KR20240099975A - Novel compound and pharmaceutical composition for treating liver disease comprising the same - Google Patents

Abstract

The present invention relates to a compound represented by the following formula (Ⅰ), an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and the compound according to the present invention can be usefully used in the prevention or treatment of liver disease (liver function damage and liver fibrosis). You can.
[Formula Ⅰ]

Description

Novel compound and pharmaceutical composition for treating liver disease comprising the same {NOVEL COMPOUND AND PHARMACEUTICAL COMPOSITION FOR TREATING LIVER DISEASE COMPRISING THE SAME}

The present invention relates to novel compounds, their optical isomers, pharmaceutically acceptable salts thereof, and their medicinal uses. Specifically, the new compound according to the present invention, its optical isomer, and its pharmaceutically acceptable salt can exhibit the effect of treating or preventing liver function damage and liver fibrosis.

In general, liver fibrosis, unlike cirrhosis, is reversible, is composed of thin fibrils, is known to have no nodule formation, and returns to normal once the cause of liver damage is eliminated. This may be possible, but if this liver fibrosis process continues repeatedly, crosslinking between the extra cellular matrix (ECM) increases and progresses to irreversible cirrhosis with nodules.

Liver fibrosis progresses further, resulting in cirrhosis. When hepatocyte necrosis occurs for some reason, liver cell regeneration and fibrous tissue increase, and when this process continues repeatedly for a long time, cirrhosis occurs.

Liver cirrhosis, which is formed by liver nodules or nodules regenerated by persistent or repetitive damage to the liver parenchyma, increased fibrous tissue, and regeneration of hepatocytes (liver function), is pathologically accompanied by necrosis, inflammation, and fibrosis. It is a chronic disease and ultimately progresses to complications such as liver cirrhosis, leading to death.

In particular, because there are no noticeable symptoms in the early stages and it is only discovered when the disease has progressed considerably, research is actively underway to develop methods to quickly diagnose and treat liver function damage or liver fibrosis.

Republic of Korea Patent No. 10-0762448 (September 20, 2007)

Accordingly, the inventors of the present invention made a lot of research and effort to develop a method to more effectively treat liver function damage or liver fibrosis, and as a result, created a new compound and when administered for medicinal purposes, liver function is damaged. Alternatively, the present invention was completed after confirming that the symptoms of liver fibrosis could be improved.

Therefore, the specific problem of the present invention is to provide a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

In order to solve the above problems, the present invention discloses the following means.

Specifically, the present invention discloses a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Additionally, the present invention discloses a pharmaceutical composition for preventing or treating liver disease comprising a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

In addition, the present invention discloses a food composition for preventing or improving liver disease comprising a compound represented by the following formula (I), an optical isomer thereof, or a foodologically acceptable salt thereof.

Additionally, the present invention discloses the use of a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the production of a medicament for the prevention or treatment of liver disease.

Finally, the present invention discloses a method for preventing or treating liver disease comprising administering a therapeutically effective amount of a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

The compound represented by Formula I of the present invention, its optical isomer, or its pharmaceutically acceptable salt has the effect of increasing the blood concentration of alanine aminotransferase (ALT) or aspartate aminotransferase (AST) in hepatocytes. It can be used as a pharmaceutical composition for preventing or treating liver disease by suppressing the increase.

Therefore, using the pharmaceutical composition for preventing or treating liver disease of the present invention, it is possible to reduce the level of markers whose expression level increases due to liver fibrosis, which is an example of liver disease, and is therefore effective in preventing liver function damage or liver fibrosis. It can be widely used for prevention or treatment.

The effects of the present invention are not limited to the effects mentioned above, and various effects may be included within the scope apparent to those skilled in the art from the contents described below.

Figure 1 shows the liver fibrosis inhibition efficacy of the compound (AP1-AL3) of Example 1 using a single-dose carbon tetrachloride (CCl ₄ )-induced liver fibrosis model (damage indicator: alanine aminotransferase (ALT)).
Figure 2 shows the liver fibrosis inhibition effect of Example 1 compound (AP1-AL3) using a single-dose carbon tetrachloride (CCl ₄ ) induced liver fibrosis model (damage indicator: Aspartate Aminotransferase (AST)) .
Figure 3 shows the liver fibrosis inhibition effect of the compound (AP1-AL3) in Example 1 using the single-dose carbon tetrachloride (CCl ₄ )-induced liver fibrosis model according to Experimental Example 1 through Hematocylin & Eosin staining and Masson's trichrome staining.

