muscle proliferation marker. Figure 6 As shown in FIG, it was confirmed that the relative expression level of Pax7 mRNA was increased when diflunisal was treated.

[0070] Experimental Example 4. Effect of Promoting Stem Cell Proliferation in a Muscle Injury Animal Model

[0071] An experiment was conducted to confirm whether diflunisal administration also has the effect of promoting stem cell proliferation in an actual animal model of muscle damage. Diflunisal was orally administered to C57BL / 6 male mice at a dose of 0.02 mg / kg for 2 weeks. In the second week, 10 μM cardiotoxin (CTX) was directly injected intramuscularly at a volume of 2 μl per gram (g) of body weight to induce muscle damage. Diflunisal was then administered at a dose of 0.02 mg / kg for 3 days. Immunofluorescence staining of the TA muscle tissue of the mice was then performed using a BrdU antibody. The results were as follows: Figure 7 As shown in Figure 2, it was confirmed that the proliferation of myogenic cells was increased in the diflunisal-treated group. Furthermore, quantitative comparison also confirmed that the proportion of BrdU-positive cells increased by more than 2.5 times when treated with diflunisal compared to the control group ( Figure 8 ).

[0072] Experimental Example 5. Muscle atrophy improvement effect

[0073] To determine whether diflunisal can restore or protect muscle in dexamethasone (DEX)-induced muscle atrophy, C2C 12 Myogenic cells were used for experiments. 12 Myogenic cells were differentiated in DM (differentiation medium) for 2 days, treated with 100 μM dexamethasone (DEX) for 4 hours, and then treated with diflunisal (1 μM) along with dimethyl sulfoxide as a vehicle, followed by additional culture in DM for 24 hours.

[0074] For dexamethasone-treated C2C 12 Myotube cells were immunostained with an MHC antibody to compare and analyze the degree of myotube formation. As shown in Figure 9 , MHC expression decreased when muscle atrophy was induced by dexamethasone, but was restored when diflunisal was administered. Furthermore, immunoblotting confirmed the expression of dystrophin-1, a muscle atrophy marker, and found that expression increased with dexamethasone administration but decreased with diflunisal ( Figure 9 ).

[0075] Furthermore, qRT-PCR analysis of the expression of MuRF1, another muscle atrophy marker, also confirmed that MuRF1 expression increased when muscle atrophy was induced, but its expression was significantly decreased when diflunisal was administered ( Figure 10 ).

[0076] Experimental Example 6. Effect of improving cachexia induced by cancer cell culture medium

[0077] To confirm the effect of diflunisal on improving cancer cachexia, cancer cachexia was induced in muscle cells using cancer cell culture medium (CM). Figure 11 top).

[0078] In the cancer cachexia-induced model, diflunisal was treated at concentrations of 10 nM, 100 nM, and 1000 nM, and the myotube regeneration and improvement effects were confirmed by MHC immunostaining. The improvement was confirmed by measuring the diameter of the myotubes. Figure 11 and Figure 12 As shown, when cultured in CM, cancer cachexia was induced, resulting in a decrease in myotube diameter. However, administration of diflunisal increased myotube diameter in a concentration-dependent manner. This suggests that diflunisal has the effect of ameliorating muscle loss and muscle atrophy caused by cancer cachexia.

[0079] Experimental Example 7. Effect of Improving Cancer Cachexia in Animal Models

[0080] 7-1) Effect of improving weight loss caused by cancer cachexia

[0081] Lewis lung carcinoma (LCC) cells (5×10 6 Lung cancer was induced in cells / mice. Diflunisal was orally administered daily at a dose of 0.02 or 0.2 mpk for 21 days starting 2 weeks after injection. Figure 13 Then, for each mouse, the changes in body weight, tumor weight, and body weight excluding tumor weight were measured. Figure 14 As shown in the results, there was no significant change in overall body weight in all experimental groups, but the weight of the tumor increased in the cancer-induced group, and the weight changes other than the tumor were improved in the group receiving diflunisal. In particular, the weight of the group receiving 0.02 mpk of diflunisal other than the tumor increased significantly compared to the cancer-induced group ( Figure 14 ).

[0082] 7-2) Improvement of reduced exercise capacity and muscle mass caused by cancer cachexia

[0083] In order to compare the exercise capacity in the cancer cachexia-induced mouse model, a grip strength test was performed on each experimental group. Figure 15 As shown, when compared with the cancer-induced group, it was confirmed that the grip strength of the diflunisal-administered group increased in a concentration-dependent manner, and the exercise capacity was significantly improved.

[0084] Furthermore, the expression levels of dystrophin-1 and MuRF-1, which are positively correlated markers of muscle atrophy, were confirmed by immunoblotting. It was confirmed that the relative mRNA expression levels increased when cancer was induced, but decreased again when diflunisal was administered ( Figure 16 ).

[0085] And, with Figure 17 The method shown here examines changes in muscle mass of the hindlimb muscles, namely the tibialis anterior (TA), extensor digitorum longus (EDL), soleus (SOL), and gastrocnemius (GA). When cancer is induced, muscle mass of the TA, EDL, SOL, and GAS is reduced. However, administration of diflunisal significantly improves muscle mass loss caused by cancer cachexia ( Figure 17 This result confirmed the effect of diflunisal in suppressing muscle loss caused by cancer cachexia ( Figure 17 ).

