Composition for preventing, ameliorating or treating obesity comprising wasabi leaf extract as active ingredient

Abstract

The present invention relates to a composition for treating obesity, comprising a wasabi leaf extract, which is a natural extract, as an active ingredient.　The present invention relates to a composition for preventing, ameliorating or treating obesity, comprising a wasabi leaf extract, wherein the wasabi leaf extract includes 0.8-1.6 mg / g of isovitexin and 4.8-9.6 mg / g of sinigrin, and the weight ratio of isovitexin to sinigrin in the composition is 1:1-1:6 (isovitexin: sinigrin). The composition of the present invention exhibits the effects of reducing body weight, body fat, and abdominal fat without causing appetite reduction.

Description

Composition for preventing, improving or treating obesity containing wasabi leaf extract as an active ingredient

[0001] This application claims priority to Korean Patent Application No. 10-2024-0004918, filed January 11, 2024, the entire disclosure of which is incorporated herein by reference. The present invention relates to a composition for preventing, improving, or treating obesity, comprising a wasabi leaf extract as an active ingredient, a natural extract.

[0002] Obesity is a risk factor for various life-threatening metabolic diseases, such as cardiovascular disease, hypertension, and Type II diabetes. In obesity, fat cells not only store lipids as a long-term energy source, but also secrete various adipokines that are involved in the metabolism and inflammation of both adipocytes and non-adipocytes, causing chronic inflammation and related metabolic diseases such as insulin resistance. Recently, the incidence of metabolic syndrome, also known as Type II diabetes, has been increasing worldwide due to obesity caused by a high-calorie diet and lack of exercise, and it is predicted that the number of patients with metabolic syndrome will double, reaching approximately 300 million by 2025. Obesity caused by abnormal localized fat accumulation, such as abdominal obesity, provides various causes that can seriously induce atherosclerotic diseases such as hypertension, hyperlipidemia, blood coagulation disorders, vascular inflammation, and insulin resistance accompanied by abnormally increased insulin secretion, ultimately contributing to the increased prevalence of life-threatening cardiovascular diseases. Excessive fatty acid intake leads to the accumulation of triglycerides (TG) in various tissues, which increases lipolysis and circulating fatty acids. This leads to insulin resistance in adipocytes, which in turn leads to fat accumulation in non-adipocytes such as muscle, pancreas, and liver. When insulin resistance is induced in adipocytes, excess fatty acid binding and transport proteins lead to increased fatty acid uptake by non-adipocytes, which negatively affects insulin-mediated glucose metabolism, particularly in muscle cells. Simultaneously, prolonged exposure to free fatty acids in the pancreas leads to impaired insulin secretion through a mechanism known as lipotoxicity, creating a vicious cycle. In this case, high concentrations of free fatty acids also accumulate in the liver, leading to insulin resistance and the release of large amounts of glucose from the liver.Accumulation of TG within hepatocytes also causes non-alcoholic fatty liver disease (NAFLD), which induces fat accumulation in liver cells primarily responsible for glucose metabolism, secondary to steatohepatitis, and ultimately induces fibrosis due to hepatocyte necrosis. Therefore, the balance between lipid synthesis and breakdown in hepatocytes has become an important therapeutic target for suppressing insulin resistance and NAFLD caused by metabolic syndrome.

[0003] Currently, various treatments for metabolic syndrome are being developed, and as appropriate control of oxidative stress, which is identified as a major cause of diabetes and related complications, along with appropriate blood sugar control, has recently emerged as the most essential method for diabetes treatment, attempts are being made to develop α-glucosidase inhibitors or antioxidants that have fewer side effects and are more effective.

[0004] Among these, metformin is a widely used oral biguanide antidiabetic agent and is well-known as an AMPK activator. Metformin is the first-line treatment for overweight and obese type II diabetes patients with normal renal function. It can reduce the incidence of cardiovascular disease, a serious side effect of diabetes, and is the only drug known to regulate the secretion of pancreatic enzyme granules directly involved in fat digestion and breakdown, as well as the activity of enzymes involved in hepatic glucose metabolism. Metformin is currently known to be the most widely prescribed antidiabetic drug worldwide. While metformin is primarily used to treat type II diabetes, its use for polycystic ovary syndrome (PCOS) and premature puberty is also on the rise. Metformin is also known to cause side effects such as lactic acidosis, which can be caused by renal impairment.

[0005] Accordingly, there is a need for the development of alternative health functional foods and medicines derived from natural products that are more effective and have fewer side effects.

