KR102092729B1 - Pharmaceutical composition for preventing or treating liver damage comprising Curcuma longa extract - Google Patents

Abstract

The present invention relates to a composition for preventing and treating liver damage, and more particularly, to a pharmaceutical composition for preventing and treating liver damage, including a turmeric extract capable of preventing and treating liver damage by improving a hepatocellular protective effect according to liver toxicity. It is about.

Description

Pharmaceutical composition for preventing or treating liver damage comprising Curcuma longa extract}

The present invention relates to a composition for preventing and treating liver damage, and more specifically, to improve the hepatocellular protective effect according to liver toxicity, to prevent and treat liver damage, turmeric extract, its fractions and / or components separated therefrom It relates to a pharmaceutical composition for the prevention and treatment of liver damage, including.

The liver is one of the most active organs in the human body, and is the largest organ in the human body. The liver is positioned between the digestive system and the systemic circulatory system and functions to protect the whole body from exogenous substances from outside. In addition, the liver is a very important organ responsible for various metabolism, detoxification, decomposition, synthesis and secretion.

More specifically, the liver has a function of managing energy metabolism, and all nutrients absorbed from food are metabolized as substances capable of producing energy in the liver and supplied or stored in the whole body, and about 2,000 enzymes, albumin, and coagulation Fats, such as serum proteins, bile acids, phospholipids, and cholesterol, are synthesized, stored, and distributed in the liver. In addition, the liver has a function of excreting various metabolites into the duodenum through the bile ducts and immune function, which plays an important role in maintaining our lives. In addition, since it detoxifies and detoxifies drugs, alcohol, and toxic substances in the liver as a function of detoxification, hepatocytes are easily damaged, and thus, drug, toxicity, and alcoholic liver disease may frequently occur. In addition, since the ex vivo substance that has entered the body once passes through the liver, the liver has a higher risk of being exposed to many toxic substances in addition to nutrients than other organs, so the probability of damage is very high.

Among the causes of liver disease, liver damage caused by drugs occupies the top. In particular, acetaminophen (AAP) is the most commonly used analgesic and anti-inflammatory agent known as Tylenol, but it has been reported to cause acute liver damage (liver necrosis, neurotoxin, liver hardening, etc.) when taken in excess. have.

Accordingly, there is a need to develop a material capable of preventing or treating liver damage caused by drugs such as chemicals, especially acetaminophen.

On the other hand, turmeric is a kind of the ginger family ( Zingiberaceae ) and the scientific name is Curcuma longa . It is used as a spice in tropical regions of Southeast Asia, and has strong antioxidant and anti-inflammatory properties, and has been widely used as a traditional medicine. Curcuma longa is antifungal activity (Apisariyakul Amphawan and 2 others, 1995, Antifungal activity of turmeric oil extracted from Curcuma longa, Journal of Ethnopharmacology, Vol. 49, No. 3, pp. 163 ~ 169), anti-inflammatory, anti-HIV , Biological activity such as antibacterial, antioxidant, anti- nematode activity, anti-cancer (CAC Araujo et al. 1, 2001, Biological activities of Curcuma longa L., MEMORIAS DO INSTITUTO OSWALDO CRUZ, pp.723 ~ 728), etc. There have been no research reports that turmeric extract is effective in preventing or treating liver damage caused by liver toxicity.

The present invention was devised in view of the above points, and it is possible to prevent and treat liver damage by improving the hepatocellular protective effect according to liver toxicity, a pharmaceutical composition for preventing and treating liver damage, including turmeric extract and functional ingredients, or The aim is to provide a food composition for preventing and improving liver damage.

In order to solve the above-described problems, the present invention provides a pharmaceutical composition for preventing and treating liver damage, including turmeric ( Curcuma longa ) extract or a fraction thereof as an active ingredient.

According to an embodiment of the present invention, the extract may be extracted through water, alcohol of C1 to C4, or a mixed solvent thereof.

In addition, the fraction may be a fraction of turmeric extract through a solvent roll containing ethyl acetate.

In addition, the extract or a fraction thereof may include a compound represented by Formula 1, a compound represented by Formula 2, or a mixture thereof.

[Formula 1]

[Formula 2]

In addition, the liver damage may be caused by acetaminophen (Acetaminophen) drug.

In addition, the present invention provides a pharmaceutical composition for preventing and treating liver damage comprising a compound represented by Formula 1 or a salt thereof, a compound represented by Formula 2 or a salt thereof, or a mixture thereof as an active ingredient.

