KR20220048568A - A composition for preventing or treating alcoholic liver disease comprising an extract mixture - Google Patents

Abstract

The present invention relates to a composition for preventing or treating alcoholic liver disease, containing an extract mixture, and a method for manufacturing the same, wherein the composition according to the present invention has an effect of improving the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, inhibiting fat accumulation, and facilitating fat decomposition, so that it is possible to be used as a therapeutic agent for alcoholic liver disease.

Description

A composition for preventing or treating alcoholic liver disease, and a method for preparing the same, comprising an extract mixture {A composition for preventing or treating alcoholic liver disease comprising an extract mixture}

The present invention relates to a composition for preventing or treating alcoholic liver disease comprising an extract mixture, and the composition according to the present invention comprises: chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; As an extraction mixture comprising at least one of at least one, it improves the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, and inhibits fat accumulation and promotes fat decomposition, thereby providing an excellent composition for preventing or treating alcoholic liver disease.

Excessive alcohol consumption is a global medical problem that causes enormous social, economic and clinical damage. In 2012, 3.3 million people died from alcohol-related diseases in one year. Excessive drinking damages almost all organs in the body, and since the liver is a major part of most metabolism, excessive drinking is known to damage the liver. When a human consumes alcohol, the human body undergoes an alcohol metabolism process that absorbs and decomposes the alcohol.

Alcohol metabolism proceeds through oxidative and non-oxidative pathways, and major processes of the oxidative pathway are caused by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). As this alcohol metabolism process progresses, hangovers and side effects from alcohol occur in the human body.

A hangover phenomenon refers to a phenomenon that persists for 1 to 2 days after waking up after drinking alcohol, such as unusual discomfort, headache, or decreased mental and physical work capacity. , vomiting, and other side effects may occur. Most of these phenomena occur in acetaldehyde accumulated in the body, and alcohol entering the body is decomposed in the liver to produce toxic acetaldehyde in the body. Acetaldehyde causes side effects to various organs in the human body through blood vessels or directly damages liver cells. Furthermore, alcohol that is not decomposed in the liver is delivered to the brain through the blood and stimulates the central nervous system, thereby impairing human judgment. In addition, when a part of acetic acid produced by alcohol metabolism is not decomposed into water and carbon dioxide, fatty acids are excessively produced and fat is excessively accumulated in the liver tissue, thereby causing alcoholic fatty liver.

Against this background, interest in substances that can suppress alcoholic liver disease is continuously increasing, and since steroid treatment was introduced for severe alcoholic hepatitis in 1971, clinical trials for the therapeutic efficacy of alcoholic liver disease for various substances have been conducted recently. Experiments have been made. However, despite continuous research, the development of a breakthrough substance for the treatment of alcoholic liver disease is still a long way off, and the necessity and importance of a new substance that can treat alcoholic liver disease is emerging.

Accordingly, the present inventors chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; An extract mixture containing at least one of them was prepared, and it was confirmed that the effect of improving the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, suppressing fat accumulation, and promoting the decomposition of fat were significantly superior.

Accordingly, an object of the present invention, chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; It is to provide a composition for preventing, alleviating, inhibiting or treating alcoholic liver disease comprising at least one of.

Another object of the present invention is to provide a method for preparing a composition for preventing or treating alcoholic liver disease.

The present invention relates to a composition for preventing or treating alcoholic liver disease, comprising an extract mixture, and a method for preparing the same, wherein the composition according to the present invention improves the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, inhibits fat accumulation and decomposes fat is excellent in promoting

The present inventors thus chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; The treatment or prevention effect of a composition comprising at least one of at least one of alcoholic liver disease was confirmed.

Hereinafter, the present invention will be described in more detail.

One aspect of the present invention, chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; It is a pharmaceutical composition for the prevention, alleviation, inhibition or treatment of alcoholic liver disease comprising at least one of.

As used herein, the term “chlorella” refers to chlorella as a single-celled green algae, and taxonomically belongs to the class Chlorophycea, the order Chlorococcum, and the genus Chlorella. Chlorella usually grows in fresh water such as ponds or lakes, and corresponds to spherical unicellular algae with a diameter of 2 μm to 10 μm. Reproduction refers to a plant that reproduces asexually and uses light and carbon dioxide to grow and reproduce independently if there are plant growth factors such as water, air, nitrogen and phosphoric acid.