Hereinafter, this specification will be described in more detail.

This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may also be applied to other descriptions and embodiments. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Additionally, the scope of the present invention cannot be considered limited by the specific description set forth below.

Expressions such as “comprising” used in this specification are understood as open-ended terms that imply the possibility of including other embodiments, unless specifically stated otherwise in the phrase or sentence containing the expression. It has to be.

Terms or words used in the description and claims of the present invention should not be construed as limited to their usual or dictionary meanings, and the inventor should appropriately use the concepts of terms to explain his invention in the best way. Based on the principle of definability, it must be interpreted with meaning and concept consistent with the technical idea of the present invention.

The terms used in this specification are defined as follows.

A compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof

The present invention provides a compound represented by the following formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula Ⅰ]

In the present invention, the compound represented by Formula I has a biphenyldimethyldicarboxylate parent nucleus. am.

In the present invention, pharmaceutically acceptable salts refer to salts commonly used in the pharmaceutical industry, for example, inorganic ionic salts made of calcium, potassium, sodium and magnesium, hydrochloric acid, nitric acid, phosphoric acid, and bromic acid. , inorganic acid salts made from iodic acid, perchloric acid, tartaric acid and sulfuric acid, acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, lactic acid, glycolic acid, gluconic acid. , organic acid salts made of galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p -In the group consisting of sulfonates made from toluenesulfonic acid and naphthalenesulfonic acid, amino acid salts made from glycine, arginine, lysine, etc., and amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc. It may be any one selected, but is not limited thereto.

In the present invention, the compound represented by Formula I can be prepared by the following method.

<Step 1>

<Step 2>

<Step 3>

Pharmaceutical composition for preventing or treating liver disease

Provided is a pharmaceutical composition for preventing or treating liver disease comprising a compound represented by the following formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula Ⅰ]

In the present invention, the liver disease may be any one or more selected from the group consisting of impaired liver function, liver failure, hepatitis, hepatotoxicity, liver fibrosis, and cirrhosis, but is not limited thereto.

In the present invention, the pharmaceutical composition may inhibit the increase in blood concentration of alanine aminotransferase (ALT) or aspartate aminotransferase (AST) in hepatocytes, but is not limited thereto. .

In the present invention, the pharmaceutical composition contains a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, and may further include a pharmaceutically acceptable carrier, and may include a conventional pharmaceutical composition. Depending on the method, it can be formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, and sterile injectable solutions.

In the present invention, the term “pharmaceutically acceptable” refers to a composition that is physiologically acceptable and does not usually cause allergic reactions such as gastrointestinal disorders, dizziness, or similar reactions when administered to humans.

In the present invention, the term “pharmaceutically acceptable carrier” typically includes the liquid or non-liquid basis of the pharmaceutical composition. If the pharmaceutical composition is provided in liquid form, the carrier typically includes pyrogen-free water; Isotonic saline solution or buffered (aqueous) solution, such as phosphate, citric acid, etc. buffer solution. The injection buffer may be hypertonic, isotonic, or hypotonic in a specific reference medium. That is, the buffer may have a high, equal or low salt content in a specific reference medium, and preferably, an appropriate salt concentration may be used within a range that does not induce cell damage due to osmotic pressure or other concentration effects. The reference medium is blood, lymph, cytoplasmic fluid, or other body fluids, or body fluids that occur in "in vivo" methods, for example, as common buffers or liquids, which can be used as reference media in "in vitro" methods. It is liquid. These general buffers or liquids are known to those skilled in the art.

The pharmaceutically acceptable carriers include those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Includes, but is not limited to, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

Additionally, the pharmaceutical composition of the present invention may contain diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and other pharmaceutically acceptable additives.

The pharmaceutical composition of the present invention may be prepared in the form of liquid, suspension, powder, granules, tablets, capsules, pills, or extract.

