Composition comprising tetraarsenic hexaoxide as active ingredient for treating or preventing metabolic dysfunction-associated fatty liver disease

Abstract

The present invention relates to a pharmaceutical composition comprising tetraarsenic hexaoxide as an active ingredient for treating or preventing metabolic dysfunction-associated fatty liver disease. The pharmaceutical composition according to the present invention significantly alleviates hepatocyte damage and inflammation and ameliorates metabolic dysfunction-associated fatty liver diseases including MASH and steatosis-related complications including obesity, thereby being able to treat and prevent metabolic dysfunction-related fatty liver diseases including MASH.

Description

A composition for the treatment or prevention of metabolic disorder-related fatty liver disease containing arsenic trioxide as an active ingredient

[0001] The present invention relates to a composition for treating or preventing metabolic disorder-related fatty liver disease comprising arsenic trioxide as an active ingredient, and more specifically, to a pharmaceutical composition capable of treating and preventing metabolic disorder-related fatty liver disease, including MASH, by significantly improving liver cell damage and inflammation and improving steatosis-related complications, including liver fibrosis and obesity.

[0002]

[0003] Non-alcoholic fatty liver disease (NAFLD) is a chronic disease that progresses from fatty liver to steatohepatitis, liver fibrosis, cirrhosis, and liver cancer, including non-alcoholic steatohepatitis (NASH), and is a major cause of liver-related morbidity and mortality. The prevalence of NAFLD is estimated to be about 25% of the global general population, and as the prevalence continues to increase, it places a significant burden on public health (Younossi ZM et al., Hepatology. 2016;64:1577-86; Younossi ZM et al., Hepatology. 2016;64:73-84). Although this disease shows a high global prevalence of 20–30%, there are currently no commercially available treatments.

[0004] Meanwhile, in the international academic community, the names of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis (NASH) have recently been changed to metabolic dysfunction-associated fatty liver disease (MAFLD) and metabolic dysfunction-associated steatohepatitis (MASH).

[0005] MAFLD is a condition caused by the excessive accumulation of fat in the liver that is not due to alcohol consumption. If the fatty liver condition persists, it progresses to the MASH stage, where inflammation develops and fibrosis or scarring occurs. If the MASH state continues, fibrosis can lead to additional complications such as liver cirrhosis, loss of liver function, liver failure, and liver cancer years later. Unlike fatty liver caused by alcohol, similar to conventional NASH, MASH refers to steatohepatitis that develops due to metabolic abnormalities such as obesity and diabetes. MASH is a serious disease that is a major cause of liver-related deaths and significantly increases the incidence of liver cirrhosis and liver cancer.

[0006] The only MASH treatment developed to date is Madrigal Pharmaceuticals' Resmetirom, which was recently approved by the FDA.

[0007] According to market research firm GlobalData, the global market for MASH treatments is projected to reach approximately 34 trillion won by 2026 once treatments are launched. The number of MASH patients is on the rise, and the Health Insurance Review and Assessment Service (HIRA) reported that as of 2021, 405,950 MASH patients were recorded in Korea alone, marking an increase of over 40% in five years. Currently, the number of MASH patients worldwide is estimated to exceed 440 million. Consequently, the development of treatments focused on the steatohepatitis stage of non-alcoholic fatty liver disease is being spearheaded by domestic and international pharmaceutical and biotech companies.

[0008] However, since there are currently no effective treatments for MASH, there is a continuous and urgent need for the development of a treatment with excellent therapeutic effects.

[0009]

[0010] Accordingly, the inventors of the present invention have made efforts to develop an excellent therapeutic agent for metabolic disorder-related fatty liver disease including MASH. As a result, they confirmed that a composition containing arsenic trioxide improves liver function and hepatic pathological characteristics in mice with obesity induced by the GAN (Gubra-Amylin NASH) diet, significantly improving not only steatosis but also hepatocyte damage and inflammation, and can treat and prevent metabolic disorder-related fatty liver disease including MASH by improving steatosis-related complications including liver fibrosis and obesity, and thus completed the present invention.

