KR101401416B1 - Composition for protecting liver - Google Patents

Abstract

The present invention relates to a composition for protecting the liver, and more particularly to a composition for protecting the liver comprising a specific compound. The composition of the present invention inhibits hepatocyte apoptosis and can lower the concentration of reactive oxygen species (ROS). The composition of the present invention is also useful as it can restore the glutathione content and restore liver damage by alcohol.

Description

{COMPOSITION FOR PROTECTING LIVER}

The present invention relates to a composition for protecting the liver, and more particularly to a composition for protecting the liver comprising a specific compound.

Liver disease is one of the major causes of death worldwide, especially in Korea and China, where liver damage due to excessive alcohol consumption is frequent and the liver market is particularly large. Examples of liver disease include not only alcoholic liver damage, but also drug-induced liver damage and liver cancer.

The inventors of the present invention confirmed that one or more compounds selected from the group consisting of the compounds represented by formulas (1) and (2) had liver protective effect when they were treated with the liver, and completed the present invention.

Kang et al, 2011, Journal of Medicinal Food 14: 1198-1207

It is an object of the present invention to provide a liver protecting composition for preventing or treating liver disease by preventing the death of hepatocytes and protecting the liver by restoring damaged liver cells and the like.

In order to accomplish the above object, the present invention provides a composition for protecting the liver comprising, as an active ingredient, one or more compounds selected from the group consisting of the following formulas (1) and (2)

R 1, R 2, R 3, R 4 or R 5 substituents of the formulas are independently hydrogen, C ₁ to C ₁₈ alkyl, C ₁ To C ₁₈ alkoxy groups, C ₁ To C ₁₈ alkenyl groups or C ₁ To C < ₁₈ > is an alkynyl group.

[Chemical Formula 1]

(2)

The composition of the present invention inhibits hepatocyte apoptosis and can lower the concentration of reactive oxygen species (ROS). The composition of the present invention is also useful as it can restore the glutathione content and restore liver damage by alcohol.

In addition, the composition of the present invention contains a natural product obtained from nature as an active ingredient, and has a low adverse effect when applied to a human body, and is less likely to cause resistance due to long-term use. In particular, since the composition of the present invention has excellent efficacy in the protection of hepatocytes, it can be usefully used for liver protection.

FIG. 1 shows the results of measuring the protective effect of Paeonia anomala extract against the death of human hepatocyte HepG2 cells by tert-butylhydroperoxide.
FIG. 2 shows the results of measuring the protective effect of Paeonia anomerale fraction on human hepatocyte HepG2 cell death by tert-butylhydroperoxide.
FIG. 3 is a graph showing the activity of 2-hydroxy-6-methoxy-4-O- (6'-O- alpha -L- arabinofuranosyl- beta -D-glucopyranosyl) D-glucopyranosyl) acetophenone represented by the following formula (3) and 3,3'-di-O- (3 "-O-Galoyl- D-glucopyranosyl) ellagic acid (Formula 4) in human hepatocyte HepG2 cell death (3, 3'-di-O-methyl-4-O- This is the result of measuring the protective effect.
Fig. 4 shows the results of measuring the effect of Paeonia anomala fruit and root extract on the production of reactive oxygen species.
FIG. 5 shows the results of measuring the effect of Paeonia anomala fruit and root extract on intracellular antioxidant glutathione content.
FIG. 6 shows the results of measurement of the effect of Paeonia anomala fruit and root extract on DNA damage caused by oxidative stress.
FIG. 7 shows the results of confirming the protective effect of the Paeonia anomerale fruit extract against alcoholic liver damage by measuring the liver damage index ALT in an animal experiment.
FIG. 8 shows the results of confirming the protective effect of Paeonia anomerale fruit extract against alcoholic liver damage by measuring AST as an index of liver damage in an animal experiment.