In the present invention, the chlorella culture may be a chlorella culture cell, but is not limited thereto.

As used herein, the term “saponin” is a glycoside of steroid, steroid alkaloid, or triterpene, and is a generic term for substances that dissolve in water and exhibit a soapy foaming action. Saponins are present in many plants and are also contained in the body of some echinoderms (starfish, sea cucumber). Saponin is known as a component that increases the immune system of the human body. In particular, ginsenoside and eleutheroside, which are saponin types contained in ginseng, are famous, and it is known that green tea contains various types of saponins.

As used herein, the term "Stevia rebaudiana" is a perennial herb belonging to the Asteraceae that can be used in herbal teas, beverages, herbal preparations, health supplements, and the like. In addition, stevia leaves contain a large amount of sweetening ingredients such as stevioside and rebaudioside, so it is also used as a natural sweetener.

As used herein, the term “Sophora Root” is a perennial plant of the Rosaceae legume, a dicotyledonous plant, also called thief’s stick, ginseng, and snake’s sperm tree, and is known to be prescribed for indigestion, neuralgia, hepatitis, jaundice, hemorrhoids, etc. there is.

The present invention is chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; It relates to a composition comprising at least one of at least one of, the 'extract' in the present specification includes a solvent crude extract, a specific solvent soluble extract (solvent fraction), and a solvent fraction of the solvent crude extract, and the extract is a solution, a concentrate Or it may be in a powder state.

In the present invention, chlorella obtained without limitation of region and country may be used, for example, it may be domestic or Japanese chlorella, but is not limited thereto.

The chlorella extract may be a crude extract obtained by extracting chlorella or concentrated chlorella with water and one or more solvents selected from the group consisting of a linear or branched alcohol having 1 to 4 carbon atoms, for example, it may be a water extract .

In the case of using a mixture of water and alcohol as one solvent used for preparing the crude extract of Chlorella, 10% or more to 100% (v/v), 20% or more to 100% (v/v), 30% or more to less than 100% (v/v), more than 40% to less than 100% (v/v), more than 50% to less than 100% (v/v), more than 60% to less than 100% (v/v), or It may be 70% or more to 100% (v/v) of a linear or branched aqueous solution of alcohol having 1 to 4 carbon atoms.

In the present invention, the alcohol aqueous solution may be at least one selected from the group consisting of methanol aqueous solution, ethanol aqueous solution, propanol aqueous solution, and butanol aqueous solution.

The stevia extract may be one selected from the group consisting of leaves, stems and roots of stevia.

The stevia extract may be a crude extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms.

In the case of using a mixture of water and alcohol as a solvent used for preparing a crude extract of stevia, 10% or more to 100% (v/v), 20% or more to 100% (v/v), 30% or more to less than 100% (v/v), more than 40% to less than 100% (v/v), more than 50% to less than 100% (v/v), more than 60% to less than 100% (v/v), 70 % or more to less than 100% (v/v), 50%, 60%, 70%, 80% or 90% may be a linear or branched alcohol solution having 1 to 4 carbon atoms.

The stevia extract according to the present invention may be a solvent fraction obtained by fractionating a crude solvent extract with an additional solvent, for example, in the crude solvent extract, one selected from the group consisting of ethanol, ethanol aqueous solution, ethyl ether, ethyl acetate, and butanol It may be a solvent fraction using more than one type of solvent.

For example, the crude solvent extract obtained by extracting the stevia with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms is obtained from the group consisting of ethyl ether, ethanol, aqueous ethanol solution, ethyl acetate, and butanol. It may be a solvent fraction using one or more solvents selected from

In one embodiment of the present invention, the stevia extract, or a solvent fraction thereof may include stevioside and/or rebaudioside.

In the present invention, stevioside and/or rebaudioside may be separated from the eluate obtained by eluting a stevia extract or a solvent fraction thereof using one selected from the group consisting of sodium hydroxide and ethanol in a column.