[0011]

[0012] [Prior Art Literature]

[0013] [Patent Literature]

[0014] Korean Registered Patent No. 10-1844049

[0015] Korean Registered Patent No. 10-2240693

[0016] Korean Patent Publication No. 2011-0085261

[0017]

[0018] [Non-patent literature]

[0019] Oh So-yeon, “Anticancer Efficacy of As4O6 in 5637 Human Bladder Cancer Cell Lines”, Master’s Thesis, Department of Global Pharmaceutical Clinical Pharmacy, Graduate School of Global Pharmaceutical Clinical Pharmacy, Ajou University, August 2019.

[0020]

[0021] The objective of the present invention is to provide a pharmaceutical composition that significantly improves not only hepatic steatosis but also hepatocyte damage and inflammation, and exhibits excellent therapeutic effects in improving metabolic disorder-related fatty liver diseases, including MASH.

[0022]

[0023] To solve the above problem, the present invention provides a pharmaceutical composition for the treatment or prevention of metabolic disorder-associated fatty liver disease (MAFLD) comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

[0024] [Chemical Formula 1]

[0025] As4O6

[0026] In another aspect, the present invention provides a pharmaceutical composition for the treatment or prevention of obesity comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0027]

[0028] The pharmaceutical composition according to the present invention significantly improves not only steatosis but also hepatocyte damage and inflammation, and improves steatosis-related complications including liver fibrosis and obesity, thereby exhibiting excellent therapeutic effects for metabolic disorder-related fatty liver diseases including MASH, and exhibits superior efficacy equivalent to or greater than that of Resmetirom, which received FDA approval as a treatment for MASH in 2024.

[0029]

[0030] Figure 1 is a diagram illustrating the effect of Chemical Formula 1 on body weight change and food intake in mice that were induced to be obese by a GAN diet according to Example 1 of the present invention.

[0031] Figure 2 is a diagram illustrating the results of improving liver function in mice that were induced to be obese by a GAN diet using Formula 1 according to Example 2 of the present invention.

[0032] Figure 3 is a diagram illustrating the results of improving liver pathological characteristics in mice that were induced to be obese by a GAN diet with treatment of Formula 1 according to Example 3 of the present invention.

[0033] Figure 4 is a diagram illustrating the results of improving liver fibrosis in mice that were induced to be obese by a GAN diet with treatment of Formula 1 according to Example 4 of the present invention.

[0034] Figure 5 is a diagram illustrating the results of inducing a reduction in body weight and fat cell size in mice in which Chemical Formula 1 was used to induce obesity with a GAN diet according to Example 5 of the present invention.

[0035]

[0036] Specific details for implementing the invention

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a skilled expert in the art to which this invention pertains. In general, the nomenclature used herein and the experimental methods described below are well known and commonly used in the art.

[0038] For convenience, specific terms are defined herein to facilitate a better understanding of the present invention. Unless otherwise defined herein, scientific and technical terms used herein shall have the meanings generally understood by those skilled in the art. The term "included" as used herein means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0039]

[0040] The inventors of the present invention have completed the present invention by discovering that when liver function in obese mice is improved using a composition containing a compound represented by Chemical Formula 1 as an active ingredient, liver function and liver pathological characteristics are improved, significantly improving not only steatosis but also hepatocellular damage and inflammation, and exhibiting excellent therapeutic effects in improving metabolic disorder-related fatty liver disease including MASH and steatosis-related complications including obesity.

[0041] Accordingly, the present invention relates, in one aspect, to a pharmaceutical composition for the treatment or prevention of metabolic disorder-associated fatty liver disease (MAFLD) comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

[0042] [Chemical Formula 1]

[0043] As4O6

[0044]

[0045] In the present invention, the fatty liver disease may be steatosis, statohepatitis, liver fibrosis, or liver cirrhosis.

[0046] In the present invention, the metabolic disorder-related fatty liver disease is preferably metabolic disorder-related steatohepatitis (MASH).

[0047]

[0048] The term "treatment" as used in the present invention refers to any act in which symptoms of metabolic disorder-associated fatty liver disease (MAFLD) or obesity are improved or beneficially altered by administering a composition containing a compound represented by Formula 1 according to the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0049] The term "prevention" as used in the present invention refers to any act of suppressing or delaying the symptoms of metabolic disorder-associated fatty liver disease (MAFLD) or obesity by administering a composition containing a compound represented by Formula 1 according to the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0050] The term "metabolic dysfunction-associated fatty liver disease (MAFLD)" as used in the present invention refers to a fatty liver disease caused by factors other than alcohol. The above-mentioned non-alcoholic fatty liver disease is a term encompassing "non-alcoholic steatosis," "non-alcoholic steatohepatitis," "non-alcoholic fatty liver-associated fibrosis," and "non-alcoholic fatty liver-associated cirrhosis," which are conditions in which fat accumulates in liver cells due to causes other than alcohol.