In one aspect, the present invention relates to an antimicrobial composition comprising, as an active ingredient, at least one compound selected from the group consisting of the following formulas (1) and (2) or a pharmaceutically acceptable salt thereof,

R 1, R 2, R 3, R 4 or R 5 substituents of the formulas are independently hydrogen, C ₁ to C ₁₈ alkyl, C ₁ To C ₁₈ alkoxy groups, C ₁ To C ₁₈ alkenyl groups or C ₁ To C ₁₈ alkynyl groups.

[Chemical Formula 1]

(2)

As used herein, the term " pharmaceutically acceptable "refers to the approval of a government or equivalent regulatory agency that can be used on animals, and more specifically on humans, by avoiding significant toxic effects when used in conventional medicinal dosage Received, approved, or listed in pharmacopoeia or other general pharmacopoeia.

As used herein, "pharmaceutically acceptable salt" means a salt according to one aspect of the present invention which is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. The salt may be (1) formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; (3-hydroxybenzoyl) benzoic acid, or an acetic acid, propionic acid, hexanoic acid, cyclopentenepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4- chlorobenzenesulfonic acid, 2- 4-methylbicyclo [2,2,2] -oct-2-en-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert Acid addition salts formed with organic acids such as butylacetic acid, laurylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid; Or (2) salts formed when the acidic proton present in the parent compound is substituted.

As used herein, the term " active ingredient "alone refers to an ingredient that exhibits the desired activity or that can exhibit activity with a carrier that is not itself active.

As used herein, the term " liver protection "is intended to include, without limitation, any use for protecting a healthy liver or a damaged liver, such as by protecting the death of liver cells or protecting liver cells from oxidative stress.

In a liver protecting composition that is one aspect of the present disclosure, the composition includes preventing or treating liver disease. In addition, in the liver protecting composition which is one aspect of the present invention, the liver disease includes alcoholic liver damage diseases. The composition of the present invention can prevent or treat the diseases associated with liver or diseases arising in the liver. Specifically, the above-mentioned "prevention or treatment of liver disease" means prevention of hepatocyte death induced by tert- butyl hydroperoxide, reduction in concentration of reactive oxygen species (ROS) But is not limited to, increasing the content or recovering liver damage caused by alcohol. As used herein, the term " alcoholic liver damage disorder "includes diseases caused by damage to the liver by alcohol.

In the liver protecting composition according to one aspect of the present invention, the active ingredient may be at least one compound selected from the group consisting of the following formulas (3) and (4) or a pharmaceutically acceptable salt thereof.

2-Hydroxy-6-methoxy-4-O- (6'-O- alpha -L-arabinofuranosyl- beta -D-glucopyranosyl) acetone -4-O- (6'-O- alpha -L-arabinofuranosyl- [beta] -D-glucopyranosyl) acetophenone)

Di-O-methyl-4-O- (3,3'-di-O- (3 "-O-galloyl- beta -D-glucopyranosyl) 3 " -O-galloyl- [beta] -D-glucopyranosyl) ellagic acid)

In the composition for protecting liver in one aspect of the present disclosure, the compounds do peoh California cyano (Peonia anomala extract.

There are about 35 species of plants in the genus Paeonia, which are known to be distributed in the Eurasia region and are used as important traditional pharmacy plants in Asia. In China and Korea, Paeonia lactiflora was mainly used as a traditional medicinal plant. Paeonia anomala is a traditional medicinal plant of Mongolia. Its roots are used for the treatment of abdominal pain, prevention of blood clotting, dyspepsia, nocturnal enuresis, exhaustion, airway abnormalities, etc., and Paonia anomala Fruit is known to be used for the treatment of kidney disease or gynecological disease.

As used herein, the term " Peonia " anomala ) may be contained in the form of an extract, or may be contained as a pulverized product of the herbal medicine itself, or as a dried pulverized product of the herbal medicine, but the present invention is not limited thereto. The Paeonia anomala used in the present invention is not limited to the method of obtaining it, and may be cultivated or used by purchasing a commercially available plant, or a part or whole root of the above-ground part of the herb may be used.

