KR101701124B1 - A composition effective in liver function with mixed extractions from moroheiya, rhubarb, coptis and Korean angelica - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a health functional food composition for preventing liver disease or improving liver function and containing a mixed extract of Moroheya, rhubarb, Rhodiola, and Angelica gigantosa as active ingredients, and relates to a pharmaceutical composition and a health functional food composition for preventing liver diseases, hepatitis, Or as an active ingredient of a pharmaceutical composition or a health functional food composition for the prevention or treatment improvement.

Description

[0001] Description [0002] The present invention relates to a composition having an effect of improving hepatic function, comprising a mixed extract of Moroheya, rhubarb, Rhodiola, and Angelica gigantosa

The present invention relates to a functional food containing a mixed extract of Moroheya, rhubarb, Rhodiola, and Angelica gigantosa, and more particularly to a functional food containing a mixture of Moroheya, rhubarb, ≪ RTI ID = 0.0 > and / or < / RTI >

In Korea, liver disease mortality rate is very high, 23.5 per 100,000 population (37.8 males and 9.0 females), and the number of deaths in South Korea is the highest (41.1 / 100,000) and the second highest among the 50 deaths (72.4 / 100,000) , The third leading cause of death in the 30s (10 people / 100,000 people), is the leading cause of death in the Korean middle-aged population. Especially alcoholic liver disease is a disease that can occur in most chronic overdoses.

The liver is responsible for blood storage and circulation, blood volume control and defense detoxification in the human body, and is closely related to mental activity. Our body is constantly exposed to various pollutants and toxic substances by industrialization, and the liver is continuously suffering from detoxification, and mental stress is also known to cause liver damage. The deterioration of defense detoxification due to liver damage causes abnormalities in the immune system of the human body and causes other diseases. Carbon tetrachloride and D-galactosamine have been commonly used as experimental substances causing hepatotoxicity. 16, 34, 1986, Butler, J. Pharmacol. Exp. Ther., 134, pp. 311-319, 1961).

The bile has components such as bile acid, phospholipid, bilirubin, GSH (Glutathione) and electrolyte, and is not only a pathway for excretion of toxic substances and fat degradation products, but also participates in digestion and absorption of fat to perform various physiological functions, It excretes endogenous waste products and drugs such as bilirubin and various toxic substances, increases normal fat absorption in the intestines, and plays a pivotal role in maintaining the balance of human cholesterol metabolism. (Ballatori N et al., Am. J. Physiol., 263, pp. G617-G624, 1992).

Galactosamine / Lipopolysaccharide is widely used as one of the representative cholestatic liver disease induction models, and galactosamine is known to be involved in UTP (Uridine 5'-triphosphate) UDP (Uridine diphosphate) - depletion of UDP-glucuronic acid to suppress protein production, thereby depleting GSH, a liver protective substance, inhibiting the inclusion process involved in foreign body excretion, causing hepatotoxicity, It is a histologic feature that infiltration of inflammatory cells is observed accompanied by tissue necrosis of the tissue. (Jonker AM et al., Hepatology, 11, pp. 622-627, 1990).

In addition, LPS (Lipopolysaccharide) increases the secretion of TNF-α (Tumor Necrosis Factor-α) due to the activation of inflammatory cells in the liver by rapidly increasing the intestinal portal inflow of intestinal toxins including LPS, The pathogenesis of acute chronic inflammation in the liver lowers the expression of hepatocellular transport proteins and the degradation of these transport proteins leads to disorders of bile excretion and is associated with hematological indices of cholestatic liver disease such as Alkaline phosphatase, -Glutamyl transpeptidase and the like. (Grun M et al., Acta Hepatogastroenterol (Stuttg), 24, pp. 64-81, 1977).

Aminotransferase, which is one of the most representative measurement items in the blood test for the diagnosis of liver disease, is contained in a large amount in the hepatocyte, but when the permeability of the cell membrane is enhanced or the hepatocyte is damaged, Since it is released into the blood, measurement of the activity change of this enzyme in the blood can be used as an index because it is suitable to estimate the degree of damage of the liver cell. In particular, the alanine aminotransferase (ALT) or glutamate pyruvate transferase (GPT) is known to be associated with the aspartate aminotransferase (AST or GOT) And the enzyme activity in blood is higher than that of AST in most liver diseases. Therefore, it may be a very important factor in the diagnosis of liver disease. The activity of ALT and AST in liver is known to be several thousand times higher than that in blood (Rej R., Clin. Chem. 24, pp. 1971-1979, 1978). Therefore, when liver tissue is damaged by various factors such as toxic substances, virus infection, alcohol, and obesity, it is released into the blood and circulates, thereby increasing the blood ALT and AST activity. Therefore, the activity of these in the blood can be measured and the diagnosis of liver disease can be inferred.

