JP7007663B2 - Kuro-moji extract - Google Patents

Description

Application of Article 30, Paragraph 2 of the Patent Law ・ October 15, 2016, Disclosure in Abstracts of the 23rd Annual Meeting of the Japanese Society for Pre-illness Systems, page 93

INDUSTRIAL APPLICABILITY The present invention has excellent effects of preventing and / or improving lifestyle-related diseases such as anti-obesity activity, muscle endurance improving activity, and liver function improving activity, and is useful as a food and drink, a drug, or a quasi-drug. Regarding naturally derived substances.

It is important to prevent obesity as it is said to be the cause of all diseases. In recent years, research on the effects of fat on the body has progressed, and it has been suggested that the accumulation of visceral fat has a high correlation not only with obesity but also with lifestyle-related diseases, hyperlipidemia, impaired glucose tolerance, and hypertension. It is believed that not accumulating fat can prevent many diseases. The pathological condition based on these ideas is metabolic syndrome. The Ministry of Health, Labor and Welfare estimates that one in two middle-aged men has metabolic syndrome, and about 20 million people fall under the category of metabolic syndrome and its preliminary group (Non-Patent Document 1).

In Japan, the population is aging rapidly, and locomotive syndrome has become a major social problem. One factor of locomotive syndrome is a decrease in muscle mass and strength with aging (Non-Patent Document 2). High-intensity strength training is generally considered to be the most effective way to prevent loss of muscle mass and strength, but there are concerns that elderly people may be accompanied by psychological stress and cardiovascular stress. Therefore, as strength training in the elderly, low-intensity strength training with less mental stress and less load on the circulatory system has been proposed. However, low intensity strength training is difficult to easily increase muscle mass and strength. Therefore, it is important to improve endurance and perform strength training for a long time.

In addition, the liver is an essential organ for life support that plays a central role in metabolism. Liver function is usually broadly divided into circulatory function, excretory function, metabolic function, protective / detoxification function, and hematological function, and when any of them is impaired, fatigue, malaise, loss of appetite, and jaundice And various symptoms peculiar to liver dysfunction such as slight fever will appear. If the state of liver dysfunction continues, it may cause hepatitis, cirrhosis, and lifestyle-related diseases (adult diseases) such as liver cancer (Non-Patent Document 3). Therefore, maintaining normal liver function is extremely important for busy modern people to spend their days refreshingly and to prevent lifestyle-related diseases.

2006 National Health and Nutrition Survey Report, Ministry of Health, Labor and Welfare (http://www.mhlw.go.jp/bunya/kenkou/eiyou08/01.html) Japanese Society of Clinical Orthopedic Surgery Homepage (http://www.jcoa.gr.jp/locomo/) Actual conditions of alcoholic liver disease in Japan From the results of the national tabulation (Journal of the Japanese Society of Gastroenterology Vol. 76 (1979) No. 11 P 2178-2185)

Currently, preventive medicine is attracting attention in Japan in order to curb the high medical costs, and there is a strong desire to develop effective and safe food and drink products, medicines, and quasi-drugs that can reduce the risk of developing diseases. It is rare.

In addition, although there are dietary supplements on the market to increase endurance, the effects of endurance-improving ingredients such as carnitine, which are generally on the market, are questionable, and locomotive syndrome and the like. There is a strong desire to develop dietary supplements to improve endurance in order to improve the condition.

In addition, with the increase in alcohol intake and high-calorie diet, liver diseases such as alcoholic hepatitis, fatty liver, and cirrhosis are steadily increasing. Of course, it is important to solve such a serious problem by the method of administration of medicines for the purpose of prevention / improvement of liver function. However, some liver diseases such as alcoholic hepatitis and fatty liver progress slowly and develop in earnest after a long period of time. In such cases, daily care is particularly important. From the aspect of long-term administration, it is also necessary to deal with it by ingesting food and drink. Under these circumstances, it is desired to develop foods and drinks having a high effect of preventing and improving liver dysfunction.

Therefore, an object of the present invention is to provide an excellent obesity improving agent, a muscle endurance enhancing agent, and / or a liver function improving agent, which are derived from natural products, are highly safe, and can be applied to food and drink. To do.

As a result of diligent studies to solve the above-mentioned problems, the present inventors have found that a plant of the genus Lindera of the family Lauraceae or an extract or purified product thereof has an effect of improving obesity, an effect of improving muscle endurance, an effect of improving liver function, and the like. We have found that it has an excellent effect of preventing and / or improving lifestyle-related diseases, and have completed the present invention.

That is, the present invention
Item 1.
A composition for preventing and / or ameliorating lifestyle-related diseases containing a plant of the genus Lindera or an extract thereof.
Item 2.
Item 2. The composition for preventing and / or improving a lifestyle-related disease according to Item 1, wherein the lifestyle-related disease is obesity, decreased muscular endurance, or decreased liver function.
Item 3.
Item 2. The lifestyle-related lifestyle according to Item 1 or 2, wherein the Lindera plant is derived from one or more selected from the group consisting of whole plants or leaves, stems, roots, seeds, and flowers of Lindera plants. A composition for disease prevention and / or amelioration.
Item 4.
Item 3. The composition for preventing and / or improving lifestyle-related diseases according to any one of Items 1 to 3, wherein the cultivar of the genus Lindera is 1 or 2 or more selected from the group consisting of Kuro-moji, Himekuromoji, and Oobakuromoji. thing.
Item 5.
Item 6. The composition for preventing and / or improving lifestyle-related diseases according to any one of Items 1 to 4, wherein the extract is a water extract, an organic solvent extract, or a water-containing organic solvent extract.
Item 6.
Item 6. The composition for preventing and / or ameliorating lifestyle-related diseases according to any one of Items 1 to 5, which is administered by oral, intravenous injection, inhalation or transdermal.
Item 7.
The composition for preventing and / or improving lifestyle-related diseases according to any one of Items 1 to 6, which is a food and drink product, a quasi-drug, or a pharmaceutical product.
Item 8.
Item 6. The composition for preventing and / or improving lifestyle-related diseases according to any one of Items 1 to 7, wherein the food and drink is a tea beverage.

The composition for preventing and / or improving lifestyle-related diseases of the present invention contains an active ingredient obtained from a naturally-derived plant of the genus Lindera, has no safety problem, and has an effect of improving obesity and an effect of improving muscle endurance. It has an excellent effect of preventing and / or improving lifestyle-related diseases such as an effect of improving liver function. Such a safe composition for preventing and / or improving lifestyle-related diseases can be used for food and drink products, pharmaceutical products or quasi-drugs with high safety and few side effects.

It is a graph which shows the triglyceride level in mouse serum. It is a graph which shows the cholesterol level in mouse serum. It is a graph which shows the result of the lipase inhibitory activity evaluation test. It is a graph which shows the time-dependent change of the body weight of a mouse. It is a graph which shows the weight of the liver of a mouse. It is a graph which shows the weight of the epididymis of a mouse. It is a graph which shows the weight around the kidney of a mouse. It is a graph which shows the weight of the mesentery of a mouse. It is a graph which shows the weight of a mouse brain. It is a graph which shows the weight of the visceral fat of the whole body of a mouse. It is a graph which shows the expression level of the GLUT4 gene in a mouse muscle cell. It is a graph which shows the expression level of a UCP-3 gene in a mouse muscle cell. It is a graph which shows the glucose level in mouse serum. It is a graph which shows the GOT value in mouse serum. It is a graph which shows the GPT value in mouse serum. It is a histological image of the right lobe section of the liver of a mouse in an alcohol loading test. It is a graph which shows the GOT value in mouse serum. It is a graph which shows the GPT value in mouse serum.