Specifically, in the present invention, stevioside and/or rebaudioside may be separated from an eluate eluted by using sodium hydroxide in a stevia ethanol extract packed in a column, and more specifically, packed in a column The eluate eluted with sodium hydroxide in the ethanol extract of Stevia may be separated from the eluate eluted with ethanol again in the column.

The old ginseng extract may be a crude extract obtained by extracting one or more selected from the group consisting of leaves, stems and roots of old ginseng with one or more solvents selected from the group consisting of water, and a linear or branched alcohol having 1 to 4 carbon atoms. there is.

In the case of using a mixture of water and alcohol as a solvent used for preparing the crude extract of ginseng, more than 10% to less than 100% (v/v), more than 20% to less than 100% (v/v), more than 30% to less than 100% (v/v), more than 40% to less than 100% (v/v), more than 50% to less than 100% (v/v), more than 60% to less than 100% (v/v), 70 % or more to less than 100% (v/v), 50%, 60%, 70%, 80% or 90% may be a linear or branched alcohol solution having 1 to 4 carbon atoms.

In one embodiment of the present invention, the saponin may be obtained from green tea extract.

The green tea extract may be one selected from the group consisting of green tea leaves, stems, seeds and roots.

The green tea extract may be a crude extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms.

In the case of using a mixture of water and alcohol as a solvent used for manufacturing crude extract of green tea, 10% or more to 100% (v/v), 20% or more to 100% (v/v), 30% or more to less than 100% (v/v), more than 40% to less than 100% (v/v), more than 50% to less than 100% (v/v), more than 60% to less than 100% (v/v), 70 % or more to less than 100% (v/v), 50%, 60%, 70%, 80% or 90% may be a linear or branched alcohol solution having 1 to 4 carbon atoms.

The green tea extract according to the present invention may be a solvent fraction obtained by fractionating the crude solvent extract with an additional solvent, for example, in the crude solvent extract, one selected from the group consisting of ethanol, aqueous ethanol solution, ethyl ether, ethyl acetate, and butanol. It may be a solvent fraction using more than one type of solvent.

For example, the crude solvent extract obtained by extracting the green tea with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms is obtained from the group consisting of ethyl ether, ethanol, aqueous ethanol solution, ethyl acetate, and butanol. It may be a solvent fraction using one or more solvents selected from

In one embodiment of the present invention, the green tea extract may contain saponins.

In the present invention, the saponin may be separated from the extract obtained by eluting green tea extract or a solvent fraction thereof using one selected from the group consisting of sodium hydroxide and ethanol in a column.

Specifically, in the present invention, the saponin may be one separated from the eluted by using sodium hydroxide in the green tea ethanol extract packed in the column, and more specifically, sodium hydroxide in the green tea ethanol extract packed in the column. The eluate eluted by using may be separated from the eluate eluted with ethanol again in the column.

In one embodiment of the present invention, the composition may include a chlorella extract and saponin.

In one embodiment of the present invention, the composition may include a chlorella extract and a green tea extract.

In one embodiment of the present invention, the composition may include a chlorella extract, a stevia extract and a saponin.

In one embodiment of the present invention, the composition may include a chlorella extract, a stevia extract and a green tea extract.

In one embodiment of the present invention, the composition may include chlorella extract, stevia extract, saponin and ginseng extract.

In one embodiment of the present invention, the composition may include a chlorella extract, a stevia extract, a green tea extract, and a ginseng extract.

In one embodiment of the present invention, the composition may include a chlorella extract, saponin, and a ginseng extract.

In one embodiment of the present invention, the composition may include a chlorella extract, saponin, and a ginseng extract.

In one embodiment of the present invention, the composition comprises chlorella, chlorella culture or chlorella extract; and saponins; may be included.

In one embodiment of the present invention, the composition comprises chlorella, chlorella culture or chlorella extract; stevia extract; and saponins; may be included.

In one embodiment of the present invention, the composition comprises chlorella, chlorella culture or chlorella extract; ginseng extract; and saponins; may be included.

In one embodiment of the present invention, the composition comprises chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; may be included.

