A composition for preventing or treating of obesity or metabolism disease comprising extracts from the leaves of Stewartia koreana Nakai

Abstract

The present invention relates to a composition for anti-obesity, weight or body fat reduction, and a composition for the prevention, improvement, or treatment of metabolic diseases, comprising a Japanese dogwood leaf extract. The Japanese dogwood leaf extract of the present invention exhibits excellent effects in reducing weight and body fat without accompanying cytotoxicity, inflammation, or immune responses. Furthermore, since it inhibits the increase of diabetes-related enzymes and the increase of cardiovascular and arteriosclerosis risk factors, as well as inhibits fat accumulation in the liver, it can be utilized in various ways for the improvement of obesity and for the prevention, improvement, or treatment of various metabolic diseases associated with obesity.

Description

Technology Field

[0001] The present invention relates to a composition for anti-obesity, weight or body fat reduction, and a composition for the prevention, improvement, or treatment of metabolic diseases, comprising a Japanese dogwood leaf extract. Background Technology

[0003] With recent economic development and changes in dietary habits, the incidence of metabolic diseases (Metabolic Disease, Metabolic Syndrome), including various conditions such as obesity, hyperlipidemia, hypertension, arteriosclerosis, hyperinsulinemia, diabetes, or liver disease, is rapidly increasing. While these diseases can occur individually, they are generally closely related and, in most cases, occur accompanied by various symptoms.

[0004] Obesity refers to a condition in which energy intake and expenditure are out of balance due to lifestyle or genetic causes, resulting in excess energy being accumulated as fat, abnormally high body fat, and metabolic abnormalities. According to a report by the International Obesity Task Force (IOTF), approximately 1.7 billion people, accounting for one-quarter of the world's population, need to lose weight, making it a serious social issue in both developed and developing countries. Obesity is becoming more prevalent among modern people due to improved living standards resulting from economic development, and it acts as a major cause that increases the risk of hypertension, hyperlipidemia, arteriosclerosis, heart disease, and diabetes. Among the obesity treatments sold domestically and internationally, there is 'Xenical,' which contains orlistat (approved by the US FDA) as its main ingredient. However, Xenical, which inhibits lipase activity, causes gastrointestinal side effects such as steatorrhea, gas production, and reduced absorption of fat-soluble vitamins.

[0005] Diabetes mellitus is a disease characterized by insufficient insulin secretion or impaired insulin function, resulting in hyperglycemia, which is an elevated concentration of glucose in the blood. This hyperglycemia causes various symptoms and signs, and leads to the excretion of glucose in the urine. Furthermore, as time progresses, it causes vascular disorders and functional abnormalities of the nerves, kidneys, and retina, ultimately leading to death.

[0006] Dyslipidemia refers to a condition in which the concentrations of total cholesterol, triglycerides, and LDL cholesterol in the blood are higher than normal levels, or the concentration of HDL cholesterol is low. When lipid components such as LDL cholesterol or triglycerides increase in the blood, blood flow becomes impaired. As lipid components adhere to the artery walls, they trigger a chronic inflammatory response, leading to arteriosclerosis where the inner walls of the arteries narrow and the blood vessels harden. In the long term, blood clots formed from this condition can block coronary arteries or cerebral blood vessels, causing myocardial infarction, stroke, or cerebral infarction.

[0007] Fatty liver disease (FLD) refers to a condition in which fat accumulates at a rate of 5% or more within liver cells. Depending on the cause, it is classified into alcoholic fatty liver disease (AFLD) and non-alcoholic fatty liver disease (NAFLD). The primary cause of alcoholic fatty liver disease is the excessive accumulation of fat in the liver due to excessive alcohol consumption. Non-alcoholic fatty liver disease, on the other hand, exhibits characteristic findings of liver fat accumulation in liver imaging or biopsy without a history of significant alcohol consumption, use of medications capable of inducing fatty liver, or viral infections. The aforementioned non-alcoholic fatty liver disease (NAFLD) occurs in association with obesity, type 2 diabetes, hyperlipidemia, and cardiovascular disease. It has increased rapidly worldwide over the past few decades and is the most common chronic metabolic disease in modern society. On average, it is reported that approximately 24% of the global population and 16–33% of Koreans suffer from NAFLD. The above NAFLD is known to vary in severity, ranging from simple fatty liver (liver steatosis without inflammation) to nonalcoholic steatohepatitis (NASH), liver fibrosis, and even liver cirrhosis.