Various radicals and reactive noxious oxygen species in the cell are rapidly reducing the amount of GSH by binding to GSH which has liver protective effects (Mitchell, JR et al., Drug Metab. Rev. 13, pp. 539-553, 1982, Mitchell, J. et al. J. Pharmacol. Exp. Ther. 187, pp. 211-117, 1973). Various radicals and reactive noxious oxygen species not associated with GSH bind to intracellular macromolecules, cause lipid peroxidation of the cell membrane, and also interfere with the regulation of intracellular calcium, resulting in cell death (Mitchell, JR Rev. 24, pp. 377-407, 1992, Cohen, SD Drug Metab., Rev. 29, pp. 187, pp. 211-117, 1973. Vermeulen, NPE Metab. 59-77,1997). Therefore, by measuring the degree of GSH and lipid peroxidation, liver damage and liver protection can be evaluated.

Recently, many studies have been conducted on the inhibitory action of free radicals through natural products such as edible and medicinal plants for preventing or treating hepatotoxicity and liver damage. J. Med Sci., 34, pp. 80-85, 1995. Middleton, E., Int. J. Pharmacology, 34 , pp 344-348, 1996). Silymarin and BDD (biphenyl dimethyl dicarboxylate) are used as typical liver disease drugs. Silymarin is a substance isolated from the fruit of Silybum marianum belonging to the Asteraceae family. BDD is a synthetic material similar to Schzandrin, an ingredient of Omija, and developed as a therapeutic agent for liver diseases derived from natural materials .

Macrophages are the effector cells that play a major role in the elimination of microorganisms invading in vivo. Infected macrophages secrete inflammatory cytokines such as IL-1, TNF-α and IL-6 to activate other macrophages and remove intracellular bacteria by stimulating other cells in the immune system , These cytokines lead to tissue damage caused by inflammation. In other words, macrophages start inflammatory reaction by producing arachidonic acid in cells by external stimuli such as viruses and bacteria, and COX-1 (Cyclooxygenase-1) and COX-2 (Cyclooxygenase-2) It secretes prostaglandin E2 by participating in acid metabolism. Prostaglandins were first known to help uterine contraction in the endometrium, but they are now attracting attention as inflammatory mediators secreted during cellular damage and are known to increase pain when prostaglandins are secreted. In addition, it is known that nitric oxide and TNF-α are involved in the inflammatory reaction. Generally, in the development of inflammatory disease agents and analgesics, a model that stimulates immune cells such as macrophages with external stimuli such as LPS to produce NO (nitric oxide), TNF-α, prostaglandin E _2, etc. A method of evaluating the anti-inflammatory action or analgesic effect of the candidate substance by confirming whether the administration of the therapeutic agent for inflammatory disease or candidate agent for analgesic inhibits the production of NO, TNF-α, prostaglandin E ₂ , . Until now, nonsteroidal antiinflammatory drugs (NSAIDs) such as indomethacin and aspirin, which inhibit prostaglandin synthesis, have been mainly used as antiinflammatory drugs, and steroid drugs such as dexamethasone have been used as second-line prescriptions. However, these drugs have various side effects, especially gastrointestinal disorders, side effects of liver and kidney dysfunction.

At present, attempts are being made to find a composition for improving the function of preventing and treating diseases associated with liver diseases inducing activity of a detoxifying enzyme system in the world, and studies are underway to separate natural compositions from plants. However, in Korea and other Northeast Asian regions, research on medicinal plants tends to be limited, and studies on wild vegetables, vegetables and fruits that are native to each country have not been reported yet. The present inventors have also made efforts to develop a substance having a purpose of preventing or treating diseases related to liver disease or improving liver function from plants which have not been known at all, and among them, they have been interested in rhubarb, .

Moroheya is a one-year-old grass originating in tropical Asia and Africa belonging to the Tiliaceae family. The official name is Corchorus olitorius L. There are about 40 genera and 400 species distributed in tropical and temperate forests. Among them, 11 genera of 3 genera are distributed in the country. The height is 50 to 120 cm (4 m in the tropics), and the oval oval leaf is distinctive and has a mount. The leaf quality is soft and is similar to jute (Corchorus capsularis L.). Flowers bloom from August to September with large, dark yellow flowers. Moroheya is characterized by a slippery, thick liquid that comes out when cutting leaves. This mucin is an enzyme that combines polysaccharide and protein, called mucin, and is a kind of mucopolysaccharide that is water-soluble vegetable fiber. This vegetable fiber contains a large amount to improve constipation, and is released in association with the bile acid secreted from the gallbladder in the small intestine, thereby lowering the cholesterol synthesis, lowering the cholesterol in the blood and preventing the bile acid and neutral fat from being reabsorbed, Lower triglycerides. And harmful to the body is absorbed, excreted, and gives vitality to the cells to increase immunity. It also works in the stomach to preserve the moisture in the tissues and maintain the elasticity of the joints and moisturizing.

