KR20230142287A - A composition for preventing or treating of non-alcoholic steatohepatis(NASH) containing compounds isolated from Albizzia julibrissin - Google Patents

Abstract

The present invention relates to a composition for preventing, improving, or treating non-alcoholic fatty liver disease, and more specifically, to a composition for preventing, improving, or treating non-alcoholic fatty liver disease comprising a compound isolated from manganese skin or a salt thereof. The compound according to the present invention acts on sterol regulator binding protein 1c (SREBP-1c), a major regulator of non-alcoholic fatty liver disease, to inhibit the influx of free fatty acids (FFA) into the liver, thereby reducing triglyceride (TG) By suppressing accumulation, it is expected that it can be widely used as medicine, food, and quasi-drugs to prevent, improve, and treat non-alcoholic fatty liver disease.

Description

A composition for preventing or treating non-alcoholic steatohepatis (NASH) compounds containing isolated from Albizzia julibrissin}

The present invention relates to a composition for preventing, improving, or treating non-alcoholic fatty liver disease, and more specifically, to a composition for preventing, improving, or treating non-alcoholic fatty liver disease comprising a compound isolated from manganese skin or a salt thereof.

Non-alcoholic fatty liver disease (NASH) is the most serious histological form of non-alcoholic fatty liver disease (NAFLD) and refers to a disease in which triglycerides (TG) accumulate in the liver without excessive alcohol consumption, and 20% of patients develop cirrhosis. The number of NASH patients is increasing day by day, but no approved drug has yet been developed for treatment. In addition, since most NASH patients have other risk factors such as diabetes, obesity, and hyperlipidemia, there is an urgent need to develop drugs that can respond to them. In the pathogenesis of NASH, inhibition of SREBP-1c reduces lipogenesis, which reduces the accumulation of liver fat. Therefore, many studies are being conducted to find components with inhibitory activity against SREBP-1c in natural products.

Meanwhile, Albizia julibrissin is a legume plant and is a flowering plant widely distributed around the world, including Africa, Central Asia, East Asia, and North America. Hapwanpi is used in traditional herbal medicine as a sedative and anti-inflammatory, and is used to treat depression and insomnia and remove carbuncles. The main components of Manghwanpi are reported to be triterpenoid saponins, lignans, flavonoids, phenolic glycosides, and other compounds, and the biochemical activities of these components are known to be cytotoxic, anticancer, and anti-inflammatory (Korea Registered Patent No. 10- No. 1790738). However, no study has been conducted on the inhibitory activity of extracts and isolated substances from Manghwan skin on SREBP-1c.

In order to improve the above problems, the present inventors studied the inhibitory activity on SREBP-1c for the treatment of non-alcoholic fatty liver disease. As a result, they isolated a compound from manghwan skin, and the compound acts on SREBP-1c and is released into the liver. It was confirmed that it can be usefully used as a composition for preventing and treating non-alcoholic fatty liver disease by suppressing the accumulation of triglyceride (TG) by suppressing the influx of fatty acids (FFA).

Accordingly, the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

Another object of the present invention is to provide a health functional food composition for preventing or improving non-alcoholic fatty liver disease, comprising the above compounds or their foodologically acceptable salts as active ingredients.

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

In addition, the present invention provides a health functional food composition for preventing or improving non-alcoholic fatty liver disease, comprising the above compounds or their foodologically acceptable salts as active ingredients.

In one embodiment of the present invention, the composition may have SREBP-1c inhibitory activity.

In another embodiment of the present invention, the compound may be extracted and separated from manghwan skin.

The compound according to the present invention acts on sterol regulator binding protein 1c (SREBP-1c), a major regulator of non-alcoholic fatty liver disease, to inhibit the influx of free fatty acids (FFA) into the liver, thereby reducing triglyceride (TG) By suppressing accumulation, it is expected that it can be widely used as medicine, food, and quasi-drugs to prevent, improve, and treat non-alcoholic fatty liver disease.