The compositions of the present invention can be administered orally or parenterally (e.g., by application or injection intravenously, subcutaneously, or intraperitoneally).

In the present invention, the term "oral administration" refers to a method of injecting a drug for improving pathological symptoms (periodontal disease) into the mouth, and in the present invention, the term "parenteral administration" refers to administration by mouth. This refers to methods of administration, excluding subcutaneous, intramuscular, intravenous, and intraperitoneal administration using a tube.

Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills, etc. Liquid preparations for oral administration include suspensions, liquids, emulsions, syrups, aerosols, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives are included. May be included.

Preparations for parenteral administration include aqueous solutions, liquids, non-aqueous solutions, suspensions, emulsions, eye drops, eye ointments, syrups, suppositories, aerosols, etc., sterilized according to conventional methods, and are formulated as external preparations and sterilized injection preparations. It can be used in combination, preferably in the form of a pharmaceutical composition such as cream, gel, patch, spray, ointment, warning agent, lotion, liniment agent, eye ointment, eye drop, paste or cataplasma agent. However, it is not limited to this. Compositions for topical administration may be in anhydrous or aqueous form depending on clinical prescription. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurel, glycerogelatin, etc. can be used.

The pharmaceutically acceptable additive according to the present invention may be included in an amount of 0.1 to 99.9 parts by weight, specifically 0.1 to 50 parts by weight, with respect to the composition, but is not limited thereto.

In the case of a pharmaceutical composition for preventing or treating liver disease comprising a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof according to the present invention, alanine aminotransferase (ALT) or It has the effect of preventing or treating liver disease by suppressing the increase in blood concentration of Aspartate Aminotransferase (AST). The pharmaceutical composition for preventing or treating liver disease comprising the compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof of the present invention may be administered orally, rectally, intravenously, intraarterially, intraperitoneally, or intramuscularly. , can be administered in a conventional manner via intrasternal, transdermal, topical, intraocular or intradermal routes, and specifically can be administered orally.

The pharmaceutical composition for preventing or treating liver disease comprising the compound represented by Formula I of the present invention, its optical isomer, or its pharmaceutically acceptable salt, when administered once or several times a day, is administered in an amount of 0.001 mg/kg to 0.001 mg/kg. It can be administered at a dose of 5 g/kg, and the administration may be administered once a day or in divided doses. Additionally, the pharmaceutical composition of the present invention may contain a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof at a weight percentage of 0.001 to 50% based on the total weight of the pharmaceutical composition.

The pharmaceutical composition of the present invention may further include one or more active ingredients showing the same or similar medicinal efficacy in addition to the compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Food composition for preventing or improving liver disease

The present invention provides a food composition for preventing or improving liver disease comprising a compound represented by Formula I, an optical isomer thereof, or a foodologically acceptable salt thereof.

In the present invention, the description of the compound represented by Formula I, its optical isomer, or liver disease is as described above.

When the composition of the present invention is used as a food composition, it may contain acceptable food auxiliary additives and may further include appropriate carriers, excipients, and diluents commonly used in the production of food.

The food composition of the present invention may include the form of pills, powders, granules, precipitates, tablets, capsules, or liquids, and may contain a compound represented by Formula I of the present invention, an optical isomer thereof, or a food-acceptable salt thereof. There are no particular restrictions on the types of foods that can be added. Examples of foods to which the above substances can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, These include alcoholic beverages and vitamin complexes.

In addition to the compound represented by Chemical Formula I, its optical isomer, or its foodologically acceptable salt, other ingredients may be added to the food composition, and the types thereof are not particularly limited. For example, like regular foods, it may contain various herbal extracts, foodologically acceptable food additives, or natural carbohydrates as additional ingredients, but is not limited thereto.

In the present invention, the term “food auxiliary additive” refers to a component that can be added to food as an auxiliary agent, and can be appropriately selected and used by a person skilled in the art as it is added to prepare each type of food. Examples of food supplements include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, and protective colloidal thickeners. , pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., but the types of food supplements of the present invention are not limited to the above examples.

Examples of the natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Flavoring agents other than those mentioned above include natural flavoring agents (such as thaumatin) and stevia extract (rebaudioside A, glycyrrhizin). hygin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be used to advantage.

