WO2022092441A1 - Anti-obesity food composition including stevia extract - Google Patents

Anti-obesity food composition including stevia extract Download PDF

Info

Publication number
WO2022092441A1
WO2022092441A1 PCT/KR2020/019029 KR2020019029W WO2022092441A1 WO 2022092441 A1 WO2022092441 A1 WO 2022092441A1 KR 2020019029 W KR2020019029 W KR 2020019029W WO 2022092441 A1 WO2022092441 A1 WO 2022092441A1
Authority
WO
WIPO (PCT)
Prior art keywords
extract
stevia extract
stevia
food composition
obesity food
Prior art date
Application number
PCT/KR2020/019029
Other languages
French (fr)
Korean (ko)
Inventor
김종덕
임병선
Original Assignee
전남대학교산학협력단
한국스테비아(주)
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 전남대학교산학협력단, 한국스테비아(주) filed Critical 전남대학교산학협력단
Publication of WO2022092441A1 publication Critical patent/WO2022092441A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/40—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by drying or kilning; Subsequent reconstitution
    • A23L3/44—Freeze-drying
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/23—Removal of unwanted matter, e.g. deodorisation or detoxification by extraction with solvents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/51—Concentration
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/332—Promoters of weight control and weight loss
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts
    • A23V2250/214—Tea
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2300/00—Processes
    • A23V2300/14—Extraction
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2300/00—Processes
    • A23V2300/31—Mechanical treatment

Definitions

  • the present invention relates to an anti-obesity food composition containing a stevia extract, and more particularly, by using a stevia extract having an angiogenesis inhibitory function alone or in combination with a green tea extract, a stevia extract having excellent anti-obesity effect and no side effects It relates to an anti-obesity food composition containing.
  • foods that target metabolic diseases include health functional foods that express functionality related to some metabolic diseases, weight control foods that target metabolic diseases, and some general foods that can reduce the incidence of metabolic diseases. do.
  • Obesity one of the causes of such metabolic diseases, is a cause that threatens the health of growing children and adolescents.
  • obesity in children and adolescents is highly likely to transition to adult obesity. It can also affect adult health by causing complications such as diseases, osteoarthritis, and insulin resistance.
  • anti-obesity drugs include agents that inhibit eating and agents that promote fat metabolism, inhibit fat synthesis, or increase metabolic activity. could be broadly divided into
  • the present invention provides an anti-obesity food composition containing a stevia extract that exhibits excellent anti-obesity effect and has no side effects by using a stevia extract having angiogenesis inhibitory function alone or mixed with green tea extract purpose is to
  • the anti-obesity food composition containing the stevia extract is characterized in that it has an angiogenesis inhibitory function.
  • the stevia extract is characterized in that it is prepared through an extraction step of extracting a stevia extract from a stevia leaf and a purification step of purifying the extracted stevia extract.
  • the extraction step is a pre-treatment step of grinding stevia leaves to prepare stevia powder; an extract extraction step of extracting a stevia extract from the stevia powder; It may include a concentration step of concentrating the stevia extract and a freeze-drying step of freeze-drying the concentrated stevia extract.
  • the pretreatment step is characterized in that the pulverized stevia powder is degreased with n-hexane and then dried.
  • the extract extraction step is characterized in that the stevia powder is dissolved in ethanol to extract the stevia extract through reflux.
  • the purification step may include: a first purification step of dissolving the extracted stevia extract in distilled water, filling a column, and adding a resin to the column to separate non-polar substances; a second purification step of eluting an acidic material by passing sodium hydroxide (NaOH) through the column; A neutralization step of neutralizing the stevia extract from which acidic substances have been removed by adding hydrogen chloride (HCl), and after the neutralized stevia extract is charged in a column, eluted with ethanol to obtain a stevia extract from which non-polar substances are removed 3 may include a purification step.
  • a first purification step of dissolving the extracted stevia extract in distilled water, filling a column, and adding a resin to the column to separate non-polar substances
  • a second purification step of eluting an acidic material by passing sodium hydroxide (NaOH) through the column
  • the ethanol in the third purification step is characterized in that 100% ethanol.
  • the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may further contain a green tea extract.
  • the green tea extract is characterized in that it contains green tea saponins.
  • the green tea saponin is characterized in that it includes green tea theasaponin-E1 (theasaponin E1).
  • the anti-obesity food composition containing the stevia extract according to the embodiment of the present invention as described above contains the stevia extract having angiogenesis inhibitory function, thereby exhibiting an anti-obesity effect and helping weight control.
  • 1 is a graph of the cytotoxicity test results according to the concentration of the stevia extract of the present invention.
  • Figure 2 is a graph of angiogenesis inhibition test results according to the concentration of the stevia extract of the present invention.
  • Figure 4 is a PCR test photograph of detecting a signal transduction molecule according to the concentration of the stevia extract of the present invention.
  • FIG. 5 is a conceptual diagram showing a signal transduction pathway for angiogenesis.
  • FIG. 7 is a graph of the analysis of the fat accumulation rate according to the concentrations of the stevia extract of the present invention and the stevia extract and green tea extract using 3T3-L1 cells.
  • FIG. 9 is a conceptual diagram showing the molecular pathways of adipogenesis and adipocyte formation in the mesentery.
  • 10 is a graph of weight loss results according to the period of 10 groups.
  • the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may further contain a green tea extract.
  • the present invention relates to an anti-obesity food composition containing a stevia extract, and it was found that the stevia extract has an angiogenesis inhibitory function. would like to provide
  • angiogenesis inhibitory function is further improved to provide a food composition excellent in anti-obesity effect.
  • angiogenesis is a term that refers to a series of processes in which new blood vessels are created from existing blood vessels. It is known to be essential for various diseases such as growth of the body, rheumatoid arthritis, and diabetic blindness. That is, in a normal state, factors that induce angiogenesis and factors that inhibit angiogenesis are in equilibrium in the living body, but in a disease-induced situation, factors that promote blood vessel growth increase or the inhibitory factors do not function properly. This self-regulation is not controlled and continues to grow and develop into a disease.
  • tissue enlargement can be prevented and adipocyte formation and fat formation can be inhibited, thereby bringing about an anti-obesity effect.
  • the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may contain a stevia extract.
  • Stevia extract is extracted from stevia and contains Stevioside and Rebaudioside, so it is generally used as a natural sweetener.
  • the extract may be used in an anti-obesity food composition to exhibit an anti-obesity effect.
  • Stevia extract may be prepared through an extraction step and a purification step.
  • the extraction step is a step of extracting a stevia extract from a stevia leaf, and may include a pre-treatment step, an extract extraction step, a concentration step, and a freeze-drying step.
  • dried stevia leaves may be pulverized to prepare stevia powder. This is to powder the dried Stevia leaves to improve the extraction efficiency.
  • the pulverized stevia powder in order to remove the fat-soluble substances from the stevia leaves, may be degreased with n-hexane and then dried.
  • stevia leaves which are plants, contain fat-soluble substances such as oils and fats, and since these components are not needed during extraction, only polar components can be extracted and extraction can be made more easily.
  • the stevia extract can be extracted from the stevia powder, the stevia powder can be dissolved in ethanol, and the stevia extract can be extracted through reflux.
  • reflux may be performed at 50 to 60° C. for 5 to 6.5 hours, and most preferably at 60° C. for 6 hours.
  • the reflux time is less than 5 hours, the extraction may not be completely completed, and if it is more than 6.5 hours, it may be inefficient because the extraction has already been completed.
  • the stevia extract extracted in the extract extraction step may be concentrated under reduced pressure.
  • the stevia extract concentrated in the concentration step may be freeze-dried to make the stevia extract in powder form.
  • freeze-drying may be performed at -70 to -85°C at 45 to 55 atm for 23 to 25 hours, and preferably at -80°C to 50 atm (atm) for 24 hours, but is not limited thereto. does not
  • the purification step is a step of purifying the stevia extract extracted through the extraction step and prepared in powder form, and through purification, impurities are removed and the concentration is high, so that the function of inhibiting angiogenesis and the like can be further improved.
  • This purification step may include a first purification step, a second purification step, a neutralization step, and a third purification step.
  • the stevia extract extracted through the extraction step is dissolved in distilled water, charged in a column, and then a resin is added to the column filled with the stevia extract to separate non-polar substances from the stevia extract. there is.
  • the non-polar material is adsorbed to the resin and separated from the stevia extract, but is still mixed in the column, and the non-polar material separated through the third purification step can be completely removed from the stevia extract.
  • non-polar macro resin it is preferable to use a non-polar macro resin as the resin, but the present invention is not limited thereto.
  • 40 to 60 parts by weight of the stevia extract may be dissolved in 90 to 110 parts by weight of distilled water, and 50 parts by weight is preferably dissolved in 100 parts by weight of distilled water, but is not limited thereto.
  • sodium hydroxide NaOH
  • NaOH sodium hydroxide
  • the sodium hydroxide (NaOH) is preferably 0.4N NaOH, but is not limited thereto.
  • the stevia extract from which the acidic material has been removed in the second purification step may be taken out from the column, put in a container, and neutralized by adding hydrogen chloride (HCl). This is neutralized by adding acidic hydrogen chloride to the stevia extract that has become basic by removing acidic substances. This is because the neutralization of the stevia extract is necessary for refining.
  • HCl hydrogen chloride
  • the stevia extract neutralized in the neutralization step is charged in a new column, and then eluted with ethanol (eluent) to obtain a stevia extract from which non-polar substances are removed.
  • the eluate is the stevia extract finally obtained.
  • the ethanol used herein is preferably 100% ethanol.
  • the third purification step is a step for completely eluting only the polar material, and when 100% ethanol is not used, the separation efficiency is lowered.
  • the stevia extract is preferably contained in the anti-obesity food composition at a concentration of 3 g/70 kg-man/day or 100 ⁇ g/ml or less.
  • the stevia extract may contain 1 to 3% by weight based on the total weight% of the anti-obesity food composition, more preferably 3% by weight.
  • the concentration or content of these stevia extracts was determined to be a safe dose based on animal experiments, and 3.0g/70Kg-man/day was found to be appropriate when substituted for the 70Kg adult male dose.
  • the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may further contain a green tea extract.
  • Green tea extract is an extract extracted from green tea seeds, contains green tea saponins, and has an anti-angiogenesis inhibitory function. and High-carbohydrate Diet, KIM, 2015, BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, 2.
  • green tea extract containing green tea saponins (theasaponin E1, etc.) together with a stevia extract, angiogenesis inhibitory function, adipocyte and adipogenesis inhibitory function can be more effectively exhibited.
  • the green tea extract may be contained in an amount of 0.005 to 0.015% by weight, more preferably 0.01% by weight, based on the total weight% of the anti-obesity food composition.
  • the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention contains a stevia extract having angiogenesis inhibitory function, thereby exhibiting an anti-obesity effect and helping weight control.
  • HUVEC lines were purchased from Korea Cell Line Bank (Seoul, Korea), and HUVECs were cultured at 37° C., 5% CO 2 in an incubator using EBM-2 medium (CloneticsTM, USA).
  • the cultured HUVECs were placed in a 96-well culture plate, treated with stevia extract by concentration (0, 5, 15, 30, 50, 100, 200, 300 and 500 ⁇ g/ml), and incubated at 37° C. for 24 hours. Then, 0.5% MTT solution (Sigma, MO, USA) was added and incubated at 37° C. for 4 hours. Absorbance was measured at a wavelength of 540 ⁇ m using a microplate reader (Biochrom Ltd., Cambridge, UK), and the viability was calculated.
  • FIG. 1 The result is shown in FIG. 1 .
  • a preferred concentration of the stevia extract was selected to be 100 ⁇ g/ml or less.
  • a 96-well culture plate was coated with Matrigel (BD Bioscience, MA, US) and then incubated at 37° C. for 1 hour, HUVEC was isolated, added to Matrigel-coated wells, and incubated at 37°C for 4 hours.
  • Matrigel BD Bioscience, MA, US
  • Stevia extracts were added to the wells at different concentrations (5, 50, 100, 150 and 200 ⁇ g/ml) and incubated at 37° C. for 4 hours, and then the length of the formed tube network was analyzed.
  • the length of the tube mesh was taken by dividing the plate well into 5 fields under a microscope (Nikon, Tokyo, Japan), and the captured images were analyzed using Scion Image software (NIH, ML, USA), each Total tube length was calculated for the treatment and control groups.
  • the stevia extract has an angiogenesis inhibitory function.
  • HUVECs were cultured in EBM-2 medium, and total RNA was extracted using TRI solution.
  • RevertAid Premium First Strand cDNA synthesis kit was used for DNA synthesis, PCR was used for gene expression, and primers as shown in Table 1 (signal transduction molecule gene primer) were used. At this time, the stevia extract was treated at different concentrations (5, 15, 30, 50 and 100 ⁇ g/ml).
  • PCR reaction was performed using Takara ExTaq Hot Start Version (Takara, Japan), and the reaction was 3 min at 94 °C, 30 sec at 94 °C, 45 sec at 60 °C, and 1 min at 72 °C, and 7 min at 72 °C. Incompletely amplified by reaction was supplemented, and after 30 cycles of reaction, it was developed with 1.5% agarose gel.
  • VEGFR-2 VEGFR-2
  • PI3K PI3K
  • ⁇ -catenin ⁇ -cat
  • VE-cadherin VE-cadherin
  • VEGFR-2 VEGFR-2
  • PI3K PI3K
  • ⁇ -catenin ⁇ -cat
  • AKT-1 Akt
  • NF-kB NF-kB
  • the Oil Red O staining method which is stained in response to fat in lipid droplets in a specific way, was performed to confirm the generation of intracellular fat cells. Analyzes were performed using absorbance.
  • Adipocyte induction of 3T3-L1 cells was performed in cell culture wells with a mixture of stevia extract (0, 5, 15, 30, 50 and 100 ⁇ g/ml) and stevia extract and green tea extract by concentration (sp + st ; Adipocytes were induced through MDI medium containing
  • PCR test was performed using mesenteric adipose tissue and liver tissue. The result is shown in FIG. 8 .
  • mice 8-week-old female ICR mice (Taconic, Chungbuk, Korea) were divided into 10 groups after a one-week acclimatization process, and 6 mice were randomly assigned to each group under standard conditions (light/dark cycle and relative conditions at 22°C for 12 hours). Humidity was 50% ⁇ 5%), and food and water were supplied ad libitum.
  • group 10 is HFC (high-fat diet), HF-100 (high-fat diet + stevia extract 100 mg/kg/mouse body weight/d), HF-300 (high-fat diet + stevia extract 300 mg/kg) /mouse body weight/d), HCC (high-carbohydrate diet), HC-100 (high-carb diet + stevia extract 100 mg/kg/mouse body weight/d), HC-300 (high-carb diet + stevia extract) 300 mg/kg/mouse body weight/d), NC (normal diet), PHF-300 (after 4 weeks of high-fat diet, high-fat diet + stevia extract 300 mg/kg/mouse body weight/d), ORL (high-fat diet+Orlistat 50 mg/kg/mouse body weight/d) and ST+SP (high-fat diet+stevia extract and green tea extract 350 mg/kg/mouse body weight/d).
  • ST+SP was composed of 300 mg/kg/mouse body weight/d of
  • mRNA such as ACC (acetyl-CoA carboxylase), GPAT (Glycerol-3-phosphate O-acyltransferase), FAS (fatty acid synthase) and MLYCD (malonyl-CoA decarboxylase) was decreased in the group treated with stevia extract.
  • ACC acetyl-CoA carboxylase
  • GPAT Glycerol-3-phosphate O-acyltransferase
  • FAS fatty acid synthase
  • MLYCD malonyl-CoA decarboxylase
  • genes involved in the lipogenesis and adipogenesis of adipose tissue in the mesenteric adipose tissue can be made as shown in FIG. 9 , Pparg (peroxisome proliferator-activated receptor gamma), Srebp (Sterol regulatory element-binding protein 1), Cebpa (CCAAT/ enhancer-binding protein alpha) was confirmed to be reduced in the group treated with the stevia extract.
  • Pparg peroxisome proliferator-activated receptor gamma
  • Srebp Sterol regulatory element-binding protein 1
  • Cebpa CCAAT/ enhancer-binding protein alpha
  • LPL lipoprotein lipase
  • AP2 adipocyte protein 2
  • leptin leptin
  • mice 8-week-old female ICR mice (Taconic, Chungbuk, Korea) were divided into 10 groups after a one-week acclimatization process, and 6 mice were randomly assigned to each group under standard conditions (light/dark cycle and relative conditions at 22°C for 12 hours). Humidity was 50% ⁇ 5%), and food and water were supplied ad libitum.
  • group 10 is HFC (high-fat diet), HF-100 (high-fat diet + stevia extract 100 mg/kg/mouse body weight/d), HF-300 (high-fat diet + stevia extract 300 mg/kg) /mouse body weight/d), HCC (high-carbohydrate diet), HC-100 (high-carb diet + stevia extract 100 mg/kg/mouse body weight/d), HC-300 (high-carb diet + stevia extract) 300 mg/kg/mouse body weight/d), NC (normal diet), PHF-300 (after 4 weeks of high-fat diet, high-fat diet + stevia extract 300 mg/kg/mouse body weight/d), ORL (high-fat diet+Orlistat 50 mg/kg/mouse body weight/d) and ST+SP (high-fat diet+stevia extract and green tea extract 350 mg/kg/mouse body weight/d).
  • ST+SP was composed of 300 mg/kg/mouse body weight/d of
  • Leptin increases as body weight increases and fat increases, and as shown in FIG. In addition, it was confirmed that leptin was lower than ORL (Orlistat) in ST+SP.
  • Adiponectin is an important factor in controlling fat. Referring to FIG. 11(b), it was confirmed that the activity of adiponectin was much higher in the group administered with the stevia extract, and stevia extract (ST) + green tea extract (SP) was found to be the highest in

