KR102229547B1 - A composition for improving, preventing and treating of liver diseases comprising Milk thistle and Taraxacum platycarpum Dahlst - Google Patents

Abstract

The present invention relates to a composition for the prevention, treatment or improvement of liver disease, by containing a mixture consisting of thistle and dandelion as an active ingredient, related to drug addiction, non-alcoholic fatty liver, liver fibrosis, cirrhosis, acute liver injury and acute hepatitis. It has excellent effect on various diseases, and because it is not toxic, it can be consumed in the form of food, so it can improve, prevent or treat liver disease through diet and lifestyle of daily life in the long term.

Description

A composition for improving, preventing and treating of liver diseases comprising Milk thistle and Taraxacum platycarpum Dahlst}

The present invention relates to a composition for preventing, treating, or improving liver disease containing a mixture consisting of thistle and dandelion as an active ingredient.

Clinically, the most important step in the patient's course in the various patterns and progression of liver disease is to suppress the occurrence of liver fibrosis or cirrhosis, or to suppress the progression of liver cirrhosis if it has already begun. . Therefore, numerous researchers studying the treatment of liver disease have been making efforts to develop a preventive or therapeutic agent for liver cirrhosis.

Non-alcholic steatohepatitis (NASH) is a chronic disease that causes persistent hepatitis, is difficult to diagnose early, and about 20% of patients develop liver fibrosis or cirrhosis after 20 years. , There is a difference in pattern pattern from conventional liver disease, and the prevalence is increasing due to various pathological causes.

When hepatocytes are damaged by various causes and the number of hepatocytes that make up hepatocytes decreases, hepatic stellate cells that store vitamin A are transformed into fibroblasts that make fibers. ), the amount of fiber in the liver tissue increases. In addition, excessive generation of reactive oxygen species (ROS) and a lack of defensive mechanisms such as the glutathione system, superoxide dismutase (SOD), and catalase are caused by liver fibrosis. It is the cause of causing.

Republic of Korea Patent No. 0626724 is the suppression of liver fibrosis extracted from mixed herbal medicines including Injin, Changwon, Hubak, Dermis, Jeoryeong, Taeksa, Baekchul, Bokryeong, Nabokja, Ginger, Banha, Daebokpi, Samneung, Bongchul, Cheongpi and Licorice. It discloses an extract and a pharmaceutical composition containing the same, and Korean Patent No. 00604354 discloses a herbal composition for preventing and improving liver disease containing extracts of Injin, Baekchul, Unji, Wasong and Jeoryeong as active ingredients. In addition, Republic of Korea Patent No. 0682045 has 3 to 5 parts by weight of Wimo, 3 to 5 parts by weight of Moryeom, 3 to 5 parts by weight of Baekryum, 3 to 5 parts by weight of manufacturing, 1 to 3 parts by weight of Injin, 1 to 3 parts by weight of Taeksa. Part, island water 1 to 3 parts by weight, sagan 1 to 3 parts by weight, single axis 0.5 to 1.5 parts by weight, Nabokja 0.5 to 1.5 parts by weight, Turmeric 0.5 to 1.5 parts by weight, Horse riding 0.5 to 1.5 parts by weight, Baekbokryeong 0.5 to 1.5 parts by weight , Loquat 0.5 to 1.5 parts by weight, licorice 0.5 to 1.5 parts by weight, 0.1 to 1 parts by weight of virility and 0.1 to 1 parts by weight of high alum and 17 kinds of complex herbal medicine powders extracted with a solvent to prevent and improve liver fibrosis Disclosing an extract. In addition, Korean Patent Registration No.0867320 describes 5 parts by weight of tortoiseshell, 5 parts by weight of Nabokja, 5 parts by weight of Injin, 3 parts by weight of Baekchul, 3 parts by weight of Baekbokryeong, 3 parts by weight of Taeksa, 3 parts by weight of Jinseong, 3 parts by weight of Dansam, 2 parts by weight, 2 parts by weight of cod, 2 parts by weight of fat, 1 part by weight of licorice and 1 part by weight of muk-hyang are prepared by water extraction of 13 kinds of complex herbal medicines for liver protection, liver fibrosis and liver cirrhosis prevention or improvement I'm doing it.

However, the composition as described above is not easy to handle and manufacture because a variety of herbal medicines are used together, and there is a problem in that the content of ingredients effective for liver diseases such as non-alcoholic fatty liver, liver protection, liver fibrosis and cirrhosis is low. Therefore, research and development of herbal extracts having a high content of ingredients that are particularly effective in liver disease are still required.

Korean Patent Registration No. 0626724 Republic of Korea Patent Registration No. 00604354 Korean Patent Registration No. 1854606

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of liver disease containing a mixture consisting of thistle and dandelion as an active ingredient.

In addition, another object of the present invention is to provide a food composition for preventing or improving liver disease containing a mixture consisting of thistle and dandelion as an active ingredient.

The pharmaceutical composition capable of preventing or treating liver disease of the present invention for achieving the above object may contain a mixture consisting of thistle and dandelion as an active ingredient.

The thistle and dandelion may be mixed in a weight ratio of 1: 0.05 to 0.5.

The mixture of thistle and dandelion may be an extract extracted by boiling water at 80 to 100°C.

The mixture of thistle and dandelion may be a dough obtained by mixing and kneading a pulverized product obtained by mixing each of the above substances with water.