[0051] As used in this invention, the term “fatty liver” refers to a case in which fat deposition is observed within the liver but there is no damage to hepatocellular cells (ballooning degeneration) or fibrosis. “Fatty liver hepatitis” or “fatty liver hepatitis” refers to a case in which fat deposition is observed within the liver and there are inflammatory findings accompanied by damage to hepatocellular cells (ballooning degeneration). Fatty liver hepatitis may also be accompanied by liver fibrosis. “Cirrhosis” refers to cirrhosis accompanied by histological findings of fatty liver or fatty liver hepatitis, or cirrhosis that occurred in patients with a history of histologically proven fatty liver or fatty liver hepatitis. In this specification, the definitions of terms related to fatty liver disease are intended only to include various pathological conditions related to fatty liver, and the patient's condition defined according to the said terms cannot always be clearly distinguished.

[0052] The term "Metabolic dysfunction-associated steatohepatitis (MASH)" used in this invention refers to a common liver disease associated with metabolic syndrome, characterized by hepatic steatosis, inflammation, and fibrosis, and developed due to abnormalities in metabolic processes such as obesity and diabetes.

[0053]

[0054] The compound of the present invention represented by Chemical Formula 1 above may be prepared into a pharmaceutically acceptable salt and a solvate according to methods conventional in the art. The compound of the present invention may be prepared using Realgar minerals as raw materials according to processing and purification methods conventionally used in the art, for example, including the method described in Korean Registered Patent No. 10-0272835. For the pharmaceutically acceptable salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The acid addition salt is prepared by a conventional method, for example, by dissolving the compound in an excess amount of an aqueous acid solution and precipitating the salt using a water-miscible organic solvent, for example, methanol, ethanol, acetone, or acetonitrile. An equal molar amount of the compound and an acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be dried by evaporation, or the precipitated salt may be filtered by suction.

[0055] In this case, organic and inorganic acids may be used as free acids; inorganic acids may include hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, hydroiodic acid, etc., and organic acids may include methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, Glucuronic acid, aspartic acid, ascorbic acid, carboxylic acid, vanillic acid, etc. can be used.

[0056] In addition, pharmaceutically acceptable metal salts can be produced using a base. Alkali metal or alkaline earth metal salts are obtained, for example, by dissolving a compound in an excess amount of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate. At this time, it is particularly pharmaceutically suitable to produce sodium, potassium, or calcium salts as metal salts, and the corresponding silver salt is obtained by reacting the alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0057]

[0058] A pharmaceutical composition comprising a compound represented by Formula 1 according to the present invention or a pharmaceutically acceptable salt thereof as an active ingredient may additionally be formulated into a formulation using one or more pharmaceutically acceptable excipients. Such excipients may include one or more selected from the group consisting of fillers (diluents), disintegrants, binders, lubricants (lubricants), preservatives, antioxidants, buffers, chelating agents, solubilizers, and sweeteners.

[0059] As a non-limiting example, the filler (diluent) included in the pharmaceutical formulation according to the present invention is microcrystalline cellulose, D-mannitol, lactose anhydrous, lactose monohydrate, lactose dihydrate, lactose trihydrate, pregelatinized starch, calcium carbonate, calcium phosphate dibasic, calcium phosphate tribasic, calcium sulfate, microcrystalline silicified cellulose, powdered cellulose, dextrates, dextrose, fructose, lactitol, One or more selected from the group consisting of sorbitol, starch, sucrose, talc, xylitol, maltose maltodextrin, and maltitol may be used, but are not limited thereto.

[0060] As a non-limiting example, a disintegrant included in a pharmaceutical formulation according to the present invention is croscarmellose sodium, crospovidone, alginic acid, carboxymethyl cellulose calcium, carboxymethyl cellulose sodium, microcrystalline cellulose, powdered cellulose, sodium docusate sodium, guar gum, hydroxypropyl cellulose, methyl cellulose, polacrilin potassium, poloxamer, povidone, sodium alginate, sodium glycine carbonate, sodium lauryl sulfate, sodium starch One or more selected from the group consisting of sodium starch glycolate, starch, and pregelatinized starch may be used, but are not limited thereto.