The composition according to the present invention has excellent effects of enhancing the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, inhibiting fat accumulation and promoting fat breakdown, and particularly, chlorella, chlorella culture or extract thereof; saponins; stevia extract; And it was confirmed that in the case of a composition comprising a ginseng extract, the effect of improving the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, suppressing fat accumulation and promoting fat decomposition were significantly superior. (See Figs. 1 to 4)

In the present invention, "alcoholic liver disease" may include, without limitation, both liver damage and liver disease caused by alcohol, for example, alcoholic fatty liver, alcoholic hepatitis, or alcoholic liver cirrhosis.

In one embodiment of the present invention, alcoholic liver disease may be one or more selected from the group consisting of alcoholic fatty liver, alcoholic hepatitis, or alcoholic liver cirrhosis.

The content of the extract as an active ingredient in the composition according to the present invention can be appropriately adjusted depending on the type and purpose of use, the patient's condition, the type and severity of symptoms, etc., and 0.001 to 99.9 wt%, or 0.1 to 99.9 wt%, based on the solids weight , preferably 0.1 to 50% by weight, or 0.1 to 40% by weight, but is not limited thereto.

The composition according to the present invention may be administered to mammals, including humans, by various routes. The administration method may be any method commonly used, for example, may be administered by routes such as oral, skin, intravenous, intramuscular, subcutaneous, etc., and preferably orally. The composition of the present invention can be administered in oral dosage forms such as powders, granules, tablets, capsules, ointments, suspensions, emulsions, syrups, and aerosols, or parenteral dosage forms in the form of transdermal preparations, suppositories, and sterile injection solutions according to conventional methods, respectively. It can be formulated and used as such.

The composition of the present invention may further contain adjuvants such as pharmaceutically suitable and physiologically acceptable carriers, excipients and diluents in addition to the mixed extract.

Carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate , cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ( sucrose) or lactose, gelatin, etc. may be mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Formulations for oral use include suspensions, solutions, emulsions, syrups, ointments, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, transdermal preparations, and the like. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

As a suppository formulation, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. can be used.

In an embodiment in which the composition of the present invention is applied to humans, the herbal extract composition of the present invention may be administered alone, but in general, it is combined with a pharmaceutical carrier selected in consideration of the administration mode and standard pharmaceutical practice. They may be mixed and administered.

For example, the composition containing the herbal extract of the present invention may be in the form of a tablet containing starch or lactose, or in the form of a capsule containing alone or an excipient, or an elixir or suspension containing a flavoring or coloring chemical. It can be administered orally, orally, or sublingually in the form of tablets. Such liquid formulations may contain suspending agents (eg, methylcellulose, semisynthetic glycerides such as witepsol or mixtures of apricot kernel oil and PEG-6 esters or PEG-8 and caprylic/capric glyceride mixtures, such as mixtures of glycerides).

The dosage of the composition containing the herbal extract of the present invention may vary depending on the patient's age, weight, sex, dosage form, health condition, and disease level, and may be administered from once a day to several times a day at regular time intervals according to the judgment of a doctor or pharmacist. It may be administered in divided circuits.

For example, based on the active ingredient content, the daily dose may be 0.1 to 500 mg/kg, preferably 0.5 to 300 mg/kg. The above dosage is an example of an average case, and the dosage may be higher or lower depending on individual differences.

If the daily dosage of the composition containing the mixed extract of the present invention is less than the above dosage, a significant effect cannot be obtained, and when it exceeds that, it is not only uneconomical but also outside the range of the usual dosage, so there is a risk of undesirable side effects. It may occur, so it is better to set it within the above range.

Another aspect of the present invention is the treatment of alcoholic liver disease, chlorella extract having inhibitory activity, chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; It is a method for preparing an extraction mixture comprising at least one or more of

In one embodiment of the present invention, the chlorella extract may be a crude extract obtained by extracting chlorella or concentrated chlorella with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms, for example For example, it may be a water extract.

The manufacturing process of the chlorella extract according to the present invention will be described in more detail as follows: After washing the concentrated chlorella with distilled water at least once, and storing it at -80 ° C. to -60 ° C. for 3 to 10 days , after ultrasonic treatment by adding secondary distilled water, it can be extracted by centrifugation.

