PHARMACEUTICAL COMPOSITION FOR THE PREVENTION OR TREATMENT OF NON-ALCOHOLIC FATTY LIVER DISEASE

Abstract

The present invention relates to a pharmaceutical composition for prevention or treatment of nonalcoholic fatty liver disease.

Description

[0001] Description of the Invention

[0002]

Title of invention

Technical field

[0003]

Background Art

[0004]

Contents of the invention

[0005]

Technical Problem

[0006]

Technical Solution

[0007] [Chemical Formula 1] In the above chemical formula 1,

[0008] A is an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group, and B is a pyrimidine group substituted with a C1-C6 straight or branched chain alkyl group.

[0009] 1 is each independently F, Cl, Br or I. In one embodiment, A of the above formula 1 is and at this time, R2 and R3 can each independently be a C1-C6 straight-chain or branched-chain alkyl group. In one embodiment, the above chemical formula 1 is and wherein R7 may be a C1-C6 straight-chain or branched-chain alkyl group. In one embodiment, each of the five groups of the above formula 1 may be. In one embodiment, the term "alkyl" means a straight-chain or branched hydrocarbon residue unless otherwise stated. Examples of the C1-C6 alkyl include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, hexyl, and the like. In one embodiment, the compound represented by the above formula 1 may be 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole. In one embodiment, the GLP-1 receptor agonist is semaglutide, albiglutide, dulaglutide, exenatide, liraglutide, or lixisenata. 0 ]de

[0010] (1 ixisenatide), orfogliprone, danugliprone or GSBR-1290, an analog thereof, or a pharmaceutically acceptable salt thereof. These may be used alone or in combination of two or more. The term “analog” in the present invention is used to mean a fragment analog of a peptide or a derivative thereof; or an analog of a single molecule compound or a derivative thereof. In one embodiment, the GLP-1 receptor agonist may include semaglutide, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the GLP-1 receptor agonist may comprise a GLP-1 analogue, an analogue thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the GLP-1 receptor agonist may comprise semaglutide, albiglutide, dulaglutide, liraglutide, an analogue thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the GLP-1 receptor agonist may be semaglutide, an analogue thereof, or a pharmaceutically acceptable salt thereof. Semaglutide may be represented by the following chemical formula 2, and N-e 26 - [2- (2 - {2 - [2- (2 - {2 - [ (S)- 4 -carboxy- 4- (17 -carboxyheptadecanoyl-amino)butyrylamino]ethoxy} ethoxy )acetylamino]ethoxy} ethoxy )acetyl] [Aib 8 , Arg3 4 ]GLP-1-(7-37) and can be prepared as described in Example 4 of W02006 / 097537.

[0011] [Chemical Formula 2]