[0061] By non-limiting example, a binder included in a pharmaceutical formulation according to the present invention is hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropylmethyl cellulose, acacia mucilage, alginic acid, carbomer, carboxymethyl cellulose calcium, carboxymethyl cellulose sodium, microcrystalline cellulose, powdered cellulose, ethyl cellulose, gelatin, liquid glucose, guar gum, maltodextrin, methyl cellulose, polydextrose, polyethylene oxide, povidone, sodium alginate One or more selected from the group consisting of alginate, starch, pregelatinized starch, and sucrose may be used, but are not limited thereto.

[0062] As a non-limiting example, a lubricant (lubricant) included in a pharmaceutical formulation according to the present invention is colloidal silicon dioxide, magnesium stearate, talc, sodium stearyl fumarate, polyethylene glycol 4000, polyethylene glycol 6000, sodium lauryl sulfate, starch, glyceryl behenate, hydrogenated castor oil, stearic acid, glyceryl palmitostearate, glyceryl monostearate, calcium silicate, powdered cellulose, and One or more selected from the group consisting of starch may be used, but are not limited thereto.

[0063] In addition, one or more selected from the group consisting of pearlitol flash, maltitol, sucralose, enzymatically modified stevia, and peppermint micron may be additionally included as additives, but are not limited thereto.

[0064]

[0065] In another aspect, the present invention provides a pharmaceutical formulation comprising a pharmaceutical composition according to the present invention. The pharmaceutical formulation may be a formulation for oral administration and a formulation for parenteral administration. Oral administration may be a solid formulation such as a tablet, pill, powder, granule, or capsule, or a liquid formulation such as a suspension, oral solution, emulsion, or syrup, and preferably may be a solid formulation. More preferably, the pharmaceutical formulation may be in the form of a tablet or capsule.

[0066] Solid formulations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid formulations are formulated by mixing at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. with the arsenic hexaoxide of the present invention. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as humectants, sweeteners, flavorings, and preservatives, may be included.

[0067] Preparations for parenteral administration include injectables, ophthalmics, and topical solutions. For injectables, aqueous solvents such as physiological saline solution and Ringer's solution, 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 pharmaceutical carriers such as stabilizers, emulsifiers, buffers, and preservatives may be used. For ophthalmics, sterile water, solubilizing agents (e.g., polyethylene glycol 400, glycerin), stabilizers (e.g., EDTA), buffers (e.g., boric acid), pH adjusters (e.g., hydrochloric acid, sodium hydroxide), preservatives, wettable powders, emulsifiers, and solubilizers may be used. For topical solutions, water, physiological saline solution, creams, lotions, various forms of gels, and short-chain alcohols and glycols (e.g., ethyl alcohol and propylene glycol) may be used as carriers or diluents.

[0068] The dosage of the above pharmaceutical composition will vary depending on the age, gender, and weight of the subject to treatment, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. The determination of the dosage based on these factors is within the level of a person skilled in the art, and generally, the dosage ranges from 0.01 mg / kg / day to 500 mg / kg / day. A more preferred dosage is from 0.1 mg / kg / day to 100 mg / kg / day. Administration may be performed once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

[0069] From the mouse 8 mg / kg in the example, the human equivalent dose can be expected to be in the range of 15 mg to 40 mg / 60 kg.

[0070] Routes of administration of the pharmaceutical composition according to the present invention include, but are not limited to: oral, intravenous, intramuscular, intra-arterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, ophthalmic, or rectal. Oral or parenteral administration is preferred. As used herein, the term "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrasternal, ophthalmic, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may be administered orally in any orally acceptable dosage form, including, but not limited to: capsules, tablets, and aqueous suspensions and solutions. For oral tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also typically added. When administered orally in capsule form, useful diluents include lactose and dried corn starch.

[0071] The pharmaceutical composition of the present invention may vary depending on several factors, including the activity of the specific compound used, age, body weight, general health, gender, diet, time of administration, route of administration, elimination rate, drug combination, and the severity of the specific disease to be prevented or treated.