Rhei Rhizoma is a perennial herbaceous plant belonging to the genus Rhizoma (Rhizoma Rhizoma) and has about 15 species of rhizomes and rhizomes of related plants. Among them, there are Rheum coreanum, Rheum undulatum Dried root and rootstocks are the origin of the plant. In China, it is originated from dried root and rhizomes such as Rheum palmatum, Rheum tanguticum and Rheum offcinale. Rhubarb is a major component of anthraquinone derivatives such as Rhein, Emodin, Aloe-emodin, Chrysophanol Physion, their glycosides and their derivatives, It is known to contain components such as Sennoside A, B, C, D, E, F and the like. The main pharmacological action of rhubarb from ancient times is subcutaneous action, and besides it shows inhibitory action of digestive enzymes, and it shows dandruff action. It has antimicrobial action, hemostatic action, anticancer action, diuretic action, protection of liver function, blood lipid lowering action, . In addition, rhubarb is clinically applied to the treatment of indigestion, constipation, acute inflammation, infectious disease, parasitic onset, bleeding, thrombocytopenia, burns and skin diseases.

Rhizome is an evergreen perennial plant of dicotyledonous plants and buttercups. It is native to China and it is cultivated in Korea, Japan, China and so on for herbal medicine. It grows in the ecological land of the shade of the mountain forest. The underground stem is thick and spreads to the side. It has many beard roots, 4 ~ 5 roots at the end of the stem, and the length is 10 ~ 27cm. The leaf is a compound leaf with three small leaves. The small leaf is a little hard, sharp and sharp. The petiole of the petiole is a sheath with sawteeth on the edge of the leaf. In April ~ April, two to three white flowers bloom on the flower stalk which grew to about 10cm in diameter. Flowers have blooming flowers and male flowers. The white calyx is five to six long and looks like a petal. Petals are small linear with 8 ~ 15 petals. There are many stamens and 9 ~ 16 pistils. It is named after the ovary sack has grown on it and the fascia is lined with an annulus, and the stem and the stem of the ground are thick yellow in color. It is said that the root of the root of the root of the root is extracted from the roots of the root of the root of the root and it is called the root of the root. Vomiting, dysentery, heart palpitations, burns, mental anxiety, throat dysentery, blood clots, nosebleeds, hemorrhages, and burns.

Angelica gigas Nakai is a perennial herb that belongs to the dragonfly family, Angelica gigas Nakai. Essential oil and decusin as a directional component, soothing beta-sitosterol, sucrose, nicotinic acid, folic acid and other ingredients are composed. It has various effects such as various female diseases, menstrual irregularities, menstrual cramps, anemia, poor uterus development, lack of postpartum blood, and menopausal disorders, and has a function to smoothly assist female hormone secretion. Recent studies have shown that Angelica gigas has an excellent effect on cellular immune responses. Oriental medicine is warm in nature, flavor is sweet and very enter the heart, liver, spleen enters. It replenishes and harmonizes the blood, regulates the physiology and makes the stools pass smoothly.

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1. Bruckner et al., Fund. Appl. Toxicology, 6, pp. 16-34, 1986; Butler, J. Pharmacol. Exp. Ther., 134, pp311-319, 1961. 2. Ballatori N et al., Am. J. Physiol., 263, pp. G617-G624, 1992. 3. Jonker AM et al., Hepatology, 11, pp. 622-627, 1990

The object of the present invention is to provide a food or pharmaceutical composition for prevention or treatment of liver diseases such as fatty liver, hepatitis, liver cirrhosis and the like, which comprises an extract obtained by extracting Moroheya, rhubarb, .

In order to solve the above-mentioned problems, the present invention provides a pharmaceutical composition for preventing or treating liver diseases, comprising a herbal medicine mixed extract of Moroheya, rhubarb, Huangliao, and Angelica gigantosa as essential active ingredients and containing a pharmaceutically acceptable carrier, excipient or diluent Lt; / RTI >

More specifically, the present invention relates to a method for producing a herbal composition, which comprises mixing a dry extract of a mixed extract of Moroheya, rhubarb, Rhodiola, and Angelica gigantis with 11-20 parts by weight, 15-30 parts by weight, 15-30 parts by weight, Which is effective for improving liver function.

The herbal medicine mixed extract may be characterized by being soluble in a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The present invention also relates to a pharmaceutical composition for prevention and treatment of liver diseases, hepatitis, liver cancer, liver cirrhosis, and liver cirrhosis, which comprises as an essential active ingredient a mixed extract of Moroheya, Rhodiola, to provide.

The pharmaceutical composition comprises 11-20 parts by weight, 15-30 parts by weight, 15-30 parts by weight, and 1 to 17 parts by weight of Angelica gigas Nakai, rhubarb, .

The herbal extracts of Moroheya, rhubarb, Huangliao and Angelica gigantosa inhibit ALT and AST activity in liver disease animal model experiments and inhibit lipid peroxidation and GSH content. A pharmaceutical composition for the prevention or treatment of fatty liver, hepatitis, liver cancer, and cirrhosis of the liver due to the absence of adverse effects or toxicity derived from natural products, confirming the effect of inhibiting the production of PGE ₂ induced by LPS in Raw264.7 cells May be used as an active ingredient of a health functional food composition.

