Composition for preventing or treating metabolic diseases comprising torsemide and cromolyn

Abstract

The present invention relates to a composition for treating metabolic diseases, comprising torsemide and cromolyn. According to the present invention, co-administration of torsemide and cromolyn significantly reduced weight loss, insulin resistance and hepatic fat accumulation in NASH animal models compared to the case of single administration of cromolyn, and thus the composition can be effectively utilized for the prevention or treatment of metabolic diseases, including diabetes, obesity, insulin resistance and metabolic liver diseases.

Description

[Technical Field]

[0001] The present invention relates to compositions for the treatment of metabolic disorders comprising torsemide and cromolyn. [Background technology]

[0002] The liver is a large organ located in the upper abdominal cavity on the right side of the body. In adults, it weighs approximately 1.5 kg. A healthy liver is reddish to the naked eye and has a smooth surface. The shape and size of the liver change when liver disease develops. For example, when heavy alcohol consumption causes fat to accumulate in the liver, the liver increases in size and takes on a yellowish hue. Conversely, as cirrhosis progresses, the liver decreases in size and its surface becomes uneven. The liver is an extremely important organ with various functions, which can be summarized as follows: First, the liver performs metabolic functions, i.e., properly processes various nutrients in the body. Food and drink absorbed from the intestines are transformed in the liver so that they can be used by various tissues in the body. After these tissues utilize the nutrients, remaining waste products are transported to the liver for further processing. Second, the liver stores several nutrients necessary for the body. Third, it produces bile acids, which are essential for nutrient absorption in the intestine, and excretes them into the intestine through the bile duct. Without the production of bile acids, various nutrients cannot be absorbed, resulting in nutritional deficiencies. Fourth, it produces substances (such as albumin, blood clotting proteins, and cholesterol) that are absolutely necessary for the proper functioning of our bodies. Finally, it detoxifies alcohol, drugs, and various toxins produced by our bodies (Eugene Brainward, et al., Harrison's Principles of Internal Medicine, 15th edition, 2001).

[0003] Metabolic Dysfunction Associated Steatotic Liver Disease (MASLD)It is defined as the accumulation of 5% or more of fatty acids in the form of triglycerides within the parenchymal cells of the liver, rather than liver damage caused by alcohol. Pathologically, it is classified into simple steatosis and steatohepatitis accompanied by inflammation. Metabolic Dysfunction Associated Steatohepatitis (MASH) is the above Metabolic dysfunction-associated fatty liver disease ( MASLD It is a disease that occurs during the worsening process of liver cirrhosis, in which fat accumulates in liver cells, causing liver cell degeneration / necrosis, which leads to inflammation and liver fibrosis, which can develop into cirrhosis and liver cancer, and is therefore recognized as a serious disease worldwide. MASH It has been suggested that excess free fatty acids (FFA) in the body, along with the accumulation of fat in the liver, directly induce liver damage, plays an important role. In obese and insulin-resistant individuals, the influx of FFA into the liver increases, and FFA accumulates in the liver through beta-oxidation or esterification of triglycerides (TG). In addition, the increased oxidative stress and activation of inflammatory signaling processes caused by excess FFA itself cause direct liver damage, and TG accumulates in the liver as a protective mechanism. This leads to hepatocyte death and inadequate recovery, which ultimately leads to liver fibrosis and disease progression, a serious lesion. Metabolic dysfunction-related steatohepatitis ( MASH ) (Ong JP et al., Obesity Surgery volume 15, pages 310-315, 2005). Metabolic dysfunction-related steatohepatitis It is divided into primary and secondary depending on the cause, and primary is caused by hyperhemostasis, diabetes, or obesity, which are characteristics of metabolic syndrome (Szczepaniak LS et al., 2005), while secondary is known to be caused by nutritional causes (rapid weight loss, starvation, intestinal bypass surgery), various drugs, toxic substances (poisonous mushrooms, bacterial toxins), metabolic causes, and other factors. Primary causes are associated with diabetes and obesity, which are important characteristics of metabolic syndrome. Metabolic dysfunction-related steatohepatitisThe incidence of steatohepatitis is known to be approximately 50% in diabetic patients, approximately 76% in obese patients, and in most obese diabetic patients (Gupte P et al., 2004). Biopsies performed on diabetic and obese patients with elevated alanine aminotransferase (ALT) levels have shown that the incidence of steatohepatitis is 18-36% (Braillon A et al., 1985).

