KR102178604B1 - Composition for preventing or treating obesity comprising complex extract of Juniperus rigida - Google Patents

Abstract

The present invention relates to a juniper complex extract, and more particularly, to a composition for preventing or treating obesity comprising the juniper complex extract as an active ingredient. The juniper complex extract according to the present invention has the effect of remarkably inhibiting the expression of proteins and RNA of adipocyte differentiation factors, and thus exhibits excellent anti-obesity effect, and the field of prevention, improvement or treatment of obesity, obesity-related diseases or complications It can be used in various ways.

Description

Composition for preventing or treating obesity, comprising complex extract of Juniperus rigida as an active ingredient {Composition for preventing or treating obesity comprising complex extract of Juniperus rigida}

The present invention relates to a juniper complex extract, and more particularly, to a composition for preventing or treating obesity comprising the juniper complex extract as an active ingredient.

Due to westernization of diet, excessive consumption of food, and lack of physical exercise, the number of obese patients is increasing rapidly every year. Obesity is a risk factor for major adult diseases, and it is involved in the development of adult diseases such as high blood pressure, diabetes, arteriosclerosis, stroke, heart attack, and various tumors, and also promotes the progression of the disease. Obesity is simply a problem of appearance. It is not a serious problem that is directly related to health.

In order to treat such obesity, many anti-obesity drugs are being developed every year, but there are not many obesity drugs currently available. In addition, most of the developed anti-obesity drugs are limited to drugs that suppress digestion or appetite. In the case of agents that control digestion or appetite, they are classified as psychotropic drugs because of their habits, and digestive inhibitors exhibit side effects such as diarrhea and constipation. Representative obesity drugs approved for long-term use by the US FDA until 2010 are sibutramine (Reductil), which inhibits the reuptake of norepinephrine and serotonin, and lipase secreted from the pancreas and digestive system. There was an orlistat (Xenical) that showed an effect by inhibiting (lipase).

However, Sibutramine was expelled from the United States and Korea in October 2010 due to common side effects such as elevated blood pressure, insomnia, dry mouth, and dizziness, and risk of cardiovascular symptoms such as myocardial infarction and stroke. In addition, Orlistat has a common side effect such as diarrhea, fat stool, and lost money, and in cases where the intake of fat is less than that of Westerners such as Koreans, the effect of the drug is not clear, so its use is limited. Therefore, there is a need for an obesity prevention or treatment that has excellent obesity suppression effect and has confirmed the safety of long-term use.

On the other hand, Juniperus rigida ) is an evergreen tall tall tree belonging to the cypress family and is distributed throughout Korea, China, and Siberia, also called juniper, needle juniper. In Korea, it is used as a windbreak forest on the coast because it grows in limestone areas, is resistant to cold, and withstands sea breezes well. In addition, the fruit of the juniper tree is called dusongsil or juniper berry, and has a strong, dehumidifying and diuretic effect, so it is used as an external medicine for customary arthritis or for swelling, gout and urinary genital diseases. In the West, the fruit of the juniper was used as a fungicide as a method for Indians to drink as tea during urinary tract infections. Recently, there are research results showing that the fruit of the juniper is effective for insulin-dependent diabetes.

Accordingly, the present inventors developed the present invention by developing a juniper complex extract that inhibits the expression of adipocyte differentiation factors PPARγ, C/EBP-α and C/EBP-β in order to solve the problems of the prior art as described above. Finished.

Accordingly, it is an object of the present invention to provide a composition for preventing, treating or improving obesity, including the juniper complex extract as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating obesity, comprising the juniper complex extract as an active ingredient.

In addition, the present invention provides a food composition for preventing or improving obesity comprising the juniper complex extract as an active ingredient.

The juniper complex extract according to the present invention has the effect of remarkably inhibiting the expression of proteins and RNA of adipocyte differentiation factors, and thus exhibits excellent anti-obesity effect, and the field of prevention, improvement or treatment of obesity, obesity-related diseases or complications It can be used in various ways.