[0012] Pharmaceutically acceptable salts of semaglutide may be acid addition salts or base addition salts. Among the pharmaceutically acceptable salts of semaglutide, acid addition salts include, for example, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate. , hydrochloride, 2-hydroxyethanesulfonate (isethionate), nicotinate, 2-naphthalenesulfonate, oxalate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, phosphate, glutamate or bicarbonate. The non-alcoholic fatty liver disease (NAFLD) may include non-alcoholic fatty liver disease, non-alcoholic steatohepatitis (NASH), liver fibrosis or cirrhosis. The non-alcoholic fatty liver disease may be non-alcoholic steatohepatitis. The non-alcoholic steatohepatitis may be a disease accompanied by fibrosis due to an inflammatory response due to oxidative stress, etc., and liver cell damage resulting therefrom. The above non-alcoholic steatohepatitis may be a disease accompanied by fibrosis. The above pharmaceutical composition may be a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease that suppresses neutral fat deposition in liver tissue.The pharmaceutical composition may be a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease that suppresses inflammation in liver tissue. The pharmaceutical composition may be a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease that suppresses fibrosis of liver tissue. When the pharmaceutical composition is described herein as containing A and B, the pharmaceutical composition is interpreted to mean a composition containing A and B; a combination containing A and B as separate preparations; or a composition containing the combination. Therefore, in the present invention, the composition may be used interchangeably with the combination. The pharmaceutical composition may contain the compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist, each as separate preparations, or may contain them all in the form of a composite preparation. The pharmaceutical composition may be a combination of separate preparations, or a composite preparation. The pharmaceutical composition may comprise a first compartment containing a first active ingredient, and a second compartment containing a second active ingredient. The first active ingredient may be a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof. The second active ingredient may comprise a GLP-1 receptor agonist. According to embodiments of the present invention, in the pharmaceutical composition, the first compartment containing the first active ingredient and the second compartment containing the second active ingredient are administered together, and the pharmaceutical composition may be a composition for combined administration. Specifically, the pharmaceutical composition may be a combination in which two components are formulated as separate unit dosage forms. In this case, the first active ingredient and the second active ingredient may be administered together in separate dosage forms.In one embodiment, when administering a pharmaceutical composition, it is confirmed that the NAFLD activity score (NAS) is significantly reduced compared to when administering a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof of a compound represented by chemical formula 1 as a GPR119 agonist; or a GLP-1 receptor agonist, respectively. In one embodiment, when administering a pharmaceutical composition, it is confirmed that the fat metabolism is significantly improved synergistically in terms of liver tissue cholesterol content, relative area of ​​liver tissue fat, and the proportion of liver cells containing fat globules (image analysis indices highly correlated with the steatosis score) compared to when administering a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof of a compound represented by chemical formula 1 as a GPR119 agonist; or a GLP-1 receptor agonist, respectively. In the present invention, non-limiting examples of pharmaceutically acceptable salts of the compound represented by the above chemical formula 1 may include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid or sulfuric acid; salts with organic carboxylic acids such as acetic acid, trifluoroacetic acid, citric acid, maleic acid, oxalic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid or malic acid, or sulfonic acids such as methanesulfonic acid or para-toluenesulfonic acid; salts with alkali metals such as sodium, potassium or lithium; or salts with various acids known to be capable of forming other pharmaceutically acceptable salts. The pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease of one embodiment may be used in the form of a general pharmaceutical formulation. The pharmaceutical formulation may be administered in various oral and parenteral dosage forms, and the dosage form may be determined in various ways depending on the method of use.When the pharmaceutical composition of one embodiment is formulated into various oral and parenteral dosage forms, it can be manufactured using excipients such as fillers, diluents, bulking agents, binders, wetting agents, disintegrants, and surfactants that are commonly used. Solid preparations for oral administration may include tablets, pills, powders, granules, and capsules, and such solid preparations can be manufactured by mixing the pharmaceutical composition with at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. In addition, liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized 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, macrogol, tween 61, cacao butter, laurin butter, and glycerol latin. In addition, the pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease according to the present invention can exhibit an effective amount in the range of about 1 to about 1,000 mg. The dosage or administration amount can be administered in various dosages and methods, such as once a day or divided into several times a day, depending on the subject's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity.In the present invention, non-alcoholic fatty liver disease includes both primary and secondary non-alcoholic fatty liver disease. Specifically, non-alcoholic fatty liver disease in the present invention includes simple fatty liver, non-alcoholic steatohepatitis, and liver fibrosis and liver cirrhosis caused by the progression of these diseases. However, it is not limited thereto. In one embodiment, non-alcoholic fatty liver disease (NAFLD) in the present invention may be substantially the same as metabolic dysfunction associated steatohepatitis (MAFLD) referred to in the United States, Europe, etc. In one embodiment, non-alcoholic fatty liver disease (NAFLD) in the present invention may be substantially the same as metabolic dysfunction-associated fatty liver disease (MAFLD). In one embodiment, non-alcoholic steatohepatitis (NASH) in the present invention may be substantially the same as metabolic dysfunction associated steatohepatitis (MASH) referred to in the United States, Europe, etc. The present invention also provides a method for preventing or treating non-alcoholic fatty liver disease, comprising administering to a subject in need of treatment a pharmaceutical composition comprising a therapeutically effective amount of a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist. As used herein, the term "subject in need of treatment" means a mammal including a human, and the term "administration" means providing a predetermined substance to the subject by any appropriate method.The term "therapeutically effective amount" means an amount of an active ingredient or pharmaceutical composition that is thought by a researcher, veterinarian, physician, or other clinician to induce a biological or medical response in an animal or human, including an amount that induces symptomatic relief of a disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dosage and frequency of administration of the active ingredient of the present invention will vary depending on the desired effect. In one embodiment, the route of administration of the pharmaceutical composition of the present invention may be administered via any conventional route as long as it can reach the target tissue. Examples include, but are not limited to, oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, intranasal administration, intrapulmonary administration, rectal administration, intracavitary administration, intraperitoneal administration, and intrathecal administration. In one embodiment, the pharmaceutical composition is administered once daily or at regular intervals once daily.