[0004] Metabolic dysfunction-related steatohepatitis Although it has been considered a benign disease that does not progress, it has been revealed that even in non-drinkers, patients exhibit inflammation and liver fibrosis similar to alcoholic hepatitis, and it is known to be a disease with a poor prognosis. In particular, in recent years, metabolic syndrome, which is caused by obesity and diabetes, has attracted attention. Metabolic Dysfunction Associated Steatohepatitis (MASH) However, its mechanism remains unknown, and no effective treatments or drugs have been established. Metabolic dysfunction-related steatohepatitis There is no established treatment for Metabolic dysfunction-related steatohepatitis This is because it has been shown to be related to various factors such as diabetes, obesity, coronary artery disease, and lifestyle habits. The effects of several diabetes or obesity treatment drugs on fatty liver disease have been reported, but in the case of Orlistat, which is used as an oral obesity treatment drug, it has been reported that it has induced histological improvement of the liver in patients with steatohepatitis (Hussein et al., 2007), and metformin has been shown to be effective in patients without diabetes. Metabolic dysfunction-related steatohepatitis It has been reported that blood liver enzyme levels, necrotic inflammation, and fibrosis of the liver were reduced in patients (Bugianesi et al., 2005). In addition, thiazolidinedione (TZD) drugs, which are agonists of the peroxisome proliferator-activated receptor (PPAR), suppressed fat accumulation in the liver and muscle. Metabolic dysfunction-related steatohepatitisIt has been reported that thrombin exerts a direct anti-fibrotic effect on the liver in animal models (Galli A et al., 2002). Metabolic dysfunction-related steatohepatitis At present, there are no compositions that can be approved as pharmaceutical treatments for the above diseases, and the development of novel therapeutic agents for the treatment or prevention of the above diseases is urgently needed. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Eugene Brainward, et al.Harrison's principles of internal medicine.15th edition. 2001 [Non-patent document 2] Ong JP et al., Obesity Surgery volume 15, pages 310-315, 2005 Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of metabolic diseases.

[0007] Another object of the present invention is to provide a health functional food for preventing or improving metabolic diseases.

[0008] It is also an object of the present invention to provide a method for treating metabolic disorders.

[0009] Another object of the present invention is to provide a use for the preparation of a pharmaceutical composition for the prevention and treatment of metabolic diseases. [Means for solving the problem]

[0010] To achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, which contains torsemide and cromolyn, or pharmaceutically acceptable salts thereof, as active ingredients.

[0011] The present invention also provides a health functional food for preventing or ameliorating metabolic diseases, which contains torsemide and cromolyn, or a pharmaceutically acceptable salt thereof.

[0012] The present invention also provides a method for treating a metabolic disorder, comprising administering to an individual having the metabolic disorder pharmaceutically effective amounts of torsemide and cromolyn, or pharmaceutically acceptable salts thereof.

[0013] In addition, the present invention provides the use of torsemide and cromolyn, or pharmaceutically acceptable salts thereof, for the manufacture of a pharmaceutical composition for the prevention and treatment of metabolic diseases. [Effects of the Invention]

[0014] According to the present invention, the combined administration of torsemide and cromolyn has a significant effect on the urinary tract ... MASH In animal models, it significantly reduced body weight, insulin resistance, and hepatic fat accumulation, suggesting that it can be useful in the prevention or treatment of metabolic diseases, including diabetes, obesity, insulin resistance, and metabolic liver disease. [Brief explanation of the drawings]

