KR20200100342A - Pharmaceutical Composition Comprising Fraction of Red Ginseng Extract and Extract of Glycyrrhiza uralensis for Preventing or Treating Obesity - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating obesity containing a mixture of a licorice extract and a fraction of a red ginseng extract. The mixture of the licorice extract and the fraction of the red ginseng extract has an effect of reducing obesity, thereby being used as an active ingredient in drugs and health functional foods for reducing obesity.

Description

Pharmaceutical Composition Comprising Fraction of Red Ginseng Extract and Extract of Glycyrrhiza uralensis for Preventing or Treating Obesity, including fractions of red ginseng extract and licorice extract

The present invention relates to a pharmaceutical composition for preventing or treating obesity comprising a fraction of red ginseng extract and licorice extract.

Obesity or obesity is a term collectively referred to when a person's weight is too much. Diagnosed with obesity. In most cases, it means that the weight is more than normal, but even if the weight is not that much, it is said to be obese if the proportion of fat among the components of the body is high. Obesity by itself may not be a problem, but social disorders caused by it and secondary complications caused by excess fat are serious. For example, obesity can significantly increase the incidence of hyperlipidemia, high blood pressure, arteriosclerosis, diabetes, fatty liver, and joint abnormalities.

Obesity is characterized by an increase in adipose tissue mass resulting from an increase in the number of fat cells and an increase in the size of fat cells. The amount of adipose tissue mass can be regulated by inhibition of adipogenesis from fibroblast adipocytes to mature adipocytes and induction of lipolysis in adipose tissue.

3T3-L1 adipocytes are cell lines used to study adipocyte formation differentiation. Adipocyte differentiation is a complex process involving changes in morphology, hormone sensitivity, and gene expression. Fusion adipocytes become adipocytes through clonal proliferation, growth arrest, and expression of adipocyte markers by hormonal induction. Several transcription factors have been shown to play an important role in adipocyte differentiation. Adipocyte fatty acid binding protein (AP) and peroxisome proliferator-activated receptor-γ (PPAR-γ) proteins, known as representative transcription factors related to adipocyte differentiation, differentiate adipocyte formation through the transcription of various genes responsible for fat transport and accumulation. Plays an essential role in It is known that leptin mainly functions to regulate food, and it is known that blood content increases significantly when obese, and obesity is caused if there is an abnormality in the leptin receptor. Therefore, suppressing an increase in the leptin content in the blood can help prevent and improve various diseases caused by obesity.

The currently used treatment methods for obesity include diet therapy, exercise therapy, behavior therapy, drug therapy, and surgery therapy. Dietary therapy limits caloric intake and helps to consume fat in the body to induce weight loss, exercise therapy increases the body’s calorie consumption, and behavior therapy improves habits to induce weight control.

Drug therapy is to lose weight with an appetite suppressant, food absorption inhibitor, heat generation accelerator, and the like, and various drugs are currently used for weight loss. Among them, appetite suppressants act and affect the central nervous system, and are classified into drugs that act on the catecholamine-based and serotonin-based drugs of the central nervous system. Food absorption inhibitors act on the stomach and reduce the absorption of nutrients, such as lipolytic enzyme inhibitors. Thermogenic inhibitors promote the body's metabolic rate to increase calorie consumption and thereby reduce weight. There are thyroid hormone drugs, appetite suppressants, and sympathetic agents. Currently, the obesity treatment market is dominated by Xenical, a food absorption inhibitor, and Reductil, an appetite suppressant.

However, although there are differences in degrees of obesity treatments currently used, all of them have side effects, so the development of a new treatment for obesity with secured stability is urgently needed. A plant-derived material is being studied as a candidate raw material for which stability is secured, and for example, Patent Document 1 discloses the use of a rape flower extract for anti-obesity purposes.

Korean Patent Application Publication No. 2016-0058203

An object of the present invention is to develop a plant-derived material that can effectively improve obesity and is safe for the human body.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating obesity, including a fraction of red ginseng extract and a licorice extract, a health food composition for improving obesity, including a fraction of red ginseng extract and a licorice extract.

The composition comprising the fraction of the red ginseng extract and licorice extract of the present invention reduces fat accumulation.

1 shows the expression level of PPARγ when treated with a fraction of red ginseng extract, licorice extract, and a mixture thereof; Normal: normal group, Control: control, P1: red ginseng extract fraction, GU: licorice extract, PG (3:1): red ginseng extract fraction and a 3:1 mixture of licorice extract.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating obesity comprising a fraction of red ginseng extract and licorice extract.

Panax ginseng is a perennial dark herbaceous plant belonging to the genus Panax of the Araliaceae family, and has been used as an important medicinal material in oriental medicine for a long time. Ginseng is generally classified into white ginseng and red ginseng according to the processing method, and white ginseng refers to unprocessed ginseng mined in the field, that is, fresh ginseng as it is dried, and red ginseng is dried by steaming fresh ginseng. Several chemical changes are involved, including amino acid changes. Red ginseng produces saponin components such as ginsenosides Rg2, Rg3, Rh1, and Rh2, which are not present in ginseng by the heat applied during the manufacturing process. , It is excellent in protecting brain nerve cells and improving learning ability, antithrombotic action, and antioxidant action.