Figure 1 schematically shows the process of extracting and fractionating manghwanpi according to the present invention.
Figure 2 is a schematic diagram showing the process of separating the compound according to the present invention from the husk.
Figure 3 shows the results of confirming the SREBP-1c inhibitory activity in the extract and fractions of Manghwanpi.
Figure 4 shows the results of confirming the SREBP-1c inhibitory activity of the compound of the present invention isolated from the Hwanhwan skin fraction.

As a result of studying the inhibitory activity on SREBP-1c for the treatment of non-alcoholic fatty liver disease, the present inventors isolated a compound from manghwan skin, and the compound acts on SREBP-1c to inhibit the influx of free fatty acids (FFA) into the liver. By doing so, it was confirmed that it can be effectively used as a composition for preventing and treating non-alcoholic fatty liver disease by suppressing the accumulation of triglyceride (TG).

Accordingly, the present invention provides a pharmaceutical composition and a health functional food composition for preventing, improving or treating non-alcoholic fatty liver disease, comprising a compound represented by the following formula (1) or a pharmaceutically or foodologically acceptable salt thereof.

[Formula 1]

The compound according to the present invention refers to a compound composed of the chemical formula C ₁₆ H ₁₂ O ₆ and named 3-O-methylfisetin.

The term “prevention” used in the present invention refers to all actions that suppress or delay the onset of non-alcoholic fatty liver disease by administering the composition according to the present invention.

The term "improvement" used in the present invention refers to any action that reduces the degree of sarcopenia or alleviates symptoms such as complications due to non-alcoholic fatty liver disease by administering the composition according to the present invention.

As used in the present invention, the term “treatment” refers to any action in which the symptoms of non-alcoholic fatty liver disease are improved or beneficially changed by administration of the composition according to the present invention.

In the present invention, the composition may have SREBP-1c inhibitory activity, but the activity of the composition according to the present invention is not limited thereto.

In the present invention, the compound may be extracted and separated from manghwan skin, but is not limited thereto.

The compound represented by [Chemical Formula 1] according to the present invention may be used in the form of a salt. As the salt, acid addition salts formed by various pharmaceutically or foodologically acceptable organic acids or inorganic acids can be used. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, as well as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanedio acids. It can be obtained from non-toxic organic acids such as ateates, aromatic acids, aliphatic and aromatic sulfonic acids. These non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, and fluoride. , acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propionic acid, oxalic acid, malonic acid, succinic acid, suberate, sebacate, fumarent, maly. ate, butyne-1,4-dioate, hexane-1,6-dioic acid, benzoic acid, chlorobenzoic acid, methylbenzoic acid, dinitro benzoic acid, hydroxybenzoate, methoxybenzoic acid, fdalic acid, terephdalate, benzene. Silphonic acid, toluenesilphonic acid, chlorobenzenesulfonic acid, xylenesulfonic acid, phenylacetic acid, phenylpropionic acid, phenylbutyrate, citrate, lactate, -Can be manufactured using hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propane sulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, trifluoroacetic acid, etc. there is.

At this time, the acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound of [Formula 1] in an excessive amount of aqueous acid, and dissolving this salt in a water-miscible organic solvent, such as methanol, ethanol, acetone, or It can be prepared by precipitation using acetonitrile. It can also be prepared by evaporating the solvent or excess acid from this mixture and drying it, or by suction-filtering the precipitated salt.

In addition, the compound represented by [Chemical Formula 1] according to the present invention can be made into a pharmaceutically or foodologically acceptable metal salt using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. At this time, it is agrochemically appropriate to produce lithium, sodium, potassium or calcium salts as metal salts. Additionally, the corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with an appropriate silver salt (eg, silver nitrate).

The present inventors have investigated the use of the compound according to the present invention for preventing, improving, and treating non-alcoholic fatty liver disease through specific examples.

Specifically, in one example of the present invention, it was confirmed that the extract and fraction of Manghwan bark exhibited SREBP-1c inhibitory activity (see Example 2).