[0013] It can be administered more than twice. The pharmaceutical composition of one embodiment can be used alone or in combination with methods using surgery, hormone therapy, drug therapy and biological response modifiers for the prevention and treatment of nonalcoholic fatty liver disease. One embodiment provides a composition for preventing or improving nonalcoholic fatty liver disease, comprising a compound represented by the above chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof or a mixture thereof; and a GLP-1 receptor agonist as active ingredients. The composition may be a food composition or a feed composition. In one embodiment, the term "improvement" means any act in which the disease is improved or beneficially changed by administration of the composition. In one embodiment, the term "food" includes dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, health foods, and health supplements, and includes all foods in the conventional sense. The "functional food" above is the same term as food for special health use (FoSHU), and refers to a food with high medical or healthcare effects that is processed to efficiently exhibit bioregulatory functions in addition to providing nutrition. Here, "functionality" means obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological functions. The "health food" above refers to a food that has a more active health maintenance or promotion effect than general foods, and "health supplement food" refers to a food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement may be used interchangeably.The food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process. Specifically, it can include protein, carbohydrate, fat, nutrients, seasoning, and flavoring agent, and examples of the carbohydrate include, but are not limited to, glucose, fructose, maltose, sucrose, oligosaccharide, dextrin, cyclodextrin, xylitol, sorbitol, erythrol, saccharin, or synthetic flavoring agent. The food composition of the present invention can be manufactured into various types of formulations without limitation as long as it is a formulation recognized as food. The term "feed" of the present invention means any natural or artificial diet, meal, etc. or ingredients of the meal for ingestion and digestion by livestock or suitable therefor. The feed may include feed additives or supplementary feed. The type of the feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant-based feeds such as grains, roots, fruits, food processing by-products, algae, fibers, pharmaceutical by-products, fats, starches, meal, or grain by-products; and animal-based feeds such as proteins, inorganic substances, fats, mineral substances, fats, single-cell proteins, zooplankton, or foodstuffs. These may be used alone or in combination of two or more. The present invention also provides a method for preventing or improving non-alcoholic fatty liver disease, comprising administering to a subject in need of improvement a food or feed composition comprising, as active ingredients, a compound represented by the above chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist.The present invention also provides a use of a pharmaceutical composition comprising a compound represented by the above chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist as an active ingredient for preventing or treating nonalcoholic fatty liver disease. The present invention also provides a use of a pharmaceutical composition comprising a compound represented by the above chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist as an active ingredient for preparing a pharmaceutical preparation for preventing or treating nonalcoholic fatty liver disease. Matters mentioned in each of the pharmaceutical composition, treatment method, food composition, improvement method, feed composition, and use according to the embodiments of the present invention are equally applicable to each embodiment unless contradictory.

[0014] (1) The present invention provides a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease, comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist.

[0015] [Chemical Formula 1] In the above chemical formula 1,

[0016] A is an oxadiazole group substituted with a C1-C6 straight-chain or branched-chain alkyl group, B is a pyrimidine group substituted with a C1-C6 straight-chain or branched-chain alkyl group, and each of the five independently represents F, Cl, Br, or I.

[0017] (2) In the above (1), the GLP-1 receptor agonist is semaglutide.

[0018] (semaglutide), albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, orforglipron, danuglipron, GSBR-1290, analogs thereof, or pharmaceutically acceptable salts thereof may be included. 0 드

[0019] (lixisenatide), orforglipron, danuglipron, GSBR-1290, analogs thereof, or pharmaceutically acceptable salts thereof may be included.