[0015] [Figure 1] 1 is a graph showing the average body weight (grams) of mice in each group measured each week (average body weight ± standard deviation). [Figure 2] The mean fasting blood glucose values (mg / dl) of mice in each group measured at week 13 of the experiment are shown below. [Figure 3] 1 shows an image and graph (staining %±standard deviation) showing the degree of neutral fat accumulation in liver tissue confirmed by Oil Red O staining. DETAILED DESCRIPTION OF THE INVENTION

[0016] Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. However, the following examples are presented as examples of the present invention, and if it is determined that a detailed description of a technology or configuration well known to those skilled in the art is unnecessary for the gist of the present invention, the detailed description may be omitted and the present invention is not limited thereby. The present invention is susceptible to various modifications and applications within the scope of the claims below and the scope of equivalents analyzed therefrom.

[0017] Furthermore, the terminology used in this specification is used to appropriately express preferred embodiments of the present invention, and may vary depending on the intentions of users and operators, or the practices in the field to which the present invention pertains. Therefore, the accuracy of the terminology should be determined based on the overall content of this specification. Throughout the specification, when a part "includes" a certain element, this does not mean that it excludes other elements, but that it may further include other elements, unless otherwise specified.

[0018] Unless otherwise specified, all technical terms used in the present invention are used in the same sense as commonly understood by a person of ordinary skill in the art in the field related to the present invention. In addition, although preferred methods and samples are described in this specification, similar or equivalent methods and samples are also included in the scope of the present invention. The contents of all publications referenced in this specification are incorporated herein by reference.

[0019] In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating a metabolic disease, which comprises torsemide and cromolyn, or a pharmaceutically acceptable salt thereof, as active ingredients.

[0020] In one embodiment, torsemide may be a compound represented by the following chemical formula 1, and cromolyn may be a compound represented by the following chemical formula 2:

[0021] [ka]

[0022] and;

[0023] [ka]

[0024] In one embodiment, the metabolic disease may be any one selected from the group consisting of diabetes, obesity, insulin resistance, and metabolic liver disease, and the metabolic liver disease may be any one selected from the group consisting of: Metabolic Dysfunction Associated Steatotic Liver Disease (MASLD) may be Metabolic dysfunction-associated fatty liver disease teeth, Metabolic Dysfunction Associated Steatohepatitis (MASH) , non-alcoholic fatty liver (NAFL) and NAFLD-associated liver fibrosis; Metabolic dysfunction-related steatohepatitis It is more preferable that:

[0025] In one embodiment, the prevention or treatment of metabolic disorders can consist of reducing insulin resistance, reducing weight, and reducing triglyceride accumulation in liver tissue.

[0026] In one embodiment, the pharmaceutically acceptable salt of cromolyn can be selected from the group consisting of alkali metal salts, alkaline earth metal salts, salts with inorganic acids, salts with organic acids, and salts with acidic amino acids, with cromolyn sodium being most preferred.

[0027] In one embodiment, the torsemide and cromolyn or pharmaceutically acceptable salts thereof of the present invention can be administered at a dose of 0.01 mg / kg to 1 g / kg of torsemide and cromolyn, respectively.

[0028] In one embodiment, torsemide and cromolyn can be administered simultaneously, separately, or sequentially.

[0029] The present invention includes torsemide and cromolyn represented by chemical formulas 1 and 2, as well as their pharmaceutically acceptable salts, and any possible solvates, hydrates, racemates, or stereoisomers that may be produced therefrom.

[0030] Torsemide and cromolyn represented by Chemical Formulas 1 and 2 of the present invention can be used in the form of pharmaceutically acceptable salts, and useful salts include acid addition salts formed with pharmaceutically acceptable free acids. Acid addition salts are obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, or phosphorous acid, and non-toxic organic acids such as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, and alkanedioates, aromatic acids, and aliphatic and aromatic sulfonic acids. Such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate or mandelate.