In addition, in another example of the present invention, it was confirmed that the compound isolated from the manghwan skin fraction according to the present invention exhibited SREBP-1c inhibitory activity (see Example 3).

Summarizing the results of the above examples, it can be seen that the following extracts and fractions of manghwan skin, and the compound of formula 1 exhibit SREBP-1c inhibitory activity, and can be used as medicines, foods, and quasi-drugs for the prevention, improvement, and treatment of non-alcoholic fatty liver disease. It is expected that it can be widely used.

When the composition according to the present invention is in the form of a pharmaceutical composition, it may contain the compound of [Formula 1] alone or one or more pharmaceutically acceptable carriers in a pharmaceutically effective amount. At this time, pharmaceutically acceptable carriers are those commonly used in preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, and microcrystalline cellulose. , polyvinyl pyrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, etc., but are not limited thereto. In addition, in addition to the above ingredients, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. may be additionally included.

The pharmaceutical composition of the present invention can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method, and the dosage is determined by the patient's condition and weight, and the severity of the disease. It varies depending on the degree, drug form, administration route and time, but can be appropriately selected by a person skilled in the art.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, “pharmaceutically effective amount” means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type of disease, severity, activity of the drug, and It can be determined based on factors including sensitivity, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on the patient's age, gender, condition, weight, absorption of the active ingredient in the body, inactivation rate and excretion rate, type of disease, and concomitant drug. In general, 1 to 500 mg per kg of body weight can be administered daily or every other day, or divided into 1 to 3 times per day. However, since it may increase or decrease depending on the route of administration, gender, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

In another aspect of the present invention, the present invention provides a method of treating sarcopenia comprising administering the pharmaceutical composition to a subject.

In the present invention, an “individual” may be a mammal such as a rat, livestock, mouse, or human, and may specifically be a pet dog, racehorse, or human in need of anticancer treatment, and preferably may be a human.

In another aspect of the present invention, the present invention provides a use of the pharmaceutical composition for treating sarcopenia.

When the composition according to the present invention is used in the form of a health functional food composition, in addition to food for specific health purposes and nutritional supply, it can be manufactured into medicine and food with high medical effectiveness processed to efficiently exhibit bioregulatory functions. In some cases, it can be used interchangeably as functional food, health food, or health supplement, and can be manufactured in various forms such as tablets, capsules, powder, granules, liquid, and pills to obtain useful effects.

The health functional food of the present invention may contain additional ingredients that are commonly used in food compositions to improve smell, taste, vision, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. Additionally, it may contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu). Additionally, it may contain amino acids such as lysine, tryptophan, cysteine, and valine. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dihydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxy toluene (BHT) ), etc.), colorants (tar colors, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, saccharin, sodium, etc.), Food additives such as flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents, improvers, etc. can be added. The additives can be selected depending on the type of food and used in an appropriate amount.

When using the health functional food of the present invention as a food additive, it can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods.

In the health functional food of the present invention, the content of the compound of [Chemical Formula 1] according to the present invention is not particularly limited and may vary depending on the condition of the administration subject, the type of specific disease, the degree of progression, etc. If necessary, it can also be included in the total amount of the food.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

[Example]

Example 1. Isolation and experimental method of compounds derived from Hwanhwan skin

1-1. Manufacture of manghwan skin extracts and fractions

We purchased and used manhwan skin from Sancheong, Jiri Mountain. 10.0 kg of manhwawanpi was extracted with methanol three times at 50°C for 3 hours each by ultrasonic extraction and concentrated in a reduced pressure concentrator to remove the solvent, yielding 418.4 g of extract. The extract was dissolved in water, fractionated into n-Hexane (n-Hex), chloroform (CHCl3), and ethyl acetate (EtOAc), and concentrated in a reduced pressure concentrator to remove the solvent, followed by n-Hex (93.7 g) and CHCl3 (4.3 g), respectively. ), EtOAc (4.5 g) and water (167.8 g) were obtained.