[0020] N:, •■•■사. I아,、,,.•■窓

[0021] (3) In the above (1) or (2), A may be 忌 or 브, and R2 and R3 may each independently be a C1-C6 straight-chain or branched-chain alkyl group. r"W w j, J

[0022] (4) In any one of the above (1) to (3), B is < ' W and R7 may be a C1-C6 straight-chain or branched-chain alkyl group.

[0023] (5) In any one of the above (1) to (4), the compound represented by Chemical Formula 1 may be 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole.

[0024] (6) In any one of (1) to (5), the GLP-1 receptor agonist may include semaglutide, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, an analog thereof, or a pharmaceutically acceptable salt thereof. (7) In any one of (1) to (6), the GLP-1 receptor agonist may include semaglutide, an analog thereof, or a pharmaceutically acceptable salt thereof.

[0025] (8) In any one of the above (1) to (7), non-alcoholic fatty liver disease may include at least one selected from the group consisting of simple fatty liver, non-alcoholic steatohepatitis, liver fibrosis, and liver cirrhosis.

[0026] (9) In any one of the above (1) to (8), the pharmaceutical composition can suppress fibrosis of liver tissue.

[0027] (10) The present invention provides a method for preventing or treating non-alcoholic fatty liver disease, comprising administering a pharmaceutical composition comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist.

[0028] [Chemical Formula 1] In the above chemical formula 1,

[0029] A is an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group,

[0030] B is a pyrimidine group substituted with a C1-C6 straight or branched chain alkyl group, and each of the five independently represents F, Cl, Br, or I.

[0031] (11) The composition according to (10) above may be a composition according to any one of (1) to (9) above. (12) In (10) or (11), the method may include a step of administering a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a step of administering a GLP-1 receptor agonist.

[0032] (13) In the above (12), after the step of administering the compound represented by the chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof, a GLP-1 receptor agonist may be administered.

[0033] (14) In the above (12), after administering a GLP-1 receptor agonist, a compound represented by chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof may be administered.

[0034] (15) The present invention provides a use of a pharmaceutical composition comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist for the prevention or treatment of non-alcoholic fatty liver disease.

[0035] [Chemical Formula 1] In the above chemical formula 1,

[0036] A is an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group,

[0037] B is a pyrimidine group substituted with a C1-C6 straight-chain or branched-chain alkyl group, and each of the above is independently F, Cl, Br or I. (16) In the above (15), the pharmaceutical composition may be a composition according to any one of the above (1) to (9).

[0038] (17) The present invention provides the use of a pharmaceutical composition comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist for the manufacture of a drug for preventing or treating non-alcoholic fatty liver disease.

[0039] [Chemical Formula 1] In the above chemical formula 1,

[0040] A is an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group,

[0041] B is a pyrimidine group substituted with a C1-C6 straight or branched chain alkyl group, and each of the five independently represents F, Cl, Br, or I.

[0042] (18) In the above (17), the pharmaceutical composition may be a composition according to any one of the above (1) to (9).

[0043] (19) The present invention provides a pharmaceutical combination comprising a compound represented by the above chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and a GLP-1 receptor agonist.

[0044] [Chemical Formula 1] In the above chemical formula 1,

[0045] A is an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group,

[0046] B is a pyrimidine group substituted with a C1-C6 straight or branched chain alkyl group, and each of the five independently represents F, Cl, Br, or I.

[0047] (20) In the above (19), the GLP-1 receptor agonist is semaglutide.

[0048] (semaglutide), albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide

[0049] ( 1 ixisenat ide) , orforgl ipron , danugl ipron ,

[0050] GSBR-1290, their analogs, or their pharmaceutically acceptable salts. Seed: ~Si W 、、、 .,.•Shu

[0051] < J <1 Orphan

[0052] (21) In the above (19) or (20), A is Hi or Eun, and R2 and R3 can each independently be a C1-C6 straight-chain or branched-chain alkyl group.

[0053] (22) In any one of the above (19) to (21), B is , and R7 can be a C1-C6 straight or branched chain alkyl group.

[0054] (23) In any one of the above (19) to (22), the compound represented by the above chemical formula 1 may be 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole.

[0055] (24) In any one of the above (19) to (23), the GLP-1 receptor agonist may include semaglutide, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, an analog thereof, or a pharmaceutically acceptable salt thereof.