Licorice (Glycyrrhiza uralensis Fischer, Glycyrrhiza glabra Linne) is a perennial herbaceous plant belonging to the legume family. The stem grows to about 1m, and the root goes deep into the ground. The leaves are alternately formed, and the leaves are brackish. There are 7-17 small leaves each, and the shape is ovate or wide ovate. The flower is male color and blooms in July and August. It is well formulated with all drugs and has a neutralizing effect. Pharmacological action is effective in detoxification, hepatitis, hives, dermatitis, eczema, etc. Jinhae·Gotdam, muscle relaxation, diuretic, anti-inflammatory, and suppresses peptic ulcer.

In the present invention, the extract is ginseng or licorice leaves, young sprouts, body, body bark, stem, stem bark, root, root bark, rhizome, seed, fruit, immature fruit, ripe fruit, pulp, pericarp, flower, It can be obtained from any one selected from the group consisting of androecium, pistil group (gynoecium), calyx, stamen, petal, calyx piece, carpel, and combinations thereof. The rhizome is also referred to as rhizome, and indicates that the stem grows underground and acts like a root. The stamen group represents all stamens in one flower, and the pistil group represents all pistils in one flower. The carpel is a component that makes pistils of flowers, and generally represents a modified form of leaves, which are collectively referred to as flower leaves. The ginseng may be a root, and the licorice may be at least one of a root and a rhizome, but is not limited thereto, and an extract obtained from other parts of ginseng or licorice may be mixed and used for the purpose of improving muscle reduction.

With respect to 30 parts by weight of the fraction of the red ginseng extract, the licorice extract is 0.1 to 25 parts by weight, 1 to 25 parts by weight, 1 to 20 parts by weight, 3 to 17 parts by weight, 6 parts by weight It may be included in to 14 parts by weight, but is not limited thereto. If the licorice extract is included below the lower limit, even if the fraction of the red ginseng extract and the licorice extract are mixed, the effect of improving obesity is low. In addition, when the licorice extract exceeds the upper limit, the synergistic effect is lowered even if the fraction of the red ginseng extract and the licorice extract are mixed.

The term "extract" as referred to in the present invention refers to a material extracted from a raw material by any method, and is used to include all of the extracted extract, the concentrate obtainable therefrom, the dried product and powder of the concentrate without limitation.

The extract may be obtained by extracting from a raw material or a dried product thereof, and the raw material of the extract may be used without limitation, such as cultivated or commercially available ones.

When the extract is obtained by extracting from the raw material, all conventionally known extraction methods such as solvent extraction method, ultrasonic extraction method, filtration method and reflux extraction method can be used as the extraction method, and it is preferably prepared by using a solvent extraction method or a reflux extraction method. can do. The extraction process may be repeated several times, and thereafter may be additionally subjected to steps such as concentration or freeze-drying. Specifically, the obtained extract is concentrated under reduced pressure to obtain a concentrate, the concentrate is freeze-dried, and then a high-concentration extract powder can be prepared using a grinder. The extract also includes a fraction obtained by further fractionating the extract.

The extract may be extracted using water, an organic solvent, or a mixture thereof as an extraction solvent. The organic solvent is an alcohol, preferably a C ₁ -C ₄ lower alcohol, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, It may be any one selected from the group consisting of benzene and a mixed solvent thereof. When a mixture of water and an organic solvent is used as an extraction solvent, the mixture of water and an organic solvent may preferably be a mixture of water and a lower alcohol of C ₁ -C ₄ , more preferably a mixture of water and ethanol. I can.

The solvent used in the preparation of the red ginseng extract may be water or an organic solvent such as alcohol.

The solvent used in the preparation of the licorice extract was 10% (v/v) to 70% (v/v) ethanol aqueous solution, 13% (v/v) to 65% (v/v) ethanol aqueous solution, 16% (v/ v) to 60% (v/v) ethanol aqueous solution, 19% (v/v) to 55% (v/v) ethanol aqueous solution, 22% (v/v) to 50% (v/v) ethanol aqueous solution, 25 % (v/v) to 40% (v/v) ethanol may be an aqueous solution, but is not limited thereto. When the ethanol aqueous solution is less than the lower limit, the active ingredient is not extracted even if the extract is prepared, so obesity cannot be improved. When the ethanol aqueous solution is at a concentration exceeding the upper limit, obesity is not improved even if the fraction of the red ginseng extract and the licorice extract are mixed and treated.