Green tea extract is a crude extract obtained by extracting one or more selected from the group consisting of leaves, seeds, stems and roots of green tea with one or more solvents selected from the group consisting of water, and linear or branched alcohols having 1 to 4 carbon atoms. can be

The manufacturing process of the green tea extract according to the present invention will be described in more detail as follows: The green tea extract is obtained by grinding one branch from the group consisting of green tea leaves, seeds, stems and roots and degreasing it by immersion with nucleic acid, This is refluxed at 20 to 100° C. for 1 to 8 hours and extracted. The extract can be extracted by filtering several times with an automatic filter to remove particles, and concentrating with a vacuum concentrator. Thereafter, the resin and the extract are reacted, the reactant is filled in a column, eluted with a solvent, and the process of reacting with the resin and eluting with the solvent is repeated 1 to 4 times, and then freeze-dried to be used as a green tea extract.

Stevia extract may be a crude extract obtained by extracting one or more selected from the group consisting of leaves, stems and roots of Stevia with one or more solvents selected from the group consisting of water, and a linear or branched alcohol having 1 to 4 carbon atoms. there is.

The manufacturing process of the stevia extract according to the present invention will be described in more detail as follows: The stevia extract is powdered by grinding one branch from the group consisting of leaves, stems and roots of stevia, immersed in nucleic acid to degrease, and 20 to 100 ℃ reflux for 1 hour to 8 hours and extracted. The extract can be extracted by filtering several times with an automatic filter to remove particles, and concentrating with a vacuum concentrator. Thereafter, the resin and the extract are reacted, the reactant is filled in a column, eluted with a solvent, and the process of reacting with the resin and eluting with the solvent is repeated 1 to 4 times, and then freeze-dried to be used as a stevia extract.

The old ginseng extract may be a crude extract obtained by extracting one or more selected from the group consisting of leaves, stems and roots of old ginseng with one or more solvents selected from the group consisting of water, and a linear or branched alcohol having 1 to 4 carbon atoms. there is.

The manufacturing process of the old ginseng extract according to the present invention will be described in more detail as follows: After grinding the old ginseng and making it into a powder, it is extracted under reflux with an extraction solvent in an amount of about 3 to 30 times by volume based on the weight of the old ginseng. After extraction, the filtrate is collected, and the resultant is concentrated. The extraction temperature is not particularly limited, but may be 40 to 120°C.

The extraction process can be repeated once or several times, and in one preferred example of the present invention, a method of re-extraction after the primary extraction can be adopted, which means that even if the herbal extract is effectively filtered, This is to prevent the loss because the water content is high, and the extraction efficiency is lowered only by the primary extraction. In addition, as a result of verifying the extraction efficiency for each step, it was found that about 80 to 90% of the total extraction amount was extracted by the secondary extraction.

In one embodiment of the present invention, when the extraction process is repeated twice, the obtained residue is subjected to reflux extraction with an extraction solvent, about 5 to 15 times by volume, preferably 8 to 12 times by volume. After extraction, it is filtered, combined with the filtrate obtained previously, and concentrated under reduced pressure to prepare an extract. In this way, extraction efficiency can be increased by mixing the filtrate obtained after the secondary extraction and each extraction, but the extract of the present invention is not limited to the number of extractions.

If the amount of the solvent used in the preparation of the extract is too small, stirring is difficult, the solubility of the extract is lowered, and the extraction efficiency is lowered. Since a phase problem may occur, it is preferable that the amount of the solvent used is within the above range.

The filtered extract thus obtained is about 10 to 30 times, preferably 15 to 25 times, more preferably about 20 times the total amount of the concentrate in order to control the content of the remaining lower alcohol to be suitable for use as a pharmaceutical raw material. It can be prepared as a powdery extract by azeotropically concentrating 1 to 5 times, preferably 2 to 3 times with pear water, adding the same amount of water again to homogeneously suspend it, and then freeze-drying it.

The extraction method used in the present invention may be any method commonly used, for example, cold extraction, hot water extraction, ultrasonic extraction, or reflux cooling extraction method, but is not limited thereto.

Another aspect of the present invention, chlorella extract chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; It is a food composition for alleviation, suppression or improvement of alcoholic liver disease comprising at least one of them.