[0006] Numerous references and citations are provided throughout this specification. The disclosures of these references are incorporated herein by reference in their entirety to further clarify the state of the art and the scope of the present invention.

[0007] The purpose of the present inventors is to provide a composition for preventing, improving or treating obesity, which contains wasabi leaf extract as an active ingredient.

[0008] Another object of the present inventors is to provide a method for producing a wasabi leaf extract that exhibits an effect of preventing, improving or treating obesity.

[0009] Another object of the present inventors is to provide a method for preventing, improving or treating obesity using wasabi leaf extract as an effective ingredient.

[0010] Other objects and advantages of the present invention will become more apparent from the detailed description, claims and drawings below.

[0011] One aspect of the present invention is to provide a composition for preventing, improving or treating obesity, comprising a wasabi leaf extract as an active ingredient.

[0012] Wasabi, or horseradish (Wasabia / Eutrema japonica Matsum.), is a native Japanese plant that has long been used as a medicinal herb. It contains large amounts of isothiocyanate compounds, such as allyisothiocyanate, which impart a unique, pungent flavor, making it a popular spice and pickle. While its antioxidant and anticancer properties are well known, the physiological activities of parts of wasabi other than the root, particularly the leaves, remain largely unknown.

[0013] The wasabi leaf extract of the present invention contains isovitexin in a content of 0.8 to 1.6 mg / g and sinigrin in a content of 4.8 to 0.6 mg / g, and has been confirmed to have at least one effect selected from the group consisting of weight loss, body fat loss, and abdominal fat loss without causing a decrease in appetite.

[0014] The composition of the present invention includes extracts of the above natural ingredients as effective ingredients. The term 'extract' used in this specification includes the extracted product obtained by juicing a raw material or treating a raw material with an extraction solvent, or a processed product formulated (e.g., powdered) thereof.

[0015] When the extract used in the composition of the present invention is obtained by treating the raw material with an extraction solvent, various extraction solvents can be used, for example, a polar solvent or a non-polar solvent can be used. Polar solvents include (i) water, (ii) alcohol (preferably, methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO), and non-polar solvents include acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, Chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF can also be used.

[0016] Preferably, the extract used in the present invention may be obtained by juicing the raw material or by using any one selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixtures thereof as an extraction solvent, but is not limited thereto.

[0017] In addition, the term 'extract' used in this specification has the meaning commonly used in the art as a crude extract as described above, but in a broad sense, it also includes a fraction obtained by further fractionating an extract. That is, it includes not only an extract obtained by juicing the raw material or using the extraction solvent described above, but also one obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, a fraction obtained through various additional purification methods such as separation by various chromatographies (designed for separation according to size, charge, hydrophobicity, or affinity), etc., are also included in the extract of the present invention.

[0018] In addition, the extract of the present invention may be obtained by removing the solvent through an additional process, such as filtration, concentration, or drying, or by performing all of filtration, concentration, and drying. Filtration may be performed, for example, using filter paper or a vacuum filter, concentration may be performed using a vacuum concentrator, and drying may be performed using spray drying or freeze drying to obtain a powdered extract.

[0019] In one embodiment, the wasabi leaf extract of the present invention is applied at a temperature of 98 to 102°C and a pressure of 0.3 to 0.5 kg / cm 2 It may be extracted using water as an extraction solvent under pressure conditions. More specifically, the wasabi leaf extract of the present invention is prepared by crushing the raw material and then extracting it using water 15 to 20 times the amount of the raw material as an extraction solvent at 98 to 102°C for 8±1 hours and a pressure of 0.3 to 0.5 kg / cm. 2It is extracted by maintaining. The extracted liquid can be concentrated after passing through a filter. The concentration temperature can be 60-65℃, and the concentration can be concentrated to 30-50 brix. The concentrated liquid can be manufactured into powder by spray drying.

[0020] In the composition of the present invention, the wasabi leaf extract, which is an effective ingredient, contains 0.8 to 1.6 mg / g of isovitexin and 4.8 to 0.6 mg / g of sinigrin, and the weight ratio of isovitexin to sinigrin in the wasabi leaf extract may be 1:1 to 1:6 (isovitexin: sinigrin). Preferably, the weight ratio of isovitexin to sinigrin may be 1:3 to 1:6 (isovitexin: sinigrin) or 1:3±0.5 (isovitexin: sinigrin). When the wasabi leaf extract contains isovitexin and sinigrin in the above-mentioned weight ratio, it is characterized in that it exhibits a more remarkable synergistic effect with respect to the prevention, improvement and / or treatment efficacy of obesity compared to when it contains them in other weight ratios.