[0031] The acid addition salts of the present invention can be prepared by conventional methods, for example, by dissolving torsemide or cromolyn in an excess amount of aqueous acid and precipitating the salt with a water-miscible organic solvent, such as methanol, ethanol, acetone, or acetonitrile. Alternatively, the mixture can be dried by evaporating the solvent and excess acid, or by suction filtering the precipitated salt. Pharmaceutically acceptable metal salts can also be prepared using a base. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving the compound in an excess amount of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, sodium, potassium, or calcium salts are preferably prepared as metal salts. The corresponding silver salts can be obtained by reacting alkali metal or alkaline earth metal salts with an appropriate silver salt (e.g., silver nitrate).

[0032] The pharmaceutical composition of the present invention may further contain a known therapeutic agent for metabolic diseases in addition to torsemide and / or cromolyn, or a salt thereof, as an active ingredient, and may be used in combination with other known therapies for the treatment of these diseases.

[0033] In the present invention, the term "prevention" means any action of inhibiting or delaying the occurrence, spread, and recurrence of metabolic diseases by administering the pharmaceutical composition of the present invention, and "treatment" means any action of improving or beneficially modifying the symptoms of metabolic diseases by administering the composition of the present invention. Those skilled in the art should be able to know the exact criteria for diseases for which the composition of the present invention is effective and judge the degree of improvement, enhancement, and treatment by referring to materials presented by the Korean Medical Association, etc.

[0034] In the present invention, the term "therapeutically effective amount" used in conjunction with an active ingredient refers to an amount effective for preventing or treating a target disease. The therapeutically effective amount of the composition of the present invention may vary depending on several factors, such as the administration method, the target site, and the patient's condition. Therefore, the appropriate dosage for human use must be determined by taking safety and efficacy into consideration. It is also possible to extrapolate the dosage for human use from the effective amount determined through animal experiments. These factors to be considered when determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; and E.W. Martin, ed., Remington's Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

[0035] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to any medical treatment and that does not cause side effects. The effective dose level can be determined based on factors including the patient's health condition, the type of metabolic disease, the cause and severity of the metabolic disease, the activity of the drug, sensitivity to the drug, the method of administration, the time of administration, the route of administration and excretion rate, the duration of treatment, coadministered or concurrently used drugs, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or multiple times. Taking all of the above factors into consideration, it is important to administer an amount that can achieve maximum effect at the minimum dose without side effects, which can be easily determined by one skilled in the art.

[0036] The pharmaceutical compositions of the present invention may contain carriers, diluents, excipients, or combinations of two or more thereof commonly used in biological preparations. The term "pharmaceutically acceptable" as used herein means that the composition is non-toxic to cells exposed to the composition or to humans. The carrier is not particularly limited as long as it is suitable for in vivo delivery of the composition. For example, compounds listed in the Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture of one or more of these components may be used. Other common additives, such as antioxidants, buffers, and bacteriostats, may be added as needed. Furthermore, the compositions may be formulated into injectable dosage forms, such as aqueous solutions, suspensions, and emulsions, as well as pills, capsules, granules, or tablets, by adding diluents, dispersants, surfactants, binders, and lubricants. Furthermore, formulations can be prepared appropriately depending on the disease or ingredients using appropriate methods in the art or the methods disclosed in Remington's Pharmaceutical Sciences (Mack Publishing Company, Easton PA, 18th, 1990).

[0037] In one embodiment, the pharmaceutical composition may be in one or more dosage forms selected from the group including oral dosage forms, topical preparations, suppositories, sterile injection solutions, and sprays, with oral or injectable dosage forms being more preferred.

[0038] The term "administration" as used herein refers to providing a predetermined substance to an individual or patient by any appropriate method. Depending on the intended method, administration can be parenteral (e.g., intravenous, subcutaneous, intraperitoneal, or topical injection) or oral. The dosage range varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. Liquid formulations for oral administration of the compositions of the present invention include suspensions, oral solutions, emulsions, syrups, etc., which may contain various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, etc. The pharmaceutical compositions of the present invention can be administered by any device capable of delivering the active substance to target cells. Preferred administration methods and formulations include intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, and infusion injections. Injectable preparations can be prepared using aqueous solvents such as physiological saline and Ringer's solution, or non-aqueous solvents such as vegetable oils, higher fatty acid esters (e.g., ethyl oleate), and alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin), and can contain pharmaceutical carriers such as stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium metabisulfite, BHA, tocopherol, EDTA), emulsifiers, buffers for pH adjustment, and preservatives to prevent microbial growth (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

[0039] The term "individual" as used in the present invention refers to all animals, including monkeys, cows, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, and guinea pigs, including humans, who have or may develop the metabolic diseases, and administration of the pharmaceutical composition of the present invention to an individual can effectively prevent or treat the diseases. The pharmaceutical composition of the present invention can be administered in parallel with existing therapeutic agents.