1 is a diagram showing a method for preparing an ethanol extract of juniper according to the present invention.
Figure 2 is a diagram showing the cytotoxicity measurement results of the juniper composite extract according to the present invention in adipocytes.
Figure 3 is a diagram showing the results of measuring the protein expression of adipocyte differentiation factor when treated with the juniper complex extract according to the present invention.
4 is a diagram showing the results of measuring the protein expression of adipocyte differentiation factor upon treatment of a control (CLS) containing chlorogenic acid, L-arginine, and synephrine.
Figure 5 is a diagram showing the result of measuring the mRNA expression of adipocyte differentiation factor when treated with the juniper complex extract according to the present invention.
6 is a diagram showing the results of measuring the antioxidant activity of the ethanol extract of juniper according to the present invention.

Hereinafter, the present invention will be described in detail.

According to an aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating obesity, comprising the juniper complex extract as an active ingredient.

According to a preferred embodiment of the present invention, the juniper complex extract includes juniper extract, chlorogenic acid, L-arginine and synephrine. The juniper complex extract was prepared by mixing chlorogenic acid, L-arginine, and synephrine in a weight ratio of 0.5-2: 0.5-2: 0.5-2 to prepare a first solution, and the first solution and the juniper extract was 0.5- It is preferable to prepare by mixing at a weight ratio of 2: 0.5-2.

In the present invention, the juniper extract may be extracted from various parts of the juniper tree, and includes a juniper extract and a fraction of the extract. The juniper extract may be preferably extracted from the leaves, stems, roots or fruits of the juniper, and is preferably extracted from the stem or leaves of the juniper.

The juniper extract may be obtained by extraction, separation and fractionation from nature using a method of extraction, separation and fractionation known in the art. The'extract' as defined in the present invention is obtained by extraction treatment from juniper using an appropriate solvent, and includes, for example, a crude extract of juniper, a polar solvent soluble extract, or a non-polar solvent soluble extract.

The juniper extract may be extracted according to various extraction solvents and extraction methods, and any suitable solvent for extracting the extract from juniper may be used as long as a pharmaceutically acceptable organic solvent, and water or an organic solvent may be used. Can be used, but is not limited thereto. For example, as the solvent, an alcohol having 1 to 4 carbon atoms such as water, methanol, ethanol, propanol, isopropanol, butanol, phosphate buffer, etc. may be used alone or It can be used by mixing two or more types. In particular, it is preferable to use methanol or ethanol as the solvent, and it is more preferable to use ethanol.

In addition, methods such as hot water extraction, cold precipitation extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, compression method, etc. may be used when preparing juniper extract, and it is more preferable to use hot water extraction or solvent extraction. Do. After obtaining the extract using water or an organic solvent as described above, a liquid product may be obtained by cooling, heating and filtering at room temperature by a conventional method known in the art, or additionally evaporating the solvent, spray drying or freeze drying. You may.

In addition, the juniper extract of the present invention may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying of the extracted and fractionated extract or fraction as described above. In addition, the extract or fraction was further purified by using various chromatography such as silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. Can also be obtained.

Therefore, the juniper extract used in the present invention is a concept including all extracts, fractions, and purified products obtained in each step of extraction, fractionation, or purification, and their dilutions, concentrates, or dried products.

In one embodiment of the present invention, the juniper complex extract according to the present invention comprises an ethanol extract of juniper.

In the present invention, it was confirmed that the juniper complex extract inhibited the expression of adipocyte differentiation factors PPARγ, C/EBP-α and C/EBP-β at the protein and RNA levels in adipocyte 3T3-L1.

Therefore, the juniper complex extract of the present invention can be used as a drug or food useful for the prevention, treatment, and improvement of obesity, obesity-related diseases or complications. In addition, the composition of the present invention can be used for the prevention, treatment and improvement of obesity, obesity-related diseases, and complications caused by obesity in not only humans but also other animals.

The composition of the present invention can also be used for the prevention, improvement or treatment of obesity or abdominal obesity in each part of the body, and the target abdominal obesity is a long time sitting in a chair, lack of exercise, alcohol (alcohol) intake, stress, etc. It includes, but is not limited to, abdominal obesity.