[0008] Although various drugs are being developed to prevent, treat, or improve obesity and metabolic diseases as described above, there has been a continuous need for new compositions using natural products that are safe for the human body and free from side effects.

[0009] Korean dogwood ( Stewartia koreanaThe Japanese pagoda tree (*Nakai*) is a deciduous tree belonging to the theaceae family. It is a native Korean species found in the Jirisan Mountain region, known by names such as *Bidannamu*, *Nogajinamu*, and *Geumsumok* due to the patterned bark. In autumn, its leaves turn yellow, and its flowers resemble camellias, so it is also planted as an ornamental tree. While the wood is used for furniture, it is currently utilized as a valuable medicinal herb. Native to the central and southern regions of South Korea and Japan, the Japanese pagoda tree grows to a height of 10 to 20 meters. Its leaves are oval-shaped with small serrations along the edges. Flowers bloom in June and July, and in October, the fruit ripens, splitting into five sections to release round seeds.

[0010] The shoots of the Korean gourd tree are made into tea and consumed; due to its pleasant taste, it is popular among enthusiasts. Korean gourd tea is known to be effective for various liver diseases, such as hepatitis and cirrhosis, and is also said to have detoxifying effects against various forms of poisoning, including alcohol, pesticides, and heavy metals. The *Compendium of Materia Medica* also describes its blood-activating properties as "relaxing muscles and invigorating blood," noting that it helps dissolve blood clots caused by contusions and is effective for pain associated with numbness. The problem to be solved

[0012] Accordingly, the inventors of the present invention [use] the leaves of the Japanese quince tree ( Stewartia koreana While researching the activity of an extract obtained from *SkLHWE* leaves hot water extract, it was confirmed that the *SkLHWE* leaf extract has excellent anti-obesity, weight or body fat reduction, and preventive, improving, or therapeutic effects on metabolic diseases, thereby completing the present invention.

[0013] Therefore, the objective of the present invention is to provide a composition comprising a Japanese dogwood leaf extract for anti-obesity, weight or body fat reduction, and prevention, improvement, or treatment of metabolic diseases. means of solving the problem

[0015] To solve the above problems, the present invention provides a health functional food composition for anti-obesity comprising a Japanese dogwood leaf extract.

[0016] In addition, the present invention provides a health functional food composition for reducing body weight or body fat, comprising a Japanese dogwood leaf extract.

[0017] In addition, the present invention provides an anti-obesity feed additive comprising a Japanese dogwood leaf extract.

[0018] In addition, the present invention provides a health functional food composition for preventing or improving metabolic diseases, comprising a Japanese dogwood leaf extract.

[0019] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of metabolic diseases comprising a Japanese dogwood leaf extract. Effects of the invention

[0021] The leaf extract of the Korean dogwood of the present invention has excellent effects in reducing body weight and body fat without accompanying cytotoxicity, inflammation, or immune response. In addition, it can inhibit the increase of diabetes-related enzymes and the increase of cardiovascular and arteriosclerosis risk factors, as well as inhibit fat accumulation in the liver, so it can be utilized in various ways for the improvement of obesity and for the prevention, improvement, or treatment of various metabolic diseases related to obesity. Brief explanation of the drawing

