KR20220103862A - Pharmaceutical composition for improving liver function or preventing and treating endocrine system diseases - Google Patents

Abstract

The present invention relates to a pharmaceutical composition, and more particularly, to a pharmaceutical composition that is extracted and fermented from natural raw materials and is effective in improving liver function or preventing and treating endocrine system diseases. The pharmaceutical composition of the present invention contains a composite fermented product obtained by fermenting a composite extract extracted from natural raw materials containing Cudrania tricuspidata, Artemisia capillaris, Curcuma longa, Crataegus pinnatifida, Salvia miltiorrhiza, Poncirus trifoliate, and Chinemys reevesii.

Description

Pharmaceutical composition for improving liver function or preventing and treating endocrine system diseases {Pharmaceutical composition for improving liver function or preventing and treating endocrine system diseases}

The present invention relates to a pharmaceutical composition, and more particularly, to a pharmaceutical composition extracted and fermented from natural raw materials to improve liver function or prevent and treat endocrine diseases.

The endocrine system is a group of glands and organs that regulate and control various human functions by producing and secreting hormones that control metabolism throughout the human body. disease will occur.

In recent years, the incidence of endocrine system diseases is increasing due to changes in lifestyle. In particular, various endocrine diseases are caused by disruption of the balance of endocrine factors due to excessive accumulation of adipose tissue, which is one of the endocrine systems. Specifically, adipose tissue is the largest endocrine tissue in the living body, and it produces various endocrine factors such as adiponectin and leptin, and is involved in maintaining body homeostasis, but excessive accumulation of visceral fat due to lifestyle changes such as eating habits causes endocrine factors A number of endocrine diseases can be invented as the secretion of is excessively secreted or decreased.

These, endocrine system diseases are typically diabetes, obesity, hypertension, etc., as is well known, endocrine system diseases are difficult to treat, and continue to be chronic and cause disorders in the metabolism of carbohydrates, lipids and proteins, retina, kidney, It can lead to complications such as nerve and cardiovascular system.

As endocrine diseases are difficult to treat with surgery, improvement of symptoms and prevention of complications are important in treatment. Although there are drug therapy, diet therapy, and exercise therapy as treatment methods, commonly used drug therapy is a constant problem with side effects and patient tolerance due to drug taking, so it is necessary to develop an endocrine therapy using natural products.

(Patent Document 1): Republic of Korea Patent Publication No. 10-2020-0018919

It is an object of the present invention to provide a pharmaceutical composition extracted and fermented from natural raw materials to improve liver function or prevent and treat endocrine diseases.

The present invention is a pharmaceutical composition for improving liver function or endocrine system disease and treatment, and a complex fermented product obtained by fermenting a complex extract extracted from natural raw materials including Cujipon fruit, Injinho, turmeric, hawthorn, ginseng, ginseng and gwipan as an active ingredient. do.

In the pharmaceutical composition for improving liver function or endocrine system disease and treatment according to an embodiment of the present invention, the natural raw material is 10 to 200 parts by weight of the Injinho, 10 to 200 parts by weight of the turmeric with respect to 100 parts by weight of the cucurbita fruit. parts, 10 to 200 parts by weight of the hawthorn, 10 to 200 parts by weight of the ginseng, 10 to 200 parts by weight of the ground ginseng, and 10 to 200 parts by weight of the ear plate.

As a pharmaceutical composition for improving liver function or endocrine system disease and treatment,

In the pharmaceutical composition for improving liver function or endocrine system disease and treatment according to an embodiment of the present invention, the natural raw material is 80 to 120 parts by weight of the Injinho, 80 to 120 parts by weight of the turmeric with respect to 100 parts by weight of the Cuji mulberry fruit parts, 80 to 120 parts by weight of the hawthorn, 80 to 120 parts by weight of the ginseng, 20 to 100 parts by weight of the ground ginseng, and 20 to 100 parts by weight of the ear plate.

In the pharmaceutical composition for improving liver function or endocrine system disease and treatment according to an embodiment of the present invention, the complex fermented product may be obtained as a freeze-dried powder after drying under reduced pressure.

In the pharmaceutical composition for improving liver function or endocrine system disease and treatment according to an embodiment of the present invention, the complex fermented product may be freeze-dried at -70°C to -90°C.

In the pharmaceutical composition for improving liver function or endocrine system disease and treatment according to an embodiment of the present invention, the complex fermented product is inoculated with a Bacillus subtilis strain to the complex extract, and after primary fermentation, lactic acid bacteria (Lactobacilus) platarum) strain may be inoculated and fermented secondarily.

In the pharmaceutical composition for improving liver function or endocrine system disease and treatment according to an embodiment of the present invention, the complex fermented product is Bacillus subtilis 3 to 7 parts by weight and lactic acid bacteria ( Lactobacilus platarum) strain 0.5 to 3 parts by weight may be inoculated and fermented.

In the pharmaceutical composition for improving liver function or endocrine system disease and treatment according to an embodiment of the present invention, the complex extract may be a reflux extraction of a natural raw material in hot water.

The present invention is a health food composition comprising, as an active ingredient, a complex fermented product obtained by fermenting a complex extract extracted from natural raw materials including Cuji mulberry fruit, Injinho, turmeric, hawthorn, ginseng, jisil and gwipan.

The pharmaceutical composition according to the present invention is effective in improving liver function and preventing and treating endocrine system diseases, and is derived from natural raw materials and has secured stability and can be usefully used as a preventive or therapeutic agent.