In the liver protecting composition which is one aspect of the present invention, the Paeonia anomala extract may contain a Paenia anomer-C ₁ -C ₆ alcohol extract, and specifically, a Paenia anomalla-methanol extract or a Paeniaanola May be a mala-ethanol extract.

The Paeonia anomerale extract herein may be a crude extract of a solvent selected from the group consisting of water, C ₁ -C ₆ alcohols, and combinations thereof. The C ₁ -C ₆ alcohol may specifically be methanol or ethanol. When extracting Paeonia anomala with a solvent, it is preferable to extract by adding a solvent corresponding to about 5 to 15 times of Paeonia anomala. Specifically, it is preferable to extract by adding about 10 times solvent, It is not. The extraction may be performed by heat extraction, cold extraction, reflux cooling extraction, ultrasonic extraction or the like, and there is no limitation as long as it is an extraction method that is obvious to a person skilled in the art. The extraction may be carried out at room temperature, but it may be carried out under heating at a temperature of about 40 to 100 ° C, more preferably about 80 ° C, for a more efficient extraction, It is not. The extraction time is preferably about 2 to 4 hours, more preferably about 3 hours, but it is not limited thereto and may be varied depending on conditions such as extraction solvent and extraction temperature. The extraction may be carried out one or more times several times to obtain a larger amount of the active ingredient, preferably 1 to 5 times, more preferably 3 times of continuous extraction.

In this specification, the Paenia anomala extract may contain crude extracts of Paeonia anomalas as described above, and may be included as a soluble fraction of the organic solvent obtained by further extracting the crude extract with an organic solvent having a low polarity. Examples of the organic solvent include, but are not limited to, hexane, methylene chloride, ethyl acetate, n-butanol, and the like. The extract or the soluble fraction of the extract may be used as it is, or may be used as an extract form by concentration after filtration, and may be used as a form of a lyophilizate by concentration and freeze-drying.

In one aspect of the present invention, the composition comprises at least one compound selected from the group consisting of Formulas (1) and (2) or pharmaceutically acceptable salts thereof in an amount of 0.001 to 99.9% by weight based on the total weight of the composition do. If the compound or its salt is contained in an amount of less than 0.001% by weight based on the total weight of the composition, it is not sufficient to protect liver cells. If it exceeds 99.9% by weight, it affects other components and is not suitable. In view of the above, the composition may contain 0.005 to 95% by weight, 0.01 to 90% by weight, 0.05 to 95% by weight, at least one compound selected from the group consisting of formulas (1) From 5 to 65% by weight, from 10 to 60% by weight, from 15 to 55% by weight, from 20 to 50% by weight, from 25 to 45% by weight, from 0 to 85% by weight, from 0.1 to 80% by weight, from 0.5 to 75% By weight or 30 to 40% by weight.

In the liver protecting composition which is one aspect of the present specification, The composition comprises a pharmaceutical composition.

In one embodiment, the pharmaceutical composition may be a pharmaceutical composition effective for liver disease, and specifically a pharmaceutical composition effective for oxidative stress-induced liver disease.

When the composition according to the present invention is applied to medicines, it may be formulated into an oral or parenteral dosage form in the form of solid, semi-solid or liquid by adding an inorganic or organic carrier to the composition as an active ingredient.

Examples of the agent for oral administration include tablets, pills, granules, capsules, powders, fine granules, powders, emulsions, syrups and pellets. Examples of the parenteral administration agent include injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, and the like. In order to formulate the active ingredient of the present invention, it can be easily formulated according to the conventional method. Surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffering agents, suspending agents and other adjuvants can be suitably used.

The pharmaceutical composition according to the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously and the like.

In addition, the dosage of the active ingredient will vary depending on the age, sex, and weight of the subject to be treated, the specific disease or condition to be treated, the severity of the disease or condition, the route of administration, and the judgment of the prescriber. Dosage determinations based on these factors are within the level of those skilled in the art. Typical doses are from 0.001 mg / day to 2000 mg / kg / day, more specifically from 0.5 mg / kg / day to 1500 mg / kg / day.