In addition, complications due to obesity include, but are not limited to, diseases caused by elevated blood levels of triglycerides, cholesterol, and low-density lipids. For example, complications due to obesity include metabolic syndrome (visceral fat syndrome, metabolic abnormality syndrome, etc.), hypertriglyceridemia, hypoHDLemia, angina, myocardial infarction, hypogonadism, sleep apnea, premenstrual syndrome, and stress urine. Urinary incontinence, including incontinence, hyperactivity disorder, chronic fatigue syndrome, osteoarthritis, weight gain-related cancer, orthostatic hypotension, pulmonary hypertension, menstrual disorder, diabetes, hypertension, impaired glucose tolerance, coronary artery thrombosis, drowsiness, depression, anxiety , Psychosis, delayed movement disorder, drug addiction, substance abuse, cognitive impairment, Alzheimer's disease, cerebral ischemia, obsessive-compulsive behavior, panic attack, social phobia, bulimia, atherosclerosis, gallbladder diseases such as cholelithiasis, loss of appetite, polycystic ovary disease and There are reproductive disorders, infections, varicose veins, skin diseases such as epidermal proliferation and eczema, insulin resistance, chronic arterial obstruction, orthopedic injury, thromboembolism, heart disease, urinary disease, lipid syndrome, hyperglycemia, stress, etc. It is not limited.

The pharmaceutical composition for preventing or treating obesity of the present invention may contain one or more pharmaceutically acceptable carriers for administration. Pharmaceutically acceptable carriers can be used by mixing saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and at least one of these ingredients, and if necessary, antioxidants and buffers. , Other conventional additives such as bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, and pills, capsules, granules, or tablets. Furthermore, it can be formulated according to each disease or component by an appropriate method in the art.

The pharmaceutical composition for preventing or treating obesity may further include additional ingredients. Examples of such additional ingredients are lactose, talc, starch, magnesium stearate, crystalline cellulose, lactose, gelatin, mannitol, peroxic acid, isomerized sugar, polyvinyl alcohol, boric acid and sodium carbonate.

The pharmaceutical composition for preventing or treating obesity may be administered orally or parenterally, and is preferably administered orally. For parenteral administration, intravenous injection, intramuscular injection, intra-articular injection, intra-synovial injection, intrathecal injection, intrahepatic injection, intralesional injection, or It can be administered by intracranial injection. The appropriate dosage of the pharmaceutical composition for preventing or treating obesity is determined by factors such as the formulation method, the mode of administration, the patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. It can be prescribed in various ways.

The composition of the present invention may further contain one or more known active ingredients having an effect of preventing or treating obesity together with the juniper complex extract.

In addition, the composition of the present invention may be used alone for the prevention or treatment of obesity, obesity-related diseases or complications, or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, or methods using a biological response modifier.

According to another aspect of the present invention, the present invention provides a food composition for preventing or improving obesity, comprising the juniper complex extract as an active ingredient.

When the juniper complex extract of the present invention is used as a food additive, the juniper complex extract may be added as it is, or may be appropriately used according to a conventional method, such as being used by mixing with other foods or food ingredients.

In addition, the mixed amount of the active ingredient, juniper complex extract can be suitably changed according to the purpose of use (prevention, health or therapeutic treatment), and the juniper complex extract is 0.01 to 95 based on the total weight of the food composition. It is preferably included in weight %, more preferably 1 to 80 weight %. If the content is less than 0.01% by weight, the dosage efficiency may decrease, and if it exceeds 95% by weight, formulation may be difficult.

As a specific example, when preparing food or beverage, the juniper composite extract of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less based on the raw material. However, in the case of long-term intake for health and hygiene purposes or for health control purposes, it may be added in an amount below the above range, and there is no problem in terms of safety, so the active ingredient can be used in an amount above the above range. have.

There is no particular limitation on the type of food. Examples of foods to which the juniper complex extract of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages , Tea, drinks, alcoholic beverages, vitamin complexes, etc., and includes all health foods in the usual sense.