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Botany (AREA)
  • Mycology (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

The present invention relates to a food composition including a stevia extract and can provide an anti-obesity food composition including a stevia extract that exhibits an anti-obesity effect through an inhibitory function against angiogenesis. In addition, the anti-obesity food composition that may further include a green tea extract to exhibit a further enhanced effect can be provided.

Description

스테비아 추출물을 함유하는 항비만 식품조성물Anti-obesity food composition containing stevia extract
본 발명은 스테비아 추출물을 함유하는 항비만 식품조성물에 관한 것으로, 더욱 자세하게는 신생혈관형성 억제기능을 가지는 스테비아 추출물 단독 또는 녹차 추출물과 혼합하여 사용함으로써, 우수한 항비만 효과를 나타내며 부작용이 없는 스테비아 추출물을 함유하는 항비만 식품조성물에 관한 것이다.The present invention relates to an anti-obesity food composition containing a stevia extract, and more particularly, by using a stevia extract having an angiogenesis inhibitory function alone or in combination with a green tea extract, a stevia extract having excellent anti-obesity effect and no side effects It relates to an anti-obesity food composition containing.
현재 저출산과 생명공학기술 발달로 평균수명이 연장되고 있으며 우리나라 고령인구 비율이 계속 증가되고 있어 건강관리에 대한 관심 또한 높아지고 있는 추세이다.Currently, the low fertility rate and the development of biotechnology are extending life expectancy, and as the proportion of the elderly population in Korea continues to increase, interest in health care is also increasing.
한편, 대사성질환을 관리하기 위해서는 여러 의약품이 사용되고 있으나 만성질환이라는 측면에서 식습관 및 운동을 통한 예방이 강조되고 있다. 이를 타깃으로 하여 여러 카테고리의 식품이 개발 유통되고 있다.On the other hand, various medicines are used to manage metabolic diseases, but prevention through diet and exercise is emphasized in terms of chronic diseases. Various categories of food products are being developed and distributed targeting this.
식품 중 대사성질환을 타깃으로 한 식품에는 일부 대사성질환과 관련이 있는 기능성을 표현하는 건강기능식품, 대사성질환을 타깃으로 하는 체중조절용식품, 대사성질환의 유발을 저감화할 수 있는 일부 일반식품 등이 해당한다.Among foods, foods that target metabolic diseases include health functional foods that express functionality related to some metabolic diseases, weight control foods that target metabolic diseases, and some general foods that can reduce the incidence of metabolic diseases. do.
국민건강영양조사 분석결과에 의하면 국내 30세 이상 성인 중 28.85가 대사증후군 유병율을 보이고 있으며, 대사증후군 5개의 구성 요소(허리둘레, 중성지방, 고밀도지방, 혈압, 공복혈당) 중 1개 이상 요소에서 기준치를 초과한 대상자는 73.7%에 해당한다고 나타났다.According to the analysis results of the National Health and Nutrition Examination Survey, 28.85 of Korean adults aged 30 years or older have a prevalence of metabolic syndrome, and at least one of the five components of metabolic syndrome (waist circumference, triglyceride, high-density fat, blood pressure, and fasting blood sugar) It was found that 73.7% of subjects exceeded the standard.
이와 같은 대사성질환을 유발하는 원인 중 하나인 비만은 성장하는 소아청소년의 건강을 위협하는 원인이며, 특히 소아와 청소년 비만은 성인 비만으로 이행할 가능성이 높고, 대사성질환 외 심혈관질환, 위장질환, 폐질환, 골관절질환, 인슐린저항성 등의 합병증을 유발하여 성인기 건강에도 영향을 미칠 수 있다.Obesity, one of the causes of such metabolic diseases, is a cause that threatens the health of growing children and adolescents. In particular, obesity in children and adolescents is highly likely to transition to adult obesity. It can also affect adult health by causing complications such as diseases, osteoarthritis, and insulin resistance.
특히 우리나라 소아청소년에서 대사증후군의 유병율은 전반전으로 6.1에서 10.4%까지 다양하게 보고되고 있으며, 30세 이상에서 고혈압, 당뇨병 그리고 고콜레스테롤 혈증 유병율 모두 매우 높게 나타나고 있는 상황이다.In particular, the prevalence of metabolic syndrome in Korean children and adolescents is reported to vary from 6.1 to 10.4% in the first half, and the prevalence of hypertension, diabetes, and hypercholesterolemia are all very high in those over 30 years of age.
이에 다양한 체지방 감소 관련 기능성 식품, 비만 치료제 등이 개발되고 있는 실정이며, 현재까지의 비만 치료제는 식용을 억제시키는 제제와 지방의 대사를 촉진시키거나 지방질의 합성을 억제시키거나 대사활성을 증가시키는 제제로 크게 나눌 수 있었다.Accordingly, various functional foods related to body fat reduction, anti-obesity drugs, etc. are being developed. To date, anti-obesity drugs include agents that inhibit eating and agents that promote fat metabolism, inhibit fat synthesis, or increase metabolic activity. could be broadly divided into
그러나, 이러한 식품과 치료제들은 합성제제로 이루어져 부작용이 많을 뿐만 아니라 습관성 문제도 대두되고 있다.However, these foods and therapeutic agents are made of synthetic formulations, and thus not only have many side effects, but also habituation problems are emerging.
이에 따라, 천연소재를 이용하여 부작용이 없으면서도 항비만 효과를 통해 체중 조절에 도움을 줄 수 있는 제제의 개발이 필요하다.Accordingly, it is necessary to develop a formulation that can help weight control through an anti-obesity effect without side effects using natural materials.
상기와 같은 문제를 해결하고자, 본 발명은 신생혈관형성 억제기능을 가지는 스테비아 추출물 단독 또는 녹차 추출물과 혼합하여 사용함으로써, 우수한 항비만 효과를 나타내며 부작용이 없는 스테비아 추출물을 함유하는 항비만 식품조성물을 제공하는 데 목적이 있다.In order to solve the above problems, the present invention provides an anti-obesity food composition containing a stevia extract that exhibits excellent anti-obesity effect and has no side effects by using a stevia extract having angiogenesis inhibitory function alone or mixed with green tea extract purpose is to
상기와 같은 과제를 해결하기 위하여, 본 발명의 실시예에 따른 스테비아 추출물을 함유하는 항비만 식품조성물을 제공할 수 있다.In order to solve the above problems, it is possible to provide an anti-obesity food composition containing a stevia extract according to an embodiment of the present invention.
여기서, 스테비아 추출물을 함유하는 항비만 식품조성물은 신생혈관억제 기능을 가지는 것을 특징으로 한다.Here, the anti-obesity food composition containing the stevia extract is characterized in that it has an angiogenesis inhibitory function.
또한, 상기 스테비아 추출물은 스테비아 잎으로부터 스테비아 추출물을 추출하는 추출단계 및 추출된 스테비아 추출물을 정제하는 정제단계를 통해 제조되는 것을 특징으로 한다.In addition, the stevia extract is characterized in that it is prepared through an extraction step of extracting a stevia extract from a stevia leaf and a purification step of purifying the extracted stevia extract.
또한, 상기 추출단계는 스테비아 잎을 분쇄하여 스테비아 분말로 제조하는 전처리단계; 상기 스테비아 분말에서 스테비아 추출물을 추출하는 추출물추출단계; 상기 스테비아 추출물을 농축하는 농축단계 및 농축된 스테비아 추출물을 동결건조시키는 동결건조단계를 포함할 수 있다.In addition, the extraction step is a pre-treatment step of grinding stevia leaves to prepare stevia powder; an extract extraction step of extracting a stevia extract from the stevia powder; It may include a concentration step of concentrating the stevia extract and a freeze-drying step of freeze-drying the concentrated stevia extract.
또한, 상기 전처리단계는 분쇄된 스테비아 분말을 n-헥산으로 탈지한 후 건조시키는 것을 특징으로 한다.In addition, the pretreatment step is characterized in that the pulverized stevia powder is degreased with n-hexane and then dried.
또한, 상기 추출물추출단계는 상기 스테비아 분말을 에탄올에 용해시켜 환류를 통해 스테비아 추출물을 추출하는 것을 특징으로 한다.In addition, the extract extraction step is characterized in that the stevia powder is dissolved in ethanol to extract the stevia extract through reflux.
또한, 상기 정제단계는 추출된 스테비아 추출물을 증류수에 용해시키고 컬럼(Column)에 충전시킨 후, 상기 컬럼에 수지를 첨가하여 비극성 물질을 분리시키는 제1 정제단계; 상기 컬럼에 수산화나트륨(NaOH)을 통과시켜 산성 물질을 용출시키는 제2 정제단계; 산성물질이 제거된 스테비아 추출물에 염화수소(HCl)을 첨가하여 중화시키는 중화단계 및 중화된 스테비아 추출물을 컬럼(Column)에 충전시킨 후, 에탄올을 통해 용출시켜 비극성 물질이 제거된 스테비아 추출물을 획득하는 제3 정제단계를 포함할 수 있다.In addition, the purification step may include: a first purification step of dissolving the extracted stevia extract in distilled water, filling a column, and adding a resin to the column to separate non-polar substances; a second purification step of eluting an acidic material by passing sodium hydroxide (NaOH) through the column; A neutralization step of neutralizing the stevia extract from which acidic substances have been removed by adding hydrogen chloride (HCl), and after the neutralized stevia extract is charged in a column, eluted with ethanol to obtain a stevia extract from which non-polar substances are removed 3 may include a purification step.
또한, 상기 제3 정제단계에서 에탄올은 100% 에탄올인 것을 특징으로 한다.In addition, the ethanol in the third purification step is characterized in that 100% ethanol.
또한, 본 발명의 실시예에 따른 스테비아 추출물을 함유한 항비만 식품조성물은 녹차 추출물을 더 함유할 수 있다.In addition, the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may further contain a green tea extract.
여기서, 상기 녹차 추출물은 녹차 사포닌을 포함하는 것을 특징으로 한다.Here, the green tea extract is characterized in that it contains green tea saponins.
또한 상기 녹차 사포닌은 녹차의 테아사포닌-E1(theasaponin E1)을 포함하는 것을 특징으로 한다.In addition, the green tea saponin is characterized in that it includes green tea theasaponin-E1 (theasaponin E1).
상기와 같은 본 발명의 실시예에 따른 스테비아 추출물을 함유하는 항비만 식품조성물은 신생혈관형성 억제기능을 가지는 스테비아 추출물을 함유함으로써, 항비만 효과를 나타내 체중조절에 도움을 줄 수 있다.The anti-obesity food composition containing the stevia extract according to the embodiment of the present invention as described above contains the stevia extract having angiogenesis inhibitory function, thereby exhibiting an anti-obesity effect and helping weight control.
특히, 녹차 추출물과 함께 사용하여 각 추출물을 단독으로 사용하는 것보다 우수한 항비만 효과를 나타낼 수 있다.In particular, when used together with green tea extract, it can exhibit superior anti-obesity effect than using each extract alone.
또한 스테비아 추출물과 같은 천연소재를 사용함으로써, 합성제제인 기존의 항비만제제의 다양한 부작용 발생, 습관성 등의 문제를 해소할 수 있다.In addition, by using natural materials such as stevia extract, it is possible to solve problems such as occurrence of various side effects and habituation of conventional anti-obesity agents, which are synthetic agents.
도 1은 본 발명의 스테비아 추출물의 농도에 따른 세포독성 시험 결과 그래프.1 is a graph of the cytotoxicity test results according to the concentration of the stevia extract of the present invention.
도 2는 본 발명의 스테비아 추출물의 농도에 따른 신생혈관 형성 억제 시험 결과 그래프.Figure 2 is a graph of angiogenesis inhibition test results according to the concentration of the stevia extract of the present invention.