1 is a TLC analysis result of the pharmaceutical composition of the present invention,
2 is a weight loss effect result of the pharmaceutical composition of the present invention;
3 is a result of the blood glucose reduction effect of the pharmaceutical composition of the present invention,
4 to 6 show the liver function improvement effect of the pharmaceutical composition of the present invention,
7 to 14 are graphs showing the results of improving the lipid metabolism function of the pharmaceutical composition of the present invention.

Unless otherwise defined in technical terms and scientific terms used in this specification, those of ordinary skill in the art to which this invention belongs have the meanings commonly understood, and in the following description and accompanying drawings, the subject matter of the present invention Descriptions of known functions and configurations that may unnecessarily obscure will be omitted.

Also, the singular forms used herein may be intended to include the plural forms as well, unless the context specifically dictates otherwise.

In addition, in the present specification, the unit used without special mention is based on the weight, for example, the unit of % or ratio means weight % or weight ratio, and weight % means any one component of the entire composition unless otherwise defined. It means % by weight in the composition.

In addition, the numerical range used herein includes the lower limit and upper limit and all values within the range, increments logically derived from the form and width of the defined range, all values defined therein, and the upper limit of the numerical range defined in different forms. and all possible combinations of lower limits. Unless otherwise defined in the specification of the present invention, values outside the numerical range that may occur due to experimental errors or rounding of values are also included in the defined numerical range.

As used herein, the term 'comprising' is an open-ended description having an equivalent meaning to expressions such as 'comprising', 'containing', 'having' or 'characterized', and elements not listed in addition; Materials or processes are not excluded.

terms in this specification. The term “endocrine system disease” refers to diabetes, obesity, hypertension, low lipid metabolism, and the like.

As used herein, the term “prevention” refers to any action that inhibits or delays the onset of endocrine diseases by administration of the composition of the present invention.

As used herein, the term "treatment" refers to any action that improves or beneficially changes the symptoms of endocrine system diseases by administering the composition of the present invention.

As used herein, the term "food" refers to a natural product or processed product containing one or more nutrients, and preferably means a state that can be directly eaten through a certain processing process, and has a conventional meaning As such, it is intended to include all foods, food additives, functional foods and beverages.

As used herein, the term "functional food" refers to a food group or food composition that has added value to act and express the function of the food for a specific purpose by using physical, biochemical, or bioengineering methods, etc. to regulate the biological defense rhythm, It refers to food that has been designed and processed to sufficiently express biological control functions related to disease prevention and recovery, etc., and includes “health functional food” in particular.

As used herein, the term "health supplement" refers to a food manufactured and processed using raw materials or ingredients having useful functions in the human body, and has a useful effect for health purposes such as regulating nutrients or physiological action with respect to the structure and function of the human body. It refers to food manufactured and processed by extracting, concentrating, refining, or mixing ingredients contained in food raw materials or using specific ingredients as raw materials for the purpose of health supplements to obtain health benefits. Conventionally, as endocrine diseases are difficult to treat with surgery, improvement of symptoms and prevention of complications are important in treatment. Although there are drug therapy, diet therapy, and exercise therapy as treatment methods, commonly used drug therapy is a constant problem with side effects and patient tolerance due to drug taking, so it is necessary to develop an endocrine therapy using natural products.

The present invention is a pharmaceutical composition for improving liver function or endocrine system disease using, as an active ingredient, a complex fermented product obtained by fermenting a complex extract extracted from natural raw materials including Cuji mulberry fruit, Injinho, turmeric, hawthorn, dandelion, ginseng and gwipan. Since it is derived from raw materials, safety is ensured and blood pressure rise is suppressed. It can reduce blood sugar and weight, and improve liver function and lipid metabolism.

Specifically, Cudrania tricuspidata is a deciduous tree belonging to the mulberry family, which is called a mulberry tree, a Gutgashi tree, and a mulberry tree. The root bark, bark, and leaves of the mulberry tree contain various components effective for the human body, so they have been used as medicines such as antihypertensive, tuberculosis treatment, antipyretic, diuretic, and terrestrial agent depending on the site.

In particular, Cuji mulberry fruit is grown in autumn, and it has a taste that combines bitterness and sweetness. Its efficacy is to lower fever, remove residual heat from blood, relax muscles and tendons, and allow meridians to flow well. It is also used as a stamina food as well as to replenish yang energy.

The present invention uses the fruit of the Cudrania mulberry tree, and the general components of the Cudrania fruit include moisture 76.0-80.1%, crude protein 2.2-3.5%, crude fat 1.7-2.9%, crude fat 0.8-1.2%, carbohydrate 14.5-16.4% Inorganic components include Ca, Fe, K, Mg, Na, P, etc., and it is known that insoluble dietary fiber is significantly more than soluble dietary fiber.

Injinho ( Artemisia capillaris Thunb . ) grows in sandy soils at the foot of mountains and streams. From late spring to early summer, cut young outposts that grow about 10 to 15 cm and dry them in the shade. The taste is bitter and bitter, and the nature is cool. Acts on the liver, nasal, and bladder. It lowers heat, removes dampness, and makes urine come out well. In pharmacological experiments, it has been found to have an antidiarrheal action, antipyretic action, blood pressure lowering action, blood sugar level lowering action, promoting recovery of damaged liver parenchyma, antibacterial action, antiviral action, and roundworm paralysis action. It is used for hepatitis, urine discomfort, itching, scabies, and scabies. When used as an external medicine, wash with decoction. In other countries, Injinho is written as Saninjin (山茵陳).

In the present invention, dried outplants of Injinho may be used.