[0023] Figure 1 shows the results of confirming DPPH free radical scavenging activity, PTP1B (protein tyrosine phosphatase 1B) inhibitory activity, α-glucosidase inhibitory activity, and lipase inhibitory activity at different concentrations in the group treated with Styrax japonica leaf extract (SKLHWE). Figure 2 shows the results of confirming cytotoxicity after treatment with Styrax japonica leaf extract (SKLHWE) at different concentrations. Figure 3a shows the results of serum biochemical analysis of GOT / AST, GPT / ALT, and glucose following treatment with Styrax japonica leaf extract (SKLHWE) in a high-fat diet-induced obese mouse model. Figure 3b shows the results of confirming changes in total cholesterol, high-density lipoprotein, triglycerides, and low-density lipoprotein following treatment with Styrax japonica leaf extract (SKLHWE) in a high-fat diet-induced obese mouse model. Figure 3c shows the results of confirming changes in cardiovascular risk factors (CRF) and atherogenic index (AI) following treatment with Styrax japonica leaf extract (SKLHWE) in a high-fat diet-induced obese mouse model. Figure 4 is a figure showing the results of confirming changes in body weight loss and weight gain rate in a high-fat diet mouse model according to treatment with the extract (SKLHWE) of the Korean dogwood leaf of the present invention at different concentrations. Figure 5 shows the results of confirming the inhibition of fat production by measuring the change in abdominal fat weight after treating a high-fat diet mouse model with Styrax japonica leaf extract (SKLHWE). Figure 6 shows the results confirming the fatty liver inhibitory effect of Styrax japonica leaf extract (SKLHWE). Figure 7 shows the results of confirming whether the spleen weight was measured to determine whether the spleen leaf extract (SKLHWE) induced immune and inflammatory responses. Specific details for implementing the invention

[0024] The present invention relates to a composition for anti-obesity, weight or body fat reduction, and a composition for the prevention, improvement, or treatment of metabolic diseases, comprising a Japanese dogwood leaf extract.

[0025] The leaf extract of the Korean dogwood of the present invention has excellent effects in reducing body weight and body fat without accompanying cytotoxicity, inflammation, or immune responses. In addition, it can inhibit the increase of diabetes-related enzymes and the increase of cardiovascular and arteriosclerosis risk factors, as well as inhibit fat accumulation in the liver, so it can be utilized in various ways for the improvement of obesity and for the prevention, improvement, or treatment of various metabolic diseases related to obesity.

[0026] The following explains this in more detail.

[0027] The present invention relates to a health functional food composition for anti-obesity containing a Japanese dogwood leaf extract, and a health functional food composition for reducing body weight or body fat.

[0028] It was confirmed that the Japanese pagoda tree leaf extract of the present invention can lower total cholesterol concentration and effectively reduce the increase in high-density and low-density lipoproteins in an obesity-induced mouse model. Therefore, the composition of the present invention may be used for cholesterol reduction.

[0029] In addition, the Japanese pagoda tree leaf extract of the present invention exhibited a concentration-dependent effect of reducing weight gain induced by a high-fat diet, thereby reducing the amount of weight gained. Therefore, in the present invention, the anti-obesity may refer to one or more activities selected from the group consisting of weight reduction, body fat reduction, and abdominal fat reduction. Accordingly, the present invention provides a health functional food composition for weight or body fat reduction comprising a Japanese pagoda tree leaf extract. The body fat reduction may be, in particular, abdominal fat reduction.

[0030] Specifically, the above-mentioned functional food is synonymous with food for special health use (FosHU) and refers to a food with high medical or therapeutic effects that is processed to efficiently exhibit bio-regulatory functions in addition to providing nutrition. Here, "functionality" means obtaining useful effects for health purposes, such as regulating nutrients or physiological actions regarding the structure and function of the human body. The food of the present invention can be manufactured by methods commonly used in the industry, and during such manufacturing, raw materials and ingredients commonly added in the industry may be added. Furthermore, the formulation of the food can be manufactured without restriction as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms of formulations; unlike general pharmaceuticals, it uses food as a raw material, offering the advantage of not having side effects that may occur with long-term use of pharmaceuticals, and possessing excellent portability.