When the food composition of the present invention is prepared as a beverage, it may include additional ingredients such as various flavoring agents or natural carbohydrates, like a conventional beverage. The natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame may be used. The natural carbohydrate is contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight, based on the total weight of the food composition of the present invention.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols , Carbonating agents used in carbonated beverages, etc. may be included. In addition, the composition of the present invention may include flesh for the manufacture of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination. The ratio of the additives is not largely limited, but is preferably contained within the range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

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

Example 1. Preparation of juniper complex extract

First, an ethanol extract of juniper was prepared. Juniper tree used for the extraction was used as an extraction sample from the dried juniper tree produced in the spring in Goesan, Chungbuk, and an ethanol extract of Juniper tree was prepared in the same manner as in FIG. 1. In more detail, 70% ethanol was added to the dried juniper, 10 times the amount of the sample, and then immersed at room temperature for 24 hours to separate the supernatant from the precipitate. The sample extract was filtered using a filter paper (Whatman No. 2), and concentrated under reduced pressure with an EYELA dryer to completely remove the solvent to prepare an ethanol extract of juniper. The ethanol extract of juniper was stored and used at 20°C.

Juniper tree complex extract (JCLS) was prepared using the prepared juniper ethanol extract. The juniper complex extract (JCLS) is a juniper ethanol extract, chlorogenic acid (cat No. C3878, Sigma aldrich), L-arginine (cat No. A5006, Sigma aldrich), and synephrine. (synephrine) (cat No. 75256, Sigma aldrich) was prepared by mixing. More specifically, the juniper complex extract is prepared by mixing chlorogenic acid, L-arginine, and synephrine in a weight ratio of 1:1:1, and the first solution and the juniper ethanol extract are 1:1 It was prepared by mixing at a weight ratio of.

In addition, the control (CLS) was prepared by mixing chlorogenic acid, L-arginine, and synephrine in a weight ratio of 1:1:1.

Example 2. Measurement of Cytotoxicity of Juniper Tree Complex Extract

2-1. Materials and reagents

Adipocyte 3T3-L1 cells were purchased and used from ATCC (USA). The hemocytometer (haemacytometer, Marienfeld, Germany) and MTT (3-[4,5-dimethylthiazol]-2-yl] -2,5-diphenyl-tetrazolium bromide) used for cytotoxicity measurement were purchased from Sigma-Aldrich. , DMSO (dimethyl sulfoxide) was purchased from BioShop (Canada) and used.

2-2. Culture and differentiation of 3T3-L1 cells

DMEM (dulbecco's modified eagle medium) medium added with 100 U/ml of 10% bovine calf serum (BCS) and 1% penicillin/streptomycin was used for the culture and maintenance of 3T3-L1 adipocytes. And, it was adapted in an incubator at 37° C. and 5% CO _{2 and} subcultured when the cells grew 60%. For cell differentiation, 1×10 ⁶ cells per well were dispensed into a 6-well plate, and the cells were cultured to be 100% dense. After 2 days, 10% FBS, 1% PS, and differentiation-inducing substance MDI (0.5 mM IBMX, 1 μM DEX, 10 μg/ml insulin) were added to DMEM medium, and then 10% FBS and 10 μg/ml insulin. It was treated with DMEM medium containing gulf for 3 days, and after that, cultured with DMEM containing 10% FBS, and adipocyte differentiation was induced based on the formation of intracellular adipocytes. The sample was treated from the time point when the differentiation inducing medium was added.

2-3. MTT Measurement of cytotoxicity through assay

Cytotoxicity of juniper complex extract in 3T3-L1 cells was determined by Carmichael's method (Carmichael J, DeGraff WG, Gazdar AF, Minna JD and Mitchell JB. Evaluation of a tetrazolium based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. ( 1987) 47 : 936-942.).

Specifically, 3T3-L1 cells were dispensed to a 96 well plate at 1×10 ⁵ cells/well. After adding 0.02 ml of juniper complex extract at concentrations of 5, 10, 50, 100, 500 and 1000 μg/ml to the dispensed cells, each was cultured in an incubator at 37° C. and 5% CO ₂ for 24 hours. After adding 0.04 ml of MTT solution prepared at a concentration of 2.5 mg/ml and incubating for 4 hours, the culture solution was removed, 0.1 ml of DMSO was added to each well and reacted at room temperature for 30 minutes, and then 540 nm using an ELISA reader. The absorbance at was measured. CLS, juniper ethanol extract (JR), synephrine (SP), chlorogenic acid (CG), and L-arginine (LA) were used as controls. Cytotoxicity was measured by the rate of decrease in absorbance of the sample solution-added group and the non-added group. The cytotoxicity measurement results are shown in FIG. 2.