[Formula 1]

[Formula 2]

According to an embodiment of the present invention, the compound represented by Chemical Formula 1 and the compound represented by Chemical Formula 2 may be derived from turmeric ( Curcuma longa ).

In addition, the present invention provides a food composition for preventing and improving liver damage, including turmeric ( Curcuma longa ) extract or a fraction thereof as an active ingredient.

According to an embodiment of the present invention, the extract or a fraction thereof may include a compound represented by Formula 1, a compound represented by Formula 2, or a mixture thereof.

[Formula 1]

[Formula 2]

In addition, the present invention provides a food composition for preventing and improving liver damage comprising a compound represented by Formula 1 or a salt thereof, a compound represented by Formula 2 or a salt thereof, or a mixture thereof as an active ingredient.

[Formula 1]

[Formula 2]

According to an embodiment of the present invention, the compound represented by Chemical Formula 1 and the compound represented by Chemical Formula 2 may be derived from turmeric ( Curcuma longa ).

Hereinafter, terms used in the present invention will be described.

As used in the present invention, the term "prevention" in the present invention means all actions that inhibit liver damage or delay the onset of disease by administration of the composition.

In the present invention, the term, "treatment" means any action that improves or beneficially alters the symptoms of liver damage by administration of the composition.

According to the composition according to the present invention, hepatotoxicity can be significantly prevented and treated or improved by protecting hepatocytes from liver damage caused by liver toxicity, in particular, liver toxicity caused by drugs, thereby preventing, treating and improving liver damage. B. It can be widely applied as functional food.

1 is a graph of liver cell viability according to an embodiment and a comparative example of the present invention treated on a human liver cell line in which liver damage is induced by acetaminophen.
Figure 2 is a graph showing the AST activity of liver cells according to an embodiment and a comparative example of the present invention treated on a human liver cell line in which liver damage has been induced by acetaminophen.
Figure 3 is a graph of the LDH concentration of liver cells according to an embodiment and a comparative example of the present invention treated in human liver cell line induced liver damage by acetaminophen.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art to which the present invention pertains can easily practice. The present invention can be implemented in many different forms and is not limited to the embodiments described herein.

First, the pharmaceutical composition for preventing and treating liver damage according to an embodiment of the present invention includes a turmeric ( Curcuma longa) extract or a fraction thereof as an active ingredient.

First, the turmeric extract will be described.

The turmeric (Cu rcuma longa) to be extracted may be purchased and used commercially, or may be used that has been harvested or grown in nature. In addition, the turmeric may use any one or more parts of roots, stems, and leaves, preferably roots. At this time, the turmeric is not limited to whether or not the object is dried or pulverized during extraction, but preferably, dried and pulverized can be used.

The turmeric extract may be extracted using a conventional extraction method in the art, for example, an ultrasonic extraction method, a filtering method, and a reflux extraction method. In a specific embodiment, the turmeric crushed product obtained by pulverizing turmeric from which foreign substances have been removed by washing and drying may be extracted through water, an alcohol of C1 to C4, or an extraction solvent that is a mixed solvent thereof. More preferably, the extraction solvent may be extracted using ethanol or an ethanol aqueous solution. At this time, the extract can be extracted for 15 to 36 hours at a temperature condition of 20 to 30 ℃. In addition, after extracting once, the extract can be filtered, and the remaining extraction target can be extracted 2 to 4 more times under the same conditions to obtain an extract.

The obtained extract may be included in the composition as it is or concentrated under reduced pressure. At this time, since the concentration under reduced pressure can be performed under conditions commonly used in the art, detailed description of the present invention is omitted.

Next, the fraction of turmeric extract will be described.

The fraction may be one obtained by fractionating the turmeric extract obtained by the above-described method through any one or more fractional solvents of water, ethyl acetate, and n-butanol, and preferably preventing liver damage. , It may be an ethyl acetate fraction for turmeric ethanol extract in terms of the content of the active ingredient for treatment.

When explaining an example of obtaining the fraction, extract the turmeric ethanol extract through distribution with water and ethyl acetate, and then extract the water layer again with n-butanol, and then concentrate each layer under reduced pressure to ethyl acetate fraction, n-butanol Fractions and water fractions can be obtained.

According to a preferred embodiment of the present invention, the extract and fraction may include a compound represented by the following formula (1), a compound represented by the formula (2), or a mixture thereof, through which the prevention and treatment of liver damage A remarkable effect can be exhibited.