The food composition according to the present invention may include chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; Since it includes at least one of at least one, the contents overlapping with the pharmaceutical composition are omitted to avoid excessive complexity of the present specification.

The content of the extract as an active ingredient contained in the food composition of the present invention is not particularly limited, as appropriate, depending on the type of food, desired use, etc., for example, it can be added as 0.01 to 15% by weight of the total food weight, health drink composition Silver may be added in a proportion of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

The health beverage composition of the present invention has no particular limitation on the liquid component other than containing the extract as an essential component in the indicated ratio, and may contain various flavoring agents or natural carbohydrates as additional components as in a conventional beverage. Examples of the above-mentioned natural carbohydrates include monosaccharides, such as disaccharides such as glucose, fructose, etc., such as maltose, sucrose, and the like, and conventional polysaccharides such as dextrin, cyclodextrin, and the like. sugar and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

The composition of the present invention comprises various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the compositions of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention relates to a composition for preventing or treating alcoholic liver disease, comprising an extract mixture, and a method for preparing the same, wherein the composition according to the present invention has an effect of enhancing the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, inhibiting fat accumulation and decomposing fat It has excellent promoting effect, so it can be used as a therapeutic agent for alcoholic liver disease.

1 is a control (CONT) and a composition according to an embodiment of the present invention (domestic chlorella extract: CHL, Japanese chlorella extract: CHL.J, green tea saponin: SP, stevia extract: STV, ginseng extract: GSM) human A graph showing the activity of alcohol dehydrogenase in hepatic stellate cells.
2 is a graph showing the activity of acetaldehyde dehydrogenase in human liver stellate cells of a control group and a composition according to an embodiment of the present invention.
3 is a graph showing the adipogenesis inhibitory activity of a control group and a composition according to an embodiment of the present invention.
4 is a graph showing the lipolysis promoting activity of the control group and the composition according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Preparation Example 1: Preparation of chlorella, stevia and crude ginseng extract

1-1. Chlorella Extract Preparation

Chlorella from Korea or Japan was purchased from Daesang Co., Ltd.

The concentrated chlorella was washed three times with distilled water, stored in a cryogenic freezer at -80 ° C. for one week, and then sonicated by adding 5 ml of secondary distilled water, and the supernatant was collected by centrifugation to sample 1 and 2 (Sample 1: Domestic chlorella extract, Sample 2: Japanese chlorella extract) was used in the experiment.

1-2. Saponin Extraction from Green Tea Seed Extract

Green tea seeds were purchased from the green tea farm of Master Shin Kwang-soo in Suncheon and used.

Green tea seeds were ground into powder, immersed in a hexane solution, degreased while sonicated, and extracted under reflux at 60 °C for 4 hours with 70% ethanol. After extraction, the particles were filtered with a filter machine and then concentrated with a concentrator. The resin was activated and reacted with the concentrated green tea seed extract at room temperature for 1 hour. Thereafter, the reaction mass was packed in a column, eluted with 0.3% sodium hydroxide, and the eluate was neutralized with hydrogen chloride. Then, it was reacted with the activated resin overnight, filled in a column, eluted with 100% ethanol, and then lyophilized for use.

1-3. Preparation of Stevia Extract

Stevia was obtained from Stevia Korea Co., Ltd. and was used.

Stevia leaves were pulverized and powdered, immersed in hexane solution, degreased while sonicated, and extracted under reflux with 70% ethanol at 60 °C for 4 hours. After extraction, the particles were filtered with a filter machine and then concentrated with a concentrator. The concentrated stevia extract was reacted with the activated porous resin at room temperature for 1 hour. After packing into a column and eluting with 0.3% sodium hydroxide, the eluate was neutralized with hydrogen chloride. Then, it was reacted with the activated porous resin overnight, filled in a column, eluted with 100% ethanol, and then lyophilized for use.

And, it was confirmed that stevioside and rebaudioside components were included in the stevia extract.

1-4. Preparation of ginseng extract

Ginseng was purchased at a herbal medicine wholesaler. 600 g of dried ginseng was pulverized into a powder, and then extracted with 70% ethanol under reflux at 60 °C for 4 hours. After extraction, the particles were filtered with a filtration machine, concentrated with a concentrator, and then lyophilized for use.