[0021] The above-mentioned effective ingredient may be included in an amount of 0.1 to 5% by volume based on the total volume of the composition of the present invention, and the prevention, improvement, or treatment of obesity may be achieved by a complex mechanism through one or more activities selected from the group consisting of weight reduction, body fat reduction, and abdominal fat reduction.

[0022] In one embodiment, the composition of the present invention may be a pharmaceutical composition. The pharmaceutical composition may include the above-described active ingredient and may be formulated into a pharmaceutical unit dosage form by adding a pharmaceutically acceptable carrier, excipient, or diluent.

[0023] The above “pharmaceutically acceptable” refers to a non-toxic composition that is physiologically acceptable and does not inhibit the action of the active ingredient when administered to humans and does not typically cause allergic reactions such as gastrointestinal upset, dizziness, or similar reactions.

[0024] Examples of the carrier, excipient or diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, the pharmaceutical composition may further include a filler, an anticoagulant, a lubricant, a wetting agent, a fragrance, an emulsifier or a preservative.

[0025] The term "pharmaceutically effective amount" means an amount that produces a response greater than that of a negative control, and preferably an amount sufficient to produce an effect in the prevention and / or treatment of obesity.

[0026] Additionally, the pharmaceutical composition of the present invention can be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to a mammal. The formulation may be, for example, a formulation selected from the group consisting of gels, pastes, ointments, powders, emulsions, sprays, and aerosols, but is not limited thereto.

[0027] In addition, the pharmaceutical composition of the present invention can be administered in combination with a known compound having an effect of preventing, improving, and / or treating obesity.

[0028] Additionally, the pharmaceutical composition of the present invention can be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to a mammal. The formulation can be in the form of a powder, granule, tablet, emulsion, syrup, aerosol, soft or hard gelatin capsule, sterile injectable solution, or sterile powder.

[0029] The route of administration of the pharmaceutical composition of the present invention is not limited to the above, but may be administered orally or parenterally. Parenteral routes of administration may include, for example, transdermal, nasal, intraperitoneal, intramuscular, subcutaneous, or intravenous routes. Furthermore, the pharmaceutical composition of the present invention may be administered in combination with a known compound that has a preventive and / or therapeutic effect on metabolic syndrome.

[0030] In another embodiment, the composition of the present invention may be a food composition.

[0031] The food composition of the present invention includes processed forms of all natural materials, such as foods, functional foods, nutritional supplements, health feeds, and food additives. The above-mentioned types of food compositions can be manufactured in various forms using conventional methods known in the art.

[0032] For example, as a health food, the extract of the present invention itself can be manufactured in the form of tea, juice, and drink and consumed, or can be granulated, encapsulated, or powdered and consumed. In addition, in addition to the extract of the present invention, white peony root, cornelian cherry, scutellaria baicalensis, reishi mushroom, tangerine peel, ginseng root, angelica root, gardenia fruit, astragalus membranaceus, malt, tangerine seed, vitamin C, fructooligosaccharide, stevioside, purified water, maltodextrin, etc. can be further included alone or in mixture within a range that does not inhibit the purpose of the present invention; however, other medicinal ingredients and / or additives that the food composition of the present invention can additionally include are not limited to the above examples.

[0033] For example, the food composition according to the present invention may include water-soluble vitamins such as thiamine (vitamin B1), riboflavin, ascorbic acid, niacin, and vitamin B6; fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid; weak acids such as glycolic acid and acetic acid; and amino acids such as eight essential amino acids, threonine, valine, methionine, isoleucine, leucine, phenylalanine, tryptophan, and lysine, as well as aspartic acid, serine, glutamic acid, proline, glycine, alanine, cysteine, tyrosine, histidine, and arginine.

[0034] The composition of the present invention can be used as an additive or therapeutic agent for the purpose of alleviating obesity in both humans and non-human animals. Accordingly, the present invention provides a composition for preventing, alleviating, or treating animal obesity, a feed additive for preventing or alleviating animal obesity comprising the composition, and a treatment for animal obesity comprising the composition.

[0035] Another embodiment of the present invention provides a method for preparing a wasabi leaf extract for preventing, improving or treating obesity.