The liver disease may be selected from drug addiction, non-alcoholic fatty liver, liver fibrosis, cirrhosis, acute liver injury and acute hepatitis.

The drug addiction may be caused by carbon tetrachloride.

The thistle may be a mixed extract in which a thistle flower, a mixture of stems and leaves, and roots are mixed in a weight ratio of 1:14-25:0.1-0.9.

In the mixture of stems and leaves, stems and leaves may be mixed in a weight ratio of 1: 1-4.

The mixed extract of thistle may be a mixture of an extract obtained by mixing the thistle flower, and a mixture of stems and leaves, and extracting the extract in the middle bath at 80 to 100°C, and the extract obtained by extracting the root in the bath at 80 to 100°C.

The powder obtained by drying and pulverizing the remaining residue after extracting the roots at 80 to 100° C. to the mixed thistle extract may be added.

In addition, the food composition capable of preventing or improving liver disease of the present invention for achieving the other object described above may contain a mixture consisting of thistle and dandelion as an active ingredient.

The composition for improving, preventing or treating liver disease of the present invention can be effectively used for the prevention and treatment of liver disease because it alleviates diseases selected from drug addiction, non-alcoholic fatty liver, liver fibrosis, cirrhosis, acute liver injury and acute hepatitis. It is not toxic, so it can be consumed in the form of food.

The present invention relates to a composition for preventing, treating, or improving liver disease containing a mixture consisting of thistle and dandelion as an active ingredient.

Hereinafter, the present invention will be described in detail.

The composition for preventing, treating or improving liver disease of the present invention contains a mixture consisting of thistle and dandelion as an active ingredient.

Thistle

The thistle used in the present invention may be white patterned thistle (Silybum marianum (L.) Gaertn.), spiny thistle (Cirsium japonicum Fisch ex DC. var. spinossimum Kitam.), or a mixture thereof, preferably white patterned thistle. It is to use thistle alone.

The white-patterned thistle (Silybum marianum (L.) Gaertn.) is a deep, shiny green plant with milky white veins, and is a long, elongated plant divided into root-leaves with wavy patterns and thorns at the edges. It is mainly found in levee fences and in abandoned land. Useful parts of plants are whole herbs, roots, leaves, seeds, and shells, and thistle seeds contain a flavonoid compound known as silymarin. Thistle seed extract contains 70-80% silymarin.

Thistle according to the present invention is a mixed extract of a thistle flower, a mixture of stems and leaves, and a mixture of roots.

The flowers of thistle are purple or red corolla and 19-24 mm in length, and it is preferable to use only those having a length of 10 to 15 mm and not use those with a length longer than that.

In addition, the thistle stem and leaves are used by mixing in a weight ratio of 1: 1-4, preferably in a weight ratio of 1: 2-3. If the content of the leaves based on the stem is less than the lower limit, the bitter taste may be strong, and if the content of the leaves is greater than the upper limit, it is not preferable for the prevention, treatment or improvement of liver disease.

In addition, the thistle root is harvested once every three years, unlike flowers, stems, and leaves that are harvested twice a year, so it is preferable to use it after drying.

The flower of thistle, a mixture of stems and leaves, and roots are mixed in a weight ratio of 1: 14-25: 0.1-0.9, preferably 1: 16-19: 0.4-0.8. If the content of stem and leaf mixture and root based on flowers is less than the lower limit, the effect on preventing, treating, or improving liver disease may not be excellent, and if it exceeds the upper limit, the bitter taste is strong and The effectiveness for prevention, treatment or improvement may be diminished.

The mixed extract of the present invention is an extract extracted in a bath for 20 to 30 hours, preferably 24 to 28 hours at 80 to 100 ℃. If the boiling water extraction temperature and time are less than the lower limit, the active ingredient may not be extracted, and if it exceeds the upper limit, the tan taste may be strong and the nutrient component may be destroyed. In addition, when the extraction is performed in other ways, such as extraction of an organic solvent in addition to the extraction of a water bath, the effect on preventing, treating, or improving liver disease may be insignificant and a strong bitter taste may occur.

Specifically, the mixed extract is mixed with a flower, and a mixture of stems and leaves to extract the middle bath, and the roots are separately extracted with the bath and mixed later. Unlike the present invention, in the case of extracting the water by mixing flowers, a mixture of stems and leaves, and roots, the bitter taste is strong and the effect on preventing, treating, or improving liver disease may be lowered.

The method for preparing such a mixed thistle extract specifically includes the steps of (A) mixing a mixture of flowers of thistle, and a mixture of stems and leaves, and extracting the extract by boiling water at 80 to 100° C. to obtain an extract; (B) extracting the roots of thistle in a bath at 80 to 100° C. to obtain an extract; And (C) lyophilizing the extracts obtained in steps (A) and (B), respectively, to powder and mix them. In addition, after the step (C), (D) drying and pulverizing the residue extracted in the step (B); And (E) mixing 100 parts by weight of the powder mixed in step (C) with 0.1 to 3 parts by weight of the powder in step (D).

Unlike the above steps (A) and (B), when the bath is extracted by mixing flowers, a mixture of stems and leaves, and roots, the bitter taste is strong and the effect on preventing, treating or improving liver disease may be lowered. There is; When the flowers, the mixture of stems and leaves, and the roots are all extracted in a bath, the yield may decrease.