FIG. 1 shows the effect of the present invention on the ALT and AST activity in hepatotoxic animal model experiments induced by carbon tetrachloride mixed rats of Moroheya, rhubarb, Huangliao and Angelicae herbicide mixed extracts and the group treated with silymarin (positive control).
FIG. 2 shows the effect of the present invention on the MDA (Malondialdehyde) level and the silymarin-administered group of 300 mg / kg mixed extract of Moroheya, rhubarb, Rhodiola, and Angelicae radix herb extracts in a hepatotoxic animal model experiment induced by carbon tetrachloride FIG.
FIG. 3A is a graph showing the effect of GOT contained in sera in hepatocytes of Moroheya, rhubarb, Huangliao, and Angelicae radix mixed extract of the present invention in an animal model of hepatotoxicity caused by carbon tetrachloride.
FIG. 3B is a graph showing the effect of GPT contained in serum in hepatocytes of Moroheya, rhubarb, Huangryang, and Angelicae radix mixed extract of the present invention in an animal model of liver damage caused by carbon tetrachloride.
FIG. 4A is a graph showing the GOT improvement effect of liver tissue in a liver cirrhosis animal model induced by the DMN-induced mixed extract of Moroheya, rhubarb, Huangryang, and Angelica gigantosa of the present invention.
FIG. 4B is a graph showing the GPT improvement effect of liver tissue in DMN-induced liver cirrhosis animal model of the mixed extract of Moroheya, rhubarb, Huangliao, and Angelica gigantosa of the present invention.
FIG. 5 is a graph showing the amount of collagen fibers in a liver cirrhosis animal model induced by DMN of the mixed extracts of Moroheya, rhubarb, Huangliao, and Angelica gigantosa of the present invention.
FIG. 6 is a graph showing the effect of inhibiting the production of prostaglandin E ₂ induced by LPS in the mixed extracts of Moroheya, rhubarb, Rhodiola, and Angelica gigantosa of the present invention.

As a result of continuous research on the hepatoprotective and hepatitis prevention and treatment effects of mixed herbal medicine extracts of Moroheiya, Daehang, Huangryun and Angelica gigantosa, herbals which have been proven to be used for a long time in clinical use for a long time, The present invention provides a method for inhibiting ALT activity and AST activity in an animal model experiment of liver disease induced by carbon tetrachloride or D-galactosamine (GalN), wherein the mixed herbal extract of Moroheya, rhubarb, The lipid peroxidation content is reduced, GSH content is not effectively depleted, and the effect of inhibiting the production of prostaglandin E ₂ derived from macrophages into lipopolysaccharide is confirmed, thereby completing the present invention.

The " extract " as defined in the present invention is obtained by extracting and isolating from a natural raw material using an extraction and separation method known in the art and using an appropriate solvent, and examples thereof include Moroheya, rhubarb, Or crude extracts of Moroheya, rhubarb, Rhodiola and Angelica gigas, polar solvent-soluble extracts or non-polar solvent-soluble extracts.

As an appropriate solvent for extracting the extract, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used. Examples of the solvent include, but are not limited to, purified water, methanol, Alcohol having 1 to 4 carbon atoms, acetone, ether, benzene, chloroform, and the like, including ethanol, propanol, isopropanol and butanol, Various solvents such as ethyl acetate, methylene chloride, hexane, and cyclohexane may be used alone or in combination.

As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. The desired extract may further be subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation on the method of preparing the extract, and it is obvious that any known method can be used.

The extract is characterized in that it is contained in the form of a pulverized product, an extract or a powder extract thereof. The composition according to the present invention includes a mixed extract comprising a single extract of each of the herbal medicines or a combined extract prepared by extracting together plants containing two or more herbal medicines.

The extract according to the present invention may be a crude extract prepared by applying a conventional water extraction method, a solvent extraction method using alcohol or the like as a solvent, or may be a fractional extract purified by column chromatography. The extraction method according to the present invention can be applied to any extraction method known to a person skilled in the art. For example, extraction with water, alcohol or mixed solvent, extraction with hot water or room temperature But is not limited thereto.

In a preferred embodiment of the present invention, the dry herbicides are washed in order to remove foreign substances present on the surface of the material, dried and dried in a drier, mixed at a proper mixing ratio, and pulverized to obtain a herb pulverized product.

In addition, distilled water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably distilled water alone or 10 to 50% by volume, preferably 5 to 20 times, preferably 10 to 15 times, It is preferable to add water or a mixed solvent of water and ethanol and to cool the solution at a temperature of 0 ° C to room temperature, preferably 4 to 6 ° C, for 12 hours to 48 hours, preferably 20 hours to 24 hours, Extracting the extract of the complex herb extract which is soluble in a polar solvent by extraction with water at a temperature of 90 ° C or higher for 1 hour to 24 hours, preferably 2 hours to 5 hours, An extract can be obtained.