Turmeric generally refers to the use of the tuberous roots of several plants belonging to Curcuma as medicines, food, and coloring agents. Plants of the ginger family include turmeric ( Curcuma longa Radix ), onion turmeric ( C. aromatica ), Kwangseo ahchul ( C. kwangsiensis ), and bon ah chul ( C.zedoaria ). Commonly compared to turmeric, turmeric is a rhizome whereas turmeric is dried using tubers. It is native to or cultivated in Southeast Asia including southern China, India, and Okinawa, and is also cultivated in the central and southern regions of Korea. Originally, turmeric was given the name because it turns yellow and golden when mixed with alcohol. Other names include magic (horse gold), hwangul (黃鬱), silver gold (乙金), geogeum (乞金), jade gold (玉金), wanggeum (王金), and turmeric (深黃).

Hawthorn refers to dried medicinal herbs of the Ripe hawthorn ( Crataegus pinnatifida Bunge var. typica Schneider ) and the ripe fruits of plants and animals of the Rosaceae family. The name Sansa is given because the fruit tastes like apples and is red in color, like small apples. Because hawthorn fruits grow in the grassy forests of the mountains, monkeys and mice eat them well, so they are also called “seosa” or “husa” by adding the letters “who” or “mouse” (鼠). Also, because the shape of the mountain temple is similar to that of a red jujube, it was also called red tide.

Salvia miltiorrhiza BUNGE means a herbaceous perennial plant belonging to the family Lamiaceae. It resembles the shape of ginseng and is red in color, so it is called Dansam. The height is 40-80 cm, and there are many hairs throughout. The leaves are opposite to ovate or lanceolate. The back side is densely hairy and has dull sawtooths. Flowers are purple and bloom from May to June and run in layers. The roots are used as herbal medicine. It contains tansinnon and vitamin E, and it has been recognized that it dilates peripheral blood vessels and lowers blood pressure in animal experiments. Yakseong (藥性) is slightly cold and bitter in taste, it has antibacterial action against Staphylococcus aureus, E. For myocardial infarction, danhyang and barn are used in combination, and for insomnia and anxiety symptoms caused by nervous breakdown, keel and moryeo are used together to treat.

In addition, dansam's efficacy is recognized for women's menstrual pain, menstrual irregularities, and postpartum pain in the lower abdomen, as well as early symptoms of chronic hepatitis, liver dysfunction, and cirrhosis. It is also used for thrombophlebitis and hypertension, but is not used for bleeding disorders.

Ponciri Fructus (枳實) is the dried unripe fruit of the Poncirus trifoliata Rafinesque of the Rutaceae family . dry at low temperature. In oriental medicine, it has the efficacy of phagisojeok and hwadamsanbi, and has been used to treat red blood cells, obesity, intestinal pain, sarcoma, and stool incontinence.

Ingredients of oilseed oil include essential oils such as limonene, linalool, camphene, etc., and pravos such as poncirin, naringin, hesperidin, neohesperidin, etc. Coumarins such as noid and umbelliferone, auraptene, and imperatorin have been reported. Ponkyrin is known to have anti-platelet action, intestinal bacteria suppression action and anti-helicobacter pylori activity, and naringin has been reported to have cholesterol suppressive effect, change leukemia cells into normal cells, and inhibit breast cancer cell proliferation.

The ear plate (龜板, Testudinis Plastrum) is a part of the tortoise's abdominal or sternum, and contains colloids and fat, and is used for bodily, chronic wasting fever in pulmonary tuberculosis, oil wells, uterine bleeding, and pain in the pelvis. It works. Bokgap is also called earplate and baegap is called gipgap. The ear plate is an oval flat plate with 12 shells joined, 7-13cm long and 3-8cm wide. . There are two types of outer surfaces: those with a yellowish-brown pattern, and those with black and yellow only on the instep. The back surface is usually grayish white, and the quality is smooth.

The complex fermented product is fermented with the above-mentioned complex extract extracted from natural raw materials including Cuji mulberry fruit, Injinho, turmeric, hawthorn, ginseng, jisil and gwipan. GABA material is produced, blood sugar lowering, weight loss, and lipid compatibility improvement has a remarkable effect on In particular, by including the ear plate, it can have a significantly high effect in improving lipid metabolism.

Specifically, the natural raw material is based on 100 parts by weight of cucurbita fruit, 10 to 200 parts by weight of Injinho, 10 to 200 parts by weight of turmeric, 10 to 200 parts by weight of hawthorn, 10 to 200 parts by weight of ginseng, 10 to 200 parts by weight of ginseng and 10 to 200 parts by weight of the ear plate may be included. Specifically, with respect to 100 parts by weight of kkujippong fruit, 80 to 120 parts by weight of injinho, 80 to 120 parts by weight of turmeric, 80 to 120 parts by weight of hawthorn, 80 to 120 parts by weight of ginseng, 20 to 100 parts by weight of ginseng, and 20 to 100 parts by weight of ginseng. It may contain parts by weight. In the above range, it can have economic efficiency by including a large amount of active ingredients having an important meaning in substantially improving liver function and preventing and treating endocrine system diseases compared to the amount of material input.

The complex extract may refer to a mixture of active ingredients separated from natural raw materials, that is, substances exhibiting a desired activity. In one embodiment, the complex extract may be an extract of water, an organic solvent, or a mixed solvent thereof, and otherwise may include a dry powder thereof or any form formulated using the same. In another aspect, the complex extract may include fractionation of the extract that has undergone the extraction process. The complex extract may be, for example, an extract extracted by an ultrasonic extraction method, a reflux cooling extraction method, a hot water extraction method, or a known method using the organic solvent as an extraction solvent.