When the root residue powder prepared in step (D) is added, the effect on diseases can be significantly improved compared to the case where the root residue powder is not added.

dandelion

The dandelion (Taraxacum platycarpum Dahlst.; Taraxacum mongolicum Hand.-Mazz) is also referred to as Geumjamcho, Jeongji, Pogongyeong, Pogongcho, Anjilbang, and also has the nickname of Sitbang. Leaves or stems evenly contain proteins, carbohydrates, fats, ash, minerals, and vitamins. Young shoots are used as herbs or soup. In oriental medicine, the roots and outposts before flowering are said to be effective in antipyretic, anti-inflammatory, diuretic, and dry stomach, and are used as a treatment for hypothyroidism, sore throat, bronchitis, lymphadenitis, eye disease, mastitis, hepatitis, cholecystitis, indigestion, dyspepsia, and constipation. Use.

These thistles and dandelions are mixed in a weight ratio of 1:0.05 to 0.5, preferably 1:0.1 to 0.4.

If the content of dandelion based on thistle is less than the above lower limit, the effect of preventing, treating, or improving liver disease may be lower than that of using each substance alone, and if it exceeds the upper limit, preventing, treating or improving liver disease The effect of improving may be reduced.

When thistle and dandelion are used alone, their efficacy is 1.5 to 20 times lower than when thistle and dandelion are used together.

The form of the composition of the present invention may be prepared in the form of granules, pills, powders, juices, tea bags, and the like.

For example, in the case of producing granules or pills, the thistle extract is lyophilized and pulverized dandelion powder is mixed with water to prepare a dough, and the prepared dough is used to form a granule or a pill.

As another example, in the case of preparing granules, powders, pills, and juice, the powder obtained by mixing the thistle and dandelion in the proportions of the present invention, filtering the extract extracted from the boiling water at 80 to 100°C, lyophilizing it, and then powdering the obtained powder Can be molded to obtain granules or pills.

In another example, in the case of granules, powders, pills, and blue, the thistle and dandelion are mixed in the proportions of the present invention, and the extract extracted in a bath for 20 to 30 hours at 80 to 100°C, and the thistle and dandelion in the proportion of the present invention The mixed and pulverized powder is mixed in a weight ratio of 4-5:1, and then the mixture is extracted with a bath for 40 to 55 hours at 60 to 70°C and concentrated to obtain blue. Further, the concentrated concentrate may be dried to obtain a powder, and the concentrate may be partially dried to form granules or rings, and then dried again to obtain complete granules or rings.

When this method is used, since raw material powder is used, it is not necessary to use substances such as excipients, and an active ingredient can be obtained in a larger amount.

As another example, in the case of manufacturing a tea bag, the powder obtained by lyophilizing the extract of thistle and dandelion is roasted at 80 to 120° C. in a tea bag.

The mixture consisting of thistle and dandelion of the present invention is broadly meant to include a mixed product of thistle and dandelion, such as a mixed powder of thistle and dandelion, formulated to administer a mixture of thistle and dandelion to animals. . Although the experiment was conducted with a mixture of thistle and dandelion in the present invention, it will be expected by those skilled in the art that the desired effect can be achieved even in the same form as a mixed product of thistle and dandelion.

Meanwhile, in the present specification, the term "containing as an active ingredient" means including an amount sufficient to achieve the efficacy or activity of thistle and dandelion. For example, the thistle and dandelion are used in a concentration of 10 to 1500 µg/ml, preferably 100 to 1000 µg/ml. Since thistle and dandelion are natural products and do not have side effects on the human body even if they are administered in an excessive amount, the upper limit of quantity of thistle and dandelion included in the composition of the present invention can be selected and carried out by a person skilled in the art within an appropriate range.

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, Lubricating agents or flavoring agents can be used.

For administration, the pharmaceutical composition may contain one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

Acceptable pharmaceutical carriers for compositions formulated as liquid solutions are sterilized and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injection formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Further, it can be preferably formulated according to each disease or ingredient using a method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration. .

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, Usually the skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person skilled in the art. Or it can be prepared by incorporating it into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

In addition, the present invention provides a food composition for preventing, treating or improving liver disease containing thistle and dandelion as active ingredients.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice creams, vitamin complexes, health supplements. Etc.

The food composition of the present invention may include not only thistle and dandelion as active ingredients, but also ingredients commonly added during food production, and, for example, include proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. . Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, various plant extracts, etc. may be additionally included in addition to thistle and hyssles of the present invention. .

The present invention provides a health functional food comprising a food composition for preventing, treating or improving liver disease comprising the thistle and dandelion as active ingredients. Health functional foods are foods prepared by adding thistle and dandelion to food ingredients such as beverages, teas, spices, chewing gum, confectionery, etc., or by encapsulation, powdering, or suspension. However, unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of thistle and wheatgrass added in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but can be added within the range that does not damage the original taste of the food. It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, usually 0.1 to 100% by weight, preferably 0.5 to 80% by weight, may be added. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, the present invention provides the use of thistle and dandelion for the manufacture of a drug or food for the prevention, treatment or improvement of liver disease. As described above, thistle and dandelion can be used for liver protection.

In addition, the present invention provides a method for improving, preventing or treating liver disease comprising administering an effective amount of thistle and dandelion to a mammal.