When extracting red ginseng, the amount of solvent added may be 5 times to 20 times, 7 times to 18 times, and 9 times to 16 times the weight of the herbal medicine, but is not limited thereto. When extracting licorice, the amount of solvent added may be 5 times to 25 times, 8 times to 20 times, and 10 times to 15 times the weight of the herbal medicine, but is not limited thereto. Extraction may be carried out at 60 ℃ to 100 ℃, 65 ℃ to 95 ℃, 70 ℃ to 90 ℃, but is not limited thereto. Extraction may be performed for 4 hours to 20 hours, 4 hours to 16 hours, 4 hours to 12 hours, but is not limited thereto. Extraction may be performed once to 8 times, 1 to 6 times, 1 to 5 times, but is not limited thereto, and the extract may be a single extract obtained from each extraction or a mixed extract of extracts obtained from each extraction. Among the extraction conditions, when the amount, temperature and time of the solvent added during extraction are less than the lower limit or the number of extractions exceeds the upper limit, even if the fraction obtained from the red ginseng extract and the licorice extract are mixed, obesity cannot be improved.

Fractions of the red ginseng extract can be obtained by column-purifying fraction of the red ginseng extract. As the stationary phase used in the column purification fractionation of red ginseng water extract, fillers such as silica gel, activated alumina, synthetic polymer, magnesium silicate, activated carbon, cellulose, and ion exchange resin can be used, and silica gel or aromatic synthetic resin is used as a filler. It is preferable to use an aromatic synthetic resin, it may be an aromatic type synthetic adsorbent column, and it is more preferable that the Diaion HP-20 synthetic adsorbent is used as a filler, but is not limited thereto. Separation using the column may be performed once to several times until a fraction having a desired purity is purified, and concentration and recrystallization may be performed as necessary.

When preparing the purified fraction, the desorption solvent may be any one selected from the group consisting of water, an organic solvent, and a mixture thereof. The organic solvent is alcohol, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, benzene, acetone, acetonitrile, and a mixed solvent thereof. It may be any one selected from the group consisting of, preferably the organic solvent may be any one selected from the group consisting of acetone, acetonitrile, C ₁ to C ₄ alcohol and a mixed solvent thereof, more preferably the The organic solvent may be ethanol, but is not limited thereto.

When the desorption solvent is ethanol when preparing the purified fraction, the desorption solvent may be ethanol or an aqueous ethanol solution, and the desorption solvent is an aqueous ethanol solution of 40% (v/v) to less than 100% (v/v), 50% (v/v) to less than 100% (v/v) aqueous ethanol solution, 60% (v/v) to less than 100% (v/v) ethanol aqueous solution, 70% (v/v) to 100% (v /v) less than an aqueous ethanol solution, but may be ethanol, but is not limited thereto. When the concentration of the ethanol aqueous solution used as the desorption solvent is less than the lower limit, there is a problem that the effect of improving the obesity of the obtained fraction is lowered.

When the aqueous ethanol solution, which is a desorption solvent, is added during the preparation of the purified fraction, the aqueous ethanol solution is flowed through the column, for example, by pressurization.

The licorice extract may additionally perform a conventional fractionation process in order to increase the effect of improving obesity, and fractions obtained through the fractionation process are also included in the present invention. When preparing the fraction, the organic solvent used for preparing the extract may be used. Fractions obtained by passing the extract according to the present invention through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity, or affinity), etc. Active fractions obtained through various purification methods are also included in the fractions of the present invention. By separating the fraction in which several components are mixed according to the properties of the active component through concentration gradient column chromatography, etc., a specific active fraction having a higher effect of improving obesity can be prepared.

The column chromatography can separate and purify the active fraction using a stationary phase selected from the group consisting of silica gel, Sephadex, LH-20, ODS gel, RP-18, polyamide, Toyopearl, and XAD resin. . Column chromatography may be performed several times by selecting an appropriate stationary phase as necessary, but is not limited thereto. In using the chromatography, the elution solvent, the elution rate, and the elution time may be applied to a solvent, speed, or time generally used in the art.

The term "obesity" of the present invention refers to a metabolic disease caused by an imbalance between intake and consumption of calories, and refers to a state in which adipose tissue is abnormally increased due to excess calories. Men are considered obese when their body fat is 25% of their body weight and women are over 30% of their body weight. Clinically, the state of BMI (Body Mass Index) of 25.0-30.0 is defined as overweight, and the case of 30.0 or more is considered obese. It is defined as

In one embodiment of the present invention, a mixture of a fraction of red ginseng extract and a licorice extract was prepared in a ratio of 3:1, 1:1, and 1:3, and treated on mice fed a high-fat diet. When the mixture of the fraction of red ginseng extract and the mixture of licorice extract was treated, the group treated with the fraction of red ginseng extract and the mixture of licorice extract had a high fat diet, but compared to the control group that did not process the sample, weight, abdominal fat, leptin, dietary efficiency, and neutral Fat decreased. Among the mixtures of the fraction of red ginseng extract and the mixture of licorice extract, when the mixture was mixed at a ratio of 3:1, there was an effect of improving obesity compared to the fraction of the red ginseng extract, licorice extract, or mixtures mixed in other ratios.