1-5. Preparation of extraction mixture

Sample 6: The domestic chlorella extract prepared in Example 1-1, the saponin extracted in Example 1-2 at a concentration of 25 ug/ml, and the stevia extract extracted in Example 1-3 at a concentration of 100 ug/ml to prepare a sample. prepared.

Sample 7: The Japanese chlorella extract prepared in Example 1-1, the saponin extracted in Example 1-2 at a concentration of 25 ug/ml, and the stevia extract extracted in Example 1-3 at a concentration of 100 ug/ml. was prepared.

Sample 8: The domestic chlorella extract prepared in Example 1-1, the saponin extracted in Example 1-2 at a concentration of 25 ug/ml, the stevia extract extracted in Example 1-3 at a concentration of 100 ug/ml, Example 1 A sample was prepared by mixing the high ginseng extract prepared in -4 at a concentration of 100 ug/ml.

Sample 9: Japanese chlorella extract prepared in Example 1-1, saponin extracted in Example 1-2 at a concentration of 25 ug/ml, and Stevia extract extracted in Example 1-3 at a concentration of 100 ug/ml, Example Samples were prepared by mixing the high ginseng extract prepared in 1-4 at a concentration of 100 ug/ml.

Example 1: Alcohol Dehydrogenase Assay (ADH)

Alcohol DH (ADH) is an enzyme that interconverts alcohol to aldehydes or ketones by reducing NAD+ to NADH. It is a seven dehydrogenase that occurs in many organisms, which can be toxic in humans and many other animals. It plays a role in breaking down alcohol.

In order to evaluate the ability of the composition according to the present invention to improve alcohol dehydrogenase activity, an experiment was conducted as follows. Human astrocytes were cultured in the astrocyte medium of a 6-well plate and cultured for 8 days by exchanging the medium while treating samples 1 to 9 with alcohol every 2 days. After 8 days, cells were collected and lysed using the Alcohol Dehydrogenase Assay Kit (Abcam, Great Britan) using the kit according to the manufacturer's protocol. The activity of alcohol dehydrogenase was analyzed by measuring absorbance at 450 nm using a 96 well plate, and the activity of alcohol dehydrogenase measured in FIG. 1 was shown.

As can be seen from the measurement results, it was confirmed that samples 1 to 9 all improved the activity of alcohol dehydrogenase compared to the control. In particular, it was confirmed that samples 7 and 8, in which chlorella extract, stevia extract, ginseng extract and saponin were combined, significantly improved the activity of alcohol dehydrogenase in hepatic stellate cells.

Example 2: Acetaldehyde Dehydrogenase Assay (ALDH)

Human astrocytes were cultured in astrocyte medium in a 6-well plate and cultured for 8 days while exchanging the medium while treating samples 1 to 9 with alcohol every 2 days. After 8 days, the cultured cells were collected and the cells were lysed using the ALDH activity assay kit (ab155893) kit according to the manufacturer's instructions. Then, the absorbance at 450 nm was measured using a 96-well plate to quantitatively analyze the activity of acetaldehyde dehydrogenase, and the results are shown in FIG. 2 .

As can be seen from the measurement results, as can be seen in FIG. 2, samples 1 to 9 all significantly improved the activity of acetaldehyde dehydrogenase compared to the control group. In particular, it was confirmed that Samples 8 and 9, in which chlorella extract, stevia extract, ginseng extract and saponin were combined, significantly improved the activity of acetaldehyde dehydrogenase in hepatic stellate cells.

Example 3: Adipogenesis inhibitory effect assay and hepatocyte triglyceride accumulation analysis

Human hepatic stellate cells (hSC) cells were cultured in each medium in a 6-well plate and cultured in a 5% CO ₂ incubator. After 2 days of culture or when cell confluency reached 70%, the old medium was replaced with a differentiation inducing medium, and the cells were maintained for 24 hours to suppress cell division. After 2 days, the culture medium was changed to a maintenance medium. The cells were cultured for 8 days while changing the medium every 2 days, and samples 1 to 9 were added every time the culture medium was cultured as an induction medium and a maintenance medium. After that, the cells were collected and the triglyceride accumulated in the cells was quantitatively measured through color and fluorescence observation with an Adipogenesis Detection Assay Kit (Adipogenesis Detection Assay Kit, abcam, USA) using a 96-well plate. 3 is shown.