The composition for preventing and / or ameliorating lifestyle-related diseases of the present invention contains a Lauraceae plant of the genus Lindera or an extract or purified product thereof. Here, the extract also includes a single component or a purified product refined to a fraction close to a single component.

Lindera plants are widely distributed in Japan, Southeast Asia, etc., and about 100 species are known. In Japan, it is often found in Honshu, Shikoku, and Kyushu. The leaves and roots are fragrant, and Kuro-moji, a plant belonging to the genus Lindera, has been decocted and drunk for a long time in the Oki Islands of Shimane Prefecture.

Lindera plants are highly regenerative plants that grow from seedlings (growing from seeds), coppicing (growing from stumps), and even from root stocks of old trees. It can be said that it is a very easy-to-use plant because it can be re-collected within 2 to 3 years after logging.

The varieties of plants of the genus Lindera are not limited as long as they are plants included in the genus Lindera, as long as the effects of the present invention are exhibited, but they are not limited to those of the genus Lindera, but they are L. strychnifolia, L. citriodora, and L. communis. ), Okinawa Kobashi (L. communis var. Okinawensis), Kanakuginoki (L. erythrocarpa), Yamakobashi (L. glauca), Honbayama Kobashi (L. communis), Hosoba Yamakobashi (L. communis) var. Lancea Mojyama, L. obtusiloba, L. obtusiloba f. Villosa, L. praecox, Hosoba abrachan (L. paraGorigaL. Praecox var. Pubescens), Kuro-moji (L. selicea), Usugekuromoji (L. selicea var. Kuro-moji (L. umbellata), Ooba kuromoji (L. umbellate var. Membrancea (Maxim) Mojama), Kimino Ova kuromoji (L. umbellata var. L. kariensis, L. megaphylla, L. methcalfiana, L. mourpinesis var. L. somsoni, L. tonkiensis, L. tonkiensis var. Subsessilis and the like can be mentioned.

From the viewpoint of remarkably exerting the effect of the present invention, it is preferable to use 1 or 2 or more selected from the group consisting of Kuro-moji, Himekuro-moji, and Ooba-kuromoji, and more preferably Kuro-moji. Among the plants of the genus Lindera, Kuro-moji is a deciduous shrub distributed in Shikoku, Kyushu, Chugoku region, etc. The new branch has a green color, and when it becomes a branch of about 1 cm, it has a lenticel. Kekuromoji grows abundantly in Shikoku such as Kochi prefecture, and leaves and branches can be collected.

In the present invention, when the plant of the genus Lindera is used as it is, the preparation method thereof is not particularly limited. Lindera plants are whole plants or parts of Lindera plants, especially leaves, stems, roots, seeds, and flowers, one type at a time or a combination of two or more of them. It can be appropriately cut to prepare a crushed or crushed powder. The dried product of the genus Lindera or the crushed product thereof can be used, for example, by boiling in boiling water. The crushed or crushed material of the genus Lindera can also be used by roasting if necessary.

When a plurality of parts of a plant belonging to the genus Lindera are combined and used as they are, the combination is not particularly limited. From the viewpoint of obtaining a higher effect of the present invention, the site of the plant of the genus Lindera is preferably a combination of a stem and a leaf. The weight ratio between the stem and the leaves is not particularly limited, but can be 1 to 10: 1 based on the dry weight from the viewpoint of obtaining a higher effect of the present invention, and 1 to 5 can be obtained. 1 is preferable, 1 to 3: 1 is more preferable, and 2 to 3: 1 is further preferable.

Roasting of plants of the genus Lindera is carried out by a known method using a rotary roasting machine or the like, and is not particularly limited. The roasting temperature can be, for example, 150 to 400 ° C, preferably 180 to 350 ° C, more preferably 200 to 330 ° C. The roasting time is appropriately determined depending on the conditions such as the roasting temperature, but can be, for example, 5 to 120 minutes, preferably 10 to 90 minutes, more preferably 15 to 60 minutes.

In the present invention, when an extract of a plant of the genus Lindera is used, the extraction method is not particularly limited. The extract of the genus Lindera is a dried product of the whole plant or a part of the genus Lindera, particularly the leaves, stems, roots, seeds, and flowers, one type at a time or a combination of two or more thereof. The dried product is appropriately cut and extracted from the crushed or crushed powder with a solvent. The plants of the genus Lindera are easily available from native plants, and commercially available products can also be used.

When the whole plant or a part of the genus Lindera is dried, it can be done by using a sun-dried, air-dried or a dryer, but not limited to. When drying in the sun, the time required for drying depends on the weather and the like, but can be, for example, 6 hours or more, preferably 1 day or more, and more preferably 2 days or more. When a dryer is used, it is generally under drying conditions of 100 ° C. or lower, preferably 40 ° C. or lower, for example, a rotary dryer, a hot air dryer, a heat transfer dryer, a vacuum dryer, a vacuum freeze dryer, etc. It is possible to use a cold air dryer, a vibration dryer, a filtration dryer, a vacuum stirring dryer, or the like.

When the extract of the genus Lindera is used as a composition for preventing and / or improving lifestyle-related diseases, the extract of the genus Lindera may be used as it is, or may be appropriately diluted or concentrated. When obtaining an extract of Lindera genus, it is possible to use a fresh plant body, and it is possible to use a genus Lindera plant which has been refrigerated, frozen, or dried and stored, or a concentrate of an extract of the genus Lindera. Can also be used after being dissolved or diluted in an appropriate solvent such as water. The concentrate of the extract of the genus Lindera is not limited, but liquid, paste-like, and muddy-like ones can be used.

Examples of the solvent used for the extraction of plants of the genus Lindera include water, organic solvents and hydrous organic solvents. By using an appropriate extraction solvent, a water extract, an organic solvent extract or a water-containing organic solvent extract of a plant of the genus Lindera can be obtained. Examples of these extraction solvents include water, alcohol, or a mixture thereof, and water, methanol, ethanol, 1,3-butylene glycol, propylene glycol, glycerin, or a mixture thereof is preferable. From the viewpoint of obtaining a higher effect of the present invention, water, methanol, ethanol or hydrous ethanol is more preferable as these extraction solvents. When methanol is used, the concentration is not limited, but from the viewpoint of obtaining higher extraction efficiency, for example, 40% to 99%, preferably 50% to 99%, more preferably 60% to 99%, 70. It can be used at concentrations of% to 95% and 80% to 95%. When hydrous ethanol is used, the ethanol concentration is not limited, but from the viewpoint of obtaining higher extraction efficiency, for example, 40% to 99%, preferably 50% to 99%, and more preferably 60% to 99%. , 70% to 95%, 80% to 95% can be used.

The site of the plant of the genus Lindera used for extraction is not limited, but examples thereof include leaves, stems, roots, seeds, flowers, etc., and leaves and stems from the viewpoint of obtaining a higher effect of the present invention. Parts or combinations thereof are preferable. As used herein, the stems of plants of the genus Lindera may contain bark and / or heartwood.

The extraction method is not particularly limited, but for example, whole or part of a plant belonging to the genus Lindera is cut, crushed by a known method such as a mixer, an extraction solvent is added and stirred, and room temperature or heating is performed for a certain extraction time. A method of separating and extracting an extract of a plant of the genus Lindera from the extract supernatant after treatment with is mentioned. Further, the extraction method may be a method in which a plant of the genus Lindera is immersed in an extraction solvent and then allowed to stand at room temperature or under heating conditions.