[0040] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable additive, such as starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, candy, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, talc, etc. The pharmaceutically acceptable additive according to the present invention is preferably included in an amount of 0.1 to 90 parts by weight of the composition, but is not limited thereto. In one aspect, the present invention relates to a health functional food containing torsemide and cromolyn, or a pharmaceutically acceptable salt thereof, for preventing and improving metabolic diseases.

[0041] In one embodiment, torsemide may be a compound represented by the following chemical formula 3, and cromolyn may be a compound represented by the following chemical formula 4:

[0042] [ka]

[0043] and;

[0044] [ka]

[0045] In one embodiment, the metabolic disease may be any one selected from the group consisting of diabetes, obesity, insulin resistance, and metabolic liver disease, and the metabolic liver disease may be any one selected from the group consisting of: Metabolic Dysfunction Associated Steatotic Liver Disease (MASLD) may be Metabolic dysfunction-associated fatty liver disease teeth, Metabolic Dysfunction Associated Steatohepatitis (MASH), non-alcoholic fatty liver (NAFL) and NAFLD-associated liver fibrosis; Metabolic dysfunction-related steatohepatitis It is more preferable that:

[0046] In one embodiment, the prevention or treatment of metabolic diseases may consist of reducing insulin resistance, reducing body weight, and reducing triglyceride accumulation in liver tissue.

[0047] In one embodiment, the pharmaceutically acceptable salt of cromolyn can be selected from the group consisting of alkali metal salts, alkaline earth metal salts, salts with inorganic acids, salts with organic acids, and salts with acidic amino acids, with cromolyn sodium being most preferred.

[0048] In one embodiment, the torsemide and cromolyn or pharmaceutically acceptable salts thereof of the present invention can be administered at a dose of 0.01 mg / kg to 1 g / kg of torsemide and cromolyn, respectively.

[0049] In one embodiment, torsemide and cromolyn can be administered simultaneously, separately or sequentially.

[0050] When the composition of the present invention is used as a food composition, the composition can be added as it is or used together with other foods or food ingredients, and can be used appropriately by a conventional method. The composition can contain a nutrient-acceptable food supplement additive in addition to the active ingredient, and the amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment).

[0051] The term "food supplement additive" as used in the present invention refers to a component that can be added supplementarily to food, and is added to prepare various forms of health functional foods, and can be appropriately selected and used by those skilled in the art. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated drinks, etc., but the types of food supplement additives of the present invention are not limited by these examples.

[0052] The food composition of the present invention may include a functional health food. The term "functional health food" as used herein refers to a food prepared and processed in the form of a refined product, capsule, powder, granule, liquid, or pill using raw materials or ingredients that have beneficial functions for the human body. Here, "functional" refers to the regulation of nutrients for the structure and function of the human body or the attainment of beneficial health effects, such as physiological effects. The functional health food of the present invention can be prepared by methods commonly used in the art, and can be prepared by adding raw materials and ingredients commonly added in the art. Furthermore, the dosage form of the functional health food can be prepared without limitation, as long as it is recognized as a functional health food. The food composition of the present invention can be prepared in various dosage forms, and unlike general pharmaceuticals, it is made from food ingredients, which has the advantage of being free of side effects that can occur with long-term drug administration, and is highly portable. The functional health food of the present invention can be taken as a supplement to enhance the effects of therapeutic agents for metabolic diseases.