Furthermore, the extract obtained from the extraction step is further filtered, and the extract obtained by concentrating the filtrate under reduced pressure at 80 to 90 ° C is azeotropically concentrated 1 to 5 times with 10 to 60 times of the total amount of X, and then subjected to hot air drying or vacuum drying , Preferably by hot air drying to obtain a powder X-ray. By the same method as described above, the present invention can contain the above-mentioned Moroheya, rhubarb, Rhodiola and Angelica gigantosa extract as essential active ingredients.

The herbal extracts for preventing and treating liver diseases of the present invention are not limited thereto, but may be selected from the group consisting of 11 to 20 parts by weight, 15 to 30 parts by weight of Moroheya, rhubarb, 15 to 30 parts by weight, and Angelica keiskei at 1 to 17 parts by weight.

The herbal medicine extracts have been used as foods for a long time, and the extract of the present invention extracted therefrom also has no toxicity and side effects.

Also, as a result of examining the effect of the composition obtained by the above method on the carbon tetrachloride-induced chronic hepatotoxicity, it was confirmed that the hepatocyte-treated hepatic fibrosis progressively improved in liver cirrhosis.

The composition for treating and preventing liver cancer according to the present invention can be prepared by a conventional method in the pharmaceutical field.

The pharmaceutical composition comprising the extract according to the present invention can be administered orally or parenterally in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories, Can be used. Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, Or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyoxyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As a suppository base, Witepsol, Macrogol, Tween 60, cacao butter, laurin, glyceroglatin and the like can be used.

The pharmaceutical composition of the present invention can be administered by any of parenteral agents such as oral agents, injections, patches, and spotting agents.

The pharmaceutical composition of the present invention is administered by a suitable route according to the preparation form. The method of administration is not particularly limited, and it can be administered by the contents, external application and injection. The injection can be administered, for example, intravenously, intramuscularly, subcutaneously, intradermally, and the like.

The dosage of the pharmaceutical composition of the present invention is suitably set according to the form of the preparation, the method of administration, the purpose of use, and the age, weight, and symptom of the patient applied to the composition, and the amount of the active ingredient For example from 10 ug to 2,000 mg / kg per adult day. However, those skilled in the art will appreciate that the pharmacokinetic properties of a particular reagent and its mode and route of administration; Age, health, weight; It will be understood that the dosage varies depending on known factors such as the nature and extent of the symptoms, the type of concurrent treatment, the frequency of treatment and the effect of the purpose.

The composition of the present invention can also be used as a health functional food for improving liver function. The health functional food may also be used as a general health food or as a functional health food having multipurpose functions such as diet, fatigue recovery, and hangover.

Examples of the functional food to which the extract of the present invention can be added include various foods, candies, chocolates, beverages, gums, tea, vitamin complexes and health supplement foods. Examples of the functional foods include powders, granules, tablets, pills, Can be used.

In the present invention, the term " food " means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be eaten directly through a certain degree of processing, Which includes food, food additives, functional foods and beverages.

The above-mentioned " functional food " refers to a food group which is imparted with added value to function and express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, Refers to a food prepared by processing a body so as to sufficiently express the body's control function on the body, such as recovery, and the like. Specifically, it may be a health functional food. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

In the present invention, the term " beverage " means a general term for drinking or enjoying a taste, and includes a functional beverage. The beverage contains, as an essential ingredient, the composition for improving liver function or hangover, in an indicated ratio, and there are no particular restrictions on other ingredients, and various flavors or natural carbohydrates such as ordinary beverages may be added as an additional ingredient can do.

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

Since the herbal extract of the present invention has little toxicity and side effects, it can be safely used for prolonged use even for prophylactic purposes.

The herbal extract of the present invention may be added to foods or beverages for the purpose of improving gastrointestinal diseases. In this case, the amount of the food composition of the present invention in the food or beverage may generally be 20 to 70% by weight of the total food, and the beverage composition may contain 0.02 to 30 g, preferably 0.3 to 1 g Of the food composition of the present invention.

The present invention will be described in more detail based on the following examples and experimental examples, but the present invention is not limited by the examples or the experimental examples.

Examples. Preparation of mixed herbal extracts of Moroheiya, rhubarb, Rhododendron and Angelica japonica

200 g, 300 g, 300 g and 150 g of each of the finely divided Moroheiya, rhubarb, Rhododendron and Angelica gigas purchased from Kyungdong market (Seoul, Korea) were mixed and then 10 L of ethanol was added. And extracted repeatedly. The supernatant obtained by repeating the cooling operation at room temperature for 3 times was added to the supernatant liquid. The supernatant liquid obtained by repeating the filtration operation was collected, and the ethanol was completely evaporated under a reduced pressure using a rotary evaporator. The concentrate was then dissolved in a small amount of distilled water, Was prepared.