[0036] The above extract is prepared by mixing wasabi leaves with an extraction solvent at a temperature of 96 to 110°C and a pressure of 0.3 to 0.7 kg / cm 2 It can be obtained by extraction under pressure.

[0037] The above wasabi leaves can preferably be dried leaves.

[0038] The above wasabi leaves can preferably be dried leaves.

[0039] One embodiment of the present invention provides a use of a wasabi leaf extract or a composition comprising a wasabi leaf extract for use in the manufacture of a food, medicine, or health functional food for treating, preventing, or improving obesity. The wasabi leaf extract comprises 0.8 to 1.6 mg / g of isovitexin and 4.8 to 0.6 mg / g of sinigrin, and the weight ratio of isovitexin to sinigrin in the wasabi leaf extract may be 1:1 to 1:6 (isovitexin: sinigrin). In order to include an effective ingredient in the extract at an optimal content, wasabi leaves are mixed with an extraction solvent and extracted at a temperature of 96 to 110°C and a weight of 0.3 to 0.7 kg / cm 2 It can be extracted by pressure.

[0040]

[0041] The composition of the present invention exhibits at least one effect selected from the group consisting of weight loss, body fat reduction, and abdominal fat reduction without causing a decrease in appetite, thereby exhibiting an effect of preventing, improving, or treating obesity.

[0042] Figure 1 shows a photograph of wasabi leaf extract obtained as deep brown powders according to one embodiment.

[0043] Figure 2(ac) shows the results of analyzing the content of standard substances (including isovitexin) in wasabi leaf extract using the HPLC method.

[0044] Figure 3 shows the results of comparing the amount of intracellular fat accumulation through the Oil Red O test after removing the culture medium for each group of cells.

[0045] Figure 4 shows the results of confirming the change in body weight over time after substance administration in each group of experimental animals.

[0046] Figure 5 shows the results of confirming the weight gain in each group of experimental animals 6 days and 84 days after substance administration.

[0047] Figure 6 shows whole-body DEXA (Dual-energy x-ray absorptiometry) images showing representative total body mass and abdominal fat pads in each group of experimental animals 84 days after substance administration. (A = Mice orally administered 10 ml / kg of vehicle (distilled water) with NFD (intact control group); B = Mice orally administered 10 ml / kg of vehicle with HFD (HFD control group); C = Mice orally administered 250 mg / kg of metformin with HFD (metformin); D = Mice orally administered 200 mg / kg of WL with HFD (WL200); E = Mice orally administered 100 mg / kg of WL with HFD (WL100); F = Mice orally administered 50 mg / kg of WL with HFD (WL50))

[0048] Figure 7 is a graph showing changes in total body and abdominal fat density in experimental animals of each group 84 days after substance administration.

[0049] Figure 8 shows representative histological images of adipocytes taken from the periovarian and abdominal wall fat pads of mice 84 days after substance administration in each group of experimental animals. (A = Mice orally administered 10 ml / kg of vehicle (distilled water) on NFD (intact control group); B = Mice orally administered 10 ml / kg of vehicle on HFD (HFD control group); C = Mice orally administered 250 mg / kg of metformin on HFD (metformin); D = Mice orally administered 200 mg / kg of WL on HFD (WL200); E = Mice orally administered 100 mg / kg of WL on HFD (WL100); F = Mice orally administered 50 mg / kg of WL on HFD (WL50))

[0050] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0051]

[0052] Example

[0053] I. Extract Preparation and Analysis

[0054] 1. Manufacturing of Wasabi Leaf Extract (Wasabi Leaf / Folium, Wasabi japonica (Miq.) Matsum extracts)

[0055] It was confirmed that the contents of isovitexin and sinigrin, the effective components of wasabi leaves, in the extract changed depending on the temperature and pressure conditions during the production of wasabi leaf extract. Therefore, in order to find the optimal extraction conditions for isovitexin and sinigrin, the effective components of wasabi leaves, when using water as a solvent, extraction was performed at various temperatures and pressures, and the results are shown in Table 1 below.