The food composition of the present invention may include health functional foods. In the present invention, the term “health functional food” refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and pills using raw materials or ingredients with functional properties useful to the human body. Here, functionality means controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes such as physiological effects. The health functional 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 components commonly added in the art. In addition, unlike general drugs, it is made from food, so it has the advantage of not having any side effects that may occur when taking the drug for a long time, and it can be highly portable.

The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing food, the active ingredient of the present invention may be added in an amount of 0.01 to 50% by weight, preferably 0.1 to 10% by weight, of the raw material composition, but is not limited thereto. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be used even below the above range.

Use for the manufacture of medicaments for the prevention or treatment of liver disease

The present invention provides the use of a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the production of a medicament for the prevention or treatment of liver disease.

In the present invention, acceptable carriers for the preparation of drugs may be mixed, and other agents may be further included.

How to prevent or treat liver disease

The present invention provides a method for preventing or treating liver disease, comprising administering a therapeutically effective amount of a compound represented by Formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

In this specification, the term “subject in need of treatment” refers to mammals, including humans, and the term “administration” refers to providing a predetermined substance to a patient by any appropriate method. In the prevention or treatment method of the present invention, the compound represented by Formula I of the present invention, its optical isomer, or its pharmaceutically acceptable salt is administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, or transdermally. , can be administered in the conventional manner via topical, intraocular or intradermal routes.

In the method for preventing or treating liver disease of the present invention, the compound represented by Formula I of the present invention, its optical isomer, or its pharmaceutically acceptable salt is administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, and sternally. It can be administered in a conventional manner via intradermal, topical, intraocular or intradermal routes, and specifically can be administered orally.

As used herein, the term “therapeutically effective amount” refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in tissue systems, animals, or humans, as considered by researchers, veterinarians, doctors, or other clinicians. This includes amounts that induce relief of symptoms of the disease or disorder being treated. It is obvious to those skilled in the art that the therapeutically effective dosage and frequency of administration for the compound represented by Formula I of the present invention, its optical isomer, or its pharmaceutically acceptable salt will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by a person skilled in the art, and can be determined based on the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of dosage form, and the patient's age, weight, and general health. It can be adjusted according to various factors, including condition, gender and diet, administration time, administration route and secretion rate of the composition, treatment period, and concurrently used drugs.

In the method for preventing or treating liver disease of the present invention, in adults, the compound represented by formula (I) of the present invention, its optical isomer or pharmaceutically acceptable salt thereof, when administered once to several times a day, is 0.001 It is preferable to administer at a dose of mg/kg to 5 g/kg.

Matters mentioned in the compound represented by Formula I of the present invention, its optical isomer or pharmaceutically acceptable salt, pharmaceutical composition, food composition, use, prevention or treatment method apply equally unless they contradict each other.

Examples and experimental examples are presented to aid understanding of the present invention. The following examples and experimental examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by the examples and experimental examples.

Example

Example 1: Preparation of Compound 1 (AP1-AL3)

¹ H NMR, ESI-MASS, and purity of Compound 1 (AP1-AL3) synthesized in Example 1 below were measured using the following methods.

^One H NMR spectrum

1) Device

Bruker Avance Neo 400 MHz

2) Details

a) NMR sample amount, NMR solvent: precursor 5 mg, NMR solvent 0.5 mL

b) NMR solvent: CDCl3-d1 (Cambridge Isotope Laboratories, Inc.)

c) Number of scanning time (CT): 24

d) Standard peak is 7.83 ppm (d, J = 8.4 Hz, 1H).

ESI-MASS

1) Device

Thermo scientific ISQ EM

2) Sample status

1 mg of the test sample was measured and dissolved in 1.0 mL acetonitrile.

3) Mobile phase

50% MeCN/purified water, flow rate: 200 μL/min

Purity (high-performance liquid chromatography; HPLC)

1) Mobile phase condition

Time [min] Solvent A [%] Solvent B [%] Flow [mL/min] 0 60 40 One 20 0 100 One

(In the table above, Solvent A is sodium carbonate buffer solution (pH = 9.2), and Solvent B is acetonitrile.)