도 3의 (a) 내지 (f)는 본 발명의 스테비아 추출물의 농도에 따른 신생혈관 형성 억제 시험 결과 사진.3 (a) to (f) are photographs of angiogenesis inhibition test results according to the concentration of the stevia extract of the present invention.
도 4는 본 발명의 스테비아 추출물의 농도에 따른 신호전달분자를 검출한 PCR 검사 사진.Figure 4 is a PCR test photograph of detecting a signal transduction molecule according to the concentration of the stevia extract of the present invention.
도 5는 신생혈관 형성을 위한 신호전달 경로를 나타낸 개념도.5 is a conceptual diagram showing a signal transduction pathway for angiogenesis.
도 6의 (a) 내지 (f)는 3T3-L1 세포를 이용하여 본 발명의 스테비아 추출물의 농도에 따른 지방 축적 정도를 분석한 Oil Red 염색 결과 사진.6 (a) to (f) are images of Oil Red staining results obtained by analyzing the degree of fat accumulation according to the concentration of the stevia extract of the present invention using 3T3-L1 cells.
도 7은 3T3-L1 세포를 이용하여 본 발명의 스테비아 추출물과 스테비아 추출물 및 녹차 추출물의 농도에 따른 지방 축적율 분석 그래프.7 is a graph of the analysis of the fat accumulation rate according to the concentrations of the stevia extract of the present invention and the stevia extract and green tea extract using 3T3-L1 cells.
도 8은 10개 그룹의 장간막 지방조직과 간 조직의 PCR 검사 사진.8 is a PCR test photograph of mesenteric adipose tissue and liver tissue of 10 groups.
도 9는 장간막의 지방형성 및 지방세포 형성 분자 경로를 나타낸 개념도.9 is a conceptual diagram showing the molecular pathways of adipogenesis and adipocyte formation in the mesentery.
도 10은 10개 그룹의 기간에 따른 체중 감소 결과 그래프.10 is a graph of weight loss results according to the period of 10 groups.
도 11의 (a) 및 (b)는 10개 그룹의 Leptin 및 adiponectin 활성 측정 결과 그래프.11 (a) and (b) are graphs of Leptin and adiponectin activity measurement results of 10 groups.
본 발명의 실시예에 따른 스테비아 추출물을 함유하는 항비만 식품조성물을 제공할 수 있다.It is possible to provide an anti-obesity food composition containing a stevia extract according to an embodiment of the present invention.
또한, 본 발명의 실시예에 따른 스테비아 추출물을 함유한 항비만 식품조성물은 녹차 추출물을 더 함유할 수 있다.In addition, the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may further contain a green tea extract.
이하, 도면을 참조한 본 발명의 설명은 특정한 실시 형태에 대해 한정되지 않으며, 다양한 변환을 가할 수 있고 여러 가지 실시예를 가질 수 있다. 또한, 이하에서 설명하는 내용은 본 발명의 사상 및 기술 범위에 포함되는 모든 변환, 균등물 내지 대체물을 포함하는 것으로 이해되어야 한다.Hereinafter, the description of the present invention with reference to the drawings is not limited to specific embodiments, and various modifications may be made and various embodiments may be provided. In addition, it should be understood that the contents described below include all transformations, equivalents, and substitutes included in the spirit and scope of the present invention.
이하의 설명에서 제1, 제2 등의 용어는 다양한 구성요소들을 설명하는데 사용되는 용어로서, 그 자체에 의미가 한정되지 아니하며, 하나의 구성요소를 다른 구성요소로부터 구별하는 목적으로만 사용된다.In the following description, terms such as 1st, 2nd, etc. are terms used to describe various components, meanings are not limited thereto, and are used only for the purpose of distinguishing one component from other components.
본 명세서 전체에 걸쳐 사용되는 동일한 참조번호는 동일한 구성요소를 나타낸다.Like reference numbers used throughout this specification refer to like elements.
본 발명에서 사용되는 단수의 표현은 문맥상 명백하게 다르게 뜻하지 않는 한, 복수의 표현을 포함한다. 또한, 이하에서 기재되는 "포함하다", "구비하다" 또는 "가지다" 등의 용어는 명세서상에 기재된 특징, 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것이 존재함을 지정하려는 것으로 해석되어야 하며, 하나 또는 그 이상의 다른 특징들이나, 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것들의 존재 또는 부가 가능성을 미리 배제하지 않는 것으로 이해되어야 한다.As used herein, the singular expression includes the plural expression unless the context clearly dictates otherwise. In addition, terms such as "comprises", "comprises" or "have" described below are intended to designate the existence of features, numbers, steps, operations, components, parts, or combinations thereof described in the specification. It should be construed as not precluding the possibility of addition or existence of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.
여기서, 반복되는 설명, 본 발명의 요지를 불필요하게 흐릴 수 있는 공지 기능, 및 구성에 대한 상세한 설명은 생략한다. 본 발명의 실시형태는 당 업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해서 제공되는 것이다.Here, repeated descriptions, well-known functions that may unnecessarily obscure the gist of the present invention, and detailed descriptions of configurations will be omitted. The embodiments of the present invention are provided in order to more completely explain the present invention to those of ordinary skill in the art.
이하, 본 발명의 실시 예를 첨부한 도 1 내지 도 11을 참조하여 상세히 설명하기로 한다.Hereinafter, an embodiment of the present invention will be described in detail with reference to the accompanying drawings 1 to 11.
본 발명은 스테비아 추출물을 함유한 항비만 식품조성물에 관한 것으로, 스테비아 추출물이 신생혈관형성 억제기능을 갖는 것을 발견하였고, 이를 통해 스테비아 추출물을 사용하여 항비만 효과를 나타내며 부작용이 없는 항비만 식품조성물을 제공하고자 한다.The present invention relates to an anti-obesity food composition containing a stevia extract, and it was found that the stevia extract has an angiogenesis inhibitory function. would like to provide
또한, 추출 방법을 통해 신생혈관형성 억제기능 등이 보다 효과적으로 나타나도록 한다.In addition, through the extraction method, the angiogenesis inhibitory function and the like appear more effectively.
또한, 스테비아 추출물과 함께 녹차 추출물을 함유함으로써, 신생혈관형성 억제기능이 보다 향상되어 항비만 효과가 우수한 식품조성물을 제공하고자 한다.In addition, by containing green tea extract together with stevia extract, angiogenesis inhibitory function is further improved to provide a food composition excellent in anti-obesity effect.
한편, 혈관신생(angiogenesis)은 기존의 혈관에서 새로운 혈관이 생성되는 일련의 과정을 일컫는 용어로서, 태아의 발생, 여성의 월경, 상처 치유과정 등의 정상적인 상황뿐 아니라 암세포의 성장과 전이, 비만세포의 성장, 류머티즘 관절염, 당뇨성 실명증 등의 여러 질병에서도 필수적으로 요구되는 것으로 알려져 있다. 즉, 정상적인 상태에서 생체는 혈관신생을 유도하는 인자들과 억제하는 인자들이 평형을 이루고 있으나, 질환이 유발되는 상황에서는 혈관성장을 촉진하는 인자들이 증가하거나, 억제하는 인자가 제대로 작용하지 못하여 혈관신생이 자율적으로 조절되지 못하고 계속 성장함으로써 질환으로 발전되게 된다. On the other hand, angiogenesis is a term that refers to a series of processes in which new blood vessels are created from existing blood vessels. It is known to be essential for various diseases such as growth of the body, rheumatoid arthritis, and diabetic blindness. That is, in a normal state, factors that induce angiogenesis and factors that inhibit angiogenesis are in equilibrium in the living body, but in a disease-induced situation, factors that promote blood vessel growth increase or the inhibitory factors do not function properly. This self-regulation is not controlled and continues to grow and develop into a disease.
따라서, 신생혈관의 형성을 억제함으로써 어떠한 조직에 새로운 혈관의 형성을 억제함으로써, 조직이 비대 해지는 것을 막을 수 있고 지방세포 형성 및 지방 형성의 억제가 가능하여, 항비만 효과를 가져올 수 있다고 사료된다.Therefore, it is thought that by inhibiting the formation of new blood vessels in any tissue by inhibiting the formation of new blood vessels, tissue enlargement can be prevented and adipocyte formation and fat formation can be inhibited, thereby bringing about an anti-obesity effect.
본 발명의 실시예에 따른 스테비아 추출물을 함유하는 항비만 식품조성물은 스테비아 추출물을 함유할 수 있다.The anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may contain a stevia extract.
스테비아 추출물은 스테비아로부터 추출된 것으로, Stevioside 및 Rebaudioside 등을 함유하고 있어 일반적으로 천연 감미료로 사용되고 있으나, 본 발명에서는 신생혈관형성 억제 기능, 지방세포 및 지방 형성 억제 기능을 갖는 것을 확인하였고, 이에 따라 스테비아 추출물은 항비만 식품조성물에 사용되어 항비만 효과를 나타낼 수 있다.Stevia extract is extracted from stevia and contains Stevioside and Rebaudioside, so it is generally used as a natural sweetener. The extract may be used in an anti-obesity food composition to exhibit an anti-obesity effect.
뿐만 아니라 본 발명의 추출방법을 통해 추출된 스테비아 추출물을 사용함으로써, 보다 우수한 신생혈관 억제 기능을 나타낼 수 있다. 이러한 스테비아 추출물의 제조방법에 대해 하기에서 자세히 설명하기로 한다.In addition, by using the stevia extract extracted through the extraction method of the present invention, it is possible to exhibit a superior angiogenesis inhibitory function. A method for preparing such a stevia extract will be described in detail below.
스테비아 추출물은 추출단계 및 정제단계를 통해 제조될 수 있다.Stevia extract may be prepared through an extraction step and a purification step.
구체적으로, 추출단계는 스테비아 잎으로부터 스테비아 추출물을 추출하는 단계로, 전처리단계, 추출물추출단계, 농축단계 및 동결건조단계를 포함할 수 있다.Specifically, the extraction step is a step of extracting a stevia extract from a stevia leaf, and may include a pre-treatment step, an extract extraction step, a concentration step, and a freeze-drying step.
먼저, 전처리단계는 건조된 스테비아 잎을 분쇄하여 스테비아 분말로 제조할 수 있다. 이는 추출 효율을 향상시키기 위해 건조된 스테비아 잎을 분말화하는 것이다.First, in the pretreatment step, dried stevia leaves may be pulverized to prepare stevia powder. This is to powder the dried Stevia leaves to improve the extraction efficiency.
또한, 전처리단계는 스테비아 잎의 지용성 물질을 제거하기 위하여, 분쇄된 스테비아 분말을 n-헥산으로 탈지한 후 건조시킬 수 있다.In addition, in the pretreatment step, in order to remove the fat-soluble substances from the stevia leaves, the pulverized stevia powder may be degreased with n-hexane and then dried.
즉, 식물인 스테비아 잎에는 오일, 지방 등 지용성 물질이 함유되어 있는데, 추출 시 이러한 성분은 필요없기 때문에 이를 제거함으로써, 극성성분만 추출하도록 할 수 있으며 추출이 더욱 용이하게 이루어지도록 할 수 있는 것이다. That is, stevia leaves, which are plants, contain fat-soluble substances such as oils and fats, and since these components are not needed during extraction, only polar components can be extracted and extraction can be made more easily.
추출물추출단계는 스테비아 분말에서 스테비아 추출물을 추출할 수 있는데, 스테비아 분말을 에탄올에 용해시키고 환류를 통해 스테비아 추출물을 추출할 수 있다.In the extract extraction step, the stevia extract can be extracted from the stevia powder, the stevia powder can be dissolved in ethanol, and the stevia extract can be extracted through reflux.
이때, 에탄올은 에탄올의 농도가 70% 미만일 경우 극성 물질이 완전히 추출되지 못하는 우려가 있고, 70% 초과일 경우 알코올의 탈수작용에 따른 성분의 구조 변화가 일어날 가능성이 있어, 70% 에탄올을 사용하는 것이 바람직하나, 이에 한정되지 않는다.At this time, in ethanol, when the concentration of ethanol is less than 70%, there is a fear that the polar material cannot be completely extracted. It is preferred, but not limited thereto.
또한, 환류는 50 내지 60℃에서 5 내지 6.5시간 동안 이루어질 수 있으며, 60℃에서 6시간 동안 이루어지는 것이 가장 바람직하다.In addition, reflux may be performed at 50 to 60° C. for 5 to 6.5 hours, and most preferably at 60° C. for 6 hours.