In the present invention, "prevention" refers to any action that suppresses or delays obesity by administration of the composition, and "treatment" refers to any action in which obesity is improved or beneficially changed by the composition.

The pharmaceutical composition of the present invention can give a synergistic effect on the effect of the fraction of the red ginseng extract and the licorice extract within a range that does not impair the effect of the present invention in addition to the fraction and licorice extract of the red ginseng extract. It may further contain other ingredients and the like. Conventional adjuvants, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, or carriers may be included, for example.

The route of administration of the pharmaceutical composition includes oral, intravenous, intramuscular, intraarterial, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal, such as topical application by application. It can be applied in a way. The parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intralesional injection or infusion techniques.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type and severity of the individual, age, sex, type of disease, It can be determined according to the activity of the drug, its sensitivity to the drug, the time of administration, the route of administration and the rate of excretion, the duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and can be easily determined by a person skilled in the art.

The pharmaceutical composition of the present invention is not particularly limited as long as it is an individual for the purpose of improving obesity, and anything can be applied. For example, non-human animals such as monkeys, dogs, cats, rabbits, mormots, rats, mice, cows, sheep, pigs, goats, etc., humans, birds and fish can be used.

The pharmaceutical composition may further contain one or more active ingredients exhibiting the same or similar function in the fraction of the red ginseng extract and the licorice extract. Each of the above compositions may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, etc., external preparations, suppositories, and sterile injectable solutions according to a conventional method.

Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills, and the like. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. have.

Formulations for parenteral administration include powders, granules, tablets, capsules, sterilized aqueous solutions, solutions, non-aqueous solvents, suspensions, emulsions, syrups, suppositories, aerosols, and other external preparations and sterile injectables according to the usual methods. It can be formulated and used in the form of a cream, gel, patch, spray, ointment, warning agent, lotion, liniment, pasta, or cataplasma. However, it is not limited thereto. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used as the non-aqueous solvent and suspension. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.

The pharmaceutical composition may additionally contain an adjuvant such as a preservative, a stabilizer, a hydrating agent or an emulsification accelerator, a salt and/or a buffering agent for controlling the osmotic pressure, and other therapeutically useful substances. It can be formulated according to the formulation or coating method.

The dosage of the pharmaceutical composition may vary according to various factors including the individual's age, weight, general health, sex, administration time, route of administration, excretion rate, drug formulation, and the severity of a specific disease.

In addition, the pharmaceutical composition of the present invention may be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and methods of using a biological response modifier. In addition, the pharmaceutical composition may be administered at a dosage within the range of 0.001 to 200 mg/kg based on an adult, and when the pharmaceutical composition is for external use, once a day in an amount of 1.0 to 3.0 ㎖ based on an adult It is recommended to apply once and continue for at least 1 month, but the dosage is not intended to limit the scope of the present invention.

When formulating the pharmaceutical composition in a unit dosage form, the fraction of the red ginseng extract and licorice extract of the present invention as an active ingredient are preferably contained in a unit dose of about 0.01 to 1,500 mg, and the dosage required for adult treatment is Depending on the frequency and intensity of administration, the range of about 1 to 500 mg per day is usually but not limited thereto, and a higher daily dose may be desirable for some patients.

In addition, the present invention provides a health food composition for improving obesity comprising a fraction of red ginseng extract and licorice extract.

The mixture of the fraction of the red ginseng extract and the licorice extract is preferably contained in an amount of 0.005 to 20.0% by weight based on the total weight of the health functional food, more preferably contained in an amount of 0.01 to 10.0% by weight, but limited thereto. It doesn't work. When the effective content of the mixture of the fraction of the red ginseng extract and the licorice extract is less than 0.005% by weight, obesity cannot be alleviated, and when it exceeds 20% by weight, the effect of increasing the efficacy due to the increase in the content decreases, which is uneconomical.

In addition to the mixture of the fraction of the red ginseng extract of the present invention and the mixture of licorice extract, the health functional food is preferably increased in the effect of the mixture of the fraction of the red ginseng extract and the mixture of licorice extract within a range that does not impair the effect of the present invention. Other ingredients that may have an effect may additionally be included. Conventional auxiliaries, such as stabilizers, solubilizers, vitamins, pigments and fragrances, or carriers may be included, for example.

In addition, it may include ingredients that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. Examples of the carbohydrate include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, a natural flavoring agent taumatin, a stevia extract, and a synthetic flavoring agent may be used. For example, when the health functional food of the present invention is manufactured as a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, natural extract, etc. may be additionally included in addition to the fraction and licorice extract of the red ginseng extract of the present invention. .

There is no particular limitation on the kind of the health functional food. Examples of foods to which the mixture of the fraction of the red ginseng extract of the present invention and the mixture of licorice extract can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , Various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and includes all health functional foods in the usual sense.

Hereinafter, the present invention will be described in detail by way of manufacturing examples and examples.