[0056] Extraction conditions (temperature, pressure (kg / cm) 2 ))Isovitexin (mg / g)Sinigrin (mg / g)Isovitexin:Sinigrin ratio 115~125℃, 1.50.253.21:12.898~102℃, 0.3~0.51.356.81:5.0395℃, 0.25~0.30.84.21:5.25

[0057] The experimental results confirmed that the effective ingredient contents of the wasabi leaf extract showing the optimal effect were 0.8-1.6 mg / g of isovitexin and 4.8-9.6 mg / g of sinigrin. In order to produce raw materials that passed this standard, experiments were conducted under three different temperature conditions. As shown in Table 1 above, it was confirmed that approximately 80% of isovitexin and sinigrin were extracted at 95℃ or lower, and the optimal content and ratio were extracted at 98-102℃. In addition, it was confirmed that when extracted at 115-125℃, the effective ingredients were denatured by heat, resulting in a rapid decrease in content. Therefore, it was confirmed that the optimal extraction conditions for isovitexin and sinigrin, which are the effective ingredients of the wasabi leaf extract, are extraction at 98-102℃. Thus, the wasabi leaf extract was prepared as follows. Add 18 times the amount of water to 150 kg of dried wasabi leaves, and heat at 100℃ for 8 hours, pressure 0.4-0.5 kg / cm 2 , and the temperature change was extracted at ±0.5℃. The extracted liquid passed through a 1um filter, was transferred to a buffer tank, and then transferred to a concentration tank in appropriate amounts to concentrate. The concentration temperature was 60-65℃, and the concentration was concentrated to 30-50 brix. The concentrate was mixed with dextrin at a ratio of 0-10%, and was produced as deep brown powders through a spray dryer under the conditions of an inlet temperature of 170-200℃ and an outlet temperature of 85-100℃ (Fig. 1).

[0058]

[0059] 2. Confirmation of the content of effective ingredients in wasabi leaf extract

[0060] The content of effective ingredients contained in wasabi leaf extract was analyzed as follows.

[0061] (1) Preparation of standard solution

[0062] Standard substance 1: Isovitexin

[0063] Molecular formula: C 21 H 20 O 10 , molecular weight: 432.38

[0064] CAS No.: 38953-85-4

[0065]

[0066] Standard substance 2: sinigrin

[0067] Molecular formula: C 10 H 17 NO9S2, molecular weight: 359.36

[0068] CAS No.: 534-69-0

[0069]

[0070] Approximately 8 mg of the standard substance was precisely weighed and dissolved in methanol to make exactly 50 mL, which was used as the standard stock solution. A solution made by dissolving 3 mL of the above standard stock solution in methanol to make exactly 50 mL was used as the standard solution.

[0071]

[0072] (2) Preparation of test solution

[0073] Approximately 330 mg of wasabi leaf extract was precisely weighed, added methanol, ultrasonically extracted for 30 minutes, diluted to exactly 50 mL with methanol, and used as the test solution. The test solution was filtered through a 0.45 μm syringe filter and placed in a brown vial for testing.

[0074]

[0075] (3) Analysis

[0076] High speed liquid chromatography (HPLC) analysis was performed under the high speed liquid chromatography conditions shown in Table 2 and the mobile phase conditions shown in Table 3, and the results are shown in Figure 2.

[0077] Injection volume 10 μL, column temperature 30°C, mobile phase A: water with 0.05% TFAB: acetonitrile, flow rate 1.0 mL / min, detector wavelength 270 nm

[0078]

[0079] Time (min)Movement (%)ABinitial86144085154559555595

[0080]

[0081] (4) Calculation

[0082] A calibration curve was created using the peak area of ​​the standard solution, and the concentration of the active ingredient in the test solution was obtained using the calibration curve equation. Then, the content of the active ingredient was calculated using the equation below.

[0083] Active ingredient content (mg / g) = (A × B × C × D) / E

[0084] A: Concentration of test solution (μg / mL)

[0085] B: Final volume (mL)

[0086] C: Dilution factor

[0087] D: Standard purity (%)

[0088] E: Sample amount (mg)

[0089] As a result of the above experiment, it was confirmed that the wasabi leaf extract manufactured in 1 above contains 0.8 to 1.6 mg / g of isovitexin and 4.8 to 9.6 mg / g of sinigrin.

[0090]

[0091] 3. Efficacy evaluation according to the content ratio of active ingredients

[0092] To evaluate the efficacy of the optimal mixing ratio of isovitexin and sinigrin, the effective ingredients of wasabi leaves, 3T3-L1 preadipocyte cells purchased from ATCC were subcultured in DMEM (containing 10% Bovine Serum, 100 units / ml penicillin, 100 ug / ml streptomycin) medium and used to induce adiocyte differentiation and adipogenesis. Preadipocyte cells were used as the normal group, and preadipocyte cells were cultured to a post-confluent state after seeding as the control group. After differentiation induction, the medium was changed every two days to maintain adipocytes. Isovitexin and sinigrin, the effective ingredients of wasabi leaf extract, were each treated at 100 ug / ml, and their complex ratios were mixed at isovitexin: sinigrin = 6:1, 3:1, 1:1, 1:3, 1:6, and 1:9, and each was added at the same concentration of 100 ug / ml for treatment.