As shown in Fig. 2, it can be seen that both the juniper complex extract (JCLS) and the control group showed close cell viability of 100% at 100 μg/ml in 3T3-L1 cells. Therefore, the following cell experiments were conducted at a concentration of 100 μg/ml or less.

Example 3. Western Blotting Protein expression of adipocyte differentiation factor

Expression of adipocyte differentiation-related factors PPARγ, C/EBP-α, and C/EBP-β in the 3T3-L1 cell line was confirmed.

Specifically, after differentiating 3T3-L1 cells in the same manner as in Example 2-2, juniper complex extract (JCLS) of 25, 50 and 100 μg/ml was treated, respectively. The 3T3-L1 cell line treated with the juniper complex extract was dissolved in 10 ml of RIPA buffer to which 1 tab of Complete mini was added, followed by centrifugation at 4° C. and 13,200 rpm for 20 minutes. The supernatant obtained by centrifugation was quantified with a BCA protein assay kit, and 20 μl of protein was electrophoresed on a 10% acryl amide gel. The protein separated by electrophoresis was transferred to a PVDF membrane (polyvinylidene fluoride membrane) using a transfer device, and then incubated in a blocking buffer (5% skim milk in TBST) at room temperature for 1 hour. The primary antibody was diluted and reacted overnight at 4° C., and then washed 3 times with TBST buffer (tris-buffered saline and tween 20 buffer) at 10 minute intervals. The secondary antibody was diluted 1:1,000 and incubated for 2 hours at room temperature, washed three times with TBST, and then the band was identified and quantified using a LAS 4,000 instrument. Western blotting was performed in the same manner as the control group (CLS). Western blotting results and graphing the results are shown in FIGS. 3 and 4.

As shown in Figures 3 and 4, as a result of measuring the protein expression activity of PPARγ, C/EBP-α and C/EBP-β after 3T3-L1 differentiation, in the juniper complex extract (JCLS) treatment group, the above concentration-dependently Protein expression was reduced, and the protein inhibition rate was confirmed to be 56.8%, 77.1%, and 64.2%, respectively, when treated with the juniper complex extract at a concentration of 100 μg/ml. On the other hand, the protein inhibition rates of the control group (CLS) with a concentration of 100 μg/ml were 70.0%, 80.6%, and 88.7%, respectively. The above results indicate that the juniper complex extract (JCLS) has an excellent effect of inhibiting the expression of adipocyte differentiation factor protein.

Example 4. RT- PCR Of adipocyte differentiation factors through mRNA Expression measurement

4-1. Total RNA isolation and cDNA synthesis

In the 3T3-L1 cell line, mRNA expression of PPARγ, C/EBP-α and C/EBP-β, which are factors related to adipocyte differentiation, were confirmed.

Specifically, after differentiating 3T3-L1 cells in the same manner as in Example 2-2, juniper complex extract (JCLS) of 25, 50 and 100 μg/ml was treated, respectively. After removing the culture medium from the 3T3-L1 cells treated with the juniper complex extract, 1 ml of trizol lysis buffer was dispensed into each well to dissolve the cells, and then 200 μl of chloroform was dispensed for 20 seconds. It shook up and down. Then, centrifugation was performed at 13,200 rpm for 20 minutes, and the supernatant was transferred to a tube containing 500 μl of isopropanol and mixed. Centrifugation was performed again at 13,200 rpm for 20 minutes, the supernatant was removed, and 1 ml of 75% DEPC water (EtOH-diethylpyrocarbonate water) was dispensed into each tube, centrifuged at 13,200 rpm for 5 minutes, and the supernatant was removed. It was dried at room temperature. After 50 μl of DEPC-treated distilled water was dispensed and dissolved, 5 μl of RNA and 195 μl of sterile water were added to a 96-well plate, and the absorbance was measured at 260 nm and 280 nm, respectively, to measure the total amount of RNA. Oligo(dT) 15 Primer (500 μg/ml) 1 μl, extracted RNA (2 μg) and nuclease free water to adjust the volume to 10 μl, reacted at 75° C. for 5 minutes and then 5X Reaction buffer, MgCl ₂ , PCR nucleotide mix, RNasin inhibitor, reverse transcriptase, and nuclease-free water were added and reacted at 25° C. for 5 minutes, 42° C. for 60 minutes, and 70° C. for 15 minutes to synthesize cDNA.