이는 환류 온도가 50℃ 미만일 경우 추출이 용이하게 이루어지지 않아 추출 효율이 저하될 수 있으며, 60℃ 초과일 경우 성분이 변형되거나 파괴될 수 있기 때문이다.This is because when the reflux temperature is less than 50 ℃, extraction is not easily made, so that the extraction efficiency may be lowered, and if it is more than 60 ℃, the component may be deformed or destroyed.
또한, 환류 시간이 5시간 미만일 경우 추출이 완전히 완료되지 못할 수 있고, 6.5시간 초과일 경우 이미 추출이 완료되었기 때문에 비효율적일 수 있다.In addition, if the reflux time is less than 5 hours, the extraction may not be completely completed, and if it is more than 6.5 hours, it may be inefficient because the extraction has already been completed.
이와 같은 환류를 통해 스테비아 분말에서 스테비아 추출물을 추출함으로써, 추출하고자 하는 극성 물질에 대한 추출이 더욱 잘 이루어질 수 있다.By extracting the stevia extract from the stevia powder through such reflux, the extraction of the polar substance to be extracted can be made better.
농축단계는 추출물추출단계에서 추출된 스테비아 추출물을 감압 농축시킬 수 있다. In the concentration step, the stevia extract extracted in the extract extraction step may be concentrated under reduced pressure.
동결건조단계는 농축단계에서 농축된 스테비아 추출물을 동결건조시켜, 스테비아 추출물을 분말 형태로 만들 수 있다.In the freeze-drying step, the stevia extract concentrated in the concentration step may be freeze-dried to make the stevia extract in powder form.
이때, 동결건조는 -70 내지 -85℃에서 45 내지 55기압(atm)으로 23 내지 25시간동안 이루어질 수 있으며, -80℃에서 50기압(atm)으로 24시간 동안 이루어지는 것이 바람직하나, 이에 한정되지는 않는다.At this time, freeze-drying may be performed at -70 to -85°C at 45 to 55 atm for 23 to 25 hours, and preferably at -80°C to 50 atm (atm) for 24 hours, but is not limited thereto. does not
정제단계는 추출단계를 통해 추출되어 분말 형태로 제조된 스테비아 추출물을 정제하는 단계로, 정제를 통해 불순물을 제거하고 고농도화시켜 신생혈관 억제 기능 등이 보다 향상되도록 할 수 있다.The purification step is a step of purifying the stevia extract extracted through the extraction step and prepared in powder form, and through purification, impurities are removed and the concentration is high, so that the function of inhibiting angiogenesis and the like can be further improved.
이러한 정제단계는 제1 정제단계, 제2 정제단계, 중화단계 및 제3 정제단계를 포함할 수 있다.This purification step may include a first purification step, a second purification step, a neutralization step, and a third purification step.
먼저 제1 정제단계는 추출단계를 통해 추출된 스테비아 추출물을 증류수에 용해시키고, 칼럼(Column)에 충전시킨 후, 스테비아 추출물이 충전된 컬럼에 수지를 첨가하여, 스테비아 추출물에서 비극성 물질을 분리시킬 수 있다. First, in the first purification step, the stevia extract extracted through the extraction step is dissolved in distilled water, charged in a column, and then a resin is added to the column filled with the stevia extract to separate non-polar substances from the stevia extract. there is.
이때, 비극성 물질은 수지에 흡착되어 스테비아 추출물에서 분리된 상태이긴 하나 아직 컬럼 안에서 섞여 있는 상태이고, 제3 정제단계를 통해 분리된 비극성 물질이 스테비아 추출물에서 완전히 제거될 수 있다.At this time, the non-polar material is adsorbed to the resin and separated from the stevia extract, but is still mixed in the column, and the non-polar material separated through the third purification step can be completely removed from the stevia extract.
이때, 수지는 non-polar macro resin를 사용함이 바람직하나, 이에 한정되지는 않는다.In this case, it is preferable to use a non-polar macro resin as the resin, but the present invention is not limited thereto.
여기서 스테비아 추출물은 40 내지 60중량부를 증류수 90 내지 110중량부에 용해시킬 수 있고, 50중량부를 증류수 100중량부에 용해시키는 것이 바람직하나, 이에 한정되지는 않는다.Here, 40 to 60 parts by weight of the stevia extract may be dissolved in 90 to 110 parts by weight of distilled water, and 50 parts by weight is preferably dissolved in 100 parts by weight of distilled water, but is not limited thereto.
제2 정제단계는 제1 정제단계에서 수지가 첨가된 컬럼에 수산화나트륨(NaOH)을 통과시켜 산성 물질을 용출시킬 수 있다. 이를 통해 스테비아 추출물에서 원하지 않는 물질인 산성 물질을 제거할 수 있다. In the second purification step, sodium hydroxide (NaOH) may be passed through the column to which the resin is added in the first purification step to elute the acidic material. Through this, it is possible to remove acidic substances, which are unwanted substances, from the stevia extract.
여기서 수산화나트륨(NaOH)은 0.4N NaOH를 사용하는 것이 바람직하나, 이에 한정되지는 않는다.Here, the sodium hydroxide (NaOH) is preferably 0.4N NaOH, but is not limited thereto.
중화단계는 제2 정제단계에서 산성물질이 제거된 스테비아 추출물을 컬럼에서 빼내 용기에 넣고 염화수소(HCl)을 첨가하여 중화시킬 수 있다. 이는 산성물질이 제거되어 염기성이 된 스테비아 추출물에 산성인 염화수소를 첨가하여 중화시키는 것이다. 재정제를 위해서는 스테비아 추출물의 중화가 필요하기 때문이다.In the neutralization step, the stevia extract from which the acidic material has been removed in the second purification step may be taken out from the column, put in a container, and neutralized by adding hydrogen chloride (HCl). This is neutralized by adding acidic hydrogen chloride to the stevia extract that has become basic by removing acidic substances. This is because the neutralization of the stevia extract is necessary for refining.
제3 정제단계는 중화단계에서 중화된 스테비아 추출물을 새 컬럼(Column)에 충전시킨 후, 에탄올(용리액)을 통해 용출시켜 비극성 물질이 제거된 스테비아 추출물을 획득할 수 있다. 용출액이 최종적으로 얻어지는 스테비아 추출물이다.In the third purification step, the stevia extract neutralized in the neutralization step is charged in a new column, and then eluted with ethanol (eluent) to obtain a stevia extract from which non-polar substances are removed. The eluate is the stevia extract finally obtained.
여기서 사용되는 에탄올은 100% 에탄올인 것이 바람직하다.The ethanol used herein is preferably 100% ethanol.
이는 제3 정제단계가 극성 물질만을 온전히 용출해내기 위한 단계인데, 100% 에탄올을 사용하지 않을 경우, 분리 효율이 저하되기 때문이다.This is because the third purification step is a step for completely eluting only the polar material, and when 100% ethanol is not used, the separation efficiency is lowered.
이러한 스테비아 추출물은 3g/70kg-man/day 또는 100㎍/ml 이하의 농도로 항비만 식품조성물에 함유되는 것이 바람직하다. 또한 스테비아 추출물은 항비만 식품조성물 전체 중량%에 대하여 1 내지 3중량%가 함유될 수 있으며, 보다 바람직하게 3중량%가 함유될 수 있다.The stevia extract is preferably contained in the anti-obesity food composition at a concentration of 3 g/70 kg-man/day or 100 μg/ml or less. In addition, the stevia extract may contain 1 to 3% by weight based on the total weight% of the anti-obesity food composition, more preferably 3% by weight.
이러한 스테비아 추출물의 농도 또는 함량은 동물 실험을 바탕으로 하여 안전 용량을 결정한 것으로, 70Kg의 성인남자의 용량으로 치환 했을 때 3.0g/70Kg-man/day가 적당한 것으로 나타났다.The concentration or content of these stevia extracts was determined to be a safe dose based on animal experiments, and 3.0g/70Kg-man/day was found to be appropriate when substituted for the 70Kg adult male dose.
또한, 본 발명의 실시예에 따른 스테비아 추출물을 함유한 항비만 식품조성물은 녹차 추출물을 더 함유할 수 있다.In addition, the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention may further contain a green tea extract.
녹차 추출물은 녹차 씨앗으로부터 추출된 추출물로써 녹차 사포닌을 포함하고, 신생혈관형성 억제 기능을 근간으로, 항비만(1. Anti-hyperlipidemic and Fat Pad Lowering Effect of Standardized Tea Seed Cake Extract in Mice Fed High-fat and High-carbohydrate Diet,KIM, 2015, BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, 2. Theasaponin E1 as an effective ingredient for anti-angiogenesis and anti-obesity effects, KIM, 2014, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY) 효과뿐만 아니라, 고지혈증(한국등록특허 제10-1854322호, 녹차씨앗 추출물을 유효성분으로 포함하는 고지혈증 치료용 조성물) 및 항암 효과(HPLC fractionation and pharmacological assessment of green tea seed saponins for antimicrobial, anti-angiogenic, and hemolytic activities, KIM, 2016, Biotechnology and Bioprocess Engineering) 등을 나타낼 수 있다.Green tea extract is an extract extracted from green tea seeds, contains green tea saponins, and has an anti-angiogenesis inhibitory function. and High-carbohydrate Diet, KIM, 2015, BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, 2. Theasaponin E1 as an effective ingredient for anti-angiogenesis and anti-obesity effects, KIM, 2014, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY Patent No. 10-1854322, Composition for treating hyperlipidemia containing green tea seed extract as an active ingredient) and anticancer effect (HPLC fractionation and pharmacological assessment of green tea seed saponins for antimicrobial, anti-angiogenic, and hemolytic activities, KIM, 2016, Biotechnology and Bioprocess Engineering) and the like.
이와 같이 녹차의 사포닌(테아사포닌-E1(theasaponin E1) 등)을 함유하는 녹차 추출물을 스테비아 추출물과 함께 사용함으로써, 신생혈관 억제 기능, 지방세포 및 지방 형성 억제 기능이 보다 효과적으로 나타나도록 할 수 있다.As such, by using a green tea extract containing green tea saponins (theasaponin E1, etc.) together with a stevia extract, angiogenesis inhibitory function, adipocyte and adipogenesis inhibitory function can be more effectively exhibited.
이러한 녹차 추출물은 항비만 식품조성물 전체 중량%에 대하여 0.005 내지 0.015중량%가 함유될 수 있으며, 보다 바람직하게 0.01중량%가 함유될 수 있다.The green tea extract may be contained in an amount of 0.005 to 0.015% by weight, more preferably 0.01% by weight, based on the total weight% of the anti-obesity food composition.
이와 같은 녹차 추출물의 함량은 항비만 효과 및 지방형성억제 작용 규명의 동물실험을 기반으로 결정한 것이다.The content of such green tea extract was determined based on animal experiments to identify anti-obesity and fat formation inhibitory effects.
상기에서 설명한 바와 같이, 본 발명의 실시예에 따른 스테비아 추출물을 함유하는 항비만 식품조성물은 신생혈관형성 억제기능을 가지는 스테비아 추출물을 함유함으로써, 항비만 효과를 나타내 체중조절에 도움을 줄 수 있다.As described above, the anti-obesity food composition containing a stevia extract according to an embodiment of the present invention contains a stevia extract having angiogenesis inhibitory function, thereby exhibiting an anti-obesity effect and helping weight control.
특히, 녹차 추출물과 함께 사용하여 각 추출물을 단독으로 사용하는 것보다 우수한 항비만 효과를 나타낼 수 있다.In particular, when used together with green tea extract, it can exhibit superior anti-obesity effect than using each extract alone.
또한 스테비아 추출물과 같은 천연소재를 사용함으로써, 합성제제인 기존의 항비만제제의 다양한 부작용 발생, 습관성 등의 문제를 해소할 수 있다.In addition, by using natural materials such as stevia extract, it is possible to solve problems such as occurrence of various side effects and habituation of conventional anti-obesity agents, which are synthetic agents.