[0093] In order to confirm the effect on fat accumulation according to the optimal ratio of the effective ingredients in the wasabi extract, the amount of fat accumulated in the cells of each group was compared through the Oil Red O test after removing the culture medium, and the results are shown in Figure 3.

[0094] The amount of intracellular fat accumulation in the positive induction control group in which differentiation was induced by MDI (3-isobutyl-1-methylxanthine, dexamethasone, insulin) was set as 100%, and the relative amount of intracellular fat accumulation according to each component treatment was expressed as a %.

[0095] As a result of the experiment, when vehicle, MDI, isovitexin alone, sinigrin alone, isovitexin (IS): sinigrin (SI) 6:1 complex, IS:SI 3:1 complex, IS:SI 1:1 complex, IS:SI 1:3 complex, IS:SI 1:6 complex, and IS:SI 1:9 complex were treated at the same concentration of 100 ug / ml, the fat accumulation ratios were confirmed to be 38%, 100%, 71%, 73%, 70%, 66%, 54%, 48%, 51%, and 68%, respectively, and the complex was confirmed to have a better fat accumulation inhibition effect than isovitexin and sinigrin alone at all ratios. In particular, it was confirmed that fat accumulation was suppressed by 17%, 23%, and 20% more in the IS:SI 1:1 complex, 1:3 complex, and 1:6 complex, respectively, than in the single administration group. Therefore, it was confirmed that excellent fat accumulation suppression activity was present when the content ratio of isovitexin and sinigrin in the wasabi leaf extract was 1:1 to 1:6, preferably 1:3 to 1:6, and most preferably 1:3.

[0096] Accordingly, an additional in vivo efficacy evaluation was performed using the 1:3 complex (a composition containing wasabi extract, in which isovitexin and sinigrin are combined in a weight ratio of 1:3 (IS:SI)) (hereinafter referred to as WL), which was confirmed to have the most remarkable effect in inhibiting fat accumulation activity.

[0097]

[0098] II. In vivo efficacy evaluation

[0099] Experimental animals and experimental methods

[0100] A total of 165 6-week-old female SPF / VAF CrljOri:CD1[ICR] mice [OrientBio, Seungnam, Korea] were acclimated to the laboratory environment for 10 days and then fed a high-fat diet (HFD) for 1 week. Only experimental animals that showed a certain weight gain were divided into 6 groups of 8 animals each and used in the experiment. All experimental animals were handled in accordance with the animal experiment ethics standards with prior approval from the Experimental Animal Ethics Committee of Daegu Haany University.

[0101] Military detachments (6 groups total; 8 animals per group):

[0102] Normal control group (Normal pellet diet; NFD feeding medium administration group)

[0103] HFD control group (45%Kcal high fat diet; HFD feeding medium administration group)

[0104] Control drug group administered with metformin 250 mg / kg and HFD supply

[0105] HFD-fed experimental group administered the highest dose of WL (200 mg / kg)

[0106] Experimental group fed HFD with WL intermediate dose (100 mg / kg)

[0107] Experimental group fed HFD with low dose of WL (50 mg / kg)

[0108]

[0109] Dose Frequency

[0110] Dosage frequency: Once daily, for 84 days

[0111] Route of administration: Oral administration

[0112] Medium: Sterile distilled water

[0113] Dosage: 10 ml / kg

[0114]

[0115] Selection of experimental substance dosage:

[0116] In this experiment, the highest dose of WL was set to 200 mg / kg based on a previous efficacy exploration experiment using a 45% Kcal HFD-fed mouse model by the present researchers et al., and the intermediate and low doses were set to 100 and 50 mg / kg, respectively, with a ratio of 2, and were dissolved in sterile distilled water and administered orally at 10 ml / kg, which is the general oral administration volume for mice [Flecknell, 1996; KFDA Guidelines, Notification No. 2017-071, 2017]. In addition, the dose of metformin used as a control drug was set to 250 mg / kg based on previous references [Jeong et al., 2013; Choi et al., 2017]. Metformin is a representative drug used as an obesity treatment through AMPK activation.