[0031] The health functional food of the present invention includes ingredients that are typically added during food manufacturing, and may include, for example, proteins, carbohydrates, fats, nutrients, and flavorings. For example, when manufactured as a drink, flavorings or natural carbohydrates may be included as additional ingredients in addition to the Japanese pagoda tree leaf extract as an active ingredient. For example, natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, etc.), and sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). Natural flavorings (e.g., taumarin, stevia extract, etc.) and synthetic flavorings (e.g., saccharin, aspartame, etc.) may be used as flavorings.

[0032] The health functional food or food of the present invention may be provided in the form of tablets, capsules, pills, or liquids, and forms to which the extract of the present invention may be added include, for example, various drinks, meat, sausages, bread, candies, snacks, noodles, ice cream, dairy products, soups, isotonic drinks, beverages, alcoholic drinks, chewing gum, tea, and vitamin complexes.

[0033] When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The amount of the active ingredient may be appropriately used according to its purpose of use (prevention or improvement). Generally, when manufacturing food or beverages, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, relative to the total raw materials. However, in the case of long-term consumption for health purposes, the above amount may be less than the above range, and since there are no issues regarding safety, the active ingredient may be used in an amount greater than the above range.

[0034] There are no special restrictions on the types of the above-mentioned health functional foods. Examples of foods to which the above-mentioned health functional food composition may be added include meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the conventional sense.

[0035] Since the leaf extract of the Korean dogwood of the present invention is a natural extract, it is not toxic to cells and can be used in various ways as a composition for health functional foods, pharmaceuticals, food, and quasi-pharmaceuticals.

[0036] For example, when the Japanese dogwood leaf extract of the present invention is administered to animals other than humans, it can be used as an anti-obesity feed or feed additive.

[0037] The above feed additive may be in the form of a concentrate, powder, or granules, and may be used together with substances such as amino acids, inorganic salts, vitamins, antibiotics, antimicrobial substances, antioxidants, antifungal enzymes, digestive and absorption enhancers, growth promoters, disease preventive agents, etc. as auxiliary ingredients.

[0038] The above feed additive may be administered to animals alone or in combination with other feed additives among edible carriers. Additionally, the above feed additive may be easily administered as a top dressing, by mixing directly into animal feed, or separately from feed, as a separate oral formulation, by injection or transdermal injection, or in combination with other ingredients. Typically, as is well known in the art, a single daily dose or a divided daily dose may be used. When the above feed additive is administered separately from animal feed, as is well known in the art, the administration form of the extract may be prepared as an immediate-release or sustained-release formulation by combining them with a non-toxic, pharmaceutically acceptable edible carrier. Such edible carriers may be solid or liquid, for example, corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. When a solid carrier is used, the administration form of the extract may be a tablet, capsule, powder, trolley, or sugar-containing tablet or a finely dispersed top dressing. When a liquid carrier is used, it may be in the form of a soft gelatin capsule, or a syrup, liquid suspension, emulsion, or solution. Additionally, the dosage form may contain adjuvants, such as preservatives, stabilizers, wetting agents or emulsifiers, solution promoters, etc.

[0039] The animal feed containing the Japanese pagoda tree leaf extract of the present invention as a feed additive may be any protein-containing organic grain flour commonly used to meet the dietary requirements of animals. Such protein-containing grain flour is typically composed mainly of corn, soybean flour, or a corn / soybean flour mix. The above feed additive may be used by adding it to the animal feed by immersion, spraying, or mixing. The veterinary composition or feed additive of the present invention may be applied to the diet of companion animals.

[0040] When a composition containing the extract of the Japanese dogwood leaves of the present invention is used as feed or feed additive for obese animals, it can regulate the animal's body weight, reduce total cholesterol, and improve various cardiovascular problems caused by obesity.

[0041] In addition, the present invention provides a health functional food composition for preventing or improving metabolic diseases and a pharmaceutical composition for preventing or treating metabolic diseases, comprising a Japanese dogwood leaf extract.