Preferably, the complex extract may be a hot water extract in which the natural raw material is extracted under reflux in hot water. Since hot water extract can contain a large amount of nutrients in natural raw materials, it can have more efficacy when used as a preventive and therapeutic agent for endocrine system diseases. For example, 100 parts by weight of a natural raw material may be put into 1000 to 2000 parts by weight of hot water and extracted under reflux for 2 to 3 hours. The hot water may be at a temperature of 40 to 70 °C, preferably 50 to 60 °C. When the hot water temperature is lower than the above range, extraction of the natural raw material is hardly made, and when the hot water temperature is higher than the above range, carbonization of the raw material occurs to obtain a very turbid complex extract.

The complex fermented product was obtained by inoculating the above-mentioned complex extract with a Bacillus subtilis strain and performing the primary fermentation, followed by inoculating the lactic acid bacteria ( Lactobacilus platarum ) strain to perform secondary fermentation, which can be obtained as a freeze-dried powder after drying under reduced pressure. have.

Specifically, the complex fermented product may be fermented by inoculating 3 to 7 parts by weight of Bacillus subtilis strain and 0.5 to 3 parts by weight of Lactobacilus platarum strain with respect to 100 parts by weight of the complex extract. Accordingly, GABA (γ-aminobutyric acid) effective for lowering blood sugar can be biosynthesized.

Hereinafter, the process of obtaining a complex fermented product by fermenting the complex extract of the present invention will be described in detail.

First, 5-7 parts by weight of MSG (L-sodium glutamate) is mixed with 100 parts by weight of the above-described complex extract and sterilized at 115-130° C. for 4-5 minutes. After the sterilization process, 2-4 parts by weight of glucose is mixed again based on 100 parts by weight of the complex extract, 3-7 parts by weight of Bacillus subtilis (Bacillus subtilis) is added and inoculated to perform the primary fermentation process. The primary fermentation process may be performed under shaking conditions at 40 to 45° C., preferably at 41° C., for 40 to 56 hours, preferably for 48 hours.

Then, 1-3 parts by weight of glucose and 4-7 parts by weight of skim milk are mixed with 100 parts by weight of the complex extract to the fermented product that has undergone the primary fermentation process, and lactic acid bacteria (lactobacillus plantarum) plantarum))) 0.5 to 3 parts by weight can be inoculated to perform the secondary fermentation process. The secondary fermentation process may be performed under shaking conditions at 28 to 33° C., preferably at 30° C., for 60 to 80 hours, preferably for 72 hours.

Thereafter, after drying under reduced pressure through a rotary vacuum evaporator, it may be freeze-dried and stored at -70°C to -90°C, specifically, at an ultra-low temperature of -75°C to -85°C. The freeze-drying pressure may be 3 to 80 torr, preferably 40 to 60 torr, and the time may be 5 to 48 hours, preferably 20 to 30 hours. Drying may proceed smoothly within the above range.

The pharmaceutical composition for the prevention and treatment of liver function and endocrine system disease of the present invention described above has safety as a natural ingredient, and exhibits liver function improvement ability, lipid metabolism activity function, and blood sugar lowering activity, thereby preventing liver function individual efficacy and endocrine system disease And it can be usefully used for treatment.

The complex fermented product of the present invention may be in various oral and parenteral formulations during actual clinical administration. When formulated, commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. can be formulated. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include one or more excipients, for example, starch, calcium carbonate, sucrose in the above herbal extract. ) or lactose (loctose), gelatin, etc. may be mixed and prepared. In addition, lubricants such as magnesium straight and talc may be further included in addition to simple excipients, and liquid formulations for oral use include suspensions, internal solutions, emulsions, and syrups. Various excipients in addition to commonly used simple diluents such as water and liquid paraffin , for example, wetting agents, sweetening agents, fragrances, preservatives, and the like may be included.

A sterile aqueous solution, a non-aqueous solvent, and a suspension for parenteral administration may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. It is not limited.

The composition according to the present invention may be formulated and used in various formulations such as tablets, capsules, granules, pills, suspensions, and syrups according to conventional methods known in the art to which the present invention pertains.

The complex fermented product of the present invention not only provides an excellent effect of improving liver function and treating or preventing endocrine system diseases, but also has no toxicity and side effects due to drugs, so it can be safely used even when taken for a long period of time.

Accordingly, the complex extract can be used as a main, auxiliary material, and food additive of food, and the present invention can be provided as a health food composition including a complex fermented product. The health food composition is to be included in health supplements, functional foods, food additives, and the like. The health supplements, functional foods, and food additives have an action for improving liver function or preventing endocrine system diseases. Functional foods and dietary supplements may further include a food supplementary additive that is pharmaceutically acceptable, and may further include suitable carriers, excipients and diluents commonly used in the manufacture of functional foods.

Foods to which the complex fermented product can be added include, for example, various foods, beverages, gum, tea, vitamin complexes, functional foods, and the like. In addition, in the present invention, food includes special nutritional foods (eg, formula milk, infant food, etc.), processed meat products, fish meat products, tofu, jelly, noodles (eg, ramen, noodles, etc.), health supplements, seasoned foods ( Ex, soy sauce, soybean paste, red pepper paste, mixed soy sauce, etc.), sauces, sweets (eg snacks), dairy products (eg fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruit and vegetable beverages, soy milk, fermented beverages, etc.) and natural seasonings (eg, ramen soup, etc.). Food, beverage or food additive may be prepared by a conventional manufacturing method.