[Formula 1]

[Formula 2]

Specifically, the compound represented by Chemical Formula 1 and the compound represented by Chemical Formula 2 have an effect of improving the cell viability of the human liver cancer cell line (HepG2) by inhibiting liver toxicity induced by acetaminophen treatment, in particular, silymarin, a positive control. It can be seen that even when treated at a lower concentration, the hepatocyte protection effect is more excellent.

In addition, as can be seen through Figure 2, the compound represented by the formula (1) and the compound represented by the formula (2) are treated with a compound represented by the formula (2) or a compound represented by the formula (1) with an increased AST activity level by acetaminophen treatment. It can be seen that it was significantly reduced. In addition, it can be seen that more effective in reducing the AST activity level than silymarin treated with a higher concentration.

In addition, as can be seen through FIG. 3, when hepatocytes are killed by damage, LDH (Lactate dehydrogenase) enzyme present in the hepatocytes is released outside the cells to increase the concentration, and the increased LDH concentration by acetaminophen treatment is represented by Formula 1 Through the treatment of the compound represented by Formula 2 or the compound represented by Formula 2, it can be confirmed that it was significantly lowered, and through this, it can be seen that there is an apoptosis inhibitory effect on hepatocytes damaged by the drug. In addition, it can be seen that more effective in reducing LDH concentration than silymarin treated at a higher concentration.

As described above, the compound represented by Chemical Formula 1 or the compound represented by Chemical Formula 2 is included in a turmeric extract or a fraction thereof, and will be said to be an active ingredient capable of preventing and treating liver damage. For example, the compound is a turmeric extract or a fraction thereof It can be obtained through the step of separation and purification through silica gel column chromatography, reverse phase silica gel column chromatography and Sephadex LH-20 column chromatography, and can be performed through one to several times until a single compound is purified. Concentration and recrystallization can be performed as needed.

As an example, the obtained ethyl acetate fraction can be fractionated using silica gel column chromatography (inner diameter 10 × 20 cm, volume ratio of elution solvent CHCl ₃ and MeOH 40-60: 1), and each aliquot is subjected to thin layer chromatography ( After confirming by thin layer chromatography (TLC), silica gel column chromatography (inner diameter 5 × 20 cm, elution solvent CHCl ₃ and volume ratio of 10 to 30: 1 of elution solvent) was performed on each of the five fractions obtained. Refractions can be obtained for each fraction. Subsequently, octadecyl column chromatograph (ODS cc), inner diameter 4 × 10 cm, volume ratio of MeOH and H ₂ O 5-20: 1) was performed on each redistribution to each redistribution. For the re-refractions (CUE-2-3-1 to CUE-2-3-10), it is possible to obtain compounds represented by the above formula (1) and compounds represented by the formula (2) among these re-fractions. At this time, the yield of the compound represented by Formula 1 and the compound represented by Formula 2 may be about 30 mg of the compound represented by Formula 1 based on 100 g of ethyl acetate fraction, and the compound represented by Formula 2 may be about 15 mg per day. Can.

The compound represented by Formula 1 or the compound represented by Formula 2 separated and purified through the above-described method may be included in the form of a pharmaceutically acceptable salt. As the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. 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 and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes It is obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulphite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfate Nate, xylene sulfonate, phenyl acetate, phenyl propionate, phenyl butyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propane sulfonate, naphthalene-1- Sulfonate, naphthalene-2-sulfonate or mandelate.

In addition, the acid addition salt is a conventional method, for example, the compound represented by the formula (1) or the compound represented by the formula (2) is dissolved in an excess of an aqueous acid solution, and the salt is water-miscible organic solvent, such as methanol, It can be prepared by precipitation using ethanol, acetone or acetonitrile.

In addition, the compound represented by the formula (1) or the compound represented by the formula (2) of the present invention may be used in the form of a pharmaceutically acceptable metal salt using a base. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the inexpensive compound salt, and evaporating and drying the filtrate. At this time, it is suitable to manufacture sodium, potassium or calcium salts as metal salts. Further, the corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

In addition, the pharmaceutical composition according to an embodiment of the present invention may include a pharmaceutically acceptable carrier. The composition comprising a pharmaceutically acceptable carrier may be various formulations, oral or parenteral.

In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in one or more compounds, such as starch, calcium carbonate, sucrose or lactose ( lactose) and gelatin. Also, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspending agents, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used as diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents, suspension solvents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

In addition, the pharmaceutical composition is selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, intravenous solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizers and suppositories. It can have either formulation.