The term "mammal" as used herein refers to a mammal that is an object of treatment, observation or experiment, and preferably refers to a human.

The term "effective amount" as used herein refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, which Includes an amount that induces relief of symptoms of the disease or disorder. It is apparent to those skilled in the art that the effective amount and the number of administrations for the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. It can be adjusted according to various factors including condition, sex and diet, time of administration, route of administration and rate of secretion of the composition, duration of treatment, and drugs used concurrently. In the prevention, treatment or improvement method of the present invention, in the case of adults, it is preferable to administer thistle and dandelion at a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.

In the treatment method of the present invention, the composition comprising thistle and dandelion as active ingredients is a conventional method through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered as.

Hereinafter, a preferred embodiment is presented to aid in the understanding of the present invention, but it is obvious to those skilled in the art that various changes and modifications are possible within the scope of the present invention and the scope of the technical idea, but the following examples are only illustrative of the present invention, It is natural that such modifications and modifications fall within the scope of the appended claims.

Example 1. Jungtang extract

Manufacture of thistle

Thistle flower 1 part by weight, a mixture of stem and leaf (stem: leaf = 1:2 weight ratio) 18.6 parts by weight and 45 parts by weight of water were mixed and extracted with water at 95° C. for 24 hours to obtain extract A, and then thistle root 0.4 Part by weight and 45 parts by weight of water were extracted in a bath at 95° C. for 24 hours to obtain extract B, and then freeze-dried to prepare and mix powder A and powder B to prepare a mixed extract powder.

Jungtang extract

After mixing 100 parts by weight of thistle, 40 parts by weight of dandelion, and 860 parts by weight of water, the mixture was heated at 90° C. for 24 hours, and then sterilized at 100° C. for 5 minutes by mixing the bathed liquid and the compressed liquid obtained by compressing the suspension. Then, it was filtered to obtain a bath extract.

Example 2. Jungtang Extract + Powder -> Jungtang Extract

In the same manner as in Example 1, but after mixing 100 parts by weight of thistle, 40 parts by weight of dandelion, and 860 parts by weight of water, the liquid was boiled at 90° C. for 24 hours and then boiled; 100 parts by weight of thistle and 40 parts by weight of dandelion were mixed, and then pulverized powder; was mixed in a weight ratio of 4:1, and the mixture was extracted in a water bath at 60° C. for 48 hours to obtain an extract.

Example 3. Addition of thistle_root residue

It was carried out in the same manner as in Example 1, but when the thistle powder was prepared, the thistle root was extracted in a bath, and the remaining residue was dried and pulverized to obtain C powder (root residue powder), and the A powder, B powder, and C powder were prepared. Mixing (0.3 parts by weight of C powder with respect to 100 parts by weight of A powder + B powder) to prepare a mixed extract powder of thistle, and then using this to obtain a water bath extract.

Example 4. Dough

It was carried out in the same manner as in Example 1, but instead of preparing a bowl extract, 100 parts by weight of thistle and 40 parts by weight of dandelion were mixed and pulverized, and then the pulverized product and water were mixed to prepare a dough.

Comparative Example 1. Omission of thistle

It was carried out in the same manner as in Example 1, except that thistle was omitted and only dandelion was used to obtain a bath extract.

Comparative Example 2. Omission of dandelion

It was carried out in the same manner as in Example 1, except that dandelion was omitted and only thistle was used to obtain a Jungtang extract.

<Test Example>

Test Example 1. Hepatoprotective effect on liver toxicity induced by carbon tetrachloride

As experimental animals, 102 males of 6-7 weeks (180-210 g) of SD (Sprague Dawley) rats were used. Before the start of administration, the body weight was measured and divided into 6 animals per group so that the average weight was constant. As a liver toxic substance, carbon tetrachloride was mixed with olive oil in a ratio of 1:3 (v/v) and administered once at a dose of 500 µl/kg of carbon tetrachloride. The test drug was administered orally once a day for 3 consecutive days, and carbon tetrachloride was administered intraperitoneally 3 hours after the last drug administration.

Administration group was control (group 1), carbon tetrachloride administration group (group 2), Example 1 70 mg/kg administration group (group 3), Comparative Example 1 70 mg/kg administration group (group 4), Comparative Example 2 70 mg /Kg administration group (group 5), Comparative Example 3 70 mg/kg administration group (group 6), Comparative Example 4 70 mg/kg administration group (group 7), Comparative Example 5 70 mg/kg administration group (group 8) , Comparative Example 6 It was orally administered to the 70 mg/kg administration group (Group 9).

The experimental animals were killed 24 hours after the last drug administration to obtain serum, and the concentration was measured using Asan Pharmaceutical's GOT and GPT kits. The results are shown in Table 1 below.

division GPT(IU/L) GOT(IU/L) Control 20.9±4.8 52.0±7.9 Carbon tetrachloride administration group 99.5±5.1 134.±26.5 Example 1 62.5±3.5 100.1±3.3 Example 2 55.1±1.8 90.3±2.1 Example 3 59.8±2.9 96.1±2.4 Example 4 65.9±1.5 102.5±4.1 Comparative Example 1 75.4±2.7 110.7±3.9 Comparative Example 2 67.1±4.0 103.8±6.8

As shown in Table 1 above, it was confirmed that the mixed extracts prepared according to Examples 1 to 4 of the present invention exhibit a hepatoprotective effect against liver toxicity induced by carbon tetrachloride (CCl4).