[0042] In the present invention, it was confirmed that the leaf extract of the Japanese dogwood tree exhibits excellent inhibitory activity against α-glucosidase, PTP1B (protein tyrosine phosphatase 1B), and lipase, which are known to be enzymes associated with obesity and diabetes. It was also confirmed that when administered to obese mice induced by a high-fat diet, it can improve cardiovascular risk factors (CRF) and atherogenic index (AI). Furthermore, the present invention confirmed that the leaf extract of the Japanese dogwood tree can effectively inhibit liver fat accumulation.

[0043] The above metabolic disease may include, without limitation, all metabolic diseases widely known in the field as being related to obesity, and preferably, the above metabolic disease may be one or more selected from the group consisting of obesity, diabetes, hypertension, arteriosclerosis, hepatic steatosis, dyslipidemia, hyperglycemia, or non-alcoholic fatty liver. More specifically, the above dyslipidemia may be one or more selected from the group consisting of hyperlipidemia, hyper-LDL cholesterolemia, hypertriglyceridemia, and low-HDL cholesterolemia. In addition, the above non-alcoholic fatty liver may be any one selected from the group consisting of simple fatty liver, nutritional fatty liver, starvation fatty liver, obese fatty liver, diabetic fatty liver, steatohepatitis, liver fibrosis, and liver cirrhosis.

[0044] Suitable dosages of the pharmaceutical composition according to the present invention can be prescribed in various ways depending on factors such as the formulation method, mode of administration, patient's age, body weight, sex, pathological condition, food, time of administration, route of administration, excretion rate, and response sensitivity.

[0045] The pharmaceutical composition of the present invention may be formulated into an oral or parenteral administration formulation depending on the route of administration. When formulating, it may be prepared using one or more buffers (e.g., saline solution or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g., aluminum hydroxide), suspenders, thickeners, wetting agents, disintegrants or surfactants, diluents or excipients.

[0046] The leaf extract of the Styrax japonica of the present invention can be formulated for oral use and can be formulated as a tablet, pill, powder, granule, liquid, gel, syrup, slurry, suspension, or capsule for oral administration. Such solid formulations may be prepared by mixing at least one excipient with the pharmaceutical composition of the present invention, for example, starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethylcellulose, or gelatin, etc. For example, tablets or sugar tablets can be obtained by combining an active ingredient with a solid excipient, grinding the mixture, adding a suitable auxiliary agent, and processing it into a granular mixture.

[0047] The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modulators for the inhibition and treatment of metabolic diseases.

[0048] The term "extract" in the present invention may include not only crude extracts obtained by treating with an extraction solvent but also processed products of Styrax japonica leaf extract, for example, a concentrate obtained by further concentrating Styrax japonica leaf extract, a dried product obtained by drying the same, or a powder obtained by grinding the same. For example, Styrax japonica leaf extract may be prepared in a powder state by additional processes such as vacuum distillation and freeze-drying or spray drying.

[0049] In addition, the Japanese pagoda tree leaf extract of the present invention, in a broad sense, includes processed products of Japanese pagoda tree leaf extract formulated for administration to animals, such as Japanese pagoda tree leaf extract powder.

[0050] In the present invention, the Japanese pagoda tree leaf extract may be an extract obtained by extracting Japanese pagoda tree leaves with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, preferably a water extract, and more preferably a hot water extract. When prepared as a hot water extract, it may be prepared by a method of hot water extraction at 80 to 120°C for 5 to 15 minutes, and the supernatant obtained by centrifuging the hot water extract at 2,000 to 4,000 rpm may be used as an active ingredient in the composition according to the present invention.

[0051] The leaf extract of the Korean dogwood of the present invention not only exhibits a high yield of 15% or more compared to extractions using other parts of the Korean dogwood, but can also exhibit superior anti-obesity, preventive or therapeutic effects for metabolic diseases.

[0052] Since the Japanese pagoda tree leaf extract of the present invention is a natural product, it has almost no toxicity or side effects and does not induce unwanted immune or inflammatory responses, so it can be taken for a long period for preventive purposes.

[0054] The present invention will be described in detail below through examples. The following examples are intended solely to explain the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention.