In addition, the pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. In the present invention, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the extent of liver damage, the cause of the liver damage, age, sex , The activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of excretion, the duration of treatment, the factors including the drugs used simultaneously and other factors well known in the medical field. However, for the desired effect, the pharmaceutical composition according to an embodiment of the present invention is preferably administered at 0.0001 to 100 mg / kg per day, preferably at 0.001 to 100 mg / kg, and the compound or formula represented by Formula 1 The compound represented by 2, or a mixture thereof, is preferably administered at 0.0001 to 100 mg / kg per day, preferably at 0.001 to 10 mg / kg per day . The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, and can be easily determined by those skilled in the art.

In addition, the pharmaceutical composition according to an embodiment of the present invention may be administered by various methods known in the art, for example, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, intranasal Administration, intrapulmonary administration, or rectal administration, but is not limited thereto.

In addition, according to another embodiment of the present invention, the pharmaceutical composition for preventing and treating liver damage is an active ingredient of a compound represented by Formula 1 or a salt thereof, a compound represented by Formula 2 or a salt thereof, or a mixture thereof. Include as.

[Formula 1]

[Formula 2]

At this time, preferably, the compound represented by Formula 1 and the compound represented by Formula 2 may be derived from turmeric (Cu rcuma longa) .

On the other hand, the description of the pharmaceutical composition such as the compound represented by the formula (1) or a salt thereof, and the compound represented by the formula (2) or a salt thereof is the same as described above and will be omitted below.

The pharmaceutical composition for preventing and treating liver damage according to the present invention described above is effective in preventing and treating liver damage due to various causes, but can exhibit excellent efficacy in liver damage caused by acetaminophen drug.

Next, the present invention includes a turmeric (Cu rcuma longa) extract or a food composition for preventing and improving liver damage comprising a fraction thereof as an active ingredient.

That is, turmeric extract or a fraction thereof may be added to the food composition for the purpose of preventing and improving liver damage. In addition, the extract and fractions thereof may include a compound represented by Formula 1, a compound represented by Formula 2, or a mixture thereof.

In this case, the extract or fraction may be added as it is or used with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the turmeric extract or fractions thereof of the present invention may be added in an amount of 0.01 to 10% by weight, preferably 0.1 to 3% by weight of the raw material composition, but is not limited thereto. It may be added by changing in consideration of the period.

There are no particular restrictions on the type of food. Examples of foods include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes. And includes all health foods in the usual sense.

In addition, in the case of a health drink composition, it may contain various flavoring agents or natural carbohydrates, etc., as additional components, like a normal beverage. The natural carbohydrates described above may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention, but is not limited thereto, and may be appropriately changed according to preference.

Also used in various other nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acids and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonated drinks It may also contain a carbonizing agent. The proportion of these additives is not critical, but is generally selected in the range of 0.01 to 0.1% by weight based on the total composition weight. In addition, it may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. The proportion of the flesh is not very important, but is generally selected in the range of 0.01 to 10% by weight based on the total composition weight. These ingredients can be used independently or in combination.

In addition, the food composition for preventing and improving liver damage according to an embodiment of the present invention includes a compound represented by Formula 1 or a salt thereof, a compound represented by Formula 2 or a salt thereof, or a mixture thereof as an active ingredient. .

[Formula 1]

[Formula 2]

At this time, preferably, the compound represented by Formula 1 and the compound represented by Formula 2 may be derived from turmeric (Cu rcuma longa) .

On the other hand, the description of the food composition such as the compound represented by the formula (1) or a salt thereof, and the compound represented by the formula (2) or a salt thereof is the same as described above and will be omitted below.

The above-described prototype composition for preventing and improving liver damage according to the present invention is effective in preventing and improving liver damage due to various causes, but can particularly exhibit excellent efficacy in liver damage caused by acetaminophen drug.

The present invention will be described in more detail through the following examples, but the following examples are not intended to limit the scope of the present invention, which should be interpreted to help understand the present invention.

<Example 1>

With respect to 5 kg of dried and ground turmeric sample, a 70% ethanol aqueous solution was used as an extraction solvent and extracted at 25 ° C. for 24 hours. After filtering the extracted sample, the remaining sample was extracted twice more in the same manner. Thereafter, the obtained filtrates were combined and concentrated under reduced pressure to obtain an ethanol extract. The obtained ethanol extract was extracted with ethyl acetate (3ℓ × 2 / H ₂ O (3ℓ)), and the H ₂ O layer was extracted with n-BuOH (2.5ℓ × 2). Concentrated to obtain ethyl acetate fraction (250 g) and n-butanol fraction (100 g) and H ₂ O fraction.