[0072] In addition, in another aspect of the present invention, the invention relates to a pharmaceutical composition for the treatment or prevention of obesity comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0073] [Chemical Formula 1]

[0074] As4O6

[0075]

[0076] In addition, in another aspect of the present invention, the invention relates to a method for treating or preventing a metabolic disorder-related fatty liver disease or obesity, comprising the step of administering to an individual a pharmaceutical composition for treating or preventing a metabolic disorder-related fatty liver disease or obesity, the composition comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0077] In addition, in another aspect of the present invention, there is a new use for a pharmaceutical composition for the treatment or prevention of metabolic disorder-related fatty liver disease or obesity, comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0078]

[0079] The present invention will be described in more detail below through examples. These examples are solely for illustrating the present invention, and it will be obvious to those skilled in the art that the scope of the present invention is not to be interpreted as being limited by these examples.

[0080]

[0081] [Example]

[0082] Example 1: Effects of Formula 1 on Body Weight Change and Food Intake in Mice Induced Obesity by GAN Diet

[0083] After modeling Gubra-Amylin NASH (GAN) diet-induced obese (DIO) mice for 33 weeks, the drug was administered for 8 weeks while maintaining the GAN diet. The superior efficacy of Formula 1 was verified using Resmetirom, which received FDA approval at the 2024 MASH, as a control drug (Fig. 1).

[0084]

[0085] Figure 1 (B) The total body weight of each group was measured, and the body weight of the group treated with Formula 1 did not increase or decreased. Figure 1 (C) The body weight before and after treatment with the drug Formula 1 was significantly reduced compared to the control (vehicle) treatment group. Figure 1 (D) The food intake of each group was checked to verify that the drug did not affect diet or appetite. This suggests that Formula 1 exhibits excellent efficacy in significantly reducing body weight without affecting diet or appetite in a mouse model of obesity induced by a GAN diet.

[0086] Figure 1 (A) shows an experimental design in which drugs were administered to a mouse model in which obesity was induced by a GAN diet.

[0087] Figure 1 (B) shows the total body weight of each group measured during the period of the efficacy experiment and plotted as a graph.

[0088] Figure 1 (C) shows a comparison of body weight before and after drug treatment during the period of conducting the efficacy experiment.

[0089] Figure 1 (D) shows the measured food intake of each group during the period of conducting the efficacy experiment.

[0090]

[0091] Example 2: Effect of improving liver function in mice with obesity induced by a GAN diet according to Formula 1

[0092] In Figure 2 (A), the effect of Formula 1 in improving fatty liver was confirmed through liver histological evaluation. In Figures 2 (B, C), Formula 1 reduced liver weight and the liver-to-body weight ratio, verifying that the hypertrophied fatty liver was improved. Figures 2 (D, E) confirmed a reduction in ALT and AST, suggesting that liver function improved due to drug treatment. Intrahepatic lipid triglyceride (TG) and total cholesterol (TC) levels improved due to drug treatment (Figures 2 (F, G)).

[0093] This demonstrates that Chemical Formula 1 significantly improves fatty liver, liver function indicators, and lipid profiles in mice with obesity induced by the GAN diet, showing excellent efficacy in restoring liver health and treating metabolic diseases.

[0094] Figure 2 (A) compares the liver morphology by group for histological evaluation of the liver.

[0095] Figure 2 (B, C) shows the liver weight and liver / body weight ratio.

[0096] Figure 2 (D, E) ALT and AST were measured for liver function tests.

[0097] Figure 2 (F, G) measured intrahepatic fat and total cholesterol.

[0098]

[0099] Example 3: Effect of improving liver pathological characteristics in obese mice fed a GAN diet according to Formula 1

[0100] Figure 3 (A) shows the results of hematoxylin-eosin (H&E) staining and lipid-globule-specific oil red O (ORO) staining performed to identify steatosis, hepatocellular ballooning, and lobular and portal vein inflammation in liver tissue sections; reduced lipid globules and improved liver histological characteristics were observed in the group treated with Formula 1. Figure 3 (B) shows that the degree of steatosis, hepatocellular ballooning, and lobular and portal vein inflammation was evaluated, and treatment with Formula 1 was significantly reduced across all criteria. Figure 3 (C) shows that the overall NAS (NAFLD Activity Score) was significantly reduced by treatment with Formula 1, reflecting the improvement in steatosis as well as ballooning, which indicates hepatocellular damage, and lobular and portal vein inflammation. Formula 1 significantly improved fatty liver, hepatocellular damage, and inflammation in a mouse model of obesity induced by the GAN diet, suggesting that it demonstrated excellent efficacy in restoring liver health.