The crushing conditions are not limited, but if the whole plant of the genus Lindera or a part of the cut material is a large scale such as 1 kg to 10 kg, a large vertical cutter mixer or the like can be used, and several tens of g to several. In the case of a small scale of 100 g, a household mixer or the like can be used. For example, when applying crushing conditions on a large scale, it is appropriately changed depending on the hardness, water content, etc. of the raw material. For example, the crushing time is several tens of seconds to several minutes, and the rotation speed of the mixer is 10 to 3000 rpm. It is possible to do.

Further, the extraction conditions can be applied to ordinary conditions, and the method of immersing the dried product of the genus Lindera in a solvent at, for example, 3 to 100 ° C., heating at reflux or microwave heating is adopted, although the extraction conditions are not limited. be able to. The extraction temperature is not limited as long as an appropriate extraction amount can be obtained, but can be, for example, 20 to 100 ° C., preferably 22 to 80 ° C., and 25 to 80 ° C. when a hydrous alcohol such as 70% ethanol is used. 60 ° C. is more preferable. The extraction temperature can be, for example, 20 to 100 ° C., preferably 50 to 100 ° C., more preferably 70 to 100 ° C. when water is used. The extraction time is not limited as long as an appropriate extraction amount can be obtained, but can be, for example, 5 minutes or more and 14 days or less, preferably 10 minutes or more and 7 days or less, and more preferably 15 minutes or more and 5 days or less. The extraction is usually carried out under normal pressure, but it can also be carried out under pressure. The amount of the solvent added can be about 1 L to 100 L with respect to 1 kg of the dry weight of the plant of the genus Lindera.

It is also possible to increase the extraction efficiency by adding an acid, alkali, or enzyme to the plants of the genus Lindera before adding the extraction solvent.

A known method can be used for the operation of adding the extraction solvent and stirring. For example, a method of rotating an extraction container containing a cut or pulverized product of a plant belonging to the genus Lindera and an extraction solvent, the above-mentioned extraction container is used. Examples thereof include a method of installing in a magnetic or mechanical stirring device for mixing, a method of shaking the extraction container, and the like.

It is also possible to increase the extraction efficiency by causing vibration in the stirring operation by ultrasonic treatment. When performing ultrasonic treatment, for example, when the whole plant or a part of the cut material of the genus Lindera is a large scale such as 1 kg to 10 kg, a commercially available ultrasonic generator can be used for the genus Lindera. An ultrasonically treated product can be obtained by installing an extraction container containing a cut product or a crushed product and an extraction solvent and treating with 26 kHz ultrasonic waves for 60 minutes. In the case of a small scale of several tens to several hundreds of g. A sonicated product can be obtained by treating with ultrasonic waves of 40 kHz for 60 minutes. The temperature condition of the ultrasonic treatment is not limited as long as an appropriate extraction amount can be obtained, but can be 2 ° C to 100 ° C, preferably 10 ° C to 70 ° C. As the ultrasonic generator for large scale, for example, UT-12 manufactured by Shinmeidai Kogyo Co., Ltd. can be used, and as the ultrasonic generator for small scale, for example, US-3R manufactured by AS ONE Co., Ltd. can be used. Can be used.

In order to increase the extraction efficiency, it is also possible to perform multi-step extraction using the same type or a plurality of types of extraction solvents. In the case of multi-step extraction, the extract of the genus Lindera is separated from the extraction supernatant after further adding the same or multiple kinds of extraction solvents to the residue obtained in the first step extraction and heating at room temperature. Can be extracted.

In the microwave heating, conditions such as the output of the microwave irradiation device, the wavelength of the microwave, and the irradiation time can be appropriately set as long as the activity of the active ingredient of the present invention is not lost. For example, when a microwave of 2450 MHz and 500 W is applied to 100 g of a dried product of the genus Lindera, it can be treated in 10 seconds to 10 minutes, preferably 30 seconds to 5 minutes.

The extract supernatant can be used as it is as an extract of a plant of the genus Lindera. Furthermore, an extract of a plant of the genus Lindera can be separated and extracted from the extract supernatant. In this separation / extraction step, a known method can be adopted, and for example, filtration, centrifugation, suction, squeezing, etc. can be performed.

In the case of separation and extraction by filtration, for example, membrane filtration can be performed, but not limited to. When performing membrane filtration, for example, the temperature condition can be 2 ° C. to 70 ° C., preferably 10 ° C. to 40 ° C., and the membrane pore diameter can be 0.1 μm to 10 μm, preferably 0.1 μm to 5 μm. Is possible.

In the case of separation and extraction by centrifugation, a known device can be used, and examples of the centrifuge include a separation plate type, a cylindrical type, and a decanter type. When centrifuging, for example, the temperature condition can be 2 ° C to 70 ° C, preferably 10 ° C to 40 ° C, and the rotation speed is 1000 rpm to 10000 rpm, preferably 1500 rpm to 8000 rpm, and more preferably 2000 rpm to 6000 rpm. The centrifugation time can be 10 seconds to 30 minutes, preferably 20 seconds to 20 minutes, and more preferably 30 seconds to 15 minutes.

For the separation and extraction by squeezing, a squeezing machine can also be used, and a known device can be used as the squeezing machine, and examples thereof include a pneumatic squeezing machine and a screw type squeezing machine.

The extract of the genus Lindera can be made into a purified product by further subjecting it to a purification treatment before and after dilution or concentration. In addition to the above-mentioned extraction with a solvent, a chromatographic method, an ion exchange chromatograph method, or the like known to those skilled in the art can be used alone or in combination for the purification treatment.

When the chromatograph method is used, for example, either column chromatography using a normal phase or reverse phase carrier or an ion exchange resin, high performance liquid chromatography, thin layer chromatography, centrifugal liquid chromatography, or the like, or A method of using them in combination can be mentioned. When the chromatograph method is used, the purification conditions such as the carrier and the elution solvent can be appropriately selected according to various chromatograph methods.

From the viewpoint of obtaining higher extraction efficiency, the ratio of the cut or crushed product of the genus Lindera to the extraction solvent is such that when a water extract is obtained, for example, 1 L of water is used to obtain a plant of the genus Lindera. The cut or crushed product of the dried product can be 5 g to 300 g, preferably 10 g to 200 g, and more preferably 20 g to 100 g. When a water-containing organic solvent extract is obtained, for example, the amount of the cut or pulverized product of the genus Lindera can be 10 g to 1 kg, preferably 20 g to 500 g, and more preferably 30 g to 200 g with respect to 1 L of the solvent. ..

In the present invention, the content of the linden plant and the linden plant extract is converted into solid content based on the total amount of the composition for preventing and / or improving lifestyle diseases from the viewpoint of obtaining a higher effect of the present invention. 0.000001 to 100% by weight, preferably 0.00001 to 90% by weight, still more preferably 0.0001 to 80% by weight, still more preferably 0.001 to 70% by weight, still more preferably 0.01 to 60% by weight. , More preferably 0.1 to 50% by weight, still more preferably 0.2 to 40% by weight, still more preferably 0.3 to 30% by weight, still more preferably 0.4 to 25% by weight, still more preferably 0. 5 to 20% by weight, more preferably 0.6 to 15% by weight, still more preferably 0.7 to 10% by weight, still more preferably 0.8 to 5% by weight, still more preferably 0.8 to 4% by weight, It can be more preferably 0.8 to 3% by weight, still more preferably 0.8 to 2% by weight.