4-2. RT- PCR

In order to examine the mRNA expression of PPARγ, C/EBP-α, and C/EBP-β, RT-PCR (Reverse transcription polymerase chain reaction) was performed. The primers used in the experiment are shown in Table 1.

gene primer Sequence (5' → 3') GAPDH sense TGA AGG TCG GTG TGA ACG GAT TTG GC anti-sense CAT GTA GGC CAT GAG GTC CAC CAC PPARγ sense CGC-TGA-TGC-ACT-GCC-TAT-GA antisense TGC-GAG-TGG-TCT-TCC-ATC-AC C/EBP-α sense GTG-TGC-ACG-TCT-ATG-CTA-AAC-CA antisense GCC-GTT-AGT-GAA-GAG-TCT-CAG-TTT-G C/EBP-β sense GTT-TCG-GGA-GTT-GAT-GCA-ATC antisense AAC-AAC-CCC-GCA-GGA-ACA-T

5X green Go Taq flexi buffer, MgCl ₂ , PCR nucleotide mix (10 mM), primer, Go Taq DNA polymerase, nuclease-free water, and cDNA synthesized in Example 4-1 were added to the PCR tube and mixed. RT-PCR was performed. More specifically, the reaction was carried out for 40 cycles at 94°C for 30 seconds, 59°C for 60 seconds, and 72°C for 1 minute each. After RT-PCR, a 1.5% agarose gel to which 0.002% EtBr (ethidium bromide) was added was subjected to electrophoresis at 100 V for 40 minutes, and then the band was analyzed and quantified using LAS 4,000. The control group (CLS) was subjected to RT-PCR in the same manner. The results of measuring mRNA expression through RT-PCR and graphing the results are shown in FIG. 5.

As shown in Figure 5, when treated with juniper complex extract (JCLS) at concentrations 25, 50 and 100 μg/ml, respectively, the expression of fat differentiation factors PPARγ, C/EBP-α and C/EBP-β It can be seen that it was suppressed.

Example 5. Measurement of antioxidant activity of ethanol extract of juniper

The antioxidant activity of the ethanol extract of juniper was measured using DPPH (2,2-diphenyl-1-picrylhydrazyl). DPPH is a relatively stable free radical and is reduced by aromatic compounds and aromatic amines to decolorize deep purple, and the electron donating ability is measured using this. Such an electron donating ability measurement method is a commonly used method because it can simply measure the antioxidant activity of a plant extract and has a very high correlation with the actual antioxidant activity.

Specifically, the electron donating ability of the ethanol extract of Juniper tree prepared in Example 1 was determined by the method of Blois described in "Blois MS. Antioxidant determination by the use of a stable free radical. Nature. (1958) 26: 1199-1120" It was measured by deformation. DPPH solution (Sigma, U.S.A) 60 μl and 120 μl of juniper ethanol extract of various concentrations (5, 10, 50, 100, 500, 1000 μg/ml) were added and mixed, and then reacted for 15 minutes. After the reaction was completed, the absorbance was measured at 517 nm using a microplate reader. The electron donating ability was calculated as shown in Equation 1.

[Equation 1]

As a comparative group, hot water extract of juniper and butylhydroxytoluen was used. The measurement results of electron donating ability are shown in FIG. 6.

As shown in Figure 6, it was confirmed that the electron donating ability of the ethanol extract of juniper is high. In particular, the concentration of 1000 μg / ml of juniper ethanol extract had an electron donating ability of 86.8%, which was higher than that of the same concentration of hot water extract of juniper (44.0%). It can also be seen that it exhibits similar effects to butylhydroxytoluene, which is known as a synthetic antioxidant.