In a composition for liver protection that is one aspect of the present disclosure, the composition comprises a health food composition.

The composition may be processed into a drink, fermented milk, cheese, yogurt, juice, probiotic agent, and health supplement food or the like, and may be used in various other food additives.

In one embodiment, the composition may contain other ingredients or the like that can give synergistic effects to the main effect within a range that does not impair the intended main effect of the present invention. For example, additives such as perfume, coloring agent, bactericide, antioxidant, preservative, moisturizing agent, thickening agent, inorganic salt, emulsifier and synthetic polymer substance may be further added for improvement of physical properties. In addition, it may further contain auxiliary components such as water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides and seaweed extract. The above components may be mixed and selected without difficulty by those skilled in the art depending on the purpose of formulation or use, and the amount thereof may be selected within a range that does not impair the objects and effects of the present invention.

The composition according to the present invention may be in various forms such as a solution, an emulsion, a viscous mixture, a tablet, a powder, etc., and may be administered by various methods such as simple drinking, injection administration, spraying or squeezing.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Isolation and Identification of Compounds

Paeonia anomala , Mongolia) were collected in Tov province, Mongolia. 3.8 kg of dried fruit of Paeonia anomala and 1.6 kg of root were extracted with 3 L of ethanol for 3 days and then distilled under reduced pressure to obtain 57 g and 26 g of extracts, respectively. The fruit extract was diluted by adding 0.9 L of water and then diluted with hexane (n-hexane, 0.9 LX 3 times), methylene chloride (CH ₂ Cl ₂ , 0.9 LX 3 times), ethyl acetate (EtOAc, 0.9 LX 3 times), butanol (BuOH, 0.9 LX 3 times).

8.3 g of the butanol fraction was fractionated into four fractions of B1-B4 using ODS reverse phase chromatography (4.5 × 50 cm, flow rate 2.5 mL / min, water-methanol gradient 80:20 → 0: 100 each 1 L). B2 fraction (2.7 g) was separated by HPLC on a Hydrosphere C18 column (YMC HPLC Column, flow rate: 1 ml / min) using 2.5 L of methanol on Sephadex LH-20 column chromatography 250 × 20 mm, S-5μm , water-acetonitrile gradient _{80:20 → 60:40, t R = 14} min) using a 2-hydroxy-6-methoxy -4-O- (6'- O-alpha-L-arabinofuranosyl- beta -D-glucopyranosyl-beta-D-glucopyranosyl) acetone (2-hydroxy-6-methoxy- ) acetophenone).

The ethyl acetate fraction, 35 g, was further fractionated sequentially through E1-E12 through silica gel chromatography (230-400 [mu] m, 6.5 x 75 cm, methylene chloride-methanol gradient 250: 1 - > 0: 100 each 1 L). 26.1 g of the E12 fraction was diluted with E12-1-E12 (1 ml) through ODS reverse phase chromatography (12 nm, S-150, 6.5 x 60, flow rate 3.5 ml / min, water- methanol gradient 70:30 → 0: -13. ≪ / RTI > 2.3 g of E12-11 fraction was separated by HPLC on a Hydrosphere C18 column (HPLC column: YMC HPLC Column) using 2.5 L of methanol, Sephadex LH-20 chromatography (18-111 μm, 3 × 40 cm, flow rate 1 mL / min) Di-O- (3 "-O-Galoyl) was synthesized by using the following method: 1) 250 x 20 mm, S-5 m, water-acetonitrile gradient 80:20 -> 50:50, t _R = 22 min) -β-D-glucopyranosyl) ellagic acid (3,3'-di-O-methyl-4-O- .

MS, IR, UV-Visible, ¹ H-NMR, and ¹³ C-NMR spectra of the separated compounds were examined to confirm the structure of the compound. The characteristics of each compound are as follows.