In the present invention, the content of the purified product of the genus Lindera is 0.000000001 in terms of solid content based on the total amount of the composition for preventing and / or improving lifestyle-related diseases from the viewpoint of obtaining a higher effect of the present invention. It can be ~ 1% by weight, preferably 0.00000001 to 0.9% by weight, and more preferably 0.000000001 to 0.8% by weight.

[Use]
The present inventors have that a plant of the genus Lindera or an extract or purified product thereof has an excellent effect on lifestyle-related diseases, but is not limited to, for example, an effect of improving obesity, an effect of improving muscle endurance, and an effect of improving liver function. Was newly found. Since the plants of the genus Lindera are naturally derived and have been edible since ancient times, they are suitable for safe and effective obesity-improving applications, muscle endurance-enhancing applications, or liver function-improving applications. Although the present invention can be used for pharmaceutical purposes, it can also be used for non-pharmaceutical applications.

As used herein, the term "improvement of obesity" means having an action of lowering blood lipids, an action of inhibiting lipase, or an action of promoting energy burning. Although not limited, blood lipids can be measured by triglyceride (neutral fat), total cholesterol, LDL-cholesterol called bad cholesterol, free fatty acid and the like. Blood lipids are preferably measured in serum. The composition for preventing and / or ameliorating lifestyle diseases of the present invention is not limited because it can lower triglyceride and cholesterol in blood, but is not limited, but has lipid dysfunction (hyperlipidemia), metabolic syndrome, and hypertriglyceridemia. It is also suitable for the prevention and / or treatment of hypercholesterolemia, hypercholesterolemia, hyper-LDL cholesterolemia, and low HDL cholesterolemia. Further, the composition for preventing and / or improving lifestyle-related diseases of the present invention is also suitable for food and drink products intended for patients with these diseases and healthy persons who are aware of the prevention of these diseases.

Lipase secreted by the pancreas (pancreatic lipase) is a digestive enzyme of lipids in the gastrointestinal tract. Of the lipids ingested by the diet, triglycerides are decomposed into fatty acids and monoglycerides by pancreatic lipase and absorbed from the small intestine. A composition that inhibits the activity of pancreatic lipase is considered to be effective not only in inhibiting lipid absorption from the gastrointestinal tract but also in suppressing lipid accumulation in the body to prevent or ameliorate hyperlipidemia and / or obesity. (Http: //wwwsoc.nii.ac.jp/jasso/topics/pdf/topics7_33.pdf "Obesity Study" V0l.7 No.3 p112-114 2001)

Excess fats and carbohydrates absorbed by the diet are synthesized into triglycerides via free fatty acids and accumulated in adipocytes. Therefore, in order to reduce the accumulation of triglycerides and eliminate obesity, it is desirable to efficiently decompose and reduce free fatty acids. As a method of decomposing and reducing free fatty acids, in addition to conversion to chemical energy (synthesis of ATP) associated with activation of oxygen respiration by exercise, conversion to heat energy by uncoupling protein (UCP). Can be mentioned. As used herein, decoupling refers to inhibiting the conjugation of energy obtained by electron transfer to an ATP synthesis reaction in an oxidative phosphorylation reaction in mitochondria. The uncoupling protein has a function of releasing energy originally coupled to the ATP synthesis reaction as thermal energy by relaxing the proton gradient in mitochondria due to oxidative phosphorylation. UCP1-5 are known as uncoupled proteins. Of these, overexpression of UCP3 in mice is known to suppress weight gain and insulin resistance induced by a high-fat diet (see (Choi, CS. et al, 2007, J. et al.). Clin. Invest., Vol. 117, p. 1995-2003)). It has also been suggested that UCP3 is also involved in the reduction and elimination of active oxygen (see (Toime, LJ. And Brand, MD., 2010, Free Rad. Biol. Med., Vol. 49, p.). 606-11)). Therefore, if the function of UCP3 can be activated, it will lead to prevention and improvement of obesity and poor circulation by promoting the decomposition of free fatty acids and converting them into heat energy, as well as various types involving type 2 diabetes and oxidative stress. It is expected to lead to improvement of the disease.

As used herein, improving muscle endurance means improving the ability of muscles to work for a certain period of time. In addition, a decrease in muscle endurance means a decrease in the ability of muscles to work for a certain period of time. Although not limited, muscle endurance can be measured using the expression level of GLUT4 (Glucose transporter 4) involved in the increase in muscle endurance as an index. The composition for preventing and / or ameliorating lifestyle-related diseases of the present invention is not limited because it can promote the increase of skeletal muscle and increase the expression level of GLUT4, but it depends on locomotive syndrome, aging and illness. It is also suitable for use in loss of muscle mass, loss of muscle mass due to lifestyle-related lifestyles such as sedentary tendency, loss of muscle mass due to excessive dieting, prevention and / or treatment of sarcopenia. Further, the composition for preventing and / or improving lifestyle-related diseases of the present invention is also suitable for food and drink products intended for patients with these diseases and healthy persons who are aware of the prevention of these diseases. Further, as another embodiment of the present invention, there is an application for promoting the expression of GLUT4 containing a plant of the genus Lindera or an extract or purified product thereof as an active ingredient, an application for promoting glucose uptake in muscles, or an in vitro promotion of glucose uptake in muscles. It is possible to provide uses.

As used herein, improving liver function means improving the circulatory function, excretion function, metabolic function, protection / detoxification function, or hematological function of the liver. In addition, the decrease in liver function means that the circulatory function, excretion function, metabolic function, protection / detoxification function, or hematological function of the liver is decreased. Liver function is not limited, but liver function is determined by blood GOT (glutamate oxaloacetate transaminase) level, GPT (glutamate pyruvate transaminase) level, enzymes such as γ-GTP (γ-glutamyl transpeptidase), or a microscope of a cell section of the liver. It can be measured by observation. The composition for preventing and / or ameliorating lifestyle diseases of the present invention is not limited because it can lower the GOT value and the GPT value, but the liver disease that causes an increase in these values, for example, fatty liver. It is also suitable for the prevention and / or therapeutic agent for alcoholic hepatitis, viral hepatitis such as A, B, and C, liver cirrhosis, and liver cancer. Further, the composition for preventing and / or improving lifestyle-related diseases of the present invention is also suitable for food and drink products intended for patients with these diseases and healthy persons who are aware of the prevention of these diseases.

Fatty liver medically means a pathological condition in which fat exceeds 5% of the total weight of the liver, but liver diseases including this are the cause of death in the adult population in the 40s and 50s of developed countries. The next most serious illness. Currently, there are few drugs that are useful for the pharmacological treatment of fatty liver, so exercise and diet are recommended, but in fact, treatment of fatty liver by such a method is not so effective. The situation is that there is an urgent need to develop effective therapeutic agents rather than the target. Pathologically, when hepatocytes are observed under a microscope, fat vacuoles are observed in 30% or more of 100 cells. When the tissue becoming fatty liver is observed under a microscope, an increase in vacuoles in hepatocytes is observed, but the composition for preventing and / or improving lifestyle-related diseases of the present invention is alcohol drinking water using mice. In the test, it is possible to suppress the formation of vacuoles in hepatocytes. Although not limited, the composition for preventing and / or improving lifestyle-related diseases of the present invention is suitable for food and drink products intended for healthy subjects who are conscious of prevention and / or improvement of fatty liver or prevention of fatty liver. ..

As another embodiment of the present invention, the composition for preventing and / or ameliorating lifestyle-related diseases of the present invention can lower blood glucose and send blood glucose to muscle cells, and thus is a plant of the genus Lindera. It is possible to provide a blood glucose level suppressing application containing the extract or purified product of the above as an active ingredient. Although not limited, the composition for preventing and / or ameliorating lifestyle-related diseases of the present invention is suitable for food and drink products directed to healthy persons who are conscious of preventing and / or ameliorating diabetes.