[0086] 7-3) Effect of improving weight loss caused by cancer cachexia

[0087] In addition, in a mouse model induced with cancer cachexia, the effects of diflunisal on tissues other than muscle, such as white fat, visceral fat, liver, heart, and spleen, were compared by measuring the weight of each tissue. Figure 18 As shown, in the cancer-induced model, weight loss occurred in all cases, but administration of diflunisal was confirmed to improve the loss of fat and organ weight caused by cancer cachexia.

[0088] Thus far, the present invention has been described in detail through preferred embodiments. It will be appreciated by those skilled in the art that the present invention may be implemented in various variations without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered illustrative rather than restrictive. The scope of the present invention is not limited to the foregoing description, but is set forth in the claims, and all differences within the scope equivalent thereto should be construed as included in the present invention.

[0089] Best Practice

[0090] One embodiment of the present invention relates to a pharmaceutical composition for improving or treating muscle diseases, comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0091] The muscle disease may be selected from the group consisting of hypotonia, muscle atrophy, muscular dystrophy, muscle degeneration, myotonia, amyotrophic lateral sclerosis, myasthenia, muscle wasting disease and sarcopenia.

[0092] The muscle disease may be caused by aging, muscle loss, muscle wasting, muscle degeneration, disuse, or muscle injury.

[0093] Yet another embodiment of the present invention relates to a pharmaceutical composition for improving or treating cachexia comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0094] The cachexia may be cancer cachexia induced by cancer.

[0095] Furthermore, the composition improves and treats symptoms caused by one or more cachexia-related symptoms selected from the group consisting of decreased appetite, weight loss, increased fatigue, decreased muscle strength, decreased exercise capacity, muscle loss, fat loss, and hematopoietic toxicity.

[0096] Another embodiment of the present invention relates to a composition for muscle strengthening or muscle strength enhancement comprising diflunisal or an acceptable salt thereof.

[0097] The composition may have an effect of promoting the proliferation or differentiation of muscle stem cells.

[0098] Furthermore, the composition can be selected from the group consisting of a pharmaceutical composition, a health functional food composition, a functional food composition or an animal feed composition.

[0099] Another embodiment of the present invention relates to a composition for adjuvant anti-cancer treatment comprising diflunisal or a pharmaceutically acceptable salt thereof.

[0100] The composition may be administered in combination with more than one anticancer agent.

[0101] Yet another embodiment of the present invention relates to the use of diflunisal or an acceptable salt thereof for treating muscle diseases.

[0102] The muscle disease is selected from the group consisting of cachexia, hypotonia, muscle atrophy, muscular dystrophy, muscle degeneration, myotonia, amyotrophic lateral sclerosis, myasthenia, muscle wasting disease and sarcopenia.

[0103] Furthermore, the muscle disease is caused by aging, muscle dysfunction, muscle wasting, muscle degeneration, disuse or muscle injury.

[0104] Yet another embodiment of the present invention relates to a method for treating cachexia, comprising the step of administering to a subject an effective amount of diflunisal or a pharmaceutically acceptable salt thereof.

[0105] Furthermore, in the methods, 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 improving or treating muscle diseases, characterized in that: Contains diflunisal or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition for improving or treating muscle diseases according to claim 1, characterized in that The muscle disease is selected from the group consisting of hypotonia, muscle atrophy, muscular dystrophy, muscle degeneration, myotonia, amyotrophic lateral sclerosis, myasthenia, muscle wasting disease and sarcopenia.

3. The pharmaceutical composition for improving or treating muscle diseases according to claim 1, characterized in that The muscle disease is caused by aging, decreased muscle function, muscle wasting, muscle degeneration, disuse or muscle injury.

4. A pharmaceutical composition for improving or treating cachexia, characterized in that: Contains 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 caused by cancer.

6. The pharmaceutical composition for improving or treating cachexia according to claim 4, characterized in that The composition improves and treats symptoms caused by one or more cachexia selected from the group consisting of decreased appetite, weight loss, increased fatigue, decreased muscle strength, decreased exercise capacity, muscle loss, fat loss and hematopoietic toxicity.

7. A composition for muscle strengthening or muscle strength enhancement, characterized in that: Contains diflunisal or an acceptable salt thereof.

8. The composition for muscle strengthening or muscle strength enhancement according to claim 7, characterized in that The composition has the effect of promoting the proliferation or differentiation of muscle stem cells.

9. The composition for muscle strengthening or muscle strength enhancement according to claim 7, characterized in that The composition is one or more selected from the group consisting of a pharmaceutical composition, a health functional food composition, a functional food composition or an animal feed composition.

10. A composition for auxiliary anti-cancer treatment, characterized in that: Contains diflunisal or a pharmaceutically acceptable salt thereof.

11. The composition for adjuvant anti-cancer therapy according to claim 10, characterized in that: The composition is administered in combination with more than one anticancer agent.

12. A use of diflunisal or an acceptable salt thereof, characterized in that: Used to treat muscle diseases.

13. The use of diflunisal or an acceptable salt thereof according to claim 12, characterized in that: The muscle disease is selected from the group consisting of cachexia, hypotonia, muscle atrophy, muscular dystrophy, muscle degeneration, myotonia, amyotrophic lateral sclerosis, myasthenia, muscle wasting disease and sarcopenia.

14. The use of diflunisal or an acceptable salt thereof according to claim 12, characterized in that: The muscle disease is caused by aging, decreased muscle function, muscle wasting, muscle degeneration, disuse or muscle injury.

15. A method for treating cachexia, characterized in that: The method comprises the step of administering to the individual an effective amount of diflunisal or a pharmaceutically acceptable salt thereof.

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

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