[Chemical Formula 3] 2- Hydroxy -6- Methoxy -4-O- (6 ' -O-a-L- Arabinofuranosyl -β-D-glucopyranosyl) acetone

Colorless amorphous powder;

[?] ²⁰ _D -60 ( c 0.034, methanol);

UV (methanol)? _Max (log?) 209 (4.14), 225 (4.08), 283 (4.13) nm;

IR (ATR) v _max 3439, 2924, 2854, 2360, 2342, 1958, 1737, 1710, 1691, 1678, 1620, 1461, 1383, 1129 cm ^-1 ;

¹ H-NMR (DMSO, 500 MHz) See Table 1;

¹³ C-NMR (DMSO, 125 MHz) See Table 1;

ESI-MS ( m / z 477.5 [M + H] ⁺ , (negative ion mode) 475.2 [MH] ^- ; HRESIMS m / z 477.4393 [M + H] ⁺ (calculated molecular formula, C ₂₀ H ₂₉ O ₁₃ , 477.4425)

3,3'-di-O- (3 "-O- Galois -D-D- Glucopyranosyl ) Ella  mountain

White amorphous powder;

UV (methanol)? _Max (log?) 206 (4.29), 215 (4.28), 246 (4.35), 369 (3.73) nm;

IR (ATR) v _max 3343, 2920, 2850, 1669, 1603, 1438, 1356, 1204, 1131, 1068, 912, 843, 799, 720 cm ^-1 ;

¹ H-NMR (MeOD, 500 MHz) See Table 2;

¹³ C-NMR (MeOD, 125 MHz) See Table 1;

ESI-MS (negative ion mode) m / z 643.19 [MH] ^- ; HRESIMS m / z 643.4889 [MH] ^- (calculated molecular formula, C ₂₉ H ₂₃ O ₁₇ , 643.4914).

[Table 1] Synthesis of Compound (3) and Compound (4) ^One H- NMR ¹³ C- NMR  data

[ Experimental Example  1] Damaged liver protection

HepG2 (American Type of Cell Culture, ATCC, USA), a human hepatocellular carcinoma cell line, was used to examine the hepatoprotective effect of a compound that is an active ingredient of the composition of the present invention. HepG2 cells were cultured in 96-well plates at 10,000 cells / ml in DMEM medium containing 10% fetal bovine serum (FBS) and antibiotics (penicillin and streptomycin) at 37 ° C under 5% well, and cultured for 24 hours. (2.5, 5, 10, 20, 40 ㎍ / ml) and Paeonia anomalaf root extract (2.5, 5, 10, 20, 40 ㎍ / ml) were added to the DMEM medium containing the FBS (2.5, 5, 10, 20 占 퐂 / ml) and the single compound of the present invention (Formula 3, Formula 4) Respectively. EGCG (2.5, 5, 10, 20 μM) and quercetin (10 μM) were used as positive control. Then, the medium was washed with DBS (Dibbecco's phosphate buffered saline), and DMEM medium containing FBS was added to 250 μM tertiary butyl hydroperoxide. After incubation for 3 hours, Causing death. The culture was terminated and hepatocyte survival rate was measured with a CCK-8 assay.

As a result, it was found that the Paenia anomerale extract (Fig. 1), Paeonia anomerale fraction (Fig. 2) and the compound contained in the composition of the present invention (Fig. 3) It has been confirmed that it has protective ability against liver damage.

[ Experimental Example  2] in hepatocytes Active oxygen species  Content measurement

HepG2 cells were seeded in a 96-well plate at a concentration of 10,000 cells / well and cultured for 24 hours. Then, the FBS-removed DMEM medium was pre-treated for 24 hours with 5, 10, 20, and 40 μg / ml of P. anomala fruit and root extract, respectively. At this time, EGCG (10 μM) and quercetin (10 μM) treatment groups were also set as positive control groups. 10 μL of 2 ', 7'-dichlorodihydrofluorescein diacetate (H ₂ -DCFDA, 10 μM) solution was added and incubated for 30 minutes. After washing twice with DPBS (Dulbecco's phosphate buffered saline), DMEM medium containing 2.5% FBS was treated with 250 μM tert-butyl hydroperoxide and incubated for 30 minutes. Fluorescence (excitation wavelength = 485 nm, emission wavelength = 530 nm) of 2 ', 7'-dichlorodihydrofluorescein (DCF) was measured.