[Food and drink]
The composition for preventing and / or improving lifestyle-related diseases of the present invention can be blended as one component of food and drink. These food and drink products are not limited, but can also be used as food and drink products used for improving obesity, enhancing muscle endurance and / or improving liver function, and are used, for example, in medical institutions such as hospitals. May be provided for the patient. Alternatively, these foods and drinks can be provided as functional beverages or functional foods, but are not limited, and these functional foods and drinks can be used in medical institutions, drug stores, convenience stores, and supermarkets. , Can be provided at department stores, etc. Functional foods and drinks are foods and drinks that have medicinal properties permitted and designated by the national government and public organizations, or foods and drinks that display functionality based on the content that the company has notified the national government of the prescribed effects. To say. Examples of functional foods include foods for specified health use, foods with nutritional function, foods with functional claims, foods for the elderly, health supplements (balanced nutritional foods, supplements) and the like. It also includes foods and drinks including packages and containers, package inserts, instruction manuals, etc. that indicate pharmaceutical efficacy or functionality. Foods and drinks that indicate pharmaceutical efficacy or functionality in the application form to the national government are also included.

In the case of labeling for the purpose of improving obesity, there is no limitation, but for example, those who are concerned about blood lipids, those who are obese, those who are concerned about body weight (BMI), those who are concerned about visceral fat, etc. Those who are concerned about subcutaneous fat, those who are concerned about body fat, those who are concerned about lifestyle diseases (adult diseases) related to obesity, those who are concerned about abdominal circumference (waist size), those who want to reduce the waist circumference , Those who have high triglyceride, those who want to suppress the rise of triglyceride, those who have high total cholesterol and LDL (bad) cholesterol, those who want to increase HDL (good) cholesterol, etc.

The display for the purpose of strengthening muscle endurance is not limited, but for example, those who want to maintain stamina of muscle strength, those who want to support the decrease in muscle strength due to aging, those who want to improve walking ability, daily life and after exercise. Those who want to reduce their fatigue, those who feel tiredness or fatigue (related to muscle mass loss), those who are concerned about coldness (related to muscle mass loss), those who are concerned about coldness (related to muscle mass loss) For those who are worried about poor posture such as stooping, those who have knee pain due to walking or running, those who want to improve aerobic exercise ability, those who want to improve their usual work ability, etc. Be done.

In the case of labeling for the purpose of improving liver function, there is no limitation, but for example, those who want to maintain healthy liver function, those who are worried about sickness or tiredness of drinking, those who are worried about GOT value or GPT value, fatigue. There are indications for those who feel a feeling, those who feel tired, those who have a loss of appetite, and those who are concerned about lifestyle diseases (adult diseases) regarding liver function.

In the case of the display for the purpose of improving the blood glucose level, for example, those who have a high blood glucose level, those who are concerned about the blood glucose level, those who are concerned about the postprandial blood glucose level, and those who are concerned about the postprandial blood glucose level, are not limited. Display for those who want to be gentle (gentle) can be mentioned.

Foods include all foods, such as grains, potatoes, fish and shellfish, meats, eggs, fats and oils, milk, vegetables, legumes, fruits, sugars, seaweeds, confectionery, seasonings. Kind, cooked processed foods and the like.

The seasoned processed foods are not limited, but for example, processed marine products such as chikuwa and kamaboko; processed livestock products such as ham and sausage; sweets such as cookies, biscuits, snacks, chocolates and cakes; soba, udon and raw noodles. , Chinese noodles, noodles such as pasta; bread such as bread and sweet bread; fermented processed foods such as natto and miso; soybean foods such as tofu and okara; pickles such as lightly pickled and rice bran pickles, marine products, processed meats, vegetables, Canned fruits and the like; dairy products such as butter, margarine, yogurt, cheese and milk; ice cream, chilled foods such as sherbet and the like.

Beverages include, but are not limited to, all beverages, such as beverages made from the genus Kuromoji plant itself, roasted dried products as a substitute for tea leaves, fruit juice beverages, 100% fruit juice beverages, and low fruit juice beverages. , Fruit beverages, vegetable juices, flavored beverages, fruit beverages for dilution, etc .; carbonated beverages; coffee, coffee beverages, soft beverages with coffee, cocoa beverages, tea, green tea, matcha, crow dragon tea, wheat tea, roasted tea, etc. Beverages; vinegared beverages; soft drinks such as sports drinks; milk; milk beverages; dairy beverages; lactic acid beverages; lactic acid bacteria beverages; soybean beverages such as soymilk and prepared soymilk; alcoholic beverages such as beer, sake, shochu, liqueurs, and wine. ; Examples include nutritional beverages containing taurine, royal jelly, amino acids, vitamins, minerals, iron and the like.

There is no limitation on the time when the present invention is applied to food and drink products, but for example, it can be applied before and after a processing step, a cooking step, a heating step, a storage step and the like in a food and drink manufacturing process. For example, in the processing step and the cooking step, the raw material may contain the composition for preventing and / or improving the lifestyle-related disease of the present invention. The application method can be appropriately changed depending on the type of food and drink, the form of the raw material, and the like, and various methods such as mixing, addition, coating, spraying, and dipping can be adopted.

When the composition for preventing and / or improving lifestyle-related diseases of the present invention is applied to a beverage product, it is preferable to use a tea beverage in one embodiment. Without limitation, the leaves and / or stems of dried Lindera plants are cut by a known method and then crushed or crushed into powder. It can be sold as a tea beverage in the form of powdered plants of the genus Lindera. If necessary, the powdered material of the genus Lindera is roasted. The powdered material of the genus Lindera can also be sold as a tea beverage in a roasted state. Powdered or roasted products of the genus Lindera can be sold, for example, in the form of a tea bag containing an appropriate amount. Roasted Lindera plants are drunk by boiling in boiling water.

When the composition for preventing and / or ameliorating lifestyle-related diseases of the present invention is applied to a tea beverage, in another embodiment, the stems and leaves of plants of the genus Lindera are 1 to 3: 1 based on the dry weight. It can be used as a tea beverage by using it in a weight ratio, drying and crushing it, and roasting it.

Further, the beverage or other tea beverage can be used as a mixture of a plant of the genus Lindera or an extract thereof, or diluted with water or the like and used as a juice or a drink. Further, it can be used as a concentrated product of the mixture, powdered tea powdered by spray drying or the like, freeze-dried instant tea or the like. In that case, for example, hot water can be poured into powdered tea or instant tea and drunk. Further, for example, a mixture of a tea preparation such as green tea or black tea and a plant of the genus Lindera or an extract thereof can be used as it is as tea leaves for drinking. In that case, hot water can be poured into the mixed tea leaves of the tea preparation and the genus Lindera plant or its extract in the same manner as general Japanese tea and black tea drinks.

When the composition for preventing and / or improving lifestyle-related diseases of the present invention is applied to food and drink, an auxiliary agent used in ordinary foods and beverages should be appropriately added as long as the effect of the present invention is not impaired. Is possible. Such auxiliaries include, for example, glucose, sucrose, fructose, oligosaccharide, water candy, maltose, multitose, xylitol, sorbitol, aspartame, sucralose, stebioside, rubusoside, corn syrup, lactose, citric acid, tartrate, malic acid. , Succinic acid, lactic acid, L-ascorbic acid, dL-α-tocophenol, sodium erythorbate, glycerin, glycerin fatty acid ester, propylene glycol, polyglycerin fatty acid ester, sucrose fatty acid ester, propylene glycol fatty acid, sorbitan fatty acid ester, ester Examples include arabic gum, casein, pectin, gelatin, agar, carrageenan, vitamin Bs, vitamin Cs, nicotinic acid amides, calcium pantothenate, calcium salts, amino acids, pigments, fragrances, preservatives and the like.