Experimental Example 1. Effect on Carbon Tetrachloride Induced Chronic Liver Injury

In order to confirm the improvement effect of the composition according to the present invention on hepatic injury, the improvement effect on rat liver injury induced by carbon tetrachloride was examined using the following experimental method with the extract prepared in the above examples.

1-1. Preparation of reagents and experimental animals

The reagents were purchased from CCl ₄ (Sigma Co.), olive oil (Sigma Co.), alanine aminotransferase (ALT, GPT, Stanbio Co.) and aspartate aminotransferase (AST, GOT, Stanbio Co.) Respectively.

Male (SD) (Sprague-Dawley) male rats weighing 200 g were used, and feed (Harlan, teklan, USA) and drinking water were freely consumed. The temperature of the cage was 21 ~ 24 ℃ and the relative humidity was 60 %. Also, light was adjusted in the breeding ground so that day and night were repeated every 12 hours.

1-2. Sample collection

Twelve-hour fasted mice were intraperitoneally administered 0.1 ml per 10 g of a mixture of carbon tetrachloride and olive oil (0.5: 9.5 (v / v)) and anesthetized with ether after 24 hours to take blood from the abdominal vena cava. Tissue was collected. The mixed extracts of Moroheya, rhubarb, Rhodiola, and Angelica gigas Herbica obtained in the above Examples were orally administered for 2 hours before the administration of carbon tetrachloride and 30, 100, 300 or 600 mg per 1 kg of body weight at 2 hours after administration. Silymarin (Sigma, S0292-50G) was used as a positive control, and its dose was 300 mg / kg.

1-3. Measure ALT and AST activity

ALT and AST activities were measured with a UV spectrophotometer (Libra S22, Biochrom, UK) using IVD LAb kit (iVidilab, Korea) from the serum obtained by centrifuging the blood of Experimental Example 1-2.

result

100, 300, and 600 mg / kg of the mixed extracts of Moroheya, rhubarb, Rhodiola and Angelicae radix obtained in the above Examples for ALT and AST activity in hepatotoxic animal model experiments induced by carbon tetrachloride and the silymarin treated group Control group) was shown in Fig. Each bar graph shows the measurement + standard deviation of 9-11 mice per experimental group. In the group of 300 mg / kg of the mixed extract of Moroheiya, Rhodophyta, Rhodiola, and Angelica japonica, the inhibition of ALT activity was significantly inhibited in the group treated with silymarin. The activity of AST was higher than that of Morohayaya, 300, and 600 mg / kg, respectively.

1-4 Measurement of lipid peroxidation

(Thibarbituric acid assay) as described below (Ohkawa et al., J. Med. Chem., 40, pp. 559-573, 1997) The product, malondialdehyde, was quantified as an indicator.

The liver tissue obtained in Experimental Example 1-2 was homogenized with a 1.15% potassium chloride solution having a volume of 10 times (v / w), and then 0.2 ml of homogenate, 0.2 ml of 8.1% SDS, 1.5 ml of 20% , 0.5 ml of 5% butylated hydroxytoluene and 1.5 ml of 0.8% thiobarbituric acid. The mixture was allowed to react for 60 minutes in a thermostat maintained at 100 ° C., allowed to stand at room temperature for 1 hour, And centrifuged for 10 min. The supernatant was taken and the absorbance was measured with a UV spectrophotometer at 532 nm.

1-5. GSH content measurement

GSH content was measured by the following method (Weiberg, JMJ et al., Clin. Invest. 80, pp. 1446-1454, 1987). The liver tissue obtained in Experimental Example 1-2 was homogenized with 1% picric acid in a volume of 2 v / w, and then 2 ml of the homogenate was centrifuged at 500 x g for 3 minutes to obtain supernatant Was diluted 50 times (v / w) with distilled water. After adding 200 μl of the diluted supernatant, 700 ml of 0.3 mM NADPH and 100 μl of 6 mM 2-nitro-6-thiobenzoic acid, the mixture was allowed to stand in a thermostatic chamber maintained at 30 ° C. for 4 minutes and 5 μl of 80 units GSH reductase / The total amount of GSH was measured by measuring the absorbance at 412 nm for 1 minute using a UV spectrophotometer. In the same manner as above, 2 μl of 2-vinylpyridine and 20 μl of 0 units GSH reductase / ml solution were added, The GSSG (Gluthathione disulfide) content was determined by measuring the absorbance increase for 1 minute. The content of GSH was calculated using the formula GSH = total GSH - 2 × GSSG.

result

The effect of the present invention of 300 mg / kg of the mixed extract of Moroheya, rhubarb, Rhodiola, and Angelicae herbicidal extracts of the present invention on the MDA (malondialdehyde) level and the silymarin-treated group in the hepatotoxic animal model experiment induced by carbon tetrachloride is shown in FIG. . Each bar graph shows the measurement + standard deviation of 9-11 mice per experimental group. As shown in FIG. 2, it was confirmed that the MDA level was significantly lowered and the depletion of GSH content was effectively inhibited in the 300 mg / kg group of the mixed extract of Moroheiya, rhubarb, Huangryun, and Angelica gigantosa.