[0056] (25) In any one of the above (19) to (24), the GLP-1 receptor agonist may include semaglutide, an analog thereof, or a pharmaceutically acceptable salt thereof.

[0057] (26) In any one of (19) to (25), the combination may be for the treatment or prevention of non-alcoholic fatty liver disease.

[0058] (27) The present invention provides a pharmaceutical composition for treating or preventing non-alcoholic fatty liver disease, comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof, which uses a GLP-1 receptor agonist in combination.

[0059] [Chemical Formula 1] In the above chemical formula 1,

[0060] A is an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group,

[0061] B is a pyrimidine group substituted with a C1-C6 straight or branched chain alkyl group, and each of the above is independently F, Cl, Br or I. (28) The present invention provides a pharmaceutical composition for treating or preventing non-alcoholic fatty liver disease, comprising a GLP-1 receptor agonist, which uses a combination of a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof or a mixture thereof.

[0062] [Chemical Formula 1] In the above chemical formula 1,

[0063] A is an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group,

[0064] B is a pyrimidine group substituted with a C1-C6 straight or branched chain alkyl group, and each of the five independently represents F, Cl, Br, or I.

[0065] (29) In the above (27) or (28), the compound represented by the chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a hydrate or solvate thereof, or a mixture thereof; and the GLP-1 receptor agonist may be according to any one of the above (2) to (9).

[0066]

Effect of the invention

[0067]

Brief description of the drawings

[0068]

Embodiment of the Invention

[0069] GPR119 is one of the G protein-coupled receptors (GPCR) that activates signal transduction through cAMP by coupling to GV. GPR119 is highly expressed in pancreatic beta cells and L cells of the small intestine, and is also present in the liver and is a regulator of fatty acid biosynthesis that occurs in the liver. In addition, when GP119 is activated, it suppresses the differentiation and activation of inflammatory cells and inhibits the activation of hepatic stellate cells, thereby controlling fatty liver, inflammation, and fibrosis in non-alcoholic fatty liver disease. Hereinafter, a pharmaceutical composition according to an embodiment of the present invention will be described in detail with reference to Examples and Comparative Examples. Hereinafter, Compound 1 means 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole. Hereinafter, “GLP-1RA” described in the experimental results refers to semaglutide as a GLP-1 Receptor Agoni st.

[0070] Improvement of NAS (NAFLD Activity Score, NAFLD Activity Score) in DIO-NASH mice

[0071] Nonalcoholic steatohepatitis was induced in 5-week-old male C57BL / 6J mice by feeding them a special diet containing high fat, high fructose, and high cholesterol (Research Diet, D09100301) for 36 weeks. Then, DIO-NASH mice were administered compound 1 (oral) and semaglutide (subcutaneous injection) at 100 mg / kg and 30 nmol / kg, respectively, once daily for 8 weeks, either alone or in combination. NAS (NAFLD activity score) before and after administration through liver tissue biopsy was compared. The results were analyzed as follows. Liver tissues isolated through biopsy and necropsy were fixed in 10% neutral buffered formalin, paraffin blocks were prepared, and 3-μm-thick tissue sections were obtained. Afterwards, Hematoxyl in and Eosin (HE) Stain was performed, and the histological characteristics of the stained liver tissue were the ratio of deposited fat globules, the degree of inflammatory cell infiltration, and the degree of cell necrosis, including fat accumulation (steatosi s), inflammation (inf laminat ion), and hepatocyte swelling (hepatocyte bal looning), abnormalities.

[0072] It is divided into three categories and marked as NAS.

[0073] The NAS scoring criteria are as shown in Table 1 below.