Example 2. Validation of efficacy against non-alcoholic fatty liver

Using an animal model in which non-alcoholic fatty liver was induced by administering Tunicamycin, the efficacy of the extracts according to Examples and Comparative Examples on non-alcoholic fatty liver was verified.

A 6-week-old male C57/BL6 mouse weighing 12-22g was purchased from Daehan Biolink (North Chungcheong Province), and a solid feed at 22±2℃, 55±10% relative humidity, and a 12-hour light and dark cycle was obtained from Daehan Biolink (North Chungcheong Province). Biolink, Chungcheong-do) and tap water were freely ingested. Rats were randomly divided into the following groups with 8-10 rats per group. The control group and the drug-administered group were intraperitoneally injected at a concentration of 1 mg tunicamycin per kg of mouse body weight to induce non-alcoholic fatty liver (NASH) mediated by drug-induced ER stress. Extracts according to Examples and Comparative Examples were orally administered under the following conditions. After the last administration, after fasting for 18 hours, tunicamycin was injected once, and after 30 hours, the animals were sacrificed and an autopsy was performed. The fasting state was maintained for 18 hours before the autopsy was performed.

Fat content (triglyceride, total cholesterol) and inflammatory cytokine concentration were evaluated in liver tissue. At autopsy, serum was separated using blood obtained from abdominal veins to reduce liver transfer enzymes and low-density cholesterol (LDH). The level was measured with an auto chemistry analyzer (Chiron Ltd).

Normal: No treatment group

Negative control group (Control): after NASH induction by tunicamycin administration, distilled water administration group for 7 days

Positive control group (DDB): After NASH induction with tunicamycin administration, dimethyldiphenyl dicarboxylate (DDB) 50 mg/kg administered once a day for 7 days

Example 1: After NASH induction by administration of tunicamycin, the extract of Example 1 once a day for 7 days 50 mg/kg, 100 mg/kg, 200 mg/kg administration group

Example 2: After NASH induction by administration of tunicamycin, the extract of Example 2 once a day for 7 days 50 mg/kg, 100 mg/kg, 200 mg/kg administration group

Example 3: After NASH induction by administration of tunicamycin, the extract of Example 3 once a day for 7 days 50 mg/kg, 100 mg/kg, 200 mg/kg administration group

Comparative Example 1: After induction of NASH by administration of tunicamycin, the extract of Comparative Example 1 was administered once a day for 7 days at 50 mg/kg, 100 mg/kg, and 200 mg/kg

Comparative Example 2: After NASH induction by the administration of tunicamycin, the extract of Comparative Example 2 was administered once a day for 7 days at 50 mg/kg, 100 mg/kg, and 200 mg/kg

Test example 2-1. Fat content evaluation

division Triglyceride (mg/g _tissue ) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 2.0 - - - Negative control 4.8 - - - Positive control - 3.6 - - Example 1 - 3.5 2.8 2.3 Example 2 - 3.0 2.3 1.7 Example 3 - 3.3 2.5 2.0 Example 4 - 3.9 3.0 2.6 Comparative Example 1 - 4.3 3.9 3.4 Comparative Example 2 - 4.1 3.5 3.1

division Total cholesterol (mg/g _tissue ) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 6.3 - - - Negative control 9.2 - - - Positive control - 10.4 - - Example 1 - 9.4 7.1 6.8 Example 2 - 8.6 6.1 6.0 Example 3 - 9.0 6.4 6.4 Example 4 - 9.7 7.9 8.9 Comparative Example 1 - 10.5 8.5 9.4 Comparative Example 2 - 9.9 8.2 9.3

As shown in Tables 2 and 3 above, the extracts of Examples 1 to 4 according to the present invention are triglycerides, which are fat components that cause arteriosclerosis in liver tissues of non-alcoholic fatty liver animal models compared to Comparative Examples 1 and 2 ( Triglycerides) and cholesterol. _{In particular, when treated at a concentration of 200 mg/kg (mouse weight)} in Examples 1 to 3, the triglyceride was reduced to a level similar to that of the normal group, and the total amount of cholesterol was 100 mg/kg and 200 mg/kg at the time of treatment. It showed a level similar to that of the normal group.

On the other hand, the positive control group injected with dimethyl diphenyl dicarboxylate (DDB), which is a representative hepatitis treatment agent, showed a similar level of effect to the extracts of Examples 1 to 3 according to the present invention in reducing triglycerides in liver tissue, but cholesterol It was found that the total amount increased rather than the negative control group. In addition, Comparative Examples 1 and 2 showed an effect of the level of administration of the drugs of Examples 1 to 3 at a concentration of 50 mg/kg, even though the drug was administered in an excessive dose at a concentration of 100 mg/kg and 200 mg/kg.

Through the above results, it was proved that the extract according to the present invention is more effective in improving non-alcoholic fatty liver.