2) Test Sample solution

1 mg of the test sample was measured and dissolved in 1 mL of Solvent A.

3) Chromatographic system

a) Device: Varian prostar

b) Column: YMC C18 (4.6 x 250 mm, 5 μm particle size)

c) Wavelength: 254 nm

d) Injection volume: 10 mL

4) Procedure : 20 mL of the prepared test sample was injected and the chromatogram was recorded. Then, the percentage of each peak was measured according to the HPLC chromatogram results.

[Step 1] AL3-TIPS compound synthesis

Dissolve AL3 (CS-21023) (600 mg, 2.34 mmol) and imidazole (383 mg, 5.62 mmol) in dimethylformamide (DMF) (5 mL, anhydrous) under argon gas. , cooled in an ice bath. TIPS-Cl (Triisopropylsilyl chloride) (445 mg, 2.80 mmol) was added and the temperature was slowly raised to room temperature. After reaction at room temperature for 3 hours, the solvent was removed under reduced pressure. The reaction mixture was dissolved in ethyl acetate, transferred to a separatory funnel, washed twice each with purified water and brine, and the organic layer was dried over sodium sulfate (Na ₂ SO ₄ ). It was purified by silica gel column chromatography (10 ~ 20% ethyl acetate / n-hexane) to obtain AL3-TIPS (yield: 834 mg, yield: 86%) as a pale yellow amorphous solid (foamy solid).

^1H NMR (400 MHz, CDCl ₃ ) d 7.84 (d, J = 8.8 Hz, 1H), 7.35 (d, J = 8.4 Hz, 2H), 6.90 (d, J = 8.8 Hz, 2H), 6.57 (dd, J = 2.0, 8.8 Hz, 4H), 6.47 (d, J = 2.4 Hz, 1H), 5.84 (br s, 1H), 5.39 (dd, J = 2.4, 13.2 Hz, 1H), 3.06 (dd, J = 13.2, 16.8 Hz, 1H), 2.80 (dd, J = 1.2, 16.8 Hz, 1H), 1.32-1.20 (m, 3H), 1.17-1.05 (m, 18H);

ESI-(+) 413.3 [M+H] ⁺ .

[Step 2] Synthesis of AP1-AL3_2-1 compound

Under argon gas, M4-3 compound (270 mg, 0.67 mmol), AL3-TIPS (Triisopropylsilyl) (230 mg, 0.56 mmol), EDC.HCl [(3-Dimethylamino-propyl)-ethyl-carbodiimide Hydrochloride] (131 mg, 0.84 mmol) and DMAP (4-Dimethylaminopyridine) (25 mg, 0.20 mmol) were added to a round flask, then dichloromethane (15 mL, anhydrous) was added and stirred at room temperature for 18 hours. The reaction mixture was transferred to a separatory funnel and washed twice with purified water, and the organic layer was dried over sodium sulfate (Na ₂ SO ₄ ). AP1-AL3_2-1 (yield: 438 mg, yield: 82%) was obtained as a white solid by silica gel column chromatography (20-30% ethyl acetate / n-hexane).

^1H NMR (400 MHz, CDCl ₃ ) d 7.82 (d, J = 8.8 Hz, 1H), 7.57 (s, 1H), 7.45 (d, J = 8.4 Hz, 2H), 7.37 (s, 1H), 7.04 ( d, J = 8.4 Hz, 2H), 6.57 (dd, J = 2.0, 8.4 Hz, 1H), 6.47 (d, J = 1.6 Hz, 1H), 6.04 (br s, 2H), 6.00 (d, J = 10.4 Hz, 2H), 5.43 (dd, J = 2.4, 13.2 Hz, 1H), 4.01 (s, 3H), 3.94 (s, 3H), 3.71 (s, 3H), 2.98 (dd, J = 13.6, 16.8 Hz, 1H), 2.79 (dd, J = 2.8, 16.8 Hz, 1H), 1.33-1.23 (m, 3H), 1.12-1.09 (m, 18H).