However, the following Preparation Examples and Examples are only for illustrating the present invention, and the contents of the present invention are not limited by the following Preparation Examples and Examples.

Preparation Example 1. Preparation of fractions of red ginseng extract

1-1. Preparation of red ginseng extract

500g of 6-year-old red ginseng ( P. ginseng CA Meyer, root 75%, residual rice 25%) was dried at 110°C for 15 minutes using a far-infrared dryer (Korea Energy Technology Co., Ltd., HKD-LAB) and then cut off. 6 times of distilled water was added to 500 g of dried red ginseng, extracted for 10 hours at 85°C, and the extract was filtered and cooled to 0-10°C. Regarding the residue, 6 times of distilled water was added and extraction was further performed 1 to 4 times for 10 hours. The extract was filtered and then combined and cooled to 0 to 10°C. The cooled extract is centrifuged at 6,000 rpm for 10 to 20 minutes at 4°C (Hanil Cymed Co., Ltd., Supra 22K) to remove the sediment, and 50 to 60 using a rotary vacuum concentrator (BUCHI, R-220). It was concentrated to 71° Brix at °C. Finally, sterilized distilled water was added to the concentrate, diluted with 15° Brix, and then freeze-dried (OPERON, FDT-12012) to obtain 230 g of red ginseng extract powder.

1-2. Preparation of fractions of red ginseng extract

HP20 (Samyang Corporation, 10 L) was charged to the column and washed with distilled water. After washing was completed, 800 g of red ginseng extract was dissolved in 8 L of distilled water and poured into the column. Next, 30 L of distilled water, 30% (v/v) ethanol aqueous solution and 100% (v/v) ethanol were sequentially eluted and fractionated. First, distilled water and a 30% (v/v) aqueous ethanol solution were each sequentially added to the column, and the eluted fraction was flowed, and then 100% (v/v) ethanol was added to obtain an eluted fraction. The obtained 100% (v/v) ethanol fraction was concentrated to 71° Brix at 50-60° C. using a rotary vacuum concentrator (BUCHI, R-220). Finally, sterilized distilled water was added to the concentrate, diluted with 15° Brix, and then freeze-dried (OPERON, FDT-12012) to obtain 750.6 g of a fraction of the red ginseng extract (KGC10P1).

Preparation Example 2. Preparation of licorice extract

To 1.0 kg of licorice (Glycyrrhiza uralensis Fischer, Glycyrrhiza glabra Linne, roots and rhizomes), 13 times the weight of 30% (v/v) ethanol aqueous solution was added, extracted twice at 80℃ for 8 hours, and the extract was filtered through a 5㎛ filter. And filtered. The filtrate is concentrated at 50~60℃ using a rotary decompression concentrator (BUCHI, R-220), and finally diluted to 15°Brix by adding sterilized distilled water to the concentrate and freeze-dried (OPERON, FDT-12012). 244 g of licorice extract powder was obtained.

Preparation Example 3. Preparation of a mixture of fractions of red ginseng extract and licorice extract

The fractions or extracts obtained in Preparation Example 1 and Preparation Example 2 were prepared by different ratios. Three kinds of mixtures were prepared, and the ratio of fractions: licorice of the red ginseng extract was 3:1 (30:10), 1:1 (30:30), and 1:3 (30:90).

Experimental Example 1. Evaluation of inhibition of expression of liposynthetic factor (PPARγ) in vitro

Since PPARγ (peroxisome proliferator-activated receptor γ) protein plays an essential role in adipocyte formation differentiation, it was confirmed that treatment with a fraction of red ginseng extract, licorice extract, or a mixture thereof affects the expression of PPARγ.

3T3-L1 adipocytes were prepared in a 60 mm dish using DMEM medium supplemented with 10% Bovine Calf Serum (10% BCS) and 1% penicillin streptomycin (1% P/S). ^The cells were cultured at a density of ⁵ cells/mL at 37°C and 5% CO ₂ . Cells proliferate and become 100% confluent in the dish, and growth ceased. Two days later, dexamethasone (DEX), 3-isobutyl-1-methylxanthine (IBMX), insulin and 10% FBS, 1% Cell differentiation was induced using DMEM medium containing P/S. The date of using the differentiation medium was set to day 0 of cell differentiation, and a fraction of red ginseng extract, licorice extract, or a mixture thereof was treated at a concentration of 100 μg/ml, respectively. Two days after the start of cell differentiation (Day 2), it was replaced with DMEM medium containing 10% FBS, 1% P/S and insulin. Four days after the start of differentiation (Day 4), the DMEM medium was replaced with 10% FBS and 1% P/S, and cultured for 3 days. Proteins of the cells in which differentiation was completed were extracted using PRO-PREP protein extraction solution (iNtRON BIOTECHNOLOGY), and proteins were quantified using BCA protein analysis kit (THERMO scientific), and then used for experiments. 12 μg of protein was mixed with 2X sample buffer (ELPIS BIOTECH) and separated using 10% SDS-PAGE. After reacting the primary antibody (PPAR gamma, Cell Signaling) overnight at 4°C, the secondary antibody (goat anti-rabbit IgG, Cell Signaling) was reacted at room temperature for 2 hours, and after color development using ECL reagent (MILLIPORE), X -ray film was sensitized. The amount of PPARγ expression is shown in Table 1 below.