[0056] Example 1. Preparation of Styrax japonica leaf extract

[0057] The leaves of the Styrax japonica used in the experiment were purchased from Mori Farm (Geochang, South Korea). 100 ml of water was added to 20 g of dried Styrax japonica leaf sample, and hot water extraction was performed at 100°C for 10 minutes. Afterward, the supernatant was separated by centrifugation at 3000 rpm for 20 minutes. The hot water extract using Styrax japonica leaves showed a high yield of over 15% compared to other parts. Subsequently, the prepared hot water extract of Styrax japonica leaves was used in the experiment.

[0059] Example 2. Confirmation of the inhibitory activity of Styrax japonica leaf extract on obesity and diabetes-related enzymes in vitro

[0060] Using the Styrax japonica leaf extract prepared in Example 1, the inhibitory activities of α-glucosidase, PTP1B (protein tyrosine phosphatase 1B), and lipase, which are known enzymes associated with obesity and diabetes, were measured. The α-glucosidase, lipase, and DPPH (2,2-diphenyl-1-picrylhydrazyl) used in the experiment were purchased from Sigma-Aldrich (St. Louis, USA), and PTP1B was purchased from Cayman Chemical (Ann Arbor, USA). Each enzyme reaction was confirmed by reacting at 37°C for 15 minutes, and the absorbance was measured using a BMG Labtech Spectra Nano (Ortenberg, Germany). In vitro reactions were carried out with Styrax japonica leaf extracts at concentrations of 10, 30, 100, and 300 μg / ml, and the absorbance was measured at 405 nm using a BMG Labtech Spectra Nano (Ortenberg, Germany). To determine antioxidant activity, DPPH free radical scavenging activity was measured at 470 nm. The results of each enzyme inhibitory activity and antioxidant activity measurement are shown in Figure 1.

[0061] As shown in Figure 1, the Japanese pagoda tree leaf extract exhibited inhibitory activities of 19.5%, 37.7%, 45.4%, and 61.6% against α-glucosidase at different concentrations, and 73.3%, 91.8%, 94.4%, and 96.2% against PTP1B. It also showed inhibitory activities of 16.7%, 16.9%, 18.8%, and 28.8% against lipase, and 2.9%, 18.2%, 47.4%, and 69.5% against DPPH free radical scavenging. Inhibitory activity increased in a concentration-dependent manner in all groups treated with the Japanese pagoda tree leaf extract, confirming its excellent ability to inhibit enzymes related to obesity.

[0063] Example 3. Confirmation of cytotoxicity of Styrax japonica leaf extract

[0064] To measure the cytotoxicity of the Styrax japonica leaf extract, PC-3 cells, a human prostate cancer cell line, were used. The cells were obtained from the American Type Culture Collection (ATCC) and cultured at 37°C under conditions where 5% CO2 was supplied and humidity was maintained. Dulbecco's modified eagle medium (DMEM) containing high-concentration glucose was used, and the cells were cultured while supplying 10% FBS and 1% penicillin / streptomycin to the medium. PC-3 cells were treated with Styrax japonica leaf extract at final concentrations of 10, 30, 100, and 300 μg / ml and cultured for 24 hours, after which cytotoxicity was measured using the MTT method. The results confirming the cytotoxicity of the Styrax japonica leaf extract are shown in Figure 2.

[0065] As shown in Figure 2, the cell viability was 95.5%, 94.42%, 92.8%, and 92.4%, respectively, confirming that the Japanese dogwood leaf extract did not have toxicity to the cells.

[0067] Example 4. Preparation of Obesity Animal Model and Experimental Design

[0068] Four-week-old female ICR mice were purchased from Orient Bio (Seongnam, South Korea), and experiments were conducted after a one-week stabilization period. During the experiments, the mice were housed at the Yangsan Annex of the Animal Center at Pusan ​​National University in a facility where the temperature was 23±2°C, relative humidity was 55±5%, and a 12-hour dark-light cycle was maintained, and sufficient water and feed were supplied. After an adaptation period, a high-fat diet (60% high-fat diet) was purchased from Samtaco (Osan, South Korea) and provided to induce an obesity model.