The content of the genus Lindera plant or its extract when the composition for preventing and / or improving lifestyle-related diseases of the present invention is applied to food and drink is not limited, but may be human or animal in terms of solid content. For example, it is generally 0.00001 to 5000 mg / kg body weight per day, preferably 0.01 to 200 mg / kg body weight, and more preferably 0.1 to 100 mg / kg body weight.

The pH when the composition for preventing and / or improving lifestyle-related diseases of the present invention is applied to a beverage product is appropriately adjusted depending on the type of the beverage product. For example, in the case of a tea beverage, the pH is 3.5 to 7. It can be 5, preferably 3.8 to 7.2, more preferably 4.0 to 7.0, and even more preferably 4.5 to 6.5. The pH of the beverage product can be adjusted by appropriately blending, for example, sodium hydrogen carbonate, citric acid, or the like.

[Pharmaceuticals, quasi-drugs]
When the composition for preventing and / or improving lifestyle-related diseases of the present invention is a drug or a quasi-drug, it can be formulated into an appropriate form and administered to humans or animals in any administration form. The administration form is not limited, and examples thereof include oral, transdermal, enteral, transmucosal, and injection. From the viewpoint of remarkably exerting the effect of the present invention, oral administration is preferable as the administration form.

When the composition for preventing and / or improving lifestyle-related diseases of the present invention is orally administered, the composition for preventing and / or improving lifestyle-related diseases of the present invention is, for example, a tablet, a capsule, a granule, or a syrup. , Powder, film, drop, jelly, semi-solid pudding, etc. are formulated by a known method. In the case of oral administration, the composition for preventing and / or ameliorating lifestyle-related diseases of the present invention may be ingested without water by a film-like, drop-like, jelly-like, semi-solid pudding-like dosage form, etc. It is possible. Alternatively, it is also possible to orally administer a dry powder of a plant of the genus Lindera or an extract or purified product thereof as a crude drug or a Chinese herbal medicine preparation.

In the case of parenteral administration, for example, it is possible to formulate a dosage form such as intravenous injection, intramuscular injection, transdermal absorbent, inhalant, suppository, eye drop, nasal drop, etc. by a known method. be.

As one aspect of the dosage form, a known drug for the purpose of enhancing the stability and bioavailability of a plant of the genus Lindera or an extract or purified product thereof, or improving the medication compliance of a patient, or a combination thereof. The delivery system can be used to deliver the composition for preventing and / or ameliorating lifestyle-related diseases of the present invention to the absorption site.

The drug delivery system utilizes, but is not limited to, polymers such as cellulose, dextran, starch, polyvinyl alcohol, acetylated or methylated polymers, polylactic acid and polyglycolic acid and their block copolymers, polyethylene glycol and the like. Methods, methods using transport proteins such as albumin, and other methods using micelles, liposomes, microspheres, nanoparticles, composite emulsions, dendrimers and the like can be mentioned. These drug delivery systems are used not only for the purpose of transporting the composition for preventing and / or improving the lifestyle-related disease of the present invention to a target site such as an absorption site, but also for preventing and / or improving the lifestyle-related disease of the present invention in a timely manner. It can also be used for the purpose of release control to release the composition for use.

The composition for preventing and / or ameliorating lifestyle-related diseases of the present invention contains liquid excipients, lubricants, binders, disintegrants and the like in solid formulations as long as the effects of the present invention are not impaired. In the pharmaceutical product, a solvent, a solubilizing agent, a suspending agent, an tonicity agent, a buffering agent, a pain-relieving agent and the like can be appropriately added.

Examples of the excipient include lactose, sucrose, D-mannitol, glucose, starch, calcium carbonate, kaolin, crystalline cellulose, light anhydrous silicic acid and the like.

Suitable examples of lubricants include, for example, magnesium stearate, calcium stearate, talc, purified talc, colloidal silica, borax, polyethylene glycol and the like.

Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, bound cellulose, sucrose, D-mannitol, dextrin, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, and hydroxypropyl starch. , Methyl cellulose, ethyl cellulose, shelac, calcium phosphate, polyvinylpyrrolidone and the like.

Examples of the disintegrant include starch, dried starch, carboxymethyl cellulose, carboxymethyl cellulose calcium, carmellose calcium, croscarmellose sodium, carboxymethyl starch sodium, sodium alginate, canten powder, sodium hydrogencarbonate, calcium carbonate, sodium lauryl sulfate, and the like. Examples thereof include stearic acid monoglyceride and lactose.

Examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil and the like.

Examples of the solubilizing agent include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like.

Examples of the suspending agent include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glycerin monostearate, and for example, polyvinyl alcohol and polyvinyl. Examples thereof include hydrophilic polymers such as pyrrolidone, sodium carboxymethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose.

Examples of the tonicity agent include sodium chloride, glycerin, D-mannitol and the like.

Examples of the buffering agent include buffer solutions such as phosphates, acetates, carbonates and citrates.

Examples of the soothing agent include benzyl alcohol and the like.

Examples of the preservative include paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like.

The effective dose of a plant of the genus Lindera or an extract thereof when the composition for preventing and / or ameliorating lifestyle-related diseases of the present invention is orally administered is not limited, but can be human or animal in terms of solid content. Generally, it is 0.00001 to 5000 mg / kg body weight per day, preferably 0.01 to 200 mg / kg body weight, and more preferably 0.1 to 100 mg / kg body weight. The number of administrations is usually about 1 to 4 times a day, but can be appropriately adjusted depending on the administration route and the like.

Next, the present invention will be specifically described with reference to Test Examples, Examples and the like, but the present invention is not limited to the following Examples.

(Example 1) Extraction method 1 from a dry pulverized product using a water-containing organic solvent
20 g each of dried pulverized leaves and stems of L. sericea was dispersed in 300 ml of a 70% ethanol aqueous solution, stirred, and extracted at room temperature for 24 hours. The extract supernatant was separated by filtration, the residue was dispersed again in a 70% aqueous ethanol solution, heated under reflux, and then the extract supernatant was filtered off. Further, the residue was dispersed in a 70% ethanol aqueous solution, and after 1 hour of ultrasonic treatment, the extraction supernatant was filtered off, and the combined supernatants were sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere, and then kekuromoji. Extract 1 (6 g) from leaves and extract 2 (6 g) from stems of kekuromoji were obtained.

(Example 2) Extraction method with water from dried pulverized product 1.5 L of water is added to 100 g of dried pulverized product of leaves of Kekuromoji (L. sericea), crushed with a mixer, stirred and ultrasonically applied, and then extracted. By filtering and dehydrating Qing, an extract (2 L) was obtained together with water. Further, the mixture was sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere to obtain Extract 3 (20 g).

(Example 3) Extraction method by squeezing 1 L of water is added to a mixture (1 kg) of fresh leaves and fresh stems of L. sericea, crushed with a mixer, stirred and ultrasonically applied, and the extracted supernatant is filtered. Separately, by washing and dehydrating, an extract (2 L) was obtained together with water. Further, the mixture was sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere to obtain Extract 4 (20 g).