[Step 3] AP1-AL3 compound synthesis

AP1-AL3_2-1 compound (438 mg, 0.55 mmol) was dissolved in tetrahydrofuran (THF) (10 mL), then TBAF solution (Tetra-n-butylammonium fluoride solution) (1.64 mL, 1M solution in THF) was added and stirred for 1 hour. After removing the solvent under reduced pressure, the reaction mixture was dissolved again in ethyl acetate. The dissolved reaction mixture was transferred to a separatory funnel and washed twice with purified water and brine, and the organic layer was dried over sodium sulfate (Na ₂ SO ₄ ). It was purified by silica gel column chromatography (30 ~ 50% ethyl acetate / n-hexane) to obtain AP1-AL3 (yield: 335 mg, yield: 95%, purity: 96.97% in 12.53 minutes) as a yellow solid.

^1H NMR (400 MHz, CDCl ₃ ) δ 7.83 (d, J = 8.4 Hz, 1H), 7.55 (s, 1H), 7.42 (d, J = 8.4 Hz, 2H), 7.37 (s, 1H), 7.03 (d, J = 8.4 Hz, 2H), 6.50 (dd, J = 8.6, 1.8 Hz, 1H), 6.40 (s, 1H), 6.28 (s, 1H), 6.04 (s, 2H), 6.00 (d, J = 9.1 Hz, 2H), 5.41 (dd, J = 13.0, 2.4 Hz, 1H), 3.99 (s, 3H), 3.92 (s, 3H), 3.72 (s, 3H), 2.97 (dd, J = 16.8 , 13.2 Hz, 1H), 2.79 (dd, J = 16.9, 2.7 Hz, 1H).

ESI-(-) 641.2 [M-Na] ^-

Experimental Example: Example 1 Compound (AP1-AL3) efficacy in inhibiting liver function damage or liver fibrosis

An animal model induced by carbon tetrachloride (CCl ₄ ) is used as an animal model for research on treatments or prevention of liver function damage or liver fibrosis. Carbon tetrachloride (CCl ₄ ) causes liver fibrosis, liver cell death, and carcinogenesis, and is a representative substance used to determine the effects of toxic substances on liver cells and their metabolic processes. It is a representative substance used to culture liver cells or directly administer them to mice or rats. It is widely used to study liver toxicity using methods such as Carbon tetrachloride (CCl ₄ ) is metabolized by metabolic enzymes such as cytochrome P450, which performs complex functions in the liver, forming highly toxic free radicals, CCl ₃ and Cl, and the free radicals, CCl ₃ , accumulate. It causes oxidation of fatty acids in liver neutral fat or membrane phospholipids, causing rancidity of lipids and combining with oxygen to promote oxidation reaction with lipids. Through these lipid and oxidation processes, fat accumulates in the liver, protein synthesis ability decreases, it is broken down into glycogen, and it affects cellular structures such as mitochondria, DNA, and cell membranes, damaging or killing cells. Additionally, it may induce cell necrosis or progress liver fibrosis by activating nitric oxide (NO) and transforming growth factor-β (TGF-β). In other words, carbon tetrachloride (CCl ₄ ) inhibits protein biosynthesis in the liver, causes an increase in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the blood, and histologically is known to cause necrosis of liver cells.

Alanine aminotransferase (ALT) is an enzyme found mostly in cells of the liver and kidneys. In healthy animals or humans, blood alanine aminotransferase (ALT) levels are low, but when the liver is damaged, alanine aminotransferase (ALT) is released into the bloodstream before clear symptoms of liver damage appear. Because of this phenomenon, alanine aminotransferase (ALT) can be used as a useful indicator for detecting liver damage. Damage to liver cells results in an increase in alanine aminotransferase (ALT) levels. This indicator is most useful in detecting hepatitis or damage caused by drugs or substances that are toxic to the liver.

Aspartate Aminotransferase (AST) is an enzyme found in cells throughout the body, but is mainly found in the heart and liver. In healthy people, the concentration of Aspartate Aminotransferase (AST) in the blood is low, but when the liver or muscles are damaged, Aspartate Aminotransferase (AST) is released from the liver or muscles into the blood. Aspartate aminotransferase (AST) concentration increased by release can be used as a useful indicator for detecting hepatocyte damage (impaired liver function). When liver cells (liver function) are damaged by many stimuli, the concentration of aspartate aminotransferase (AST) in the blood increases. Aspartate Aminotransferase (AST) is one of the most useful indicators for detecting liver cell damage (impaired liver function) caused by hepatitis, liver-toxic drugs, cirrhosis, and alcoholism.