[0069] The control group consisted of 4 animals fed a normal diet and 4 animals fed a high-fat diet. The experimental group was provided with a high-fat diet at all times and consisted of 5 animals per sample concentration, treated with the samples at three different concentrations. The test groups were divided into a normal fat diet (NFD), a high-fat diet (HFD), and a high-fat diet supplemented with 4, 20, and 100 mg / kg of Styrax japonica leaf extract (high-fat diet + 4, 20, 100 mg / kg SKLHWE). The Styrax japonica leaf extract was prepared at a concentration of 20 mg / ml, diluted to 4, 20, and 100 mg / kg, and orally administered to each test group once every two days, while 100 µl of PBS was administered to the control group. Each sample was administered once every two days for a total of 10 doses. Body weight was measured immediately before the start of extract administration and every two days thereafter at the time of administration; the same scale was always used to minimize error. The experimental animals were sacrificed after being fed and raised for 20 days.

[0070] The results of all experiments using the obesity model were expressed as mean ± standard deviation (mean ± SD), and the difference in means between the control group and each experimental group was analyzed by ANOVA, and a p-value of less than 0.05 was determined to be statistically significant.

[0072] Example 5. Confirmation of the inhibitory effect of Styrax japonica leaf extract on obesity and diabetes-related enzymes in vivo

[0073] The obesity-induced experimental animals prepared in Example 4 were fasted for 24 hours after the last sample treatment and then humanely sacrificed in accordance with animal ethics. After sacrifice, blood was collected from the abdominal aorta, and serum was obtained by centrifuging at 5000 rpm for 10 minutes after a coagulation reaction for 30 minutes. Serum biochemical analysis was then performed via enzymatic reaction using a FUJI DRI-CHEM 4000i (Tokyo, Japan) for aspartate aminotransferase (GOT / AST), alanine aminotransferase (GPT / ALT), glucose, total cholesterol (TCHO), high-density lipoprotein (HDL), and triglyceride (TG). Subsequently, low-density lipoprotein (LDL) was calculated using the values ​​of TCHO, HDL, and TG. The results of the serum biochemical analysis are shown in Figures 3a and 3b.

[0074] As shown in Figures 3a and 3b, it was confirmed that serum GPT / ALT, GOT / AST, glucose, TCHO, TG, and LDL levels increased in the experimental group fed a high-fat diet compared to the control group fed a regular diet, and that levels were significantly inhibited in a concentration-dependent manner in the experimental group fed a high-fat diet together with the extract of Styrax japonica leaves. This is considered to be a result induced by the increased concentration of the extract intake.

[0075] Changes in cardiovascular risk factors (CRF) and atherogenic index (AI) were cross-validated according to treatment with different concentrations of Styrax japonica leaf extract. CRF is an indicator representing the degree to which cholesterol in the blood is transported to the liver, and a decrease in CRF may indicate preventive and therapeutic effects for cardiovascular-related diseases. The results of CRF and AI measurements are shown in Figure 3c.

[0076] As shown in Figure 3c, CRF and AI also showed a concentration-dependent decrease, confirming that the Japanese dogwood leaf extract of the present invention can have preventive and therapeutic effects on cardiovascular diseases, such as arteriosclerosis.

[0078] Example 6. Confirmation of weight loss effect of the group treated with Styrax japonica leaf extract

[0079] An experiment was conducted to confirm the body weight reduction effect of Styrax japonica leaf extract (SKLHWE). Specifically, Styrax japonica leaf extract was orally administered to obese ICR mice induced by a high-fat diet at doses of 4, 20, and 100 mg / kg every 2 days, and body weight was measured at each administration after a total of 10 administrations. The results of confirming the change in body weight and the rate of body weight gain for each group are shown in Figure 4.