As a result of the measurement, as can be seen in FIG. 3 , it was confirmed that the group administered with samples 1 to 9 significantly reduced the amount of fat accumulated compared to the control group. In particular, samples 6 and 7 in which chlorella extract and stevia extract were combined, and samples 8 and 9 in which chlorella extract, stevia extract, ginseng extract and saponin were combined were excellent in reducing fat accumulation in hepatic stellate cells compared to the control group. , among them, samples 8 and 9 showed the most excellent effect on reducing fat accumulation.

Example 4: Analysis of lipolysis efficacy

Lipolysis is the hydrolysis of intracellular triglycerides into glycerol and free fatty acids, which are released into the bloodstream or culture medium. Fat breakdown occurs in all cells, but occurs most often in white and brown adipose tissue. When lipolysis is not performed smoothly, intracellular lipid accumulation is increased, resulting in abnormal cell physiology, hyperlipidemia, and insulin resistance.

Accordingly, the present inventors measured the lipolysis efficacy of Samples 1 to 9 according to the present invention using a colorimetric method for glycerol released from cells after induction of lipolysis.

Human hepatic astrocyte cells were cultured in each medium in a 6-well plate and cultured in a 5% CO ₂ incubator, after 2 days of culture or when the cells reached 70% confluency, the medium was replaced with a differentiation inducing medium, , cells were maintained in this state for 24 h to inhibit cell division. After 2 days, the culture medium was changed to a maintenance medium, and the cells were cultured for 8 days while changing the medium every 2 days. Samples 1 to 9 were added at each culture period with an induction medium and a maintenance medium, and then cells were collected and the induction and lipolysis of lipolysis were performed with a lipolysis kit (Adipogenesis Assay Kit, Biovision, USA) using a 96-well plate. This was measured and shown in FIG. 4 .

As a result of the measurement, as can be seen in FIG. 4 , the amount of fat decomposed into glycerol was significantly increased in the group administered with samples 1 to 9 compared to the control group. In particular, samples 6 and 7 in which chlorella extract and stevia extract were combined, and samples 8 and 9 in which chlorella extract, stevia extract, ginseng extract and saponin were combined were superior in the effect of decomposing fat into glycerol in hepatic stellate cells compared to the control. Among them, samples 8 and 9 showed the most excellent lipolysis effect.

Claims (12)

chlorella, chlorella culture or chlorella extract; saponins; stevia extract; and ginseng extract; A pharmaceutical composition for preventing, alleviating, inhibiting or treating alcoholic liver disease, comprising at least one of. The composition of claim 1, wherein the composition comprises a chlorella extract, a stevia extract and a saponin. The composition of claim 1, wherein the composition comprises a chlorella extract, a saponin, a stevia extract, and a ginseng extract. The composition of claim 1, wherein the chlorella extract is a crude extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms. The composition of claim 1, wherein the high ginseng extract is a crude extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms. The composition of claim 5, wherein the solvent is an aqueous alcohol solution or anhydrous alcohol having an alcohol content of 50% or more to less than 100% (v/v). The composition according to claim 6, wherein the solvent is an aqueous alcohol solution having an alcohol content of 70%. The composition of claim 1, wherein the saponin is obtained from green tea extract. The composition of claim 1, wherein the alcoholic liver disease is at least one selected from the group consisting of alcoholic fatty liver, alcoholic hepatitis, or alcoholic cirrhosis. chlorella, chlorella cultures or chlorella extracts; saponins; stevia extract; and ginseng extract; A food composition for alleviation, suppression or improvement of alcoholic liver disease, comprising at least one of. The composition of claim 10, wherein the composition comprises a chlorella extract, a stevia extract and a saponin. The composition of claim 10, wherein the composition comprises a chlorella extract, a saponin, a stevia extract and a ginseng extract.

Images