In the present invention, functional beverage means a generic term for drinking to quench thirst or enjoy taste, and is intended to include functional beverages. The beverage is not particularly limited in other ingredients except for including the complex fermented product as an active ingredient as an essential ingredient in the indicated ratio, and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. Examples of natural carbohydrates include monosaccharides, such as glucose, fructose, etc. disaccharides, such as maltose, sucrose, etc. and polysaccharides, such as conventional sugars such as dextrin, cyclodextrin, and the like, and xylitol, sugar alcohols such as sorbitol and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatine, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. In addition, the composition of the present invention may further contain pulp for the production of natural fruit juice, fruit juice beverage, and vegetable beverage.

The fermented product of the present invention is a natural food and has almost no toxicity and side effects, so it can be safely used for long-term use for preventive purposes.

Hereinafter, the present invention will be described in more detail through Examples and Comparative Examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples.

(Example 1)

Cudrania mulberry fruit (Hongik Herb Farming Corporation), Injinho (Omni Herb Co., Ltd.), Turmeric (Omni Herb Co., Ltd.), Sansa (Omni Herb Co., Ltd.), Dansam (Omni Herb Co., Ltd.) 10 g each, and ginseng (Omni Herb Co. After mixing 7 g of Omni Herb Co., Ltd.), 700 ml of distilled water was added, extracted under reflux for 3 hours, and filtered through a filter. About 5% of MSG was mixed with respect to the filtrate (500 ml) extracted under reflux after filtration and sterilized at 121°C for 5 minutes. About 3% of glucose was mixed with respect to the sterilized filtrate, and 5% of Bacillus subtilis was inoculated, and the primary fermentation was performed while mixing at 42° C. for 48 hours.

After that, 1~1.5% (25~38g) of glucose and 4~5% (100~115g) of skim milk are mixed with the primary fermented product and inoculated with about 0.5~1% (15~25g) of Lactobacillus plantarum . The secondary fermentation was carried out while mixing at 30° C. for 72 hours.

The second fermented secondary fermented product was concentrated under reduced pressure with a rotary vacuum evaporator, and the concentrated solution was freeze-dried at -80° C. with a freeze dryer to obtain 12.21 g of powdered complex fermentation composition (fermentation mixed herbs extract or less, FMH) powder (yield 30.27) %) was obtained. The obtained complex fermentation composition was stored in a cryogenic freezer (-80° C.), and diluted in distilled water to prepare a concentration of 200 mg/kg. Hereinafter, it is denoted as FHE 200 in the drawings.

(Example 2)

It was prepared in the same manner as in Example 1, except that it was prepared at a concentration of 400 mg/kg rather than a concentration of 200 mg/kg in Example 1. Hereinafter, it is denoted as FHE 400 in the drawings.

(Comparative Example 1)

In Example 1, it was prepared in the same manner as in Example 1, except that the filtrate extracted under reflux was not fermented.

(Comparative Example 2)

In Example 1, it was prepared in the same manner as in Example 1, except that the ear plate was not added to the raw material.

Hepatic function and endocrine system disease prevention and treatment evaluation experiment.

1. Experiment preparation

C57BL/6, db/db mouse (5 weeks old, male, 20-27 g) used for this experiment was purchased from Samtaco (Korea) and used. Experimental animals were acclimatized while having a stable period of 2 weeks, and 300 g of general feed (ENVIGO CO., U.K.) and sufficient water were supplied to all experimental groups every week during the stabilization period and experimental period. After the induction period of 1 week, animal experiments were conducted from 7 weeks of age. Experimental animals were divided into groups after the blood glucose was checked in the control group and the experimental group. The conditions of the animal breeding room were 22±2℃ with a conventional system, 200-300 Lux for 12 hours a day, and all light was blocked for 12 hours. This experiment was approved by the Animal Experimentation Ethics Committee of Daejeon University (approval number DJUARB2020-023) and was conducted in accordance with the animal ethics rules.

2. Reagents and Instruments

The reagents used were blood glucose check strip (Roshe CO., Switzerland), Morinaga ultra sensitive mouse insulin ELISA kit (MioBS CO., Japan), Mouse/Rat Leptin Quantikine ELISA Kit (R&D system, U.S.A.), Mouse Adiponectin/Acrp30 Quantikine ELISA Kit (R&D system, U.S.A.), Mouse Resistin Quantikine ELISA Kit (R&D system, U.S.A.), formaldehyde (Sigma Co., U.S.A.), etc. were used.

The instruments used were Autoclave (Sanyo Co., Japan), Vortex mixer (Vision scientific Co., Korea), Centrifuge (Sigma Co., U.S.A.), Deep-freezer (Sanyo Co., Japan), Ice-maker (Vision scientific) Co., Korea), plate shaker (Lab-Line Co., U.S.A.), ELISA reader (Molecular Devices Co., U.S.A.), blood glucose checker (Roshe CO., Switzerland), automatic biochemical analyzer (Hitachi, Japan), etc. did.

3. Experiment

C57BL/6 mouse, known as the origin of db/db mouse, was classified as a normal group, and db/db mouse, known as a diabetes development model, was divided into a control group and an experimental group. The control group and the experimental group checked the blood glucose of 7-week-old mice, calculated the average, and divided 6 mice per group. From the 7th week of the experiment to the 11th week of the experiment, the normal group was not treated with anything, and the negative control group was orally administered with distilled water. The positive control group was administered glimepiride, a sulfonylurea-based diabetes treatment, at a concentration of 1 mg/kg, and the experimental group set the FHE at 200 and 400 mg/kg, once a day, orally at 10 am for 4 weeks. administered.

4. Statistical processing

Experimental results were expressed as mean±standard error of mean using SPSS 21.0, multiple comparisons were performed using ANOVA, and significance was tested at p<0.05, p<0.01, and p<0.001 levels through Tukey's HSD test.