Experimental Example 1. Single carbon tetrachloride (CCl) ₄ ) Inhibitory efficacy of hepatocyte damage (liver function impairment) of Example 1 compound (AP1-AL3) using induced hepatocyte damage model

Experiment method

The carbon tetrachloride (CCl ₄ ) single-dose administration model is the simplest animal testing method to confirm hepatocyte damage (impaired liver function), and the animal testing was conducted in accordance with the guidelines of the Animal Experiment Ethics Review Committee of the Dongguk University Preclinical Efficacy Evaluation Center. Specifically, 6-week-old male C57BL/6 mice were purchased from Charles River Orient (Seoul, Korea), their body weight was measured before the start of administration, divided into 4 to 5 mice per group to maintain a constant average weight, and incubated at 20 ± 2°C for 12 hours. They were stored under conditions of a light/dark cycle, relative humidity of 50 ± 5%, filtered, pathogen-free air, and food and water available ad libitum. As a liver toxicant, carbon tetrachloride (CCl ₄ ) is mixed with corn oil (Sigma, C8267) at a ratio of 18:82 (v/v) _and injected intraperitoneally (IP) at a dose of 0.6 ml/kg. It was administered as a single dose (Reference: EBioMedicine. 2019 Feb;40:488-503. doi: 10.1016/j.ebiom.2018.12.056. Epub 2019 Jan 9. PMID: 30638865; PMCID: PMC6413675).

Six hours after a single administration of carbon tetrachloride (CCl ₄ ) by intraperitoneal injection (IP), the drugs Example 1 compound (AP1-AL3) and obeticholic acid (OCA) were each administered orally, and administered orally for 24 hours. Blood and liver tissue were later extracted.

<Order summary>

<Administration group>

Vehicle: Corn Oil

Control group: Carbon tetrachloride (CCl ₄ ) administration group

Positive control: Obeticholic acid (OCA) 30 mg/kg

Experimental group 1: Example 1 compound (AP1-AL3) administration group 10 mg/kg and 30 mg/kg

Experiment result

In the case of liver function damage caused by a single administration of carbon tetrachloride (CCl ₄ ), the compound of Example 1 (AP1-AL3) was confirmed to partially inhibit the production of liver function damage indicators.

Specifically, when confirmed based on alanine aminotransferase (ALT) levels, the compound of Example 1 (AP1-AL3) was about 38.22% (about 38.22%) at 10 mg/kg compared to the carbon tetrachloride (CCl ₄ ) control (Control). p=0.057), showing an inhibitory effect of about 38.30% (p<0.05) at 30 mg/kg (see Figure 1 and Table 2).

In addition, when confirmed based on the Aspartate Aminotransferase (AST) level, the compound of Example 1 (AP1-AL3) was about 19.46% at 10 mg/kg compared to the carbon tetrachloride (CCl ₄ ) control (Control). It showed an inhibitory effect of (p<0.05), showed an inhibitory effect of about 23.23% (ns) at 30 mg/kg, and showed a significant inhibitory effect on liver cell damage (liver function damage) (see Figure 2 and Table 3) .

In addition, similar results to hematological analysis were confirmed through Hematocylin & Eosin staining, and hepatocyte damage (liver function damage) was significantly suppressed in the 10 mg/kg and 30 mg/kg experimental groups of Example 1 compound (AP1-AL3). The effect was confirmed (see Figure 3).

Claims (4)

Compound represented by the following formula (Ⅰ), its optical isomer or its pharmaceutically acceptable salt:
[Formula Ⅰ]
A pharmaceutical composition for preventing or treating liver disease comprising a compound represented by the following formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula Ⅰ]
According to paragraph 2,
A pharmaceutical composition wherein the liver disease is at least one selected from the group consisting of impaired liver function, liver failure, hepatitis, hepatotoxicity, liver fibrosis, and cirrhosis. Food composition for preventing or improving liver disease comprising a compound represented by the following formula (Ⅰ), an optical isomer thereof, or a foodologically acceptable salt thereof:
[Formula Ⅰ]