100 g of the obtained ethyl acetate fraction (CUE) was subjected to silica gel column chromatography (inner diameter 10 × 20 cm, volume ratio of elution solvent CHCl ₃ and MeOH 50: 1), and each aliquot (200 ml) was subjected to thin layer chromatography (thin layer). Chromatography, TLC) to obtain 5 fractions (CUE-1 ~ CUE-5). Among them, 12 fractions (CUE-2-1 to CUE-2-12) were subjected to silica gel column chromatography (inner diameter 5 × 20 cm, 20: 1 volume ratio of CHCl ₃ and MeOH) for 10 g of CUE-2 fractions. Divided into 10 fractions by performing octadecyl column chromatograph (ODS cc), inner diameter 4 × 10 cm, volume ratio of MeOH and H ₂ O 10: 1) for 1 g of CUE-2-3 fraction. (CUE-2-3-1 to CUE-2-3-10) were obtained, among which compounds represented by the following formula (CUE-2-3-1-5, 30mg) and compounds represented by the formula (2) (CUE-2-3-1-5, 15mg).

[Formula 1]

[Formula 2]

<Comparative Example 1>

The turmeric, n-butanol and water fractions obtained in Example 1 were carried out in the same manner as in Example 1 to separate and purify each fraction. As a result, the compounds represented by Chemical Formulas 1 and 2 were not detected.

<Experimental Example 1>

First, in order to confirm the hepatocellular protective activity against hepatotoxicity caused by acetaminophen using a human liver cancer cell line (HepG2 cell), 10% (v / v) fetal bovine serum (HepG2 cell) FBS), penicillin (100 U / mL) and streptomycin (100 μg / mL) were added to the DMEM medium and cultured in a cell incubator where 5% CO ₂ was maintained at a temperature of 37 ° C.

Thereafter, after culturing the human liver cancer cell line at a concentration of 2 × 10 ⁵ cells / ml for 24 hours, the hepatocytes were treated with 10 μM of the compound represented by Formula 1 according to Example 1 and the compound represented by Formula 2 according to Example 2 After treatment at a concentration of 2 hours before, treatment with 10 mM of acetaminophen was conducted to induce liver toxicity for 24 hours, and then cell viability was measured.

At this time, as a normal control (Control), acetaminophen-free liver cancer cell line, and as a negative control, hepatic cancer cell line treated with acetaminophen alone was used. As a positive control, silymarin, which is currently used as an auxiliary treatment for liver disease and cirrhosis, was used. A liver cancer cell line treated with acetaminophen after pretreatment for 2 hours at a concentration of 20 μM, 50 μM, and 100 μM was used. On the other hand, MTT assay was used to measure cell viability, and the cells after the reaction were treated with Thiazolyl blue tetrazolium bromide (MTT) reagent at a final concentration of 1 mg / ml and reacted for 1 hour. After removal of the cell culture, the formazan formed by adding DMSO was dissolved, and the absorbance at 540 nm was measured with a Microplate reader (Synergy H1, BioTek), and the cell viability was the absorbance of the sample treatment group compared to the normal control group without treatment. It was expressed as a percentage of the ratio, the results are shown in Figure 1 below.

As can be seen through Figure 1, the compound represented by Formula 1 (Sample 1) and the compound represented by Formula 2 (Sample 2) inhibit the liver toxicity induced by acetaminophen treatment to cells of the human liver cancer cell line (HepG2). It has the effect of improving the survival rate, and it can be seen that the hepatocyte protection effect is more excellent, especially when treated at a lower concentration than the positive control silymarin.

<Experimental Example 2>

Experimented in the same manner as in Experimental Example 1, but not the cell viability, AST activity level was evaluated. Specifically, the AST activity value was measured at 450 nm using an Aspartate Aminotransferase (AST) activity assay kit (Sigma). Thereafter, it was quantified by the following AST activity calculation method, and the results are shown in FIG. 2 below.

* B = amount of glutamate produced (nmole)

As can be seen through Figure 2, the compound represented by Formula 1 (Sample 1) and the compound represented by Formula 2 (Sample 2) have an elevated AST activity level by acetaminophen treatment, or a compound represented by Formula 2 It can be confirmed that the treatment was markedly lowered. In addition, it can be seen that more effective in reducing the AST activity level than silymarin treated with a higher concentration.