Normal Control Fraction treatment of red ginseng extract Licorice extract
process Fractions of red ginseng extract, and mixture of licorice extract (3:1) PPARγ/β-actin 0.2 1.0 0.81 1.3 0.49 ratio(%) 20 100 81 130 49

As shown in Table 1 and Figure 1, the mixture of the fraction of the red ginseng extract and the mixture of the licorice extract at 3:1 decreased PPARγ expression compared to the case where the fraction of the red ginseng extract or the licorice extract was treated alone.

Experimental Example 2. Evaluation of the degree of improvement in obesity in vivo

In order to evaluate the degree of improvement in obesity, the experimental groups were separated as shown in Table 2 below.

Group name Sample throughput Pay method Normal group (N) - General feed Control (C) - High fat feed Fraction of red ginseng extract (P1) KGC10P1 400 mg/kg High fat feed + P1 Licorice extract (gu) KGC10 gu 400 mg/kg High fat feed + gu Mixture of red ginseng extract fraction and licorice extract (PG) 3:1 KGC10P1 + g u (3:1) 400 mg/kg High Fat Feed + PG (3:1) Mixture of fractions of red ginseng extract and licorice extract (PG) 1:1 KGC10P1 + g u (1:1) 400 mg/kg High Fat Feed + PG (1:1) Mixture of fractions of red ginseng extract and licorice extract (PG) 1:3 KGC10P1 + g u (1:3) 400 mg/kg High Fat Feed + PG (1:3)

6-week-old C56BL/6 mice were purchased from Central Experimental Animal Co., Ltd. (Korea) and acclimatized for 7 days while maintaining appropriate temperature (25±2°C) and humidity (50±10%), controlling the contrast at a 12-hour cycle. C56BL/6 mice were each used for the normal group and the control group, and 6 mice were used for each group in the experimental group. Obesity was induced by providing a normal diet (normal control, D12450B) to the normal group, and a high fat diet (HF control, 60% fat calories, D12492, Research Diet, Canada) to the control group and the experimental group for 2 weeks. Table 3 shows the difference in components between the general diet and the high fat diet.

Regular diet (D12450B) High Fat Diet (D12492) protein 20 Kcal% 20 Kcal% carbohydrate 70 Kcal% 20 Kcal% Fat 10 Kcal% 60 Kcal%

After inducing obesity for 2 weeks, the group was separated, and a single extract (fraction of red ginseng extract, licorice extract) and composite material (fraction of red ginseng extract / licorice extract) were mixed in a ratio of 400 mg/kg in the material administration group (experimental group). Was administered orally once daily for 8 weeks.

2-1. Weight measurement

After the experiment was over, the animals were weighed. The measurement results are shown in Table 4 below.

Group Weight (g) ratio(%) Normal 26.1 60.3 Control 43.3 100.0 P1 39.9 92.1 PG 3:1 36.2 83.6 PG 1:1 38.7 89.4 PG 1:3 37.5 86.6 gu 38.4 88.7

The group treated with the mixture of the fraction of red ginseng extract and licorice extract had a high fat diet, but showed a tendency to lose weight compared to the control group that did not process the sample. Among the mixtures of the fraction of red ginseng extract and the mixture of licorice extract, when the mixture was mixed at a ratio of 3:1, there was a weight loss effect compared to the fraction of the red ginseng extract, licorice extract, or mixtures mixed with other ratios.

2-2. Measure the weight of abdominal fat

After the experiment was over, the animals were sacrificed and abdominal fat was removed and the weight was measured. The measurement results are shown in Table 5 below.

Group Abdominal fat (g) ratio(%) Normal 1.6 23.9 Control 6.7 100.0 P1 5.7 85.1 PG 3:1 4.8 71.6 PG 1:1 5.5 82.1 PG 1:3 4.9 73.1 gu 5.3 79.1

The group treated with the mixture of red ginseng extract fraction and licorice extract had a high fat diet, but showed a tendency to decrease abdominal fat compared to the control group that did not process the sample. Among the mixtures of the fraction of red ginseng extract and the mixture of licorice extract, when the mixture was mixed at a ratio of 3:1, there was an effect of reducing abdominal fat compared to the fraction of red ginseng extract, licorice extract, or mixtures mixed in other ratios.