(Example 4) Extraction method by squeezing 1 L of water was added to a mixture (1 kg) of fresh leaves and stems of L. sericea, crushed with a mixer, stirred and ultrasonically applied, and the extracted supernatant was filtered off. And after dehydration, it was squeezed to obtain an extract (1.5 L). Further, the mixture was sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere to obtain Extract 5 (25 g).

(Example 5) Method for producing dry powder Fresh leaves and stems (1 kg) of L. sericea are dried overnight in a 40 ° C. cold air dryer, finely pulverized in a crusher, and dried powder (100 g). Got

(Example 6) Extraction method using Oobakuromoji 10 mL of water was added to 1 g of dried pulverized stem of Oobakuromoji (L. umbellate var. Membrancea (Maxim) Mojama), and extraction was performed for 10 minutes. An extract was obtained by filtration and dehydration. To the residue, 10 mL of water was added again, extraction was performed for 10 minutes, and then the supernatant was filtered off and dehydrated. Extract 6 was obtained by mixing with the previously obtained extract and formulating to 20 mL with water (50 mg equivalent / ml).

(Example 7) Extraction method by microwave Add 20 mL of water to 1 g of dried ground leaves and stems of Kekuromoji (L. sericea), seal in a polypropylene container, and heat in a microwave oven (2450 MHz, 500 W) for 1 minute. By filtering and dehydrating the extracted supernatant, Extract 7 (20 mL) was obtained together with water. Extract 8 was obtained by using the same method and setting the heating time to 5 minutes.

(Example 8) Extraction method with hot water from dried pulverized product 1
1.5 L of hot water was added to 100 g of dried pulverized leaves and stems of Kekuromoji (L. sericea), and the extract supernatant was filtered off to obtain an extract (2 L) together with water. Further, the mixture was sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere to obtain Extract 9 (15 g).

(Example 9) Extraction method with hot water from fresh leaves and stems Water is added by adding 1 L of hot water to a mixture (1 kg) of fresh leaves and stems of Kekuromoji (L. sericea) and filtering the extraction supernatant. And an extract (1.5 L) was obtained. Further, the mixture was sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere to obtain Extract 10 (10 g).

(Example 10) Extraction method 2 from a dry pulverized product using a water-containing organic solvent
20 g of a dried pulverized product of the stem (including bark and heartwood) of L. sericea and 20 g of a dried pulverized product of leaves were dispersed in 300 ml of 80% methanol, stirred, and extracted at room temperature for 24 hours. After the extract supernatant was filtered off, the residue was dispersed again in 80% methanol, heated under reflux, and then the extract supernatant was filtered off. Further, the residue was dispersed in a 70% ethanol aqueous solution, and after 1 hour of sonication, the extraction supernatant was filtered off, and the combined supernatants were sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere, and then kekuromoji. Extract 11 (1.5 g) from the stem and extract 12 (5 g) from the leaves of Kekuromoji were obtained. These extracts 11 and 12 were mixed in a weight ratio of 7: 3 based on the dry weight to obtain the extract 13.

(Example 11) Extraction method with hot water from dried pulverized product 2
Add 1.5 L of hot water to 100 g each of dried crushed stalks (including bark and heartwood) and leaves of L. sericea, and filter out the extract supernatant to make an extract (2 L) together with water. Got Further, the mixture was sequentially concentrated under reduced pressure, dried under reduced pressure and freeze-dried under a nitrogen gas atmosphere to obtain an extract 14 (6 g) from the stem of Kekuromoji and an extract 15 (6 g) from the leaves of Kekuromoji. These extracts 14 and 15 were mixed to give the extract 16.

(Animal and diet)
A 7-week-old naturally weight-gaining mouse (C57BL / 6J male mouse, average weight 23.1 g) was purchased from Charles River. Animals were bred at room temperature 24 ± 3 ° C. and 12 / 12h (AM 7:00-PM 7:00) light-dark cycle conditions. Mice were given free access to water and food. Then, the mice were assigned to 3 groups (low-fat diet group, high-fat diet group, or high-fat Kuro-moji diet group) consisting of 10 mice according to their body weight, and the following diets were freely supplied for 36 days (test). period).

The details of the diet in each group are shown in Table 1. The unit of the numerical values in Table 1 is% by weight. As the Kuro-moji extract in Table 1, the extract 13 of the genus Lindera obtained in Example 10 was prepared so as to contain 1% in terms of solid content.

(Test Example 1) Evaluation test of obesity improving effect After the above test period (animal and diet) was completed, serum triglyceride and cholesterol were evaluated. After the end of the test period, blood was collected under ether anesthesia and sacrificed, and the weights of various organs were measured. In addition, serum was prepared and stored at -30 ° C until the time of analysis (Test Example 1-1) Measurement of serum triglyceride For triglyceride in serum, triglyceride E-test Wako kit (manufactured by Wako Pure Chemical Industries, Ltd.) was used. , Quantified according to the manufacturer's instructions. The results of blood triglyceride are shown in FIG. The vertical axis of the graph shows the concentration per serum in mg / dL.

As shown in FIG. 1, as a result of measuring the triglyceride in the serum, it was found that the triglyceride in the serum was significantly reduced in the high-fat Kuro-moji diet group as compared with the high-fat diet group. Surprisingly, it was found that the serum triglycerides in the high-fat Kuro-moji diet group were even lower than those in the low-fat diet group.

(Test Example 1-2) Measurement of Serum Cholesterol Cholesterol in serum was quantified using the Cholesterol E-Test Wako Kit (manufactured by Wako Pure Chemical Industries, Ltd.) according to the manufacturer's instructions. The results of blood cholesterol are shown in FIG. In FIG. 2, the vertical axis of the graph shows the concentration per serum in mg / dL.

As shown in FIG. 2, as a result of measuring the cholesterol in the serum, it was found that the cholesterol in the serum was significantly reduced in the high-fat Kuro-moji diet group as compared with the high-fat diet group. Surprisingly, it was found that serum cholesterol in the high-fat Kuro-moji diet group was reduced to the same extent as in the low-fat diet group.

(Test Example 1-3) Lipase inhibitory activity evaluation test Extracts A, B, C of the genus Lindera obtained in Example 8-2, and extracts 11 and 12 of the genus Lindera obtained in Example 10. , 13 was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 10 mg / mL and subjected to a lipase inhibitory activity evaluation test. Pig pancreatic lipase (manufactured by Sigma-Aldrich) and lipase kit S (manufactured by Dainippon Pharmaceutical Co., Ltd.) were used in the evaluation test. The measuring method was carried out by partially modifying the method of the kit and using a 96-well microplate. That is, 15 μL of the sample sample solution containing the extract, 4 μL of the enzyme solution (porcine pancreatic lipase 0.05 mg / mL, 125 mmol / L Tris-hydrochloric acid (pH 7.5)), and the coloring solution (0.1 mg / mL 5,5'-. After adding 50 μL of dithiobis (buffer solution containing 2-nitrobenzoic acid) and mixing, preheat at 30 ° C. for 5 minutes, and then preheat the substrate solution (6.69 mg / mL dimercaprol tributyrate + 5.73 mg / mL sodium dodecyl sulfate). ) After adding 5 μL and mixing, the mixture was heated at 30 ° C. for 30 minutes under shading, a reaction stop solution was added, and then the absorbance at 412 nm was measured with a microplate reader. DMSO was added as a sample solution to the control of each sample, and the same operation was performed. The results of the lipase inhibitory activity evaluation test are shown in FIG.