[0117]

[0118] Administration of experimental substances:

[0119] After a one-week HFD adaptation period, an appropriate amount of WL was dissolved in sterile distilled water and administered orally at a dose of 10 ml / kg using a 1 ml syringe with a metal sonde. That is, WL was dissolved in distilled water at concentrations of 20, 10, and 5 mg / ml, respectively, and administered orally once daily at a dose of 10 ml / kg (200, 100, and 50 mg / kg) for 84 days. In addition, metformin was also dissolved in sterile distilled water at a concentration of 25 mg / ml and administered orally once daily at a dose of 10 ml / kg (250 mg / kg) for 84 days. In the normal and HFD control groups, sterile distilled water was administered orally in the same manner for the same period instead of the test substance in order to apply the same administration and correction stress.

[0120]

[0121] Experimental results

[0122] 1. Weight changes

[0123] Since only experimental animals showing a constant weight gain after 1 week of HFD feeding were selected, in the HFD control group, a significant (p<0.01) weight gain compared to the normal control group was recognized from 6 days of high-fat feed feeding, and the weight gain during the 1-week HFD adaptation period and the 84-day experimental substance administration period also showed a significant (p<0.01) increase compared to the normal control group. Meanwhile, in all three doses of WL 200, 100, and 50 mg / kg groups, a significant (p<0.01 or p<0.05) decrease in body weight compared to the HFD control group began to be recognized from 14 days after the start of administration, and in the metformin 250 mg / kg group, from 21 days after the start of administration, respectively. A significant (p<0.01) decrease in body weight during the 84-day experimental substance administration period compared to the HFD control group was also recognized in all experimental substance administration groups. In particular, in the WL 50 mg / kg administration group, an effect of suppressing the increase in body weight and weight gain of HFD feeding comparable to that of the metformin 250 mg / kg administration group was recognized (Table 4, Figures 4 and 5).

[0124]

[0125] During the 84-day administration period, the weight gain in the HFD control group showed a change of 421.73% compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, the changes were -56.09, -75.20, -68.22, and -59.03%, respectively, compared to the HFD control group.

[0126]

[0127] 2. Changes in average feed intake

[0128] A significant (p<0.01) decrease in average feed intake was observed in the HFD control group compared to the normal control group, but no significant change in average feed intake was observed in any of the experimental substance administration groups, including the metformin 250 mg / kg administration group (Table 4). The average feed intake in the HFD control group showed a change of -16.99% compared to the normal control group, but in the metformin 250 mg / kg, 200, 100, and 50 mg / kg administration groups, changes of 0.32, -0.14, -0.04, and 0.71% were observed, respectively, compared to the HFD control group.

[0129]

[0130] 3. Changes in body fat and abdominal fat

[0131] In the HFD control group, a significant increase (p<0.01) in body fat and abdominal fat accumulation was observed compared to the normal control group, whereas in all three doses of WL administration groups, a significant decrease (p<0.01) in body fat and abdominal fat accumulation was observed in a dose-dependent manner compared to the HFD control group, and in particular, the WL 50 mg / kg administration group showed an inhibitory effect on HFD-induced body fat and abdominal fat accumulation comparable to the metformin 250 mg / kg administration group (Figs. 6 and 7).

[0132] Body fat mass showed a change of 210.56% in the HFD control group compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, it showed a change of -38.91, -59.03, -49.78, and -40.70%, respectively, compared to the HFD control group. Abdominal fat mass showed a change of 221.25% in the HFD control group compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, it showed a change of -38.33, -59.65, -49.18, and -39.24%, respectively, compared to the HFD control group.

[0133]

[0134] 4. Changes in fat weight

[0135] In the case of the HFD control group, a significant (p<0.01) increase in the relative and absolute weight of periovarian and abdominal wall accumulated fat was observed compared to the normal control group, but all three doses of WL administration groups showed a significant (p<0.01) decrease in the weight of periovarian and abdominal wall accumulated fat in a dose-dependent manner compared to the HFD control group, and in particular, the WL 50 mg / kg administration group showed an inhibitory effect on the increase in the relative and absolute weight of periovarian and abdominal wall accumulated fat induced by HFD comparable to that of the metformin 250 mg / kg administration group (Fig. 6).