(Experimental Example 1) TLC analysis

For qualitative analysis of MSG and GABA, a silica gel TLC plate was cut into a size of 10 × 20 cm, and TLC development was performed in a square chamber (30 × 25 × 10 cm). MSG 0.5% and GABA 0.5% were used as standards for comparing MSG residual amount and GABA content. As a developing solvent, acetic acid glacial: n-butylalcohol: distilled water was mixed in a ratio of 1:3:1 (v/v) and saturated at room temperature for 3 hours or more. After diluting the fermented product two-fold with distilled water, 2 μl of each sample and standard solution was added dropwise to a position 15 mm below the TLC plate, and the interval was maintained at 10 to 15 mm. After dripping, the sample of the TLC plate was dried and then developed, and the TLC plate after development was dried in a vacuum dryer at 50°C. A 0.2% ninhydrin solution, a color developing reagent, was sprayed on the dried TLC plate, and the color was developed in a vacuum dryer at 100° C. for 5 to 10 minutes, and then glutamic acid and GABA spots of the fermented product were confirmed.

1 shows the results of confirming the production of GABA through TLC analysis. Referring to FIG. 1 , it was found that GABA of about 0.2% was produced by consuming MSG as lactic acid fermentation was performed for 72 hours through fermentation.

(Experimental Example 2) Weight evaluation

Body weight was measured using a g-unit scale before blood glucose measurement, and the results are shown in FIG. 2 below. Referring to FIG. 2 , the normal group was 24.79±1.08 g, the negative control group was 38.28±1.52 g, and the positive control group was 31.69±1.44 g, and the low and high FHE concentration groups were 34.69±1.44 g and 32.77±1.69 g, respectively appeared as The FHE administration group (Example 1 (200) and Example 2 (400)) showed a significant (**: p < 0.01, ***: p < 0.001) reduction compared to the negative control group.

(Experimental Example 3) Blood sugar analysis

Blood glucose was measured by fasting for 12 hours before autopsy, and using blood collected from the tail vein and a blood glucose monitor. The blood glucose measurement results are shown in FIG. 3 , and referring to FIG. 3 , the blood glucose measurement results showed that the normal group had 146.80±26.94 mg/dL. The negative control group showed 509.20±34.30 mg/dL, the positive control group showed 259.60±26.59 mg/dL, and the low and high FHE concentration group showed 415.80±31.25 mg/dL and 319.80±49.85 mg/dL, respectively. The FHE-administered group showed a significant (**: p<0.01, ***: p<0.001) decrease compared to the negative control group.

(Experimental Example 4) liver function improvement evaluation

After completion of the experiment, blood was collected using cardiac puncture, hardened at room temperature for 30 minutes, centrifuged at 3000 rpm for 15 minutes, and serum was separated to measure hepatic function indicators (AST, ALP, LDH), and then shown in FIGS. 4 to 6 shows the results, respectively.

1)AST analysis

4 is a graph of AST analysis results. Referring to FIG. 4 , as a result of analyzing AST in the blood, the normal group was 57.40±7.13 U/L, the negative control group was 113.00±10.65 U/L, and the positive control group was 86.80±6.91 U/L, and low FHE concentration. , the high concentration group showed 104.60±4.39 U/L and 67.40±6.19 U/L, respectively. The FHE high concentration group showed a significant (***: p<0.001) decrease compared to the negative control group.

2) ALP

5 is a graph showing the results of ALP analysis. Referring to FIG. 5 , as a result of analyzing ALP in the blood, the normal group was 274.80±36.53 U/L, the negative control group was 685.80±24.28 U/L, and the positive control group was 463.40±34.62 U/L, and low FHE concentration. , the high concentration group showed 611.20±44.87 U/L and 544.80±33.12 U/L, respectively. The FHE high concentration group showed a significant (***: p<0.001) decrease compared to the negative control group.

3)LDH analysis

6 is a graph showing the results of ALP analysis. 6 , as a result of analyzing LDH in the blood, the normal group was 86.80±6.87 U/L, the negative control group was 347.00±11.58 U/L, and the positive control group was 230.80±17.74 U/L, and low FHE concentration, The high concentration group showed 319.40±16.61 U/L and 291.40±9.66 U/L, respectively. The FHE-administered group showed a significant (*: p<0.05, ***: p<0.001) decrease compared to the negative control group.

(Experimental Example 5) Lipid metabolism evaluation

After the type of experiment, blood was collected using cardiac puncture, hardened at room temperature for 30 minutes, centrifuged at 3000 rpm for 15 minutes, and serum was separated, and lipid metabolism indicators (total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides) were measured using an automatic biochemical analyzer. was used for analysis.

1) Total cholesterol

7 shows a graph of cholesterol content results. 7, as a result of measuring the total cholesterol content in the blood, the normal group showed 120.40±10.09 mg/dL, the negative control group showed 214.00±11.83 mg/dL, and the positive control group showed 152.60±8.41 mg/dL, FHE low and high concentration groups were 194.20±7.33 mg/dL and 171.80±7.79 mg/dL, respectively. The FHE-administered group showed a significant (*: p<0.05, ***: p<0.001) decrease compared to the negative control group.

2) LDL-cholesterol

8 is a graph showing the LDL-cholesterol content results. Referring to FIG. 8 , as a result of measuring the low-density lipoprotein (LDL)-cholesterol content in the blood, the normal group was 61.60±2.41 mg/dL, the negative control group was 118.80±4.09 mg/dL, and the positive control group was 87.80±2.17 mg /dL, and the low and high FHE concentrations were 112.80±3.49 mg/dL and 100.20±4.97 mg/dL, respectively. The FHE-administered group showed a significant (*: p<0.05, ***: p<0.001) decrease compared to the negative control group.