이하 본 발명의 내용을 실시예, 비교예 및 시험예를 통하여 구체적으로 설명한다. 그러나, 이들은 이해를 돕기 위하여 제시되는 것으로서, 본 발명을 예시하는 것일 뿐 본 발명의 권리범위가 이들에 의해 한정되는 것은 아니다.Hereinafter, the contents of the present invention will be described in detail through Examples, Comparative Examples and Test Examples. However, these are presented to aid understanding, and are merely illustrative of the present invention, and the scope of the present invention is not limited thereto.
[[ 실험예Experimental example 1] 세포독성 시험 1] Cytotoxicity test
본 발명의 추출방법을 통해 추출된 스테비아 추출물의 안전한 농도 범위를 결정하기 위하여, 세포독성 시험을 실시하였다.In order to determine the safe concentration range of the stevia extract extracted through the extraction method of the present invention, a cytotoxicity test was performed.
HUVEC 라인을 한국 세포 라인 은행(서울, 한국)에서 구매하여, HUVEC를 EBM-2 배지(Clonetics™, USA)를 사용해 37℃, 5% CO2 인큐베이터(incubator)에서 배양하였다. 배양된 HUVEC를 96-웰 배양 플레이트에 넣고 스테비아 추출물을 농도별(0, 5, 15, 30, 50, 100, 200, 300 및 500㎍/ml)로 처리하고 37℃에서 24시간 동안 배양하였다. 그 후, 0.5%MTT 용액(Sigma, MO, USA)을 첨가하여 37℃에서 4시간동안 배양하였다. 마이크로플레이트 리더기(Biochrom Ltd., Cambridge, UK)를 이용하여 540㎛의 파장에서 흡광도를 측정하고, 생존율을 계산하였다.HUVEC lines were purchased from Korea Cell Line Bank (Seoul, Korea), and HUVECs were cultured at 37° C., 5% CO 2 in an incubator using EBM-2 medium (Clonetics™, USA). The cultured HUVECs were placed in a 96-well culture plate, treated with stevia extract by concentration (0, 5, 15, 30, 50, 100, 200, 300 and 500 μg/ml), and incubated at 37° C. for 24 hours. Then, 0.5% MTT solution (Sigma, MO, USA) was added and incubated at 37° C. for 4 hours. Absorbance was measured at a wavelength of 540 μm using a microplate reader (Biochrom Ltd., Cambridge, UK), and the viability was calculated.
그 결과는 도 1과 같다.The result is shown in FIG. 1 .
도 1을 보면 알 수 있듯이, 스테비아 추출물 150㎍/ml부터 세포 생존율이 떨어지는 것을 확인할 수 있었다.As can be seen from FIG. 1 , it was confirmed that the cell viability decreased from 150 μg/ml of stevia extract.
이에 따라, 스테비아 추출물의 바람직한 농도는 100㎍/ml 이하인 것으로 선정하였다.Accordingly, a preferred concentration of the stevia extract was selected to be 100 μg/ml or less.
[[ 실험예Experimental example 2] 신생혈관 형성 억제 시험 2] Angiogenesis inhibition test
본 발명의 추출방법을 통해 추출된 스테비아 추출물의 신생혈관 억제 기능을 확인하기 위하여, 96- 웰 배양 플레이트에 Matrigel(BD Bioscience, MA, US)로 코팅 한 다음 37℃에서 1시간 동안 배양시키고, HUVEC를 분리하여 Matrigel- 코팅 된 웰에 첨가하고 37℃에서 4시간 동안 재배양 하였다. In order to confirm the angiogenesis inhibitory function of the stevia extract extracted through the extraction method of the present invention, a 96-well culture plate was coated with Matrigel (BD Bioscience, MA, US) and then incubated at 37° C. for 1 hour, HUVEC was isolated, added to Matrigel-coated wells, and incubated at 37°C for 4 hours.
스테비아 추출물을 농도별(5, 50, 100, 150 및 200㎍/ml)로 웰에 첨가하고 37℃에서 4 시간 동안 배양한 후, 형성된 튜브 망의 길이를 분석하였다.Stevia extracts were added to the wells at different concentrations (5, 50, 100, 150 and 200 μg/ml) and incubated at 37° C. for 4 hours, and then the length of the formed tube network was analyzed.
여기서, 튜브 망의 길이는 현미경(Nikon, Tokyo, Japan)하에서 플레이트의 웰을 불규칙적으로 5분야로 나누어 촬영하였고, 촬영된 이미지를 Scion Image 소프트웨어(NIH, ML, USA)를 사용하여 분석하였고, 각각의 처리 및 대조군에 대해 총 튜브 길이를 계산 하였다.Here, the length of the tube mesh was taken by dividing the plate well into 5 fields under a microscope (Nikon, Tokyo, Japan), and the captured images were analyzed using Scion Image software (NIH, ML, USA), each Total tube length was calculated for the treatment and control groups.
그 결과는 도 2 및 도 3과 같다.The results are shown in FIGS. 2 and 3 .
도 2 및 도 3을 보면 알 수 있듯이, 스테비아 추출물 농도가 증가함에 따라 튜브 망의 길이가 짧은 것을 확인할 수 있었다. 이에 따라 스테비아 추출물은 신생혈관 형성 억제 기능을 가지는 것으로 판단된다.As can be seen from FIGS. 2 and 3 , it was confirmed that the length of the tube network was shorter as the concentration of the stevia extract increased. Accordingly, it is determined that the stevia extract has an angiogenesis inhibitory function.
[[ 실험예Experimental example 3] 신생혈관 형성의 신호전달 확인 3] Confirmation of signal transduction of angiogenesis
신생혈관형성억제의 기전을 파악하기 위하여, HUVECs을 EBM-2 배지에 배양한 후, 총 RNA의 추출은 TRI 용액을 사용하여 추출하였다. To understand the mechanism of angiogenesis inhibition, HUVECs were cultured in EBM-2 medium, and total RNA was extracted using TRI solution.
DNA의 합성은 RevertAid Premium First Strand cDNA synthesis Kit를 사용하였고, 유전자의 발현은 PCR을 사용하였으며, 하기 표 1(신호전달분자의 유전자 primer)과 같은 primer를 사용하였다. 이때 스테비아 추출물을 농도별(5, 15, 30, 50 및 100㎍/ml)로 처리하였다.RevertAid Premium First Strand cDNA synthesis kit was used for DNA synthesis, PCR was used for gene expression, and primers as shown in Table 1 (signal transduction molecule gene primer) were used. At this time, the stevia extract was treated at different concentrations (5, 15, 30, 50 and 100 μg/ml).
또한, PCR 반응은 Takara ExTaq Hot Start Version (Takara, Japan)을 사용하였고, 반응은 94℃에서 3min, 94℃에서 30sec, 60℃에서 45sec, 그리고 72℃에서 1 분간, 또한, 72℃에서 7 분간 반응시켜 불완전하게 증폭된 것을 보완하였으며, 30cycle을 반응시킨 후 1.5% agarose gel로 전개하였다. In addition, the PCR reaction was performed using Takara ExTaq Hot Start Version (Takara, Japan), and the reaction was 3 min at 94 °C, 30 sec at 94 °C, 45 sec at 60 °C, and 1 min at 72 °C, and 7 min at 72 °C. Incompletely amplified by reaction was supplemented, and after 30 cycles of reaction, it was developed with 1.5% agarose gel.
그 다음 신호전달분자의 PCR 검사를 진행하였다. 그 결과는 도 4와 같다.Then, PCR test of the signal transduction molecule was performed. The result is shown in FIG. 4 .
Figure PCTKR2020019029-appb-T000001
Figure PCTKR2020019029-appb-T000001
한편, 도 5의 세포 표면의 VEGFR-2(VEGFR2), PI3K, β-catenin(β-cat) 및 VE-cadherin의 분자들의 신호가 핵내까지 신호가 전달되면 신생혈관이 형성되지만 도중에 신호가 차단되면 NF-kB까지 신호 전달이 일어나지 않아 신생혈관이 형성되지 못한다.On the other hand, when the signal of molecules of VEGFR-2 (VEGFR2), PI3K, β-catenin (β-cat) and VE-cadherin on the cell surface of FIG. 5 is transmitted to the nucleus, angiogenesis is formed, but when the signal is blocked in the middle Signal transduction does not occur up to NF-kB, so angiogenesis is not formed.
도 4에 나타난 바와 같이, 스테비아 추출물의 농도 증가에 따라 VEGFR-2(VEGFR2), PI3K, β-catenin(β-cat), AKT-1(Akt), NF-kB가 감소되는 것으로 나타났다.As shown in FIG. 4 , VEGFR-2 (VEGFR2), PI3K, β-catenin (β-cat), AKT-1 (Akt), and NF-kB were decreased as the concentration of the Stevia extract increased.
즉, NF-kB의 발현이 감소됨에 따라 신생혈관 형성이 억제되는 것으로 판단된다.That is, as the expression of NF-kB is reduced, it is determined that angiogenesis is suppressed.
[실험예 4] 3T3-L1 세포를 이용한 지방 축적 억제 효과[Experimental Example 4] Fat accumulation inhibitory effect using 3T3-L1 cells
3T3-L1 세포를 분화 촉진한 후, 세포내 지방구 생성을 확인하기 위하여 지방적(lipid droplets)의 지방에 특이적으로 반응하여 염색되는 Oil Red O 염색법을 수행하여 축적 중인 지방을 염색된 지방구를 흡광도를 이용하여 분석하였다.After promoting the differentiation of 3T3-L1 cells, the Oil Red O staining method, which is stained in response to fat in lipid droplets in a specific way, was performed to confirm the generation of intracellular fat cells. Analyzes were performed using absorbance.
3T3-L1 세포의 지방세포 유도는 세포 배양 웰(well)에 농도별 스테비아 추출물(0, 5, 15, 30, 50 및 100㎍/ml)과 농도별 스테비아 추출물 및 녹차 추출물의 혼합물(sp+st, 0, 5, 15, 30, 50 및 100㎍/ml)을 첨가한 후 100% 융합성(confluent)시키고 이틀 후 100μM 3-isobutyl-1-methylxanthine, Sigma I5879, 250μM dexamethasone, Sigma D4902, 170μM insulin 을 함유한 MDI 배지를 통해 지방세포로 유도하였으며, 3일 후에 10% FBS와 170nM insulin, Sigma I9278을 함유한 DMEM 배지로 각 실험 날짜까지 이틀에 한번 교환하였다.Adipocyte induction of 3T3-L1 cells was performed in cell culture wells with a mixture of stevia extract (0, 5, 15, 30, 50 and 100 μg/ml) and stevia extract and green tea extract by concentration (sp + st ; Adipocytes were induced through MDI medium containing
3T3-L1 세포 배양 중 각각 3, 6, 9일에 배양액 성분을 완전히 제거하기 위하여 phosphate buffered saline(PBS, pH 7.0)로 2번 세정하고 0.6%의 Oil Red O 용액을 준비한 후 약 2시간 동안 염색하고, PBS로 3번 세정한 후 isopropanol 1.5 mL을 넣어 oil red를 용출시켰다. 용출시킨 용액을 관찰하였으며 500nm에서 흡광도를 측정하여 지방 축적율을 계산하였다. 그 결과는 도 6 및 도 7과 같다.During 3T3-L1 cell culture, wash twice with phosphate buffered saline (PBS, pH 7.0) to completely remove the culture medium on days 3, 6, and 9, respectively, and prepare a 0.6% Oil Red O solution and then stain for about 2 hours. After washing 3 times with PBS, 1.5 mL of isopropanol was added to elute oil red. The eluted solution was observed and the absorbance was measured at 500 nm to calculate the fat accumulation rate. The results are shown in FIGS. 6 and 7 .
도 6은 지방 형성 억제 결과로, 5㎍/ml, 30㎍/ml, 50㎍/ml, 100㎍/ml의 스테비아 추출물과 스테비아 추출물(ST) 100㎍/ml 및 녹차 추출물(SP) 25㎍/ml를 혼합한 혼합물(ST+SP)을 처리한 oil red 염색 결과 사진이다.6 is a result of suppression of fat formation, 5㎍/ml, 30㎍/ml, 50㎍/ml, 100㎍/ml of stevia extract and stevia extract (ST) 100㎍/ml and green tea extract (SP) 25㎍/ This is a picture of the result of oil red staining treated with a mixture of ml (ST+SP).
도 6의 (a) 내지 (f)는 3T3-L1 세포를 이용하여 본 발명의 스테비아 추출물의 농도에 따른 지방 축적 정도를 분석한 Oil Red 염색 결과 사진.6 (a) to (f) are images of Oil Red staining results obtained by analyzing the degree of fat accumulation according to the concentration of the stevia extract of the present invention using 3T3-L1 cells.
도 6과 도 7을 보면 알 수 있듯이, 스테비아 추출물의 농도 증가에 따라 지방체가 현저히 감소되는 것을 확인할 수 있었다. 특히 도 6에서 혼합물(ST+SP)을 처리한 군에서는 거의 지방체가 나타나지 않아 스테비아 추출물 단독보다는 녹차 추출물과 혼합하여 사용하는 것이 보다 좋은 결과를 나타내는 것을 확인할 수 있었다.As can be seen from FIGS. 6 and 7 , it was confirmed that the fat body was significantly reduced as the concentration of the stevia extract increased. In particular, in the group treated with the mixture (ST + SP) in FIG. 6 , almost no fat was observed, so it was confirmed that using a mixture with green tea extract rather than stevia extract alone showed better results.
또한 도 7의 지방 축척율, 즉 지방 축적 억제 효과도 혼합물이 보다 높은 효과를 보여주는 것을 확인할 수 있었다.In addition, it was confirmed that the mixture showed a higher effect on the fat accumulation rate of FIG. 7 , that is, the fat accumulation inhibitory effect.