As a result (FIG. 4), it was confirmed that the concentration of active oxygen species in the hepatocytes increased by tert-butylhydroperoxide was significantly reduced by Paenonia anomalla extract.

[ Experimental Example  3] Hepatocyte Total glutathione   ( total glutathione ) Content measurement

HepG2 cells were seeded in a 96-well plate at a concentration of 10,000 cells / well and cultured for 24 hours. After that, the cells were pretreated with DMEM medium containing Fiona anomalafil or root extracts at 5, 10, 20 and 40 μg / ml, respectively, for 24 hours. At this time, EGCG (10 μM) and quercetin (10 μM) treatment groups were also set as positive control groups. Then, the cells were washed twice with DPBS (Dulbecco's phosphate buffered saline) and then treated with 250 μM of tert-butyl hydroperoxide for 3 hours. The cells were then lysed with lysis buffer (143 mM sodium phosphate buffer, 0.1% Triton X-100, 8 mM EDTA) and protein concentration determined by the Bradford method. 5% sulfosalicylic acid was added and the mixture was allowed to stand for 5 minutes to precipitate the protein, and then the total glutathione content was measured. The 25 μL supernatant was mixed with 120 μL of a reaction solution (68 mM 5,5'-dithiobis- (2-nitrobenzoic acid), 3.33 U / mL glutathione reductase) and then 60 μL of β-NADPH mu] M solution was added to initiate the reaction. The total glutathione content was then calculated by measuring the absorbance (450 nm).

As a result (FIG. 5), it was confirmed that the total glutathione concentration of hepatic cells in which tert-butyl hydroperoxide (t-BHP) was reduced significantly recovered or increased by Paenonia anomalla extract.

[ Experimental Example  4] Hepatocyte DNA  For damage Liver protection  effect

Comet assay was performed to verify the DNA damage protection effect. An experimental kit from Trevigen (Gaithersburg, MD, USA) was used for this experiment. HepG2 cells were seeded in a 6-well plate at a concentration of 300,000 cells / well and cultured for 24 hours. Then, 20 μg / ml of each of P. anomala fruit and root extract was added to the DMEM medium in which FBS was removed, and the mixture was pretreated for 24 hours. At this time, EGCG (10 μM) treatment group was also set as a positive control group. Then, the cells were washed with DPBS (Dulbecco's phosphate buffered saline), treated with 250 μM of tert-butyl hydroperoxide, incubated for 1 hour, harvested and suspended in ice cold cold DPBS. Subsequently, 6000 cells were mixed with 100 μL of low-melting agarose, placed on a slide glass, and then solidified for 45 minutes at 4 ° C. in a dark state. Slides were placed in ice-cooled Lysis buffer and lysed in darkness at 4 ° C for 40 min, then in alkaline solution (pH> 13) and incubated at room temperature for 40 min. Then, it was electrophoresed in a low temperature room at an intensity of 1 V / cm under alkaline conditions. The slides were washed once with distilled water twice with 70% ethanol, stained with propidium solution (2.5 μg / mL aqueous solution), and then Comet images were taken using a Nikon TE2000U fluorescence microscope.

As a result of the experiment (FIG. 6), the tertiary butyl hydroperoxide significantly induced DNA damage in the hepatocytes, resulting in a thick, coarse and long Comet tail. On the other hand, when the Paeonia anomerale extract was pretreated, the Comet tail was significantly weaker than that of the tert-butyl hydroperoxide alone, indicating that it had a protective effect against DNA damage.