Test example 2-2. Inflammatory cytokine evaluation

division lL-1β (mg/g _protein ) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 20.0 - - - Negative control 38.4 - - - Positive control - 25.7 - - Example 1 - 25.8 24.1 22.4 Example 2 - 23.1 21.0 20.1 Example 3 - 24.0 22.3 20.8 Example 4 - 31.8 29.1 28.9 Comparative Example 1 - 37.1 35.6 34.5 Comparative Example 2 - 35.2 33.7 31.2

division TNF-α (mg/g _protein ) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 18.4 - - - Negative control 59.1 - - - Positive control - 12.9 - - Example 1 - 10.2 12.5 21.8 Example 2 - 8.4 9.0 16.8 Example 3 - 9.0 10.1 18.5 Example 4 - 23.7 25.0 30.6 Comparative Example 1 - 48.2 51.4 57.5 Comparative Example 2 - 35.9 39.6 45.1

division lL-6 (mg/g _protein ) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 3.9 - - - Negative control 7.2 - - - Positive control - 5.9 - - Example 1 - 4.5 4.1 5.4 Example 2 - 3.9 3.8 4.3 Example 3 - 4.1 3.9 4.9 Example 4 - 5.3 5.0 6.2 Comparative Example 1 - 6.2 5.8 7.0 Comparative Example 2 - 6.0 5.5 6.8

As shown in Tables 4 to 6 above, treatment with DDB increased the inflammatory cytokine factors IL-1β and IL-6 levels by more than twice, and increased TNF-α levels by more than 3 times.

Administration of the extracts of Examples 1 to 3 according to the present invention in the liver tissue of the non-alcoholic fatty liver animal model inflammatory cytokine factors IL-1β, TNF-α and IL-6 to the level before DDB is added (normal). It decreased excellently, and in particular, the effect of the section administered at a concentration of 100 mg/kg was the most excellent.

On the other hand, the extracts of Example 4 and Comparative Examples 1 to 2 also had the effect of lowering the levels of IL-1β, TNF-α, and IL-6, but it was confirmed that the recovery level was lower than that of the extracts of Examples 1 to 3 . That is, the extract of the complex extract (Example 1-3) rather than the single extract (Comparative Example 1-2) reduced the inflammatory cytokine level at a significant level.

Test example 2-3. In serum Liver metastasis Enzymes and LDH Level evaluation

division AST(IU/dL) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 51.4 - - - Negative control 77.2 - - - Positive control - 51.8 - - Example 1 - 60.7 52.1 53.9 Example 2 - 54.1 48.8 51.9 Example 3 - 55.5 50.1 52.4 Example 4 - 63.9 60.8 52.1 Comparative Example 1 - 75.5 70.0 73.8 Comparative Example 2 - 66.8 65.5 67.2

division ALT(IU/dL) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 38.4 - - - Negative control 57.1 - - - Positive control - 39.9 - - Example 1 - 43.5 40.6 43.1 Example 2 - 41.1 38.6 41.2 Example 3 - 42.8 39.2 41.6 Example 4 - 48.7 46.5 49.2 Comparative Example 1 - 55.8 54.6 55.0 Comparative Example 2 - 52.1 50.7 53.8

division LDH(IU/dL) 0 mg/kg 50 mg/kg 100 mg/kg 200 mg/kg Normal 200.4 - - - Negative control 397.8 - - - Positive control - 198.5 - - Example 1 - 210.6 201.4 208.7 Example 2 - 199.4 197.5 198.1 Example 3 - 200.1 198.6 199.4 Example 4 - 280.2 271.4 274.5 Comparative Example 1 - 350.6 340.5 341.8 Comparative Example 2 - 300.1 294.7 309.6

As shown in Tables 7 to 9 above, treatment with tunicamycin induces fatty liver production and liver failure with elevated aspartate transaminase (AST) and alamine transaminase (ALT).

Administration of the extracts of Examples 1 to 3 according to the present invention effectively reduced the concentrations of AST, ALT and low-density cholesterol (LDH) in the serum of a non-alcoholic fatty liver animal model, and recovered to a normal state (normal group level). Made it.

In contrast, in Comparative Examples 1 and 2, the concentration of AST and ALT in the serum was restored to a low level.

Test Example 2-4. Lipid blockage assessment

Lipid blockage in liver tissue was measured using the thiobarbituric acid reactive substances (TBARS) method. The concentration of TBARS was expressed as μM of malondialdehyde (MDA) per gram of liver tissue using 1,1,3,3-tetraethoxypropane (TEF) as a labeling material. 200 mg liver tissue homogeneous in 2 ml of 11.5 g/L KCl and 0.13 ml of homogenate was mixed with 0.08 ml phosphoric acid and 0.26 ml of 0.67% thiobabituric acid (TBA). After the mixture was heated at 100° C. for 45 minutes, 1.03 ml n-butanol was added, stirred, and centrifuged for 15 minutes. The supernatant was measured for absorbance at 535 and 520 nm with a spectrophotometer.

Treatment with DDB remarkably caused lipid mass oxidation. On the other hand, administration of the extracts of Examples 1 to 3 significantly reduced lipid mass oxidation. On the other hand, Comparative Examples 1 to 2 showed an effect on reducing lipid mass oxidation, but the effect was only 30 to 50% of that of Example 2.

Test Example 2. Verification of the effect on liver fibrosis

Using an animal model inducing liver fibrosis by administering dimethylnitrosamine (DMN), the efficacy of the extracts according to Examples and Comparative Examples on liver fibrosis was verified.

