KR20190017394A - A composition for treating or improving hepatic fibrosis comprising Seahorse extract - Google Patents

Abstract

The present invention relates to a composition for treating or alleviating hepatic fibrosis disease containing Hippocampus coronatus extract as an active component. The composition of the present invention has an effect of treating or alleviating hepatic fibrosis by checking an inhibitory effect of hepatic fibrosis by using a hepatic stellate cell hepatic fibrosis model which causes hepatic fibrosis.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for treating or improving liver fibrosis,

The present invention relates to a composition for treating or improving liver fibrosis including hippocampus extract.

The liver is located between the digestive system and the systemic circulatory system, so it is a very important organ that performs the function of defending the whole body from the exogenous substances that enter the digestive system. Since the in vivo exogenous material passes through the liver, the liver has a higher risk of exposure to many toxic substances than other organs, and the probability of damaging the liver is higher than other organs.

Among liver tissues, hepatocytes that perform liver functions such as nutrient absorption and metabolism, storage and plasma protein synthesis, Kupffer cells which are macrophages in the liver, Hepatic stellate cells (HSC) and endothelial cells and pit cells, which cause liver fibrosis, are present. Hepatocyte is a parenchymal cell that accounts for more than 90% of the total cells constituting the liver tissue and performs the main functions of the liver.

Chronic liver injury caused by various causes commonly leads to liver fibrosis. When hepatocytes are damaged by various causes and Cooper cells are activated and secrete several cytokines, hepatic cells are activated to form connective tissues. When liver damage is sustained, damaged liver cells are replaced with connective tissues, resulting in repair of epilepsy, resulting in excessive scarring of connective tissues in liver tissue, resulting in hepatic fibrosis and liver cirrhosis due to hepatic cell damage (Friedman SL et al., J. Hepatol., 38 (1), pp. 38-53, 2003).

Hepatic fibrosis treatment has been studied mainly on hepatic stellate cells (HSCs), which produce excessive connective tissue which is the most problematic in liver fibrosis. (Boker et al., J. Hepatol., 13 (3), pp. 35-40, 2002). In addition, in order to develop liver fibrosis therapeutic agents, 1991, Bataller R et al., Semin. Liver Dis., 21, pp437-451, 2001).

[Prior Patent Literature]

Korean Patent Publication No. 1020150111710

The present invention has been made in view of the above needs, and an object of the present invention is to provide a novel composition for treating or improving liver fibrosis.

In order to accomplish the above object, the present invention provides a composition for treating or improving liver fibrosis comprising an extract of hippocampus as an active ingredient.

In one embodiment of the present invention, the composition preferably contains, as an active ingredient, an extract of hippocampus, rhubarb, scabbard, chewy, It is more preferable that the composition ratio of phytase, horsetail, and phaeton is equal to or less than 0.5 weight, but not limited to, hippopotamus, rhubarb, hippopotamus, chewy,

In another embodiment of the present invention, it is preferable that the composition contains, as an active ingredient, hippocampus, rhubarb, scabbard, chewy, hibiscus, radish, angelica, More preferably, 0.5 weight, but not limited to, the same weight of the composition, the composition ratio of the water-in-oil, water-in-oil, water-in-oil, water-in-oil, water-in-oil No.

The present invention can provide a composition capable of preventing or treating liver fibrosis diseases, including hippocampus extract and the like.

The extract according to the present invention can be obtained by extracting and isolating from nature using an extraction and separation method known in the art, and the 'extract' defined in the present invention can be obtained by using an appropriate solvent in the following examples It is extracted from the material described.

As a suitable solvent for extracting the extract from the materials described in the following examples, any of pharmaceutically acceptable solvents may be used, and water or an organic solvent may be used, and for example, Alcohol having 1 to 4 carbon atoms, acetone, ether, benzene, and the like, including methanol, ethanol, propanol, isopropanol, butanol, Various solvents such as chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane may be used alone or in combination. More preferably, ethanol (alcohol) can be used.

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. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. The method for producing the extract of the present invention is not limited, and any known method can be used.

Therefore, the extract in the present invention is a concept including all the extract, fraction and purified product obtained in each step of extraction, fractionation or purification, and a diluted solution, a concentrate or a dried product thereof.