[ Experimental Example  6] Using animal experiments Paonia Anomalla  Alcoholicity of the extract To liver damage  Protection against

Five-week-old rats (Spargue-Dawley rats) were free-fed for one week and then fed with a liquid diet containing alcohol or Feonia anomerale extract for 21 days. Serum ALT (alanine aminotransferase) and AST (aspartate aminotransferase) activities were measured using an enzyme coloring kit (Youngdong Pharma, Yongin, Korea) to compare liver damage. Blood was centrifuged at 2,580 xg for 10 min and measured using supernatant alone.

① Group 1: Control group, liquid diet only ingested

② Group 2: Ethanol intake group

③ Group 3: 50 mg / kg / day of ethanol + Feonia anomalafum extract

The results (Figs. 7 and 8) show that the levels of alanine alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in Group 3 were lower than that of Group 2.

Formulation examples of the composition are described below, but are not intended to limit the invention, but merely to illustrate the invention.

[ Manufacturing example  One] Powder

At least one compound selected from the group consisting of formulas (1) and (2): 2 g

Lactose 1 g

After mixing the above components, the mixture was packed in an airtight container to prepare a powder.

[ Manufacturing example  2] Refining

100 mg of at least one compound selected from the group consisting of formulas (1) and (2)

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

[ Manufacturing example  3] Capsule

100 mg of at least one compound selected from the group consisting of formulas (1) and (2)

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

[ Manufacturing example  4] ring

1 g < / RTI > of at least one compound selected from the group consisting of formulas (1) and (2)

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

[ Manufacturing example  5] granule

150 mg of at least one compound selected from the group consisting of formulas (1) and (2)

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

[ Manufacturing example  6] Health food

1000 mg of at least one compound selected from the group consisting of formulas (1) and (2)

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

0.13 mg of vitamin

0.15 mg of vitamin B ₂

0.5 mg of vitamin B ₆

Vitamin B ₁₂ 0.2 g

10 mg vitamin C

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

24.8 mg of magnesium chloride

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

[ Manufacturing example  7] Health drinks

1000 mg of at least one compound selected from the group consisting of formulas (1) and (2)

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered and sterilized in a 2-liter sterile container. The resulting solution was refrigerated, And used in the preparation of the composition.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

[ Manufacturing example  8] Beverages

Honey 522 mg

5 mg < RTI ID = 0.0 >

Nicotinic acid amide 10 mg

3 mg of sodium riboflavin hydrochloride

Pyridoxine hydrochloride 2 mg

Inositol 30 mg

Orthoic acid 50 mg

8 mg of at least one compound selected from the group consisting of formulas (1) and (2)

200 ml of water

A beverage was prepared using the above-mentioned composition and content by a conventional method.

Those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

And at least one compound selected from the group consisting of the following formulas (3) and (4) or a pharmaceutically acceptable salt thereof as an active ingredient.
2-Hydroxy-6-methoxy-4-O- (6'-O- alpha -L-arabinofuranosyl- beta -D-glucopyranosyl) acetone -4-O- (6'-O- alpha -L-arabinofuranosyl- [beta] -D-glucopyranosyl) acetophenone)

Di-O-methyl-4-O- (3,3'-di-O- (3 "-O-galloyl- beta -D-glucopyranosyl) 3 " -O-galloyl- [beta] -D-glucopyranosyl) ellagic acid)

A health food composition for preventing or ameliorating liver disease comprising, as an active ingredient, at least one compound selected from the group consisting of the following formulas (3) and (4) or a pharmaceutically acceptable salt thereof.
2-Hydroxy-6-methoxy-4-O- (6'-O- alpha -L-arabinofuranosyl- beta -D-glucopyranosyl) acetone -4-O- (6'-O- alpha -L-arabinofuranosyl- [beta] -D-glucopyranosyl) acetophenone)

Di-O-methyl-4-O- (3,3'-di-O- (3 "-O-galloyl- beta -D-glucopyranosyl) 3 " -O-galloyl- [beta] -D-glucopyranosyl) ellagic acid)

3. The method according to claim 1 or 2,
Wherein said compounds are separated from Peonia anomala extract.
delete 3. The method according to claim 1 or 2,
Wherein the liver disease is an alcoholic liver damage disorder.
delete delete

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