[0101] In Figure 3 (A), representative micrographs of liver tissue sections from the control group and the group treated with Formula 1 were obtained, and hematoxylin-eosin (H&E) and oil red O (ORO) staining were performed.

[0102] Figure 3 (B) shows the results of a quantitative evaluation of the degree of steatosis, hepatocellular ballooning, and lobular and portal vein inflammation for the NAS score.

[0103] Figure 3 (C) evaluated the NAS score as the sum of the three indicators measured in Figure 3 (B).

[0104]

[0105] Example 4: Effect of improving liver fibrosis in obese mice fed a GAN diet according to Formula 1

[0106] Figure 4 (A) shows liver sections of the control group (vehicle) and the Formula 1 treatment group stained with Sirius Red, demonstrating that liver fibrosis was significantly reduced in the Formula 1 treatment group. Quantification analysis of liver fibrosis before and after drug administration showed a significant reduction in the fibrotic regions of the liver (Figure 4 (B)). Immunohistochemical staining results for α-SMA and Galectin-3, markers of liver fibrosis, showed that the expression of α-SMA and Galectin-3 in the Formula 1 treatment group was significantly reduced compared to the control group, suggesting an improvement in liver fibrosis (Figure 4 (C)). This demonstrates that Formula 1 has proven an excellent therapeutic effect in improving liver fibrosis in GAN diet-induced obese rats through histological and molecular markers.

[0107] Figure 4 (A) shows liver sections of the control group (vehicle) and the chemical formula 1 treatment group stained with Sirius Red to evaluate liver fibrosis.

[0108] Figure 4 (B) shows the liver fibrosis score before and after drug administration.

[0109] In Figure 4 (C), immunohistochemical staining was performed for α-SMA and Galectin-3, markers of liver fibrosis.

[0110]

[0111] Example 5: Effect of inducing reduction in body weight and fat cell size in obese mice fed a GAN diet according to Formula 1

[0112] Figure 5 (A) shows that the total white adipose tissue mass was significantly reduced in the treatment group of Formula 1, based on the weight measurements of glycosylated red white adipose tissue (eWAT), subcutaneous white adipose tissue (sWAT), and thigh white adipose tissue (iWAT). Figure 5 (B) shows that the lipid droplet size was significantly reduced based on the H&E staining results of white adipose tissue (WAT) for morphological analysis of adipocytes. These results demonstrate the potential of Formula 1 to alleviate complications related to steatosis and suggest its therapeutic efficacy in addressing obesity-related pathologies.

[0113] Figure 5 (A) compares the weights of the glycolytic red-white adipose tissue (eWAT), subcutaneous white adipose tissue (sWAT), and thigh white adipose tissue (iWAT).

[0114] Figure 5 (B) shows the results of hematoxylin-eosin (H&E) staining of white adipose tissue (WAT) to evaluate the morphology and size of lipid globules.

[0115]

[0116] Foregoing, specific parts of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the claims and their equivalents.

Claims

1. A pharmaceutical composition for the treatment or prevention of metabolic disorder-associated fatty liver disease (MAFLD) comprising a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical Formula 1] As4O6 2. A pharmaceutical composition according to claim 1, characterized in that the fatty liver disease is steatosis, statohepatitis, fibrosis, or cirrhosis.

3. A pharmaceutical composition according to claim 1, characterized in that the metabolic disorder-related fatty liver disease is metabolic disorder-related steatohepatitis (MASH).

4. A pharmaceutical composition for the treatment or prevention of obesity comprising a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical Formula 1] As4O6 5. A pharmaceutical formulation comprising a pharmaceutical composition according to any one of paragraphs 1 to 4.

6. A pharmaceutical formulation according to claim 5, characterized as being a solid or liquid formulation for oral administration.

7. A pharmaceutical formulation according to claim 6, characterized in that the solid formulation is a tablet, pill, powder, granule, or capsule.

8. A pharmaceutical formulation according to claim 6, characterized in that the liquid formulation is a suspension, an oral solution, an emulsion, or a syrup.

9. A pharmaceutical formulation according to claim 5, characterized as being a formulation for parenteral administration.

10. A pharmaceutical formulation according to claim 9, characterized in that the preparation is an injectable, an ophthalmic, or a topical solution.

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