[0136] The absolute and relative weights of periovarian fat accumulation showed changes of 916.67 and 528.77%, respectively, compared to the normal control group in the HFD control group, but the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups showed changes of -61.56, -80.90, -70.07, and -61.85% in the absolute weight of periovarian fat accumulation, and -49.83, -72.52, -57.89, and -49.24% in the relative weight of periovarian fat accumulation, respectively, compared to the HFD control group. The absolute and relative weights of abdominal wall fat accumulation in the HFD control group showed changes of 1091.25 and 635.54%, respectively, compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, the absolute weights of abdominal wall fat accumulation showed changes of -61.28, -77.33, -70.93, and -62.71%, respectively, compared to the HFD control group, and the relative weights of abdominal wall fat accumulation showed changes of -49.72, -67.46, -59.71, and -50.42%, respectively.

[0137]

[0138] 5. Histopathological changes in periovarian and abdominal wall fat accumulation

[0139] In the HFD control group, significant hypertrophy of fat cells was observed, and a significant (p<0.01) increase in the thickness and fat cell diameter of the periovarian and abdominal wall accumulated fat tissues was observed compared to the normal control group, but all three doses of WL showed a significant (p<0.01) decrease in the thickness and fat cell diameter of the periovarian and abdominal wall accumulated fat tissues in a dose-dependent manner compared to the HFD control group, and in particular, the WL 50 mg / kg administration group showed an inhibitory effect on the increase in the thickness and fat cell diameter of the HFD-induced periovarian and abdominal wall accumulated fat tissues comparable to the metformin 250 mg / kg administration group (Fig. 8).

[0140] The thickness of the accumulated fat tissue around the ovary showed a change of 286.18% in the HFD control group compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, the changes in the thickness of the accumulated fat tissue around the ovary showed -49.84, -66.48, -61.14, and -51.62%, respectively, compared to the HFD control group. The average adipocyte diameter of the accumulated fat tissue around the ovary showed a change of 235.21% in the HFD control group compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, the changes in the average adipocyte diameter of the accumulated fat tissue around the ovary showed -39.34, -63.31, -54.43, and -40.47%, respectively, compared to the HFD control group. The thickness of abdominal wall accumulated fat tissue showed a change of 454.29% in the HFD control group compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, it showed a change of -52.59, -73.83, -67.58, and -53.85%, respectively, compared to the HFD control group. The average adipocyte diameter of abdominal wall accumulated fat tissue showed a change of 181.53% in the HFD control group compared to the normal control group, but in the metformin 250 mg / kg, WL 200, 100, and 50 mg / kg administration groups, it showed a change of -36.84, -56.22, -49.75, and -38.48%, respectively, compared to the HFD control group.

[0141] The present invention can be provided as a composition for preventing or improving obesity, preferably as a food or health functional food. The present invention can also be provided as a pharmaceutical for treating obesity.

Claims

1. A composition for preventing, improving or treating obesity containing wasabi leaf extract, A composition characterized in that the wasabi leaf extract contains 0.8 to 1.6 mg / g of isovitexin and 4.8 to 9.6 mg / g of sinigrin, and the weight ratio of isovitexin to sinigrin in the composition is 1:1 to 1:6 (isovitexin: sinigrin).

2. A composition according to claim 1, characterized in that the extract is extracted using any one selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixtures thereof as an extraction solvent.

3. A composition according to claim 1, characterized in that the weight ratio of isovitexin to sinigrin in the composition is 1:3 to 1:6 (isovitexin: sinigrin).

4. A composition according to claim 1, characterized in that the weight ratio of isovitexin to sinigrin in the composition is 1:3±0.5 (isovitexin: sinigrin).

5. In the first paragraph, the wasabi leaf extract is applied at a temperature of 98 to 102°C and a weight of 0.3 to 0.5 kg / cm 2 A composition characterized by being extracted using water as an extraction solvent under pressure conditions.

6. A composition according to claim 1, characterized in that it has at least one effect selected from the group consisting of weight loss, body fat loss, and abdominal fat loss, without causing a decrease in appetite.

7. A composition according to any one of claims 1 to 6, characterized in that the composition is for preventing or improving obesity and is a food composition.

8. A composition according to any one of claims 1 to 6, characterized in that the composition is a pharmaceutical composition.

9. Health functional food containing the composition according to Article 7.

10. A composition for preventing, improving or treating obesity in mammals, wherein the composition is for mammals in the first paragraph.

11. Feed additive for preventing or improving mammalian obesity, comprising a composition according to Article 10.

12. A mammalian obesity treatment agent comprising a composition according to Article 10.

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