[0080] As shown in Figure 4, it was confirmed that the experimental group fed a high-fat diet showed increased body weight compared to the control group fed a regular diet, and concentration-dependent effects of weight loss and reduced weight gain were confirmed in the experimental group fed a high-fat diet together with the Korean dogwood leaf extract. This indicates that the Korean dogwood leaf extract has an effect of suppressing obesity by reducing weight gain induced by a high-fat diet.

[0082] Example 7. Confirmation of changes in abdominal fat and liver weight in the group treated with Styrax japonica leaf extract

[0083] Immediately after humanely sacrificing experimental animals and collecting blood, the liver, which is related to abdominal fat and lipid metabolism, and the spleen, which is related to immune responses, were collected. After washing with PBS and removing residual moisture through filter paper, each organ was weighed, and the average weight was calculated for each experimental group. After photography, each organ was stored in a -80°C deep freezer.

[0085] 7.1 Checking for changes in abdominal fat

[0086] To determine whether the extract of Styrax japonica leaves acts on the process of abdominal fat formation and inhibits fat production in ICR mice induced with obesity through a high-fat diet, the weight of abdominal fat was measured after human sacrifice, and the results are shown in Figure 5.

[0087] As shown in Figure 5, abdominal white fat significantly increased in mice fed a high-fat diet, but it was confirmed that the amount and weight of abdominal fat decreased significantly in a concentration-dependent manner in the group treated with the Japanese dogwood leaf extract.

[0089] 7.2 Confirmation of fatty liver inhibitory effect

[0090] Obesity caused by a high-fat diet not only increases fat but also induces fatty liver, in which fat accumulates in the liver due to the increase in fat. To determine whether the leaf extract of Styrax japonica has an inhibitory effect on fatty liver, livers collected from each experimental group were photographed, and the weight of the livers was measured and compared. The results of confirming the changes in the external appearance and weight of the livers are shown in Figure 6.

[0091] As shown in Figure 6, obese ICR mice showed distinct fatty liver lesions in color and shape, but in the experimental group treated with the Japanese dogwood leaf extract, it was confirmed that the liver weight decreased in a concentration-dependent manner and the change in color also improved in a concentration-dependent manner.

[0093] 7.3 Confirmation of whether Styrax japonica leaf extract induces immune and inflammatory responses

[0094] When an immune response is induced or an inflammatory response is increased in mice following treatment with Styrax japonica leaf extract, the weight of the spleen may increase. To confirm this, the spleens of obese ICR mice were isolated, and changes in their weight were examined, and the results are shown in Figure 7.

[0095] As shown in Figure 7, in obesity-induced ICR mice, the weight of the spleen increased due to increased immune and inflammatory responses, but no significant change in spleen weight was observed in the non-obesity-induced control group (NFD), the high-fat diet-induced obesity control group (HFD), or the entire experimental group of Styrax japonica leaf extract. This indicates that Styrax japonica leaf extract inhibits the increase in body fat in high-fat diet-induced obesity models, as previously confirmed, but does not participate in immune responses.

Claims

Claim 1 Anti-obesity health functional food composition comprising Japanese dogwood leaf extract. Claim 2 In claim 1, the above composition is a health functional food composition for reducing cholesterol. Claim 3 A health functional food composition for weight or body fat reduction comprising a Japanese dogwood leaf extract. Claim 4 Anti-obesity feed additive containing Styrax japonica leaf extract. Claim 5 A health functional food composition for the prevention or improvement of metabolic diseases comprising a Japanese dogwood leaf extract. Claim 6 A pharmaceutical composition for the prevention or treatment of metabolic diseases comprising a Japanese dogwood leaf extract. Claim 7 A composition according to claim 6, wherein the metabolic disease is obesity, diabetes, hypertension, arteriosclerosis, hepatic steatosis, dyslipidemia, hyperglycemia, or non-alcoholic fatty liver. Claim 8 In paragraph 6, the above pharmaceutical composition is an oral pharmaceutical composition. Claim 9 A composition according to any one of claims 1 to 3 and 5 to 6, wherein the extract is an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Claim 10 In paragraph 4, the feed additive is an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

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