[[ 실험예Experimental example 5] 5] mRNAmRNA 발현에 관여하는 효소 및 전사인자에 대한 스테비아 추출물의 영향 Effects of Stevia Extract on Enzymes and Transcription Factors Involved in Expression
지방 및 에너지 대사에 관련되는 경로의 biomaker에 스테비아 추출물이 어떠한 영향을 미치는가를 평가하기 위하여, 장간막 지방조직과 간 조직을 사용하여 PCR 검사를 진행하였다. 그 결과는 도 8과 같다.In order to evaluate the effect of stevia extract on the biomaker of pathways related to fat and energy metabolism, PCR test was performed using mesenteric adipose tissue and liver tissue. The result is shown in FIG. 8 .
실험은 8주령 암컷 ICR 마우스(Taconic, Chungbuk, Korea)는 1주간 환경 순응 과정 후 10 그룹으로 나누어 각 그룹별로 6 마리씩 무작위로 할당한 후 표준 조건(22℃에서 12시간 동안 광/암 주기 및 상대 습도 50%±5%)하에 관리되었고 임의로 먹이와 물을 공급하였다.For the experiment, 8-week-old female ICR mice (Taconic, Chungbuk, Korea) were divided into 10 groups after a one-week acclimatization process, and 6 mice were randomly assigned to each group under standard conditions (light/dark cycle and relative conditions at 22°C for 12 hours). Humidity was 50%±5%), and food and water were supplied ad libitum.
여기서 10그룹은 HFC(고-지방 식이), HF-100(고-지방 식이+스테비아 추출물 100 mg/kg/mouse body weight/d), HF-300(고-지방 식이+스테비아 추출물 300 mg/kg/mouse body weight/d), HCC(고-탄수화물 식이), HC-100(고-탄수화물 식이+스테비아 추출물 100 mg/kg/mouse body weight/d), HC-300(고-탄수화물 식이+스테비아 추출물 300 mg/kg/mouse body weight/d), NC(정상 식이), PHF-300(4주간 고-지방 식이 후, 고-지방 식이+스테비아 추출물 300 mg/kg/mouse body weight/d), ORL(고-지방 식이+ Orlistat 50 mg/kg/mouse body weight/d), ST+SP(고-지방 식이+스테비아 추출물 및 녹차 추출물 350 mg/kg/mouse body weight/d)으로 이루어진다. 이때, ST+SP은 고-지방 식이에 스테비아 추출물 300 mg/kg/mouse body weight/d과 녹차 추출물 50 mg/kg/mouse body weight/d로 구성하였다.Here, group 10 is HFC (high-fat diet), HF-100 (high-fat diet + stevia extract 100 mg/kg/mouse body weight/d), HF-300 (high-fat diet + stevia extract 300 mg/kg) /mouse body weight/d), HCC (high-carbohydrate diet), HC-100 (high-carb diet + stevia extract 100 mg/kg/mouse body weight/d), HC-300 (high-carb diet + stevia extract) 300 mg/kg/mouse body weight/d), NC (normal diet), PHF-300 (after 4 weeks of high-fat diet, high-fat diet + stevia extract 300 mg/kg/mouse body weight/d), ORL (high-fat diet+Orlistat 50 mg/kg/mouse body weight/d) and ST+SP (high-fat diet+stevia extract and green tea extract 350 mg/kg/mouse body weight/d). In this case, ST+SP was composed of 300 mg/kg/mouse body weight/d of stevia extract and 50 mg/kg/mouse body weight/d of green tea extract in a high-fat diet.
간 조직에서 ACC(acetyl-CoA carboxylase), GPAT(Glycerol-3-phosphate O-acyltransferase), FAS(fatty acid synthase) 및 MLYCD(malonyl-CoA decarboxylase) 등의 mRNA는 스테비아 추출물을 처리한 군들에서 감소되는 것을 확인할 수 있었다.In liver tissue, mRNA such as ACC (acetyl-CoA carboxylase), GPAT (Glycerol-3-phosphate O-acyltransferase), FAS (fatty acid synthase) and MLYCD (malonyl-CoA decarboxylase) was decreased in the group treated with stevia extract. could confirm that
장간막 지방조직의 지방조직의 lipogenesis 및 adipogenesis에 관여하는 유전자의 발현은 도 9와 같이 이루어질 수 있는데, Pparg(peroxisome proliferator-activated receptor gamma), Srebp(Sterol regulatory element-binding protein 1), Cebpa(CCAAT/enhancer-binding protein alpha) 등은 스테비아 추출물을 처리한 군들에서 감소되는 것을 확인할 수 있었다.Expression of genes involved in the lipogenesis and adipogenesis of adipose tissue in the mesenteric adipose tissue can be made as shown in FIG. 9 , Pparg (peroxisome proliferator-activated receptor gamma), Srebp (Sterol regulatory element-binding protein 1), Cebpa (CCAAT/ enhancer-binding protein alpha) was confirmed to be reduced in the group treated with the stevia extract.
또한 장간막 WAT에 있어 lipogenesis 및 adipogenesis에 관련된 LPL(lipoprotein lipase), aP2(adipocyte protein 2) 및 leptin은 스테비아 추출물의 처리 농도 증가에 따라 감소되는 것으로 나타났다.In addition, in mesenteric WAT, lipoprotein lipase (LPL), adipocyte protein 2 (aP2) and leptin, which are involved in lipogenesis and adipogenesis, were found to be decreased with increasing concentration of stevia extract.
[실험예 6] 항비만 효과[Experimental Example 6] Anti-obesity effect
스테비아 추출물의 항비만 효과를 확인하기 위하여 하기와 같이 실험을 진행하였고, 2일 마다 마우스의 체중을 확인하였다. 그 결과는 도 10과 같다.In order to confirm the anti-obesity effect of the stevia extract, the experiment was conducted as follows, and the weight of the mouse was checked every 2 days. The result is shown in FIG. 10 .
실험은 8주령 암컷 ICR 마우스(Taconic, Chungbuk, Korea)는 1주간 환경 순응 과정 후 10 그룹으로 나누어 각 그룹별로 6 마리씩 무작위로 할당한 후 표준 조건(22℃에서 12시간 동안 광/암 주기 및 상대 습도 50%±5%)하에 관리되었고 임의로 먹이와 물을 공급하였다.For the experiment, 8-week-old female ICR mice (Taconic, Chungbuk, Korea) were divided into 10 groups after a one-week acclimatization process, and 6 mice were randomly assigned to each group under standard conditions (light/dark cycle and relative conditions at 22°C for 12 hours). Humidity was 50%±5%), and food and water were supplied ad libitum.
여기서 10그룹은 HFC(고-지방 식이), HF-100(고-지방 식이+스테비아 추출물 100 mg/kg/mouse body weight/d), HF-300(고-지방 식이+스테비아 추출물 300 mg/kg/mouse body weight/d), HCC(고-탄수화물 식이), HC-100(고-탄수화물 식이+스테비아 추출물 100 mg/kg/mouse body weight/d), HC-300(고-탄수화물 식이+스테비아 추출물 300 mg/kg/mouse body weight/d), NC(정상 식이), PHF-300(4주간 고-지방 식이 후, 고-지방 식이+스테비아 추출물 300 mg/kg/mouse body weight/d), ORL(고-지방 식이+ Orlistat 50 mg/kg/mouse body weight/d), ST+SP(고-지방 식이+스테비아 추출물 및 녹차 추출물 350 mg/kg/mouse body weight/d)으로 이루어진다. 이때, ST+SP은 고-지방 식이에 스테비아 추출물 300 mg/kg/mouse body weight/d과 녹차 추출물 50 mg/kg/mouse body weight/d로 구성하였다.Here, group 10 is HFC (high-fat diet), HF-100 (high-fat diet + stevia extract 100 mg/kg/mouse body weight/d), HF-300 (high-fat diet + stevia extract 300 mg/kg) /mouse body weight/d), HCC (high-carbohydrate diet), HC-100 (high-carb diet + stevia extract 100 mg/kg/mouse body weight/d), HC-300 (high-carb diet + stevia extract) 300 mg/kg/mouse body weight/d), NC (normal diet), PHF-300 (after 4 weeks of high-fat diet, high-fat diet + stevia extract 300 mg/kg/mouse body weight/d), ORL (high-fat diet+Orlistat 50 mg/kg/mouse body weight/d) and ST+SP (high-fat diet+stevia extract and green tea extract 350 mg/kg/mouse body weight/d). In this case, ST+SP was composed of 300 mg/kg/mouse body weight/d of stevia extract and 50 mg/kg/mouse body weight/d of green tea extract in a high-fat diet.
도 10을 보면 알 수 있듯이, 스테비아 추출물을 처리한 군에서 모두 체중이 감소되는 것을 확인할 수 있었다. 이것은 지방세포의 생성 및 지질 생성의 억제에 기인되는 것으로 판단된다.As can be seen from FIG. 10 , it was confirmed that all of the groups treated with the stevia extract lost weight. This is thought to be due to the suppression of the production of adipocytes and the production of lipids.
또한 HF-300 및 HC-300의 체중은 양성 대조군 ORL(Orlistat)과 거의 동일하게 나타났으며, 인위적으로 고지방 식이로 체중을 증가 시킨 후에 스테비아를 투여한 경우인 PHF에서도 현저한 체중 감소 효과를 보이는 것을 확인할 수 있었다.In addition, the body weights of HF-300 and HC-300 were almost the same as those of the positive control ORL (Orlistat), and it was found that the PHF, which was administered after artificially increasing body weight with a high-fat diet, also showed a significant weight loss effect in PHF. could check
특히 스테비아 추출물(ST)+녹차 추출물(SP)에서 체중이 현저하게 감소되는 것을 확인할 수 있었으며, 양성대조군인 ORL(Orlistat)보다 스테비아 추출물(ST)+녹차 추출물(SP)가 현저한 효과를 나타내는 것을 확인할 수 있었다.In particular, it was confirmed that the body weight was significantly reduced in the stevia extract (ST) + green tea extract (SP), and it was confirmed that the stevia extract (ST) + green tea extract (SP) showed a significant effect than the positive control ORL (Orlistat). could
[실험예 7] 렙틴(Leptin) 및 아디포넥틴(adiponectin) 활성 평가[Experimental Example 7] Leptin and adiponectin activity evaluation
각 그룹(10개 그룹)에서 혈액 샘플을 채취하여, ELISA 키트(Abcam USA)를 사용하여 렙틴과 아디포넥틴의 혈청 농도를 측정하였다. 그 결과는 도 11과 같다.Blood samples were collected from each group (10 groups), and serum concentrations of leptin and adiponectin were measured using an ELISA kit (Abcam USA). The result is shown in FIG. 11 .
각 그룹 샘플을 원심 분리하고 100㎕ 상등액을 키트 지침에 따라 모든 그룹의 샘플을 혼합하고, 키트 표준을 플레이트에 첨가한 후, 실온에서 30분 동안 배양한 다음, 96-웰 마이크로 플레이트 판독기를 사용하여 450nm에서 흡광도를 측정하였다.Centrifuge each group sample and add 100 μl supernatant according to kit instructions to mix samples from all groups, add kit standards to the plate, incubate for 30 min at room temperature, then use a 96-well microplate reader to Absorbance was measured at 450 nm.
렙틴은 체중이 증가하고 지방이 많을수록 증가는데, 도 11의 (a)에 나타난 바와 같이, 스테비아 추출물을 투여한 그룹들에서 렙틴의 활성이 낮게 나타나는 것을 확인할 수 있었다. 또한 ST+SP에서 ORL(Orlistat) 보다 렙틴이 낮게 나타나는 것을 확인할 수 있었다.Leptin increases as body weight increases and fat increases, and as shown in FIG. In addition, it was confirmed that leptin was lower than ORL (Orlistat) in ST+SP.
아디포넥틴은 지방을 제어하는 중요한 인자로써, 도 11의 (b)를 보면, 스테비아 추출물을 투여한 그룹들에서 아디포넥틴의 활성이 훨씬 높게 나타나는 것을 확인할 수 있었고, 스테비아 추출물(ST)+녹차 추출물(SP)에서 가장 높게 나타나는 것을 확인할 수 있었다.Adiponectin is an important factor in controlling fat. Referring to FIG. 11(b), it was confirmed that the activity of adiponectin was much higher in the group administered with the stevia extract, and stevia extract (ST) + green tea extract (SP) was found to be the highest in
이상으로 첨부된 도면을 참조하여 본 발명의 실시예를 설명하였으나, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고 다른 구체적인 형태로 실시할 수 있다는 것을 이해할 수 있을 것이다. 따라서 이상에서 기술한 실시예는 모든 면에서 예시적인 것이며 한정적이 아닌 것이다.Although the embodiments of the present invention have been described above with reference to the accompanying drawings, those of ordinary skill in the art to which the present invention pertains can practice the present invention in other specific forms without changing the technical spirit or essential features of the present invention. you will be able to understand Accordingly, the embodiments described above are illustrative in all respects and not restrictive.