[0053] Furthermore, there is no limit to the type of health food in which the composition of the present invention can be used. A composition containing the composition of the present invention as an active ingredient can be prepared by mixing appropriate supplementary ingredients and known additives that can be included in health functional foods, according to the selection of a person skilled in the art. Examples of foods to which the composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, energy drinks, alcoholic beverages, and vitamin supplements. The extract of the present invention can also be added to soups, teas, jellies, and juices made primarily with the extract of the present invention.

[0054] In one aspect, the present invention relates to a method for treating a metabolic disorder comprising administering to an individual having the metabolic disorder pharmaceutically effective amounts of torsemide and cromolyn, or pharmaceutically acceptable salts thereof.

[0055] In one aspect, the present invention relates to the use of torsemide and cromolyn, or pharmaceutically acceptable salts thereof, for the manufacture of a pharmaceutical composition for the prevention and treatment of metabolic diseases. [Example]

[0056] The present invention will be described in more detail with reference to the following examples, which are provided for illustrative purposes only and are not intended to limit the scope of the present invention.

[0057] Example 1 Metabolic dysfunction-related steatohepatitis Confirming weight change in animal models Metabolic Dysfunction Associated Steatohepatitis (MASH) To prepare an animal model and confirm the effect of the drug of the present invention on it, a high fat diet model was used in which a high fat diet with a fat content of 60% was ingested for 12 weeks. Metabolic Dysfunction Associated Steatohepatitis (MASH)Specifically, 21 6-week-old ICR mice were pre-fed with a normal diet for 7 days prior to the start of the experiment to allow them to adapt to the rearing environment, and then they were divided into a control group (n = 3, normal fat diet with a 10% fat content, NFD), a high-fat diet control group (n = 9, high-fat diet, HFD), a high-fat diet experimental group (n = 6, cromolyn treat), and a high-fat diet experimental group (n = 6, cromolyn + torsemide treat) ( MASH The mice were divided into two groups (inducible group and induced group). The total feeding period was 13 weeks. Experimental diet and drinking water were available ad libitum, and all experimental diets were kept refrigerated during the feeding period. Drug administration began 6 weeks after the start of the high-fat diet and continued for 6 weeks. The control group received PBS (Phosphate Buffered Saline), while the experimental groups received cromolyn sodium 10 mg / kg or cromolyn 10 mg / kg + torsemide 10 mg / kg intraperitoneally three times a week. The mice in these groups were weighed at regular intervals once a week during the experimental period to monitor weight changes.

[0058] As a result, after 12 weeks, the high-fat diet experimental group (HFD) had a mean body weight of 61.3g, which was significantly higher than the normal diet group (NFD), whose mean body weight was 50.8g. MASH We confirmed that the animal model was well established (Figure 1A). Furthermore, the cromolyn-administered high-fat diet experimental group (HFD cromolyn) did not show significant changes in body weight compared to the high-fat diet control group (HFD PBS). However, the high-fat diet experimental group administered cromolyn and torsemide in combination (HFD cromolyn + torsemide) showed a significant decrease in body weight from 9 weeks of age, reaching 57.8 g at 12 weeks, a statistically significant decrease compared to the mean weight of the high-fat diet experimental group (HFD) ( * p-value<0.05) (Figure 1B).

[0059] Example 2. Metabolic dysfunction-related steatohepatitis Confirmation of insulin resistance in animal models Cromolyn and / or torsemide MASHTo confirm insulin resistance in the animal model, blood glucose levels were monitored in each group of mice at week 13 of the experiment. Specifically, after 13 weeks of diet and drug administration as in Example 1, the experimental animals were fasted for 16 hours, and fasting blood samples were collected from the peripheral vein of the tail. Each group received an intraperitoneal injection of 1.5 mg / kg glucose solution (Sigma Aldrich, St. Louis, MO, USA), and blood samples were collected from the tail vein at 15, 30, 45, 60, 90, and 120 minutes, respectively, to measure changes in blood glucose levels.