<Experimental Example 3>

The experiment was conducted in the same manner as in Experimental Example 1, but the change in LDH concentration, not the cell viability, was measured. That is, when the liver cells are killed by the damage, the concentration of LDH (Lactate dehydrogenase) present in the liver cells is released out of the cells, thereby increasing the concentration.

Specifically, the LDH activity was measured by absorbance at 490 nm using LDH Cytotoxicity Detection Kit (TAKARA), and expressed as a percentage of the absorbance ratio of the sample treatment group compared to the normal control group without sample treatment. It is shown in 3.

As can be seen through Figure 3, it can be seen that the LDH concentration increased by acetaminophen treatment was significantly lowered through the treatment of the compound represented by Formula 1 (Sample 1) or the compound represented by Formula 2 (Sample 2). , Through this, it can be seen that there is an apoptosis inhibitory effect on hepatocytes damaged by drugs. In addition, it can be seen that more effective in reducing LDH concentration than silymarin treated at a higher concentration.

[Production Example 1]-Medicine

<Production Example 1-1>-Powder

After mixing 50 mg of turmeric ethanol extract of Example 1 and 2 g of crystalline cellulose, a powder was prepared by filling in an airtight bag according to a conventional powder manufacturing method.

<Production Example 1-2>-Tablet

Tablets were prepared by mixing 50 mg of turmeric ethyl acetate fraction of Example 1, 400 mg of crystalline cellulose, and 5 mg of magnesium stearate, followed by tableting according to a conventional tablet manufacturing method.

<Production Example 1-3>-Capsule

After mixing 30 mg of the compound represented by Formula 1 of Example 2, 100 mg of whey protein, 400 mg of crystalline cellulose, and 6 mg of magnesium stearate, the capsules were prepared by filling into gelatin capsules according to a conventional capsule preparation method.

[Production Example 2]-Food

<Production Example 2-1>-Preparation of healthy food

Turmeric ethyl acetate fraction of Example 1 1000 mg, vitamin A acetate 70 μg, vitamin E 1.0 mg, vitamin B1 0.13 mg, vitamin B2 0.15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 μg, vitamin C 10 mg, biotin 10 μg , 1.7 mg nicotinamide, 50 ug folic acid, 0.5 mg calcium pantothenate, 1.75 mg ferrous sulfate, 0.82 mg zinc oxide, 25.3 mg magnesium carbonate, 15 mg potassium phosphate, 55 mg calcium phosphate, 90 mg potassium citrate , 100 mg of calcium carbonate, 24.8 mg of magnesium chloride may be prepared by mixing, and the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method, and then granules are prepared.

<Production Example 2-2>-Preparation of health drinks

After adding purified water to 1000 g of turmeric ethyl acetate fraction of Example 1, 1000 mg of citric acid, 100 g of oligosaccharides, 2 g of plum concentrate, and 1 g of taurine, the total of 900 ml was mixed with the above components according to a conventional method for preparing a healthy beverage, After stirring and heating at 85 ° C for about 1 hour, the resulting solution was filtered to obtain a sterilized 2L container, sealed and sterilized, and then refrigerated to prepare a health drink.

Although one embodiment of the present invention has been described above, the spirit of the present invention is not limited to the embodiments presented herein, and those skilled in the art to understand the spirit of the present invention may add elements within the scope of the same spirit. However, other embodiments may be easily proposed by changing, deleting, adding, or the like, but it will also be considered to be within the scope of the present invention.

Claims (11)

delete delete delete delete delete A pharmaceutical composition for the prevention and treatment of liver damage comprising a compound represented by Formula 1 or a salt thereof, a compound represented by Formula 2 or a salt thereof, or a mixture thereof as an active ingredient.
[Formula 1]

[Formula 2]

The method of claim 6,
The compound represented by Formula 1 and the compound represented by Formula 2 are pharmaceutical compositions for preventing and treating liver damage, characterized in that it is derived from turmeric (Cu rcuma longa) . delete delete A food composition for preventing and improving liver damage comprising a compound represented by Formula 1 or a salt thereof, a compound represented by Formula 2 or a salt thereof, or a mixture thereof as an active ingredient.
[Formula 1]

[Formula 2]

The method of claim 10,
The compound represented by Chemical Formula 1 and the compound represented by Chemical Formula 2 are food compositions for preventing and improving liver damage, characterized in that it is derived from turmeric (Cu rcuma longa) .

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