[0074] [Table 1] Comparing the NAS before and after administration, the NAS before and after administration were improved (decreased) by 2 or more points (U 2-point improvement), the NAS after administration was improved (decreased) by 1 or more point (U 1-point improvement), the NAS had no change (no change), and the NAS score after administration was increased (worsened) (worsened) are shown as a percentage of the total number of subjects in Figure 1. The NAS before and after administration are shown in Figure 2. The results were analyzed by one-sided Fisher's exact test with Bonferroni correction. *** indicates P < 0.001 compared to vehicle (control) for individuals with 1 or more point improvement, and ### indicates P < 0.001 compared to vehicle (control) for individuals with 2 or more point improvement. (**: P < 0.001 compared to vehicle for > 1 point improvement. ###: P < 0.001 compared to vehicle for > 2 point improvement.) Individuals with 1 or more point improvement are the sum of individuals with 1 point improved NAS and individuals with 2 or more point improvement (n = 14 to 15 individuals in each group). Referring to Figures 1 and 2, it can be confirmed that all subjects (100%) in the combination group administered with Compound 1 and semaglutide showed improvement in symptoms, with a decrease of 1 point or more compared to before administration. Even 80% of all subjects showed an improvement of 2 points or more in NAS. In the combination administration group, it can be confirmed that at least one item among fat accumulation, hepatocyte expansion, and inflammation improved by 2 points or more; or at least two or more of these items improved by 1 point or more. In contrast, only 6.67% of the control group, 21.43% of the compound 1 monotherapy group, and 21.43% of the semaglutide monotherapy group showed an improvement of 2 points or more.It was confirmed that it was only 43%, and that when administered together, it showed results that were approximately 4 times better.

[0075] Improved lipid metabolism in the liver of DIO-NASH mice

[0076] In DI0-NASH mice, compound 1 and semaglutide were administered alone or in combination once daily for 8 weeks at the highest effective dose. Liver tissue biopsies were analyzed for liver tissue cholesterol content, relative area of ​​liver tissue fat, and the proportion of liver cells containing fat globule (an image analysis index highly correlated with steatosis score). Liver tissue cholesterol content was measured using a commercially available kit for an automatic analyzer after cholesterol extraction, and the relative area of ​​liver tissue fat was quantified through image analysis of HE-stained slides. The results are shown in Figs. 3 , 4 , and 5 , respectively. In Figs. 3 to 5, the values ​​expressed as the mean of the number of individuals n = 14 to 15 + SEM in each group (Values ​​expressed as mean of n = 14-15 + SEM) were analyzed according to Dunnett's test one-factor 1 linear model, and *** means P < 0.001 compared to the vehicle (control group). Referring to Figs. 3 to 5, when compound 1 and semaglutide were administered together, it was confirmed that fat metabolism was improved synergistically compared to each administration alone. Efficacy in improving fibrosis in a liver fibrosis mouse model

[0077] After inducing liver fibrosis with GAN feed and CCh, compound 1 was administered orally once daily for 4 weeks, and semaglutide was injected subcutaneously twice a week. The doses were 100 mg / kg and 250 nmo 1 / kg, respectively. After staining with Masson's trichrome (MT) staining, the stained fibrotic area was quantified. In addition, changes in liver weight were also measured. The results are shown in Figs. 6, 7, and 8. In Figs. 6 to 8, the values ​​expressed as mean of n = 18 (NASH-fibrosis) + SEM of the normal population n = 4 and the number of individuals in each group n = 18 + SEM were analyzed by T-test or Dunnett's test, and ** indicates P < 0.01 compared to the vehicle (control group). In Figs. 6 to 8, “normal” refers to the normal mouse group, “fibrosis” refers to the fibrosis control group, “semaglutide” or “GLP1RA” refers to the group administered semaglutide alone, “compound 1” refers to the group administered compound 1 alone, and “combination” refers to the group administered semaglutide and compound 1 together. In Fig. 6, the areas including the fibrotic area are indicated with dotted ovals so that the fibrotic area can be identified separately from other areas. The quantitative results in Fig. 7 are the results of quantifying only the fibrotic area. Referring to Figs. 6 and 7, it can be confirmed that fibrosis was significantly increased in the fibrotic control group compared to the normal mouse group, and it can be confirmed that fibrosis was relatively less in the group administered semaglutide or compound 1 alone compared to the fibrotic control group. Compared to these, a synergistic drug effect in which fibrosis was reduced was confirmed in the combination administration group.Referring to Fig. 8, it can be confirmed that the change in relative liver weight also showed a synergistic decrease when co-administered compared to the group administered with compound 1 or semaglutide alone. In a mouse model of liver fibrosis, the mRNA expression of Hedgehog interacting protein (Hhip), which inhibits the activation of hepatic stellate cells and the effect of inhibiting the expression of fibrotic genes, was evaluated. The results are shown in Fig. 9. In Fig. 9, the values ​​expressed as the mean of n = 11-18 (NASH-fibrosis) + SEM of the normal population n = 4 and the number of individuals in each group n = 11 to 18 + SEM were analyzed according to the T-test or Dunnett's test, and ** indicates P < 0.05 between two groups. ## indicates P < 0.01 compared to vehicle (control group). Referring to Figure 9, mRNA expression was decreased in fibrotic mice (vehicle (control group)) compared to normal mice, and mRNA expression was increased compared to vehicle (control group) by administration of compound 1 or semaglutide alone. On the other hand, it was confirmed that mRNA expression was significantly increased above the normal level by co-administration of these. This means that co-administration of compound 1 and semaglutide exhibits a synergistic effect in inhibiting hepatic stellate cell activation. Collagen gene expression in liver tissue was evaluated by reverse transcription-polymerase chain reaction.Ribonucleic acid (RNA) was extracted from liver tissue using TRIzol reagent and isolated. Complementary DNA (cDNA) was synthesized using reverse transcriptase. The expression of fibrosis-related genes {Col lai, Col3al, Col5al, Co^al} was analyzed by real-time polymerase chain reaction. The results are shown in Fig. 10 and Table 2 below.