30 SPF (Specific Pathogene Free) grade 6-week-old male Sprague-Dawley (SD) rats weighing 190-210g were purchased from Daehan Biolink (North Chungcheong Province) and adjusted for 1 week at 22±2℃, 55±10% relative Solid feed (Daehan Biolink, Chungcheong-do) and tap water were freely ingested at humidity and a 12-hour light and dark cycle. Randomly divided into 8 animals per group. Liver fibrosis was induced over a total of 12 times by intraperitoneal injection three times in a row at a concentration of 10 mg of DMN per kg of rat body weight. The extracts of Examples and Comparative Examples were orally administered under the following conditions. After the last administration, fasting was performed for 18 hours to maintain an empty stomach, and then the animals were sacrificed to perform autopsy.

In liver tissue, collagen content was measured in liver tissue by fibrotic tissue staining technique, and oxidative damage and changes in the concentration of prefibrogenic cytokines were confirmed. At the time of autopsy, platelet levels were checked using blood obtained from the abdominal aorta, and serum was isolated to determine the level of liver transfer enzyme, an indicator of hepatitis, with an auto chemistry analyzer (AU400, Olympus, Japan).

Normal: No treatment group

Negative control group (Control): after induction of liver fibrosis by DMN administration, group administered with distilled water for 28 days

Positive control group (DDB): dimethyldiphenyl dicarboxylate (DDB) 50 mg/kg administered once a day for 28 days after induction of liver fibrosis by DMN administration

Example 1: After induction of liver fibrosis by DMN administration, the extract of Example 1 once a day for 28 days 50 mg/kg, 100 mg/kg administration group

Example 2: After induction of liver fibrosis by DMN administration, the extract of Example 2 once a day for 28 days 50 mg/kg, 100 mg/kg administration group

Example 3: After induction of liver fibrosis by DMN administration, the extract of Example 3 once a day for 28 days 50 mg/kg, 100 mg/kg administration group

Comparative Example 1: After induction of liver fibrosis by DMN administration, the extract of Comparative Example 1 once a day for 28 days 50 mg/kg, 100 mg/kg administration group

Comparative Example 2: After induction of liver fibrosis by DMN administration, the extract of Comparative Example 2 once a day for 28 days 50 mg/kg, 100 mg/kg administration group

Test example 3-1. Platelet count evaluation in blood and serum

division Normal Negative control Positive control Example 1 Example 2 Example 3 50 mg/kg 100 mg/kg 50 mg/kg 100 mg/kg 50 mg/kg 100 mg/kg Weight(g) 333.5 248.0 287.0 270.5 270.9 270.8 270.9 271.4 271.4 Liver weight (g) 10.5 9.8 11.4 9.4 10.6 9.7 10.7 9.6 10.7 Spleen weight (g) 0.9 1.7 1.9 1.9 1.9 1.9 1.9 1.9 1.9 Total bilirubin (mg/dL) 0.1 2.3 0.7 1.2 0.9 0.9 0.8 1.0 0.9 Platelets
(k/uL) 935.4 226.1 298.1 249.8 256.7 328.9 361.8 287.9 307.5

division Normal Negative control Positive control Example 4 Comparative Example 1 Comparative Example 2 50 mg/kg 100 mg/kg 50 mg/kg 100 mg/kg 50 mg/kg 100 mg/kg Weight(g) 333.5 248.0 287.0 271.1 271.8 272.0 272.3 271.4 271.8 Liver weight (g) 10.5 9.8 11.4 9.3 10.5 9.4 9.9 9.3 10.0 Spleen weight (g) 0.9 1.7 1.9 1.8 1.8 1.8 1.8 1.8 1.8 Total bilirubin (mg/dL) 0.1 2.3 0.7 1.5 1.3 2.1 1.8 1.9 1.6 Platelets (k/uL) 935.4 226.1 298.1 235.4 246.1 231.8 240.5 236.2 241.4

As shown in Tables 10 and 11 above, treatment with DMN significantly reduced the number of platelets in blood and serum. In addition, when administering the extracts of Examples 1 to 3 according to the present invention, it was confirmed that the lowered platelet count was significantly increased, and the total amount of bilirubin was effectively reduced.

Test example 3-2. Evaluation of liver fibrosis inhibition

division Inflammation level (X100) 0 mg/kg 50 mg/kg 100 mg/kg Normal 0.81 - - Negative control 2.46 - - Positive control - 1.25 - Example 1 - 1.73 1.37 Example 2 - 1.51 1.22 Example 3 - 1.69 1.30 Example 4 - 1.81 1.46 Comparative Example 1 - 2.19 1.88 Comparative Example 2 - 2.09 1.76

division Liver fibrosis (X100) 0 mg/kg 50 mg/kg 100 mg/kg Normal 0.25 - - Negative control 2.81 - - Positive control - 2.34 - Example 1 - 2.05 1.49 Example 2 1.70 1.01 Example 3 - 1.84 1.33 Example 4 - 2.47 1.98 Comparative Example 1 - 2.77 2.54 Comparative Example 2 - 2.61 2.48

As shown in Tables 12 and 13 above, when the extracts of Examples 1 to 3 according to the present invention are administered, compared to the case of administration of the extracts of Comparative Examples 1 and 2, inflammation (inflammation score) and liver fibrosis (Metavir score) It was confirmed that was significantly further reduced.