The composition of the present invention containing the above extract as an active ingredient may be a pharmaceutical composition or a food composition.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into 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. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles 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 that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

In one embodiment of the present invention, the extract of the present invention may be contained in an amount of 0.1 to 10% by weight based on the total weight of the composition.

The composition of the present invention may also be a food composition. In addition to containing an extract as an active ingredient, such a food composition may contain various flavors or natural carbohydrates as an additional ingredient such as a conventional food composition.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

The food composition of the present invention can be formulated in the same manner as the above 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, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .

In addition, the food composition may contain various additives such as various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate, etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

Since the extract as an active ingredient of the present invention is a natural substance with little toxicity and side effects, it can be safely used for long-term administration for the purpose of prevention or treatment of liver fibrosis.

The present invention also provides a health functional food for improving liver function comprising the extract of the present invention as an active ingredient.

The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and rings for the purpose of improving liver function.

In the present invention, the term "health functional food" refers to foods manufactured and processed using raw materials or ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods, Or to obtain a beneficial effect in health use such as physiological action.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the items listed in the above-mentioned 'food additives' include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, the health functional food in the form of tablets may be prepared by granulating a mixture obtained by mixing the extract of the present invention as an excipient, a binder, a disintegrant and other additives in a usual manner, , The mixture can be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule type health functional food can be prepared by filling a normal hard capsule with a mixture of an extract of the present invention and an additive such as an excipient and the soft capsule is mixed with an excipient such as an excipient Can be prepared by filling a mixture into a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of the extract of the present invention with an excipient, a binder, a disintegrant, and the like by a conventionally known method, and if necessary, Or the surface may be coated with a material such as starch, talc.

The granular health functional food may be prepared by granulating a mixture of an extract of the present invention with an excipient, a binder, a disintegrant and the like in a granular form by a conventionally known method and, if necessary, adding a flavoring agent, ≪ / RTI >

The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.

The present invention also provides the use of a composition comprising an extract for the manufacture of a medicament for the prevention or treatment of liver fibrosis disease or food, as an active ingredient. The composition of the present invention comprising the above extract as an active ingredient can be used for the manufacture of medicines or foods for the prevention or treatment of liver fibrosis diseases.

The present invention also provides a method of preventing or treating liver fibrosis diseases comprising administering an extract to a mammal.

As used herein, the term " mammal " refers to a mammal that is the subject of treatment, observation, or experimentation, preferably a human.

As used herein, the term " therapeutically effective amount " refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, The amount that induces the relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the therapeutic method of the present invention, in the case of an adult, it is preferable to administer the extract of the present invention at a dose of 1 mg / kg to 250 mg / kg once to several times a day.

In the therapeutic method of the present invention, the composition comprising the extract of the present invention as an active ingredient may be administered orally, rectally, intravenously, intraarterally, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, ≪ / RTI >

As can be seen from the present invention, the composition of the present invention has the effect of treating or improving liver fibrosis by confirming the effect of inhibiting hepatic fibrosis using a hepatic fibroblast model which induces liver fibrosis.

1 shows the cytotoxic effect of the composition of the present invention on human interleukin
Figure 2 shows the appearance of cells after treatment with the composition of the present invention in human trunk
FIG. 3 is a graph showing the expression amount of alpha-SMA in trunk cells after treatment with the composition of the present invention in HepG2 cells induced by OA and the culture medium cultured in Hepatocyte was treated with human liver astrocytes.

The present invention will now be described in more detail by way of non-limiting examples. The following examples are intended to illustrate the present invention and the scope of the present invention is not to be construed as being limited by the following examples.

Example 1: cell  toxicity

(0, 3.125, 6.25, 12.5, 25, 50, 100, 200, ug / ml) after seeding in a 96 well plate at 1 × 10 ⁴ cells / well using human trunk line (LX-2) ) For 24 hours. After 24 hours, 50ul / well MTT reagent was added and reacted. After 4 hours, absorbance was measured at 570nm using ELISA.