2-3. Leptin analysis

After the experiment was over, the animals were sacrificed, blood was collected, and the amount of leptin (ELISA Kit, R&D Systems) was measured. Blood leptin was measured using an ELISA kit (R&D System) according to standard reagents and analysis procedures provided by the manufacturer. Serum was diluted at a ratio of 1:20 using a Calibrator diluent, and 50 μL each was dispensed into the prepared microplates, followed by reaction at room temperature for 2 hours. After washing 5 times using a washing buffer, 100 μL of leptin conjugate was dispensed into each well, followed by further reaction at room temperature for 2 hours. After washing 5 times with the washing buffer, add 100 μL of the substrate solution, react at room temperature for 30 minutes while blocking the light, and dispense 100 μL of the stop solution, and absorb the absorbance using a 450 nm wavelength within 30 minutes. Was calculated after measurement. The results of the leptin measurement are shown in Table 6 below.

Group Leptin(mg/dl) ratio(%) Normal 4 3.6 Control 112 100.0 P1 125 111.6 PG 3:1 59 52.7 PG 1:1 85 75.9 PG 1:3 84 75.0 gu 86 76.8

Among the mixtures of the red ginseng extract fraction and the licorice extract, when the mixture was mixed in a ratio of 3:1, there was a leptin reduction effect compared to the red ginseng extract fraction, the licorice extract, or the mixtures mixed in another ratio.

2-4. Dietary efficiency evaluation

Dietary intake efficiency was calculated by measuring body weight and feed intake every week during the 8-week material administration period and dividing the weight gain by the diet intake amount. Dietary intake efficiency was calculated by the following equation. Table 7 shows the dietary efficiency results.

Dietary intake efficiency = (weight gain/feed intake) × 100

Group Dietary efficiency (%) ratio(%) Normal 4.1 37.6 Control 10.9 100.0 P1 8.7 79.8 PG 3:1 5.0 45.9 PG 1:1 6.5 59.6 PG 1:3 6.0 55.0 gu 6.0 55.0

Dietary efficiency increased in the high-fat diet group with high energy density. The group treated with the mixture of the fraction of red ginseng extract and the licorice extract had a high fat diet, but the dietary efficiency tended to decrease compared to the control group that did not process the sample. Among the mixtures of the fraction of red ginseng extract and the mixture of licorice extract, when the mixture was mixed at a ratio of 3:1, there was an effect of reducing dietary efficiency compared to the fraction of red ginseng extract, licorice extract, or mixtures mixed with other ratios. From these results, it can be seen that a 3:1 mixture of a fraction of red ginseng extract and a licorice extract has an anti-obesity effect to inhibit weight gain by affecting dietary efficiency.

2-5. Biochemical Analysis-Triglyceride, Liver Level

Since obesity is a state in which more than necessary energy sources are accumulated in the body in the form of triglycerides due to excessive energy intake, it was evaluated whether fractions of red ginseng extract, licorice extract, and mixtures thereof affect triglycerides. ALT is an enzyme found in the liver, kidneys, heart, and muscles, and the largest amount is present in the liver. In particular, it is used as an indicator of liver disease because it increases in response to hepatocyte damage. Mainly, when the liver is damaged for various reasons, it is released into the blood, increasing blood levels.

After the experiment was over, the animals were sacrificed and blood was collected to measure the amount of ALT. The ALT measurement method was measured after putting a standard reagent suitable for each indicator into a biochemical analyzer (HITACHI, HITACHI 7100), and dispensing 250 μL of serum into a tube for biochemical analysis. The triglyceride measurement results are shown in Table 8 below, and the ALT measurement results are shown in Table 9 below.

Group Triglycerides (mg/dl) ratio(%) Normal 54.0 108.0 Control 50.0 100.0 P1 47.0 94.0 PG 3:1 42.0 84.0 PG 1:1 47.0 94.0 PG 1:3 45.0 90.0 gu 50.0 100.0

Group ALT(U/L) ratio(%) Normal 22.0 25.0 Control 88.0 100.0 P1 50.0 56.8 PG 3:1 31.0 35.2 PG 1:1 34.0 38.6 PG 1:3 43.0 48.9 gu 44.0 50.0

The group treated with the mixture of the red ginseng extract fraction and the licorice extract had a high fat diet, but showed a tendency to decrease triglycerides compared to the control group that did not process the sample. Among the mixtures of the fraction of red ginseng extract and the mixture of licorice extract, when the mixture was mixed at a ratio of 3:1, there was an effect of reducing triglycerides compared to the fraction of red ginseng extract, licorice extract, or mixtures mixed in other ratios.

The group treated with the mixture of the fraction of red ginseng extract and the licorice extract had a high fat diet, but showed a tendency of decreasing ALT compared to the control group that did not process the sample. Among the mixtures of the fraction of red ginseng extract and the mixture of licorice extract, when the mixture was mixed at a ratio of 3:1, there was an effect of reducing ALT compared to the fraction of red ginseng extract, licorice extract, or mixtures mixed in other ratios. From the above results, it can be seen that a 3:1 mixture of a fraction of red ginseng extract and a licorice extract can improve liver damage.