Claims (11)

  1. 스테비아 추출물을 함유하는 항비만 식품조성물.An anti-obesity food composition containing stevia extract.
  2. 제1항에 있어서,According to claim 1,
    상기 스테비아 추출물은,The stevia extract is
    신생혈관억제 기능을 가지는 것을 특징으로 하는 항비만 식품조성물.Anti-obesity food composition, characterized in that it has an angiogenesis inhibitory function.
  3. 제1항에 있어서,According to claim 1,
    상기 스테비아 추출물은,The stevia extract is
    스테비아 잎으로부터 스테비아 추출물을 추출하는 추출단계 및Extraction step of extracting stevia extract from stevia leaf and
    추출된 스테비아 추출물을 정제하는 정제단계를 통해 제조되는 것을 특징으로 하는 항비만 식품조성물.An anti-obesity food composition, characterized in that it is prepared through a purification step of purifying the extracted stevia extract.
  4. 제3항에 있어서,4. The method of claim 3,
    상기 추출단계는,The extraction step is
    스테비아 잎을 분쇄하여 스테비아 분말로 제조하는 전처리단계;A pre-treatment step of pulverizing stevia leaves to prepare stevia powder;
    상기 스테비아 분말에서 스테비아 추출물을 추출하는 추출물추출단계;an extract extraction step of extracting a stevia extract from the stevia powder;
    상기 스테비아 추출물을 농축하는 농축단계 및a concentration step of concentrating the stevia extract; and
    농축된 스테비아 추출물을 동결건조시키는 동결건조단계를 포함하는 항비만 식품조성물.An anti-obesity food composition comprising a freeze-drying step of freeze-drying the concentrated stevia extract.
  5. 제4항에 있어서,5. The method of claim 4,
    상기 전처리단계는,The pre-processing step is
    분쇄된 스테비아 분말을 n-헥산으로 탈지한 후 건조시키는 것을 특징으로 하는 항비만 식품조성물.An anti-obesity food composition, characterized in that the pulverized stevia powder is degreased with n-hexane and then dried.
  6. 제4항에 있어서,5. The method of claim 4,
    상기 추출물추출단계는,The extract extraction step is,
    상기 스테비아 분말을 에탄올에 용해시켜 환류를 통해 스테비아 추출물을 추출하는 것을 특징으로 하는 항비만 식품조성물.Anti-obesity food composition, characterized in that the stevia powder is dissolved in ethanol to extract the stevia extract through reflux.
  7. 제3항에 있어서,4. The method of claim 3,
    상기 정제단계는,The purification step is
    추출된 스테비아 추출물을 증류수에 용해시키고 컬럼(Column)에 충전시킨 후, 상기 컬럼에 수지를 첨가하여 비극성 물질을 분리시키는 제1 정제단계;A first purification step of dissolving the extracted stevia extract in distilled water, filling a column, and adding a resin to the column to separate non-polar substances;
    상기 컬럼에 수산화나트륨(NaOH)을 통과시켜 산성 물질을 용출시키는 제2 정제단계;a second purification step of eluting an acidic material by passing sodium hydroxide (NaOH) through the column;
    산성물질이 제거된 스테비아 추출물에 염화수소(HCl)을 첨가하여 중화시키는 중화단계 및Neutralization step of neutralizing by adding hydrogen chloride (HCl) to the stevia extract from which acidic substances have been removed, and
    중화된 스테비아 추출물을 컬럼(Column)에 충전시킨 후, 에탄올을 통해 용출시켜 비극성 물질이 제거된 스테비아 추출물을 획득하는 제3 정제단계를 포함하는 항비만 식품조성물.An anti-obesity food composition comprising a third purification step of filling a neutralized stevia extract in a column, and then eluting it with ethanol to obtain a stevia extract from which non-polar substances are removed.
  8. 제7항에 있어서,8. The method of claim 7,
    상기 제3 정제단계에서 에탄올은,In the third purification step, ethanol is
    100% 에탄올인 것을 특징으로 하는 항비만 식품조성물.Anti-obesity food composition, characterized in that 100% ethanol.
  9. 제1항에 있어서,According to claim 1,
    녹차 추출물을 더 함유하는 항비만 식품조성물.Anti-obesity food composition further containing green tea extract.
  10. 제9항에 있어서,10. The method of claim 9,
    상기 녹차 추출물은,The green tea extract,
    녹차 사포닌을 포함하는 것을 특징으로 하는 항비만 식품조성물.Anti-obesity food composition comprising green tea saponin.
  11. 제10항에 있어서,11. The method of claim 10,
    상기 녹차 사포닌은,The green tea saponins are
    녹차의 테아사포닌-E1(theasaponin E1)을 포함하는 것을 특징으로 하는 항비만 식품조성물.Anti-obesity food composition comprising green tea theasaponin-E1 (theasaponin E1).
PCT/KR2020/019029 2020-11-02 2020-12-23 Anti-obesity food composition including stevia extract WO2022092441A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020200144730A KR102603687B1 (en) 2020-11-02 2020-11-02 Food Composition with Anti-obesity activity containing Stevia extract
KR10-2020-0144730 2020-11-02