Example  2: Preparation of the composition of the present invention

SH1: Seahorse 8g, Rhubarb 8g, Slut 8g, Shame 8g, Walnut 4g, Fado 4g

SH2: Seahorse 8g, Rhubarb 8g, Crusher 8g, Cheongpyige 8g, Angelica 8g, Kwangju 8g, Tassa 8g, Walnut 4g, Fado 4g

SH3: Seahorse 8g, Rhubarb 8g, Crusher 8g, Cheongpyee 8g, Sukjihwang 8g, Tag 8g, Walnut 4g, Fado 4g

SH4: Seahorse 8g, Rhubarb 8g, Crusher 8g, Cheongpyige 8g, Angelica 8g, Walnut 4g, Fado 4g

Each of the above materials was pulverized with a mixer, and then 10 times of 50% fermented juice was added. After extraction at 50 ° C for 4 hours, filtration, concentration and freeze-drying were carried out to obtain a powdery state at a concentration of 200 mg / ml in DMSO Respectively.

Example  3: Western blot analysis

The cells were washed three times with PBS after discarding the cells of the cells treated with hippocampus and hippocampus for 24 hours. Cells were harvested using a scraper. Cells were harvested by centrifugation, and RIPA reagent was added to dissolve the cells. . The separated proteins were quantified using a protein quantification kit, and 30 ug of proteins were electrophoresed on SDS polyacrylamide gels. The proteins were separated in order of size, and the proteins in the gel were transferred to the nitrocellulose membrane. The primary antibody was reacted for one day, and the secondary antibody was reacted for 1 hour, and the specific protein reacted with the antibody was detected using an ECL detection reagent.

 The results of the above embodiment are as follows.

Cytotoxicity results

Human hepatocellular carcinoma cells are located between hepatocytes. When hepatocyte is abnormal, hepatocyte subpopulation becomes active. When activated, its shape changes to myofibroblast type and the expression of factors inducing fibrosis is high. Thereby increasing the activity and distribution of the surrounding connective tissue.

However, the liver becomes more and more fibrous. Since the hepatic cell itself should have no toxicity during the treatment of the drug, the cell viability was measured by the MTT method. As shown in the graph in FIG. 1, the composition (1) and other compositions At the concentration of 100 ug / ml, the survival rate was 80% for SH 1, 100% for SH 2, 140% for SH 3, and 78% for SH 4. The survival rate of% SH 3 and SH 4 was 67% and 44%, respectively, and the survival rate was less than 50% at the composition excluding SH 2 and 3 at 200 ug / ml. It was confirmed that the cell survival rate was not affected as in Fig.

OA-induced HepG2 cells were treated with hippocampus and hippocampus, and then cultured in Hepatocyte,

OA-induced HepG2 is a typical fatty liver model with fat, hepatocarcinoma itself is hepatocarcinoma, hepatocyte is becoming progressive hepatocyte, so it can sufficiently activate normal hepatocellular tissue and may cause fibrosis Type cells. ≪ / RTI > It has been reported in many literature that factors such as TGF beta-1 TNFalpha, EGF, and IGF of hepatocyte-bearing hepatocytes are released from the cell and converted to an active state in which the trunk cell is fibrified. OA-induced HepG2 cells were treated with the composition of the present invention for 48 hours. Then, only the culture medium was collected by centrifugation, and treated with LX-2 cells (5X10 ⁵ cells / well) seeded in ⁶ wells for 24 hours, And markers of fibrosis. As a result, the stem cell specimens cultured with the culture medium of HepG2 cells induced with OA expressed more alpha-SMA than the stem cells treated with HepG2 culture medium induced with OA, and the compositions of the present invention were alpha-SMA Lt; / RTI >

Claims (5)

A composition for treatment or improvement of liver fibrosis comprising hippocampus extract as an active ingredient. The composition for treatment or improvement of liver fibrosis according to claim 1, wherein the composition comprises hippocampus, rhubarb, scabbard, chewy gum, scarlet fever, taxa, wheat germ, and peach extract as an active ingredient. [3] The method according to claim 2, wherein the composition ratio of the hippocampus, rhubarb, scabbard, cheongpiju, taxa, taxa, Is 0.5 wt.%. The composition for treatment or improvement of liver fibrosis according to claim 1, wherein the composition comprises hippocampus, rhubarb, scabbard, chewy, moss, persimmon, angelica, rhododendron and phytase extract as active ingredients. The method according to claim 4, wherein the composition ratios of the hippocampus, rhubarb, rhizome, cheongpiju, angelica, rhizopus, phlox, , Wort, and flax are 0.5 wt.%.

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