1-6. Effect on Carbon Tetrachloride Induced Chronic Liver Injury

Only normal saline was administered for 4 days. In the control group, a solution prepared by mixing olive oil and carbon tetrachloride in a ratio of 1: 1 (v / v) was intraperitoneally administered to each rat at a dose of 1.0 mg / kg per kg of body weight (kg) for 3 days, followed by administration of saline . In the experimental group, a solution prepared by mixing olive oil and carbon tetrachloride at a ratio of 1: 1 (v / v) was intraperitoneally administered to each rat at a dose of 1.0 mg / kg per kg body weight for 3 days to induce acute hepatotoxicity. The extracts were suspended in 1% carboxymethylcellulose and orally administered at 6 mg / kg, 30 mg / kg and 60 mg / kg for 1 day after the administration of the carbon tetrachloride solution for 4 days. On the last day of the test, animals were anesthetized and 1.5 ml of blood was collected and perfused. The collected blood was centrifuged at 3,000 rpm for 20 minutes, and GOT and GPT were measured using CH100 ⁺ (SEAC Co.). The experimental group setting and the experimental schedule are shown in Table 1.

group Treatment group (days) One 2 3 4 Normal group Saline + saline Saline solution Control group CCl ₄ - olive oil (1: 1) + saline solution Saline solution Moroheiya, rhubarb, Huangliao and Angelica herbal medicine mixed extract × 1 CCl ₄ - Olive oil (1: 1) + Moroheya, Rhubarb, Rhodiola and Angelicae herbal medicine mixed extract Moroheiya, rhubarb, Huangliao and Angelica herbal medicine mixed extract × 5 × 10

result

As a result of measurement of GOT and GPT activity contained in serum from hepatocyte in hepatocellular carcinoma model animals, the herbicide composition of herbal medicine combination extract decreased in concentration-dependent activity. mg / kg dose in the test group. (P < 0.05, P < 0.01) (Figs. 3A and 3B). Therefore, it was confirmed that the composition of the present invention had an excellent effect on improving liver function.

Experimental Example 2. Collagen distribution and improvement effect of GOT and GPT in liver tissue

In order to confirm the improvement effect on the liver cirrhosis of the composition according to the present invention, the improvement effect on rat liver cirrhosis was examined using the following experimental method with the extract prepared in the above examples.

2-1. Preparation of reagents and experimental animals

The reagents were purchased from DMN (Dimethylnitrosamine, Sigma Co.), alanine aminotransferase (ALT, GPT, Stanbio Co.) and aspartate aminotransferase (AST, GOT, Stanbio Co.).

Wistar rats weighing 270g or less were used. Harlan, teklan USA and drinking water were freely consumed. The temperature of the kennel was 21 ~ 24 ℃ and the relative humidity was 60%. Respectively. Also, light was adjusted in the breeding ground so that day and night were repeated every 12 hours.

2-2. Collagen distribution and improvement of GOT and GPT in liver tissue in liver cirrhosis

Oral saline alone was administered to the normal group for 4 weeks for 3 days per week. On the other hand, in the control group, saline was administered after intraperitoneal administration of 1% DMN at a dose of 10 μl / kg / day to induce cirrhosis, and in the experimental group, 30 mg / kg / day, 60 mg / kg / day of the extract was orally administered for 4 weeks. On the last day of the test, animals were anesthetized, and 1.5 ml of blood was collected, fixed by perfusion, and liver tissues were separated. The collected blood was centrifuged at 3,000 rpm for 20 minutes, and then GOT and GPT were measured using CH100 ⁺ (SEAC co.). (See Figs. 4A and 4B)

On the other hand, Masson trichrome staining was performed to grasp the distribution of the collagen fibers in the isolated liver tissue. Separated liver tissues were embedded in paraffin embedded tissue using an automatic tissue processor (Citadel 2000, Shandon Co.) and cut into 4 μm thickness using a tissue cutter (LEICA RM2145). Using an image analysis system Five parts were selected for each tissue, and the ratio of cellulose was measured, and the average value was obtained and expressed as the amount of the collagen fibers. (See Fig. 5)

result

In DMN - induced cirrhosis model animals, the extract of herbal medicines lowered the activity of GOT and GPT compared to the control group. In addition, the number of collagen fibers was not significantly changed to normal range of 0.53 in normal group and 2.93 in control group, but 1.05 in moroheya, rhubarb, And inhibited the progression.

Experimental Example 3: Toxicity test

In order to investigate the toxicity of the composition of the above example by oral administration to rats, male and female rats of the SD strain were divided into three groups of 1 g / kg, 2 g / kg and 5 g / kg, Oral administration, 14 days of death, general symptoms, weight change and autopsy findings were observed. The results of the test are as follows.

result

(1) No mortality was observed during the test period in all the administration groups of the composition of the present invention during the test period, and there was no change in the general symptoms observed specifically compared with the control group.