Overall, the present inventors confirmed that the complex extract of juniper, including ethanol extract, chlorogenic acid, L-arginine and synephrine, inhibits the expression of proteins and RNA of adipocyte differentiation factors in adipocytes. This means that the juniper composite extract has an excellent anti-obesity effect, and the juniper composite extract of the present invention can be variously used in the field of preventing, improving or treating obesity, obesity-related diseases or complications.

Hereinafter, the present invention will be described in more detail through formulation examples. Formulation examples are for illustrative purposes only, and the scope of the present invention is not construed as being limited by formulation examples.

Formulation example 1. Preparation of pharmaceutical composition for preventing or treating obesity

1-1. Powdery Produce

Juniper tree complex extract 20 mg

100 mg lactose

10 mg of talc

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

1-2. Manufacture of tablets

10 mg of juniper complex extract

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

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

1-3. Preparation of capsules

10 mg of juniper complex extract

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

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

1-4. Preparation of injections

10 mg of juniper complex extract

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO ₂ H ₂ O 26 mg

It is prepared with the above ingredients per ampoule (2 ml) according to a conventional injection preparation method.

1-5. Liquid Produce

Juniper tree complex extract 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of the liquid formulation, add and dissolve each component in purified water, add an appropriate amount of lemon flavor, mix the above ingredients, add purified water, add purified water, adjust the total to 100 ml, and fill in a brown bottle. It is sterilized to prepare a solution.

Formulation example 2. Preparation of food composition for preventing or improving obesity

2-1. Manufacture of health food

Juniper tree complex extract 100mg, vitamin mixture appropriate amount, vitamin A acetate 70g, vitamin E 1.0mg, vitamin B1 0.13mg, vitamin B2 0.15mg, vitamin B6 0.5mg, vitamin B12 0.2g, vitamin C 10mg, biotin 10g, nicotinic acid Amide 1.7 mg, folic acid 50 g, calcium pantothenate 0.5 mg, inorganic mixture appropriate amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 Mg, 100 mg of calcium carbonate and 24.8 mg of magnesium chloride were mixed, and then granules were prepared and health food was prepared according to a conventional method. At this time, the composition ratio of the vitamin and mineral mixture is a mixture of ingredients suitable for a relatively healthy food in a preferred embodiment, but the mixing ratio may be arbitrarily modified.

2-5. Manufacturing of health drinks

According to the general health drink manufacturing method, 100 mg of juniper complex extract, 15 g of vitamin C, 100 g of vitamin E (powder), 19.75 g of iron lactate, 3.5 g of zinc oxide, 3.5 g of nicotinic acid amide, 0.2 g of vitamin A, 0.25 g of vitamin B1 , Vitamin B2 0.3g and quantitative water were mixed, and then stirred and heated at 85° C. for about 1 hour, and the resulting solution was filtered, obtained in a sterilized 2L container, sealed and sterilized, and stored in a refrigerator to prepare a health drink. At this time, the composition ratio is a mixture of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

As described above, specific parts of the present invention have been described in detail, and for those of ordinary skill in the art, it is obvious that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. something to do. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (6)

A pharmaceutical composition for preventing or treating obesity, comprising as an active ingredient a complex juniper extract containing chlorogenic acid, L-arginine, synephrine and juniper extract.
delete The method of claim 1,
The juniper complex extract was prepared by mixing chlorogenic acid, L-arginine, and synephrine in a weight ratio of 0.5-2: 0.5-2: 0.5-2 to prepare a first solution, and the first solution and the juniper extract was 0.5- 2: A pharmaceutical composition for preventing or treating obesity, which is prepared by mixing in a weight ratio of 0.5-2.
The method of claim 1,
The juniper extract is extracted with one or more solvents selected from water, an alcohol having 1 to 4 carbon atoms, a phosphate buffer, and a mixed solvent thereof, a pharmaceutical composition for preventing or treating obesity.
The method of claim 4,
The juniper extract is a pharmaceutical composition for preventing or treating obesity, characterized in that the ethanol extract.
A food composition for preventing or improving obesity, comprising as an active ingredient a complex juniper extract containing chlorogenic acid, L-arginine, synephrine and juniper extract.

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