3) HDL-cholesterol

9 is a graph showing the LDL-cholesterol content results. 9, as a result of measuring the high-density lipoprotein (HDL)-cholesterol content in the blood, the normal group was 6.60±0.89 mg/dL, the negative control group was 4.80±0.84 mg/dL, and the positive control group was 6.20±0.84 mg /dL, and the low and high FHE concentrations were 6.60±0.89 mg/dL and 7.40±0.89 mg/dL, respectively. The FHE-administered group showed a significant (*: p<0.05, **: p<0.01) increase compared to the negative control group.

4) Triglycerides

10 is a graph showing the result of the triglyceride content. Referring to FIG. 10 , as a result of measuring the triglyceride content in the blood, the normal group was 37.80±6.26 mg/dL, the negative control group was 170.80±7.85 mg/dL, and the positive control group was 114.80±5.50 mg/dL, and FHE The low and high concentration administration groups showed 181.60±4.39 mg/dL and 131.60±3.65 mg/dL, respectively. The FHE-administered group showed a significant (*: p<0.05) decrease compared to the negative control group.

(Experimental Example 6) Evaluation of lipid metabolism-related biomarker production in blood

1) Insulin measurement.

To measure the insulin content, blood was collected by cardiac puncture after the end of the experiment. Thereafter, the blood was hardened at room temperature for 30 minutes, centrifuged at 3,000 rpm for 15 minutes, and the serum was separated and measured using the Morinaga ultra sensitive mouse insulin ELISA kit as follows.

After putting 95 μl of sample diluent, 5 μl of standard and the sample in the coated insulin plate, the reaction was carried out at 4° C. for 2 hours. After the reaction, washing was performed using washing buffer solution, 100 μl of anti-insulin enzyme conjugate was added, and reacted for 30 minutes. After washing using washing buffer, 100 μl of enzyme substrate solution was added and reacted for 40 minutes. . After terminating the reaction by adding 100 μl of enzyme reaction stop solution, absorbance was measured in an ELISA reader 450 nm.

11 is a graph showing the result of measuring the insulin content. 11 , as a result of measuring the insulin content in the blood, the normal group showed 12.36±3.37 μg/mL, the negative control group showed 52.84±3.13 μg/mL, and the positive control group showed 32.03±2.19 μg/mL, and FHE The low and high concentration administration groups showed 43.72±2.30 μg/mL and 38.54±1.40 μg/mL, respectively. All experimental groups showed a significant (**: p<0.01, ***: p<0.001) decrease compared to the negative control group.

2) Resistin measurement

To measure the resistin content, blood was collected by cardiac puncture after the end of the experiment, and then the serum was separated by centrifugation at 3,000 rpm for 15 minutes. The resistin concentration through the separated serum was measured as follows using the Mouse Resistin Quantikine ELISA Kit. After each 50 μl of RD1W reagent was put into a 96 well plate, an additional 50 μl of standard, control, and 30-fold diluted serum were added and mixed using a plate shaker for 2 hours. Thereafter, 400 μl of wash buffer was added and washing was performed. Then, 100 μl of Mouse Resistin Conjugate was added and mixed again on a plate shaker for 2 hours. After washing again, 100 μl of Substrate Solution solution was added, reacted at room temperature for 30 minutes while blocking light, and 100 μl of Stop Solution was added, followed by measurement at a wavelength of 450 nm in an ELISA reader.

12 shows a result of resistancein measurement. 12, as a result of measuring the resistin content in the blood, the normal group was 302.55±48.51 ng/mL, the negative control group was 775.52±40.46 ng/mL, and the positive control group was 464.00±41.45 ng/mL, FHE. The low and high concentration administration groups showed 660.43±17.86 ng/mL and 518.42±48.91 ng/mL, respectively. All experimental groups showed a significant (**: p<0.01, ***: p<0.001) decrease compared to the negative control group.

3) Leptin measurement

To measure the leptin content, blood was collected by cardiac puncture after the end of the experiment, and then the serum was separated by centrifugation at 3,000 rpm for 15 minutes. The leptin concentration through the separated serum was measured as follows using the Mouse/Rat Leptin Quantikine ELISA Kit. After each 50 μl of RD1W reagent was added to a 96 well plate, an additional 50 μl of standard, control, and 50-fold diluted serum were added and mixed using a plate shaker for 2 hours. Then, 400 μl of wash buffer was added and washing was performed. Then, 100 μl of Mouse/Rat Leptin Conjugate was added and mixed again on a plate shaker for 2 hours. After washing again, 100 μl of Substrate Solution solution was added, reacted at room temperature for 30 minutes while blocking light, and 100 μl of Stop Solution was added, followed by measurement at a wavelength of 450 nm in an ELISA reader.

13 shows the measurement results of leptin. Referring to FIG. 13, as a result of measuring the leptin content in the blood, the normal group showed 2372.73±95.08 ng/mL, the negative control group was 1292.33±70.08 ng/mL, and the positive control group showed 2201.34±177.26 ng/mL, FHE. The low and high concentration groups were 1677.70±119.76 ng/mL and 1897.82±61.09 ng/mL, respectively. All experimental groups showed a significant (***: p<0.001) increase compared to the negative control group.