(2) Body weight change showed significant weight loss at 5 g / kg dose group on the 3rd day after administration, but there was no difference between the other groups in all other treatment groups.

As a result, no specific clinical symptoms were observed compared to the death or control group in all test substance administration groups, and no abnormal symptoms were observed in weight changes and anatomical findings on autopsy. In addition, the composition showed no significant toxicity at a dose of 5 g / kg or less, so that the LD ₅₀ (Lethal dose) in rats by oral administration is estimated to be 5 g / kg or more.

Experimental Example 4 Inhibitory Effect of LPS-Induced Prostaglandin E ₂ Production

To determine the effect of inhibiting the production of prostaglandin E ₂ , a Parameter ^TM PGE ₂ assay kit (R & D Systems) was used. Raw 264.7 cells (1 × 10 ⁵ cells) were treated with LPS at a concentration of 1 μg / μl to induce an inflammatory response. After the reaction mixture was treated with 20 μg / ml of the mixed extract of Moroheya, rhubarb, Rhodiola and Angelica gigantosa obtained in Example 1, 80 μl of supernatant was taken and diluted with 160 μl of diluent buffer, , And competitively reacted with prostaglandin E ₂ conjugate in wells coated with goat anti-mouse polyclonal antibody. Antibody to prostaglandin E ₂ contained in the Parameter ^TM PGE ₂ assay kit was added thereto, followed by antigen-antibody reaction for 24 hours, and the degree of color development at 450 nm was measured and compared. As the control group, LPS treatment group, LPS treatment group and indomethacin (7 ㎍ / ml concentration) treatment group were used.

result

In the experimental model in which the production of prostaglandin E ₂ was induced by LPS in Raw264.7 cells, the 20 ㎍ / ml-administered group of Moroheya, rhubarb, Huangliao and Angelicae radix mixed extracts obtained in the above example, LPS alone treatment group (LPS + ), Negative control group (LPS-) and indomethacin treatment group (Ind: indomethacin) inhibition of prostaglandin E ₂ production is shown in FIG. As shown in FIG. 6, the production of prostaglandin E ₂ at a level similar to that of the negative control (LPS-) in the 7 ㎍ / ml-treated group of the mixed extracts of Moroheya, rhubarb, The effect of inhibition was confirmed.

The extract of the present invention can be administered in the following formulations, and the following formulation examples are illustrative of the present invention, and the contents of the present invention are not limited thereto.

Preparation Example 1. Preparation of powder

Herbal medicine complex extract 200 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation Example 2. Preparation of tablets

Herbal medicine complex extract 200 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation Example 3. Preparation of capsules

Herbal medicine complex extract 200 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation Example 4. Preparation of soft capsule (content in one tablet of soft capsule)

Herbal medicine complex extract 200 mg

Polyethylene glycol 400 400 mg

Concentrated glycerin 55 mg

Purified water 35 mg

Polyethylene glycol and concentrated glycerin were mixed and then purified water was added. The mixture was kept at about 60 ° C, and flavon was added thereto. The mixture was stirred uniformly at about 1,500 rpm with a stirrer, cooled gradually to room temperature while being stirred, Remove the bubbles and use as a soft capsule contents.

The capsule of soft capsule was prepared by softening gelatin and plasticizer, which are generally known soft gelatin, plasticizer, in which 132 mg of gelatin, 52 mg of concentrated glycerin, 6 mg of diisovitol solution and 70% of ethyl vanillin as a flavoring agent, By the usual preparation method.

Formulation Example 4. Preparation of injection

Herbal medicine combination extract 100 mg

180 mg mannitol

Sterile sterilized water for injection 2,974 mg

Na ₂ HPO _{4 12} H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

Formulation Example 5. Preparation of a liquid preparation

Herbal medicine complex extract 200 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added to purified water according to the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed and then purified water was added thereto to adjust the total volume to 100 ml. The solution was filled in a brown bottle and sterilized to prepare a liquid preparation do.

Preparation Example 6. Preparation of Health Functional Foods

Estuarine Complex Extract 200 mg

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

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

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a component suitable for a health functional food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above components may be mixed , Granules may be prepared and used in the manufacture of a health functional food composition according to a conventional method.

Formulation Example 7. Preparation of health drink

Herbal medicine complex extract 2,000 mg

Citric acid 1,000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Total purified water 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the health beverage composition of the invention.

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

none

Claims (4)

Characterized in that it contains 11 to 20 parts by weight, 15 to 30 parts by weight, 15 to 30 parts by weight, and 1 to 17 parts by weight of mixed extract of Moroheiya, rhubarb, Health functional foods that are effective for improving function
The method of claim 1, wherein the herbal medicine mixed extract is soluble in a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
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