1-2. Isolation of compounds from chive extract

Among the fractions obtained in Example 1-1, the EtOAc fraction was subjected to silica gel column chromatography using EtOAc - Acetone - water (2: 1: 0.1, v/v/v) to obtain three small fractions, of which AJB6 The fraction was again subjected to YMC reverse-phase column chromatography with MeOH Water (1.5:1, v/v) and then purified with 20% aq. Preparation HPLC was performed with MeCN to obtain a compound of the following formula (1.4 mg) (Pale-yellow amorphous solid Molecular formula: C ₁₆ H ₁₂ O ₆ HR-ESI-MS m/z 323.0514 (calcd for C16H12O6Na, 323.0532) ¹ H (MeOD-d4, 600 MHz): δ 7.93 - 7.86 (m, H-4), 7.57 (d, J = 2.1 Hz, H-2'), 7.47 (dd, J = 8.4, 2.1 Hz, H-6' ), 6.85 - 6.81 (m, H-5, H-5', H-7), 3.69 (OCH3) ¹³ C NMR (MeOD-d4, 150MHz): δ 176.7 (C-3), 165.1 (C-6) ), 158.8 (C-8), 158.0 (C-1), 149.9 (C-4'), 146.6 (C-3'), 141.3 (C-2), 128.0 (C-4), 123.4 (C- 1'), 122.4 (C-6'), 117.8 (C-9), 116.7 (C-5), 116.6 (C-2'), 116.4 (C-5'), 103.3 (C-7), 60.5 (OCH3)) was isolated.

[Formula 1]

1-3. AML12 cell preparation and SREBP-1c inhibitory activity assay

The rat AML12 cell line was used, and serum starvation was performed for 6 hours after cell culture. The extract and fraction extracted in Example 1-1 and the compound isolated in Example 1-2 were treated, and 1 hour later, LXR agonist, T0901317, an SREBP-1c inducer, was added, reaction was induced for 12 hours, and mRNA analysis was performed. proceeded.

Example 2. Confirmation of SREBP-1c inhibitory activity of Manghwanpi extract

AML12 cells were treated with the extract and fractions isolated in Example 1-1, and SREBP-1c inhibitory activity was confirmed by the method described in Example 1-3. As a result, as shown in Figure 3, it was confirmed that the extract and fraction exhibited SREBP-1c inhibitory activity in both low concentration (100 μg/ml) and high concentration (250 μg/ml) experiments. The extract and CHCl ₃ fractions showed similar activity at low and high concentrations, while the Hex, EtOAc and water fractions showed that high concentrations showed more powerful activity than low concentrations.

Example 3. Confirmation of SREBP-1c inhibitory activity of compounds according to the present invention

AML12 cells were treated with the compound of Formula 1 isolated in Example 1-2, and the SREBP-1c inhibitory activity of Example 1-3 was evaluated, specifically at low (10 μM), medium (50 μM), and high concentrations of the compound. Activity was evaluated at (100 μM). As a result, it was confirmed that activity was observed at high concentrations, as shown in Figure 4.

From the above results, it can be seen that the extract and fractions of Manghwan bark according to the present invention, and the compound of Formula 1 exhibit SREBP-1c inhibitory activity, and thus, it can be inferred that they can be effective in preventing and treating non-alcoholic fatty liver disease. Could.

The description of the present invention described above is for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive.

Claims (6)

A pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
[Formula 1]

According to paragraph 1,
A pharmaceutical composition, characterized in that the composition has SREBP-1c inhibitory activity.
According to paragraph 1,
A pharmaceutical composition, characterized in that the compound is extracted and separated from manghwan skin.
A health functional food composition for preventing or improving non-alcoholic fatty liver disease, comprising a compound represented by the following formula (1) or a foodologically acceptable salt thereof.
[Formula 1]

According to paragraph 4,
A health functional food composition, characterized in that the composition has SREBP-1c inhibitory activity.
According to paragraph 4,
A health functional food composition, characterized in that the compound is extracted and separated from manghwan skin.