Publications (1)

Publication Number Publication Date
WO2022092441A1 true WO2022092441A1 (en) 2022-05-05

Family

ID=81382799

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2020/019029 WO2022092441A1 (en) 2020-11-02 2020-12-23 Anti-obesity food composition including stevia extract

Country Status (2)

Country Link
KR (1) KR102603687B1 (en)
WO (1) WO2022092441A1 (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101617590B1 (en) * 2015-07-14 2016-05-02 (주)보뚜슈퍼푸드 Antiobesity Food Composition Comprising Extract of Aronia, Acaiberry and Stevia
KR20180070066A (en) * 2016-12-16 2018-06-26 강원블루베리 영농조합법인 A product method of using blueberry concentrate extract

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101617590B1 (en) * 2015-07-14 2016-05-02 (주)보뚜슈퍼푸드 Antiobesity Food Composition Comprising Extract of Aronia, Acaiberry and Stevia
KR20180070066A (en) * 2016-12-16 2018-06-26 강원블루베리 영농조합법인 A product method of using blueberry concentrate extract

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
KATARZYNA GAWEł-BęBEN, TOMASZ BUJAK, ZOFIA NIZIOł-ŁUKASZEWSKA, BEATA ANTOSIEWICZ, ANNA JAKUBCZYK, MONIKA KARA&: "Stevia Rebaudiana Bert. Leaf Extracts as a Multifunctional Source of Natural Antioxidants", MOLECULES, MDPI AG, CH, vol. 20, no. 4, 1 April 2015 (2015-04-01), CH , pages 5468 - 5486, XP055667897, ISSN: 1420-3049, DOI: 10.3390/molecules20045468 *
KHAN MUHAMMAD IMRAN, MUHAMMAD ZUBAIR KHAN , MIN YONG KIM, JONG DEOG KIM: "P0610: Anti-hyperlipidemic and Fat Pad Lowering Effects of Standardized Formulation of Rebaudioside A, Sativoside and Theasaponin E1 in Mice Fed with High-fat and High-carbohydrate Diet", 2020 KOREAN SOCIETY OF BIOLOGICAL ENGINEERING FALL CONFERENCE AND INTERNATIONAL SYMPOSIUM: HYBRID-CONFERENCE, KOREAN SOCIETY OF BIOLOGICAL ENGINEERING, KOREA, 1 October 2020 (2020-10-01) - 23 October 2020 (2020-10-23), Korea, pages 377 - 377, XP055925658 *
KHAN MUHAMMAD IMRAN, ZUBAIR KHAN MUHAMMAD, YONG MIN, CHO, YONG KIM MIN, SUNG CHANG, KIM, DEOG JONG, KIM: "P0115: Anti-Obesity Effects of Stevia Standardized Steviosides and Rebaudiosides Mixture by Attenuating Adipogenesis and Lipogenesis via Downregulation and Suppression of the Obesity Promoting Signaling Molecules", 2020 KOREAN SOCIETY OF BIOLOGICAL ENGINEERING SPRING CONFERENCE AND INTERNATIONAL SYMPOSIUM, KOREAN SOCIETY OF BIOLOGICAL ENGINEERING, KOREA, 1 June 2020 (2020-06-01) - 26 June 2020 (2020-06-26), Korea, pages 156 - 156, XP055925636 *
KIM JONG DEOG, HYO JIN SEO; TAI SUN SHIN: "Composition for Anti-Angiogenesis containing Tea-Saponin E 1 as an Effective Ingredient", KOREAN SOCIETY OF BIOLOGICAL ENGINEERING CONFERENCE; 2012 FALL CONFERENCE, KOREAN SOCIETY OF BIOLOGICAL ENGINEERING, 1 September 2012 (2012-09-01), pages 43 - 43, XP055925632 *
PARK JEONG-EUN, KEE HEEJIN, CHA YOUN-SOO: "Effect of Stevia rebaudiana Bertoni Leaf Extract on Antiobesity in C57BL/6J Mice", KOREAN JOURNAL OF FOOD SCIENCE AND TECHNOLOGY., SEOUL, KR, vol. 42, no. 5, 31 October 2010 (2010-10-31), KR , pages 586 - 592, XP055925622, ISSN: 0367-6293 *

Also Published As

Publication number Publication date
KR20220060044A (en) 2022-05-11
KR102603687B1 (en) 2023-11-20

Similar Documents

Publication Publication Date Title
WO2018012834A1 (en) Akkermansia muciniphila strain having effect of preventing or treating degenerative brain diseases or metabolic diseases, and use thereof
WO2021261632A1 (en) Novel faecalibacterium prausnitzii strain eb-fpdk11 and use thereof
WO2015056982A1 (en) Method for inducing pluripotent stem cells and pluripotent stem cells prepared by said method
WO2019135637A1 (en) Method for preparing gynostemma pentaphyllum leaf extract containing increased quantity of low molecular effective saponin and decreased quantity of benzopyrene, and gynostemma pentaphyllum leaf extract prepared thereby
WO2020076136A2 (en) Akkermansia muciniphila strain and use thereof
WO2019059606A1 (en) Composition containing amomum villosum extract for prevention, alleviation, or treatment of obesity
WO2014058142A1 (en) Pharmaceutical composition containing aster glehni extract as active ingredientfor preventing or treating obesity or metabolic diseases
WO2022131428A1 (en) Composition comprising orlistat and eb-amdk19 strain of akkermansia muciniphila
WO2016032249A1 (en) Pharmaceutical composition containing vaccinium bracteatum thunb. extract or fraction thereof as active ingredient for preventing or treating neuroinflammation or neuro-degenerative diseases
WO2022092441A1 (en) Anti-obesity food composition including stevia extract
WO2020209690A1 (en) Pharmaceutical composition containing wild-walnut extract, fraction thereof, or physiologically active substance derived therefrom as active ingredient, and having effects of inhibiting production of advanced glycation end products and decomposing advanced glycation end products
WO2011122879A2 (en) Composition comprising coumestrol or a bean extract containing coumestrol
WO2021261631A1 (en) Novel picalibacterium prosnich eb-fpdk9 strain and uses thereof
WO2013012117A1 (en) Pharmaceutical compositions for preventing or treating inflammatory diseases, comprising phytosterol compound
WO2022139489A1 (en) Pharmaceutical composition for preventing or treating alcoholic and nonalcoholic fatty liver disease
WO2021020923A1 (en) Composition and method for preventing, alleviating, or treating liver injury
WO2015072667A1 (en) Pharmaceutical composition for preventing or treating degenerative cranial nerve diseases
WO2021033995A1 (en) Composition comprising amomum tsaoko extract for prevention, alleviation, or treatment of sarcopenia-related disease
WO2011043564A2 (en) Composition containing black ginseng extracts for preventing or treating liver cancer
JP2005503381A (en) Sesquiterpenoid derivatives with adipocyte differentiation inhibitory action
WO2021096134A1 (en) Moisturizing or anti-atopic composition containing fatty acids or fatty acid derivatives
WO2020067699A1 (en) Composition for alleviating fatigue or enhancing exercise capability comprising angelica gigas extract, cnidii rhizoma extract, and paeonia japonica extract
WO2024038980A1 (en) Lactobacillus plantarum ku15120 strain or use thereof
WO2023229282A1 (en) Composition for preventing, treating, or improving metabolic diseases, comprising lactobacillus kunkeei nchbl-003 strain or culture medium thereof
WO2023229263A1 (en) Composition for preventing, treating, or improving metabolic diseases comprising lactobacillus plantarum nchbl-004 strain or culture medium thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20960057

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20960057

Country of ref document: EP

Kind code of ref document: A1