[0078] [Table 2] Referring to Figure 10 and Table 2, it can be confirmed that the expression of all four types of collagen increased in the fibrosis control group compared to the normal group, and the reduction rate increased synergistically when administered together compared to alone. This demonstrates the effect of reducing fibrosis gene expression due to inhibition of hepatic stellate cell activation.

Claims

1. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease, comprising a compound according to formula 1, its pharmaceutically acceptable salt, their optical isomers, their hydrates or solvates, or mixtures thereof; and a GLP-1 receptor agonist: [Formula 1] , where in the above formula 1 A is an oxadiazole group substituted with a C1-C6 linear or branched alkyl group, B is a pyrimidine group substituted with a C1-C6 linear or branched alkyl group, and each X independently represents F, Cl, Br, or I.

2. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein the GLP-1 receptor agonist comprises semaglutide, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, orfoglipron, danuglipron, GSBR-1290, their analogues or pharmaceutically acceptable salts thereof.

3. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein A is or , and each of R2 and R3 independently represents a C1-C6 linear or branched alkyl group.

4. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein B is , and R7 is a C1-C6 linear or branched alkyl group.

5. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein the compound corresponding to formula 1 is 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole.

6. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein the GLP-1 receptor agonist comprises semaglutide, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, their analogues or their pharmaceutically acceptable salts.

7. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein the GLP-1 receptor agonist comprises semaglutide, its analogue or pharmaceutically acceptable salts thereof.

8. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein the non-alcoholic fatty liver disease comprises at least one selected from the group consisting of simple fatty liver disease, non-alcoholic steatohepatitis, liver fibrosis and liver cirrhosis.

9. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, wherein the pharmaceutical composition inhibits liver tissue fibrosis.

10. A method for preventing or treating non-alcoholic fatty liver disease, comprising administering a pharmaceutical composition according to claim 1.

11. A method for preventing or treating non-alcoholic fatty liver disease according to claim 10, comprising: administration of a compound according to formula 1, its pharmaceutically acceptable salt, their optical isomers, their hydrates or solvates, or mixtures thereof; and administration of a GLP-1 receptor agonist.

12. Use of a pharmaceutical composition according to claim 1 for the treatment or prevention of non-alcoholic fatty liver disease.

13. Use of a pharmaceutical composition according to claim 1 for the preparation of a medicinal product for the treatment or prevention of non-alcoholic fatty liver disease.

14. A pharmaceutical combination comprising a compound according to formula 1 below, its pharmaceutically acceptable salt, their optical isomers, their hydrates or solvates, or mixtures thereof and a GLP-1 receptor agonist: [Formula 1] where in formula 1 above, A is an oxadiazole group substituted with a C1-C6 linear or branched alkyl group, B is a pyrimidine group substituted with a C1-C6 linear or branched alkyl group, and each X independently represents F, Cl, Br, or I.

15. The pharmaceutical combination according to claim 14, wherein the combination is intended for the prevention or treatment of non-alcoholic fatty liver disease.

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