[0060] The GTT (Glucose Tolerance Test) results showed that at 90 minutes, the control group (HFD PBS) had an average fasting blood glucose level (mg / dl) of 509.6 mg / dl, while the experimental group (cromolyn + torsemide treat) had an average fasting blood glucose level (mg / dl) of 319.4 mg / dl. At 120 minutes, these levels were 486 mg / dl and 267.4 mg / dl, respectively, demonstrating a significant reduction in blood glucose levels with the combined treatment with cromolyn and torsemide (Figure 2). Additionally, the ITT (Insulin Tolerance Test) results showed that at 120 minutes, the control group (HFD PBS) had an average fasting blood glucose level of 260 mg / dl, while the experimental group (cromolyn + torsemide treat) had an average fasting blood glucose level of 158.6 mg / dl, demonstrating an even lower blood glucose level in the experimental group. This confirmed that the control group had higher insulin resistance than the experimental group, and that insulin, which lowers blood glucose, was working well in the experimental order, resulting in a significant reduction in insulin resistance ( * p-value<0.05) (Fig. 2).

[0061] Example 3. Confirmation of neutral fat in liver tissue Cromolyn and / or torsemide MASHOil Red O staining analysis was performed to confirm changes in the level of fat accumulation in the animal model. Specifically, after 13 weeks of diet and drug administration as described in Example 1, the experimental animals in each group were sacrificed, their livers were removed, and cryo-treated tissue blocks were sectioned at a thickness of 12 μm. The sectioned tissues were treated with Oil Red O working solution (oil red O:DW 3:2) and incubated at room temperature for 5 minutes. After 20 minutes of washing in running water, the tissues were mounted on cover slips using water-soluble mounting medium. After 10 minutes of incubation at room temperature, the tissues were observed under a microscope to confirm changes in the level of triglyceride accumulation in the liver tissue due to drug treatment.

[0062] The results showed that the amount of triglycerides in the liver tissue increased in the HFD PBS group compared to the control groups (NFD, HFD PBS), but was relatively reduced in the experimental groups (HFD cromolyn, HFD cromolyn + torsemide), and that the reduction was particularly significant in the group treated with both cromolyn and torsemide ( * p-value<0.05) (Fig. 3).

Claims

1. A pharmaceutical composition for preventing or treating metabolic dysfunction associated steatohepatitis (MASH), comprising torsemide or a pharmaceutically acceptable salt thereof, and cromolyn or a pharmaceutically acceptable salt thereof as active ingredients.

2. The pharmaceutical composition for preventing or treating metabolic dysfunction-associated steatohepatitis according to claim 1, wherein torsemide is a compound represented by the following chemical formula 1: 【Chemical 1】 。

3. The pharmaceutical composition for preventing or treating metabolic dysfunction-associated steatohepatitis according to claim 1, wherein cromolyn is a compound represented by the following chemical formula 2: 【Chemistry 2】 。

4. The pharmaceutical composition for preventing or treating metabolic dysfunction-associated steatohepatitis according to claim 1, which reduces insulin resistance.

5. The pharmaceutical composition for preventing or treating metabolic dysfunction-associated steatohepatitis according to claim 1, which reduces body weight.

6. The pharmaceutical composition for preventing or treating metabolic dysfunction-associated steatohepatitis according to claim 1, which reduces neutral fat accumulation in liver tissue.

7. The pharmaceutical composition for preventing or treating metabolic dysfunction-associated steatohepatitis according to claim 1, wherein the salt of cromolyn is cromolyn sodium.

8. A health functional food for preventing or ameliorating metabolic dysfunction-associated steatohepatitis, comprising torsemide or a pharmaceutically acceptable salt thereof, and cromolyn or a pharmaceutically acceptable salt thereof.

9. The health functional food for preventing or ameliorating metabolic dysfunction-related steatohepatitis according to claim 8, wherein torsemide is a compound represented by the following chemical formula 3: 【Chemistry 3】 。

10. The health functional food for preventing or ameliorating metabolic dysfunction-related steatohepatitis according to claim 8, wherein cromolyn is a compound represented by the following chemical formula 4: 【Chemistry 4】 。

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