4) Adiponectin measurement

To measure adiponectin content, blood was collected using cardiac puncture after the end of the experiment, and then serum was separated by centrifugation at 3,000 rpm for 15 minutes. The adiponectin concentration through the separated serum was measured as follows using the Mouse Adiponectin/Acrp30 Quantikine ELISA Kit. After each 50 μl of RD1W reagent was added to a 96 well plate, an additional 50 μl of standard, control, and 2000-fold diluted serum were added and mixed using a plate shaker for 2 hours. Thereafter, 400 μl of wash buffer was added and washing was performed. Then, 100 μl of Mouse adiponectin Conjugate was added and mixed again on a plate shaker for 1 hour. After washing again, 100 μl of Substrate Solution solution was added, reacted at room temperature for 30 minutes while blocking light, and 100 μl of Stop Solution was added, followed by measurement at a wavelength of 450 nm in an ELISA reader.

14 shows adiponectin measurement results. Referring to FIG. 14 , as a result of measuring the adiponectin content in the blood, the normal group was 2.73±0.30 ng/mL, the negative control group was 1.87±0.15 ng/mL, and the positive control group was 1.95±0.18 ng/mL, and FHE The low and high concentration administration groups showed 1.95±0.17 ng/mL and 2.44±0.21 ng/mL, respectively. The FHE high concentration group showed a significant (**: p<0.01) increase compared to the negative control group.

(Experimental Example 7) Comprehensive evaluation

In order to compare the effects of the pharmaceutical compositions prepared in Examples 1 and 2 and Comparative Examples 1 and 2 as improving liver function and preventing or treating endocrine system diseases, a comprehensive evaluation was performed. In Experimental Examples 1 to 6, the pharmaceutical compositions of Comparative Examples 1 and 2 were used as test groups instead of the pharmaceutical compositions of Examples, respectively, and tested, and then the pharmaceutical compositions of Examples and efficacy were compared. Efficacy was evaluated by a five-point scale method, and efficacy results are shown in Table 4 below. The higher the efficacy, the closer to 5 points.

test substance Fermentation or not With or without earplates Comprehensive Assessment Score Example 1 (low concentration) O O 4 Example 2 (High Concentration) O O 5 Comparative Example 1 X O One Comparative Example 2 O X 3 physiological saline - - 0 Glimepiride - - 4

Looking at the results in Table 1, it was confirmed that Examples 1 to 2 containing the complex fermented product of the present invention had an effect as a preventive and therapeutic agent for liver function and endocrine system disease.

In particular, in the case of Comparative Example 1, which did not go through the fermentation process, almost no GABA production was able to determine the hypoglycemic effect, and in the case of Comparative Example 2, the content of the lipid metabolism-related biomarker was sufficiently reduced or increased compared to the Example. I could confirm that it didn't work.

As described above, the present invention has been described with specific matters and limited examples and drawings, but these are only provided to help a more general understanding of the present invention, and the present invention is not limited to the above embodiments, and the present invention is not limited to the above embodiments. Various modifications and variations are possible from these descriptions by those of ordinary skill in the art.

Therefore, the spirit of the present invention should not be limited to the described embodiments, and not only the claims to be described later, but also all those with equivalent or equivalent modifications to the claims will be said to belong to the scope of the spirit of the present invention. .

Claims (9)

A pharmaceutical composition for improving liver function or preventing and treating endocrine system diseases, using a complex fermented product obtained by fermenting a complex extract extracted from natural raw materials including Cuji mulberry fruit, Injinho, turmeric, hawthorn, ginseng, jisil and gwipan as an active ingredient.
According to claim 1,
The natural raw material is based on 100 parts by weight of the cucurbita fruit, 10 to 200 parts by weight of the Injinho, 10 to 200 parts by weight of the turmeric, 10 to 200 parts by weight of the hawthorn, 10 to 200 parts by weight of the ginseng, 10 to 200 parts by weight of the ginseng. A pharmaceutical composition for preventing and treating liver function improvement or endocrine system disease, comprising 10 to 200 parts by weight of the ear plate and 10 parts by weight.
3. The method of claim 2,
The natural raw material is based on 100 parts by weight of the cucurbita fruit, 80 to 120 parts by weight of the injinho, 80 to 120 parts by weight of the turmeric, 80 to 120 parts by weight of the hawthorn, 80 to 120 parts by weight of the ginseng, 20 to 100 parts by weight of the ginseng. A pharmaceutical composition for improving liver function or preventing and treating endocrine system diseases, comprising 20 to 100 parts by weight of the ear plate and 20 parts by weight.
According to claim 1,
The complex fermented product is a pharmaceutical composition for preventing and treating liver function improvement or endocrine system disease, which is obtained as a freeze-dried powder after drying under reduced pressure.
4. The method of claim 3,
The complex fermented product is a pharmaceutical composition for preventing and treating liver function improvement or endocrine disease that is lyophilized at -70 ℃ to -90 ℃.
According to claim 1,
The complex ferment is inoculated with a Bacillus subtilis strain to the complex extract and fermented first, and then inoculated with a lactic acid bacteria ( Lactobacilus platarum ) strain for secondary fermentation to improve liver function or prevent and treat endocrine system diseases pharmaceutical composition.
5. The method of claim 4,
The complex fermented product was fermented by inoculating 3 to 7 parts by weight of Bacillus subtilis strain and 0.5 to 3 parts by weight of Lactobacillus platarum strain with respect to 100 parts by weight of the complex extract to improve liver function or prevent endocrine system diseases and therapeutic pharmaceutical compositions.
According to claim 1,
The complex extract is a pharmaceutical composition for preventing and treating liver function improvement or endocrine system disease, which is a natural raw material extracted with reflux in hot water.
A health food composition comprising, as an active ingredient, a complex fermented product obtained by fermenting a complex extract extracted from natural raw materials including Cuji mulberry fruit, injinho, turmeric, hawthorn, ginseng, jisil and gwipan.

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