Test Example 3-3. Quantification of fiber in liver tissue

To quantify fiber in liver tissue, the concentration of hydroxyproline was measured. 200 mg of liver tissue stored at -70°C was homogenized with 2 ml of 6N hydrochloric acid and then incubated at 110°C for 24 hours. After filtering the acid hydrolyzate using a filter paper, 50 µl of a sample and standard hydroxyproline in 6N HCl were incubated to remove HCL, and 50 µl of methanol, 1.2 ml of 50% isopropanol and 200 ml chloramine T solution were sequentially added. After adding to, it was incubated for 10 minutes at room temperature. Thereafter, 3 ml of an Ehrlich solution was added to the mixture, followed by incubation at 50° C. for 90 minutes, and the absorbance of the reaction mixture was measured at 558 nm. The standard curve was prepared using 0-1.0 mg/50 µl of hydroxyproline solution.

As a result, treatment with DMN rapidly increased hydroxyproline, but it was confirmed that administration of the extracts of Examples 1 to 3 according to the present invention significantly inhibited hydroxyproline in a concentration-dependent manner.

Hereinafter, examples of the formulation of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit this, but is intended to be described in detail.

Formulation Example 1. Preparation of powder

500 mg of mixed extract powder obtained in Example 1

100 mg lactose

10 mg of talc

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2. Preparation of tablets

300 mg of mixed extract powder obtained in Example 1

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet manufacturing method.

Formulation Example 3. Preparation of Capsule

200 mg of mixed extract powder obtained in Example 1

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

Formulation Example 4. Preparation of injection

600 mg of mixed extract powder obtained in Example 1

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO _4, 12H ₂ O 26 mg

It is prepared in the amount of the above ingredients per ampoule according to a conventional injection preparation method.

Formulation Example 5. Preparation of liquid formulation

4 g of mixed extract powder obtained in Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid formulations, add and dissolve each component in purified water, add an appropriate amount of lemon scent, mix the above ingredients, add purified water and add purified water to adjust the total to 100 g, then fill in a brown bottle for sterilization. To prepare a liquid formulation.

Formulation Example 6. Preparation of granules

1,000 mg of mixed extract powder obtained in Example 1

The right amount of vitamin mixture

Vitamin A acetate 70 ㎍

1.0 mg of vitamin E

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

Vitamin B12 0.2 ㎍

10 mg of vitamin C

Biotin 10 ㎍

1.7 mg of nicotinic acid amide

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

90 mg of potassium citrate

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules, but it is possible to arbitrarily modify the mixing ratio. After mixing the above ingredients according to a conventional granule preparation method, the granules And, according to a conventional method, it can be used for preparing a health functional food composition.

Formulation Example 7. Preparation of functional beverage

1,000 mg of mixed extract powder obtained in Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

900 mL total by adding purified water

After mixing the above ingredients according to the normal health drink manufacturing method, stirring and heating the mixture at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and then stored in a refrigerator. It is used to prepare the functional beverage composition of the present invention.

The composition ratio is a mixture of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

Claims (9)

It contains a mixture of thistle and dandelion in a weight ratio of 1: 0.05 to 0.5 as an active ingredient,
The thistle includes a mixture of flowers of thistle, a mixture of stems and leaves, and a mixed extract of a mixture of roots and root residue powder,
The mixed extract is a thistle flower, a mixture of stems and leaves, and roots are mixed in a weight ratio of 1: 14-25: 0.1-0.9, and the root residue powder is included in an amount of 0.1 to 3 parts by weight based on 100 parts by weight of the mixed extract. A pharmaceutical composition for preventing or treating liver disease, characterized in that it is. The pharmaceutical composition for preventing or treating liver disease according to claim 1, wherein the mixture of thistle and dandelion is an extract extracted from water at 80 to 100°C. The pharmaceutical composition for preventing or treating liver disease according to claim 1, wherein the liver disease is selected from drug addiction, non-alcoholic fatty liver, liver fibrosis, cirrhosis, acute liver injury and acute hepatitis. The pharmaceutical composition for preventing or treating liver disease according to claim 3, wherein the drug addiction is caused by carbon tetrachloride. delete The pharmaceutical composition for preventing or treating liver disease according to claim 1, wherein the mixture of stems and leaves is a mixture of stems and leaves in a weight ratio of 1: 1-4. The method according to claim 1, wherein the mixed extract is a mixture of a thistle flower, and a mixture of stems and leaves, and the extract obtained by mixing the extract in the middle bath at 80 to 100°C, and the extract obtained by extracting the root in the bath at 80 to 100°C. A pharmaceutical composition for preventing or treating liver disease. The pharmaceutical composition for preventing or treating liver disease according to claim 1, wherein the root residue powder is obtained by drying and pulverizing the remaining residue after extracting the root at 80 to 100°C in a bath. It contains a mixture of thistle and dandelion in a weight ratio of 1: 0.05 to 0.5 as an active ingredient,
The thistle includes a mixture of flowers of thistle, a mixture of stems and leaves, and a mixed extract of a mixture of roots and root residue powder,
The mixed extract is a thistle flower, a mixture of stems and leaves, and roots are mixed in a weight ratio of 1: 14-25: 0.1-0.9, and the root residue powder is included in an amount of 0.1 to 3 parts by weight based on 100 parts by weight of the mixed extract. Food composition for preventing or improving liver disease, characterized in that the.