Preparation Example 4. Preparation of pharmaceutical composition

4-1. Preparation of powder

2 g of a mixture of a fraction of the red ginseng extract of the present invention and a licorice extract

1 g lactose

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

4-2. Manufacture of tablets

100 mg of a mixture of a fraction of the red ginseng extract and licorice extract of the present invention

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

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

4-3. Preparation of capsules

100 mg of a mixture of a fraction of the red ginseng extract and licorice extract of the present invention

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, the gelatin capsules were filled according to a conventional capsule preparation method to prepare a capsule formulation.

4-4. Manufacture of pills

1 g of a mixture of a fraction of the red ginseng extract of the present invention and a licorice extract

1.5 g lactose

1 g glycerin

0.5 g xylitol

After mixing the above components, it was prepared so as to be 4 g per pill according to a conventional method.

4-5. Preparation of granules

150 mg of a mixture of the red ginseng extract fraction and licorice extract of the present invention

Soybean extract 50 mg

200 mg of glucose

Starch 600 mg

After mixing the above ingredients, 100 mg of 30% ethanol was added, dried at 60°C to form granules, and then filled into a cloth.

Manufacture Example 5. Manufacture of health food

5-1. Manufacture of flour food

0.5 to 5.0 parts by weight of a mixture of the fraction of the red ginseng extract of the present invention and the licorice extract were added to flour, and bread, cakes, cookies, crackers and noodles were prepared using this mixture.

5-2. Preparation of soups and gravies

0.1 to 5.0 parts by weight of a mixture of the fraction of the red ginseng extract of the present invention and the licorice extract were added to soups and juices to prepare health-promoting meat products, noodle soups, and juices.

5-3. Preparation of ground beef

Ground beef for health promotion was prepared by adding 10 parts by weight of a mixture of the fraction of the red ginseng extract of the present invention and the licorice extract to ground beef.

5-4. Manufacture of dairy products

5 to 10 parts by weight of a mixture of the fraction of the red ginseng extract of the present invention and the licorice extract were added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

5-5. Manufacture of wire

Brown rice, barley, glutinous rice, and adlay were gelatinized by a known method, dried, and then roasted, and then prepared into a powder having a particle size of 60 mesh with a grinder.

Black soybeans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, and then roasted, and then prepared into a powder having a particle size of 60 mesh with a grinder.

The mixture of the fraction of the red ginseng extract and the licorice extract of the present invention was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by spraying and drying with a hot-air dryer was pulverized to a particle size of 60 mesh with a grinder to obtain a dry powder.

The grains, seeds, and a mixture of the fractions of the red ginseng extract of the present invention and licorice extract were mixed in the following ratio to prepare.

Cereals (30 parts by weight of brown rice, 15 parts by weight of barley, 20 parts by weight of barley), seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame), a mixture of a fraction of the red ginseng extract and licorice extract of the present invention (3 parts by weight), Ganoderma lucidum (0.5 parts by weight), Rehmannia (0.5 parts by weight)

5-6. Manufacturing of health drinks

Homogenize 5 g of a mixture of subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%), and a fraction of the red ginseng extract of the present invention and licorice extract After mixing and sterilizing instantaneously, it was prepared by packaging it in a small container such as a glass bottle or a plastic bottle.

5-7. Manufacture of vegetable juice

Vegetable juice was prepared by adding 5 g of a mixture of the red ginseng extract fraction and licorice extract of the present invention to 1,000 ml of tomato or carrot juice.

5-8. Manufacture of fruit juice

Fruit juice was prepared by adding 1 g of a mixture of the fraction of the red ginseng extract of the present invention and the licorice extract to 1,000 ml of apple or grape juice.

Claims (10)

A pharmaceutical composition for preventing or treating obesity comprising a fraction of red ginseng extract and licorice extract. The method according to claim 1,
With respect to the fraction of the red ginseng extract 30 parts by weight, the licorice extract is a pharmaceutical composition containing 0.1 to 25 parts by weight. The method according to claim 2,
With respect to the fraction of the red ginseng extract 30 parts by weight, the licorice extract is a pharmaceutical composition containing 1 to 20 parts by weight. The method according to claim 1,
The solvent used in the preparation of the red ginseng extract or licorice extract is any one selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof. The method of claim 4,
A pharmaceutical composition in which the solvent used to prepare red ginseng extract is water. The method of claim 4,
The solvent used to prepare the licorice extract is a pharmaceutical composition of 10% (v/v) to 70% (v/v) ethanol aqueous solution. The method according to claim 1,
The fraction of red ginseng extract is obtained by column-purifying fractionation of red ginseng extract,
The fixed phase of the column is an aromatic synthetic resin pharmaceutical composition. The method of claim 7,
When the column is purified, the desorption solvent is any one selected from the group consisting of water, an organic solvent, and a mixture thereof. The method of claim 8,
The desorption solvent is 40% (v / v) or more to less than 100% (v / v) ethanol aqueous solution or ethanol pharmaceutical composition. Health food composition for improving obesity comprising a fraction of red ginseng extract and licorice extract.

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