In FIG. 3, the vertical axis shows the average inhibition rate (%). Three samples were tested for each extract, and the inhibition rate (%) was calculated by the following formula 1.
[Equation 1]
Inhibition rate (%) = 100- (test group-blank) / control x 100

As shown in FIG. 3, as a result of measuring the lipase-inhibiting activity of the linden extract, the sample group of the linden plant extract showed higher lipase-inhibiting activity as compared with the negative control containing no linden extract. As a positive control, the same degree of inhibition as compared with the sample (Camelia extract in FIG. 3) in which green tea extract (Sanphenon 90S, Taiyo Kagaku Co., Ltd.) was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 2.5 mg / mL. It became clear that it showed activity.

(Test Example 2) Evaluation test of muscle endurance effect (Test Example 2-1) Measurement of body weight During the above test period (animal and diet), the body weight was measured weekly. The measurement result of the body weight is shown in FIG. In FIG. 4, the vertical axis of the graph shows the average body weight of each group in g.

As shown in FIG. 4, the body weight of the low-fat diet group, the high-fat diet group, and the high-fat Kuro-moji diet group increased over time. The weight gain of mice was less affected by the addition of the extract of the genus Lindera, and the high-fat Kuro-moji diet group tended to increase slightly compared to the high-fat diet group.

(Test Example 2-2) Measurement of visceral fat mass The visceral fat mass was measured by the gravimetric method. For each organ of the liver, epididymis, kidney, mesentery, and brain, the amount of fat produced inside and / or around them and the total amount of visceral fat in the whole body were measured. The result of measuring the fat mass of the liver by measuring the weight of the liver is shown in FIG. The result of measuring the fat mass of the epididymis by measuring the weight of the epididymis is shown in FIG. The result of measuring the fat mass around the kidney by measuring the weight of the kidney is shown in FIG. The result of measuring the fat mass of the mesentery by measuring the weight of the mesentery is shown in FIG. FIG. 9 shows the results of measuring the amount of fat in the brain by measuring the weight of the brain. The results for the total amount of visceral fat in the whole body are shown in FIG. In FIGS. 5 to 10, the vertical axis of the graph shows the average fat weight in g.

As shown in FIGS. 5 to 10, when the visceral fat mass of each organ and the whole body was compared, no significant difference was observed between the high-fat Kuro-moji diet group and the high-fat diet group, and the cause of weight gain was the fat mass. It was suggested that it was due to an increase in skeletal muscle, not an increase.

(Test Example 2-3) Measurement of expression levels of GLUT4 gene and UCP-3 gene Therefore, the extract 11 or 12 of the genus Chromodi plant obtained in Example 10 was administered to C2C12 cells, which are mouse muscle cells. The expression levels of the GLUT4 (glucose transporter 4) gene involved in muscle endurance and the UCP-3 gene (uncoupling protein-3) involved in energy production were quantified by the qPCR method.

Specifically, the cells used were mouse-derived skeletal muscle cells (C2C12). The cells were cultured in Dulbecco's modified Eagle's medium DMEM (4.5 g / L glucose concentration) at 37 ° C. and 5% CO2 concentration. The cells were cultured to ⁵ / mL. The medium was changed every 2 days, and the cells were cultured until they became confluent (about 5 days). Then, after culturing in DMEM (1.0 g / L glucose concentration) for 3 days and confirming differentiation, the cells were replaced with a medium containing Kuro-moji extract and cultured for 2 days. Kuro-moji extract-added medium was prepared by dissolving Kuro-moji extract (extract 11 or 12) in dimethyl sulfoxide at 10 μg / mL, and adding 0.5% to the medium.

Then, cells were suspended using a cytolytic solution (CellAmp ^TM Direct RNA Prep Kit for RT-PCR: manufactured by Takara Bio Inc.), and RNA was extracted. From this, the cDNA was adjusted (PrimeScript ^TM RT gene Kit: manufactured by Takara Bio Co., Ltd.), and further by the TaqMan probe method (Probe qPCR Mix: manufactured by Takara Bio Co., Ltd.), Glucose transporter-4 (Glut-4) and Uncoupling protein-3. The expression level of (UCP-3) was quantified, β-actin was used as a housekeeping gene, and the ratio of the expression level of each gene to the expression level of the β-actin gene (indicated by "C" in the figure) was shown. .. The result of the expression level of the GLUT4 gene or the result of the expression level of the UCP-3 gene are shown in FIGS. 11 or 12, respectively.

From the results shown in FIGS. 11 and 12, it was clarified that the expression levels of both genes were significantly increased in the high-fat Kuro-moji diet group as compared with the high-fat diet group. The protein produced by GLUT4 is a channel involved in the intracellular transport of glucose, and moves to the cell surface by insulin stimulation or muscle contraction to send glucose in blood to muscle cells. Therefore, when GLUT4 increases, the amount of glucose taken up into muscle cells increases, and ATP in mitochondria increases, leading to an increase in muscle endurance. In addition, UCP-3 has the effect of inhibiting the coupling of energy obtained by electron transfer to the ATP synthesis reaction in the oxidative phosphorylation reaction in mitochondria, so that the proton gradient in mitochondria due to oxidative phosphorylation is relaxed. By doing so, it has a function of releasing energy originally coupled to the ATP synthesis reaction as heat energy. This is thought to have the effect of preventing obesity due to excessive energy uptake. In other words, it was clarified that the Kuro-moji extract exerts the effect of anti-obesity, improvement of endurance, and eventually increase of muscle mass by activating the expression of these genes.

(Test Example 2-4) Measurement of glucose in serum After the test period of the above (animal and diet) was completed, glucose in serum was evaluated. Glucose in serum was quantified using a glucose CII-Test Wako Kit (manufactured by Wako Pure Chemical Industries, Ltd.) according to the manufacturer's instructions. The results of serum glucose are shown in FIG. In FIG. 13, the vertical axis of the graph shows the concentration per serum in mg / dL.

As shown in FIG. 13, as a result of measuring glucose in serum, it was found that glucose in serum was significantly reduced in the high-fat Kuro-moji diet group as compared with the high-fat diet group. It is considered that this is due to the increase in the amount of glucose taken up into the muscle cells.

(Test Example 3) Evaluation test of liver function improving effect (Test Example 3-1) Measurement of GOT value and GPT value in serum After the test period of the above (animal and diet) is completed, the liver weight of mice and GOT in serum The values and GPT values were evaluated. As mentioned above, the measurement results of liver weight are shown in FIG. The GOT value and GPT value in serum were quantified using Transaminase CII Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.) according to the manufacturer's instructions. The results of GOT value and GPT value in serum are shown in FIG. 14 or FIG. 15, respectively. In FIG. 14 or 15, the vertical axis of the graph indicates the unit (U).

As shown in FIG. 14 or FIG. 15, as a result of measuring the GOT value and the GPT value, which are indicators of liver function, both the GOT value and the GPT value are significantly reduced in the high-fat Kuro-moji diet group as compared with the high-fat diet group. Was recognized.

(Test Example 3-2) Evaluation of fatty liver in alcohol loading test In order to investigate in detail the effect of improving liver function, a feeding test in which 10% alcohol was drunk was conducted. In the group labeled as the control diet group, the high-fat diet and water shown in Table 1 were given. In the group labeled alcohol intake group, the high-fat diet shown in Table 1 and 10% alcohol were given. In the group labeled alcohol intake Kuro-moji diet group, the high-fat Kuro-moji diet shown in Table 1 and 10% alcohol were given. Other conditions are the same as above (animals and diet). After the end of the test period, a section of the right lobe of the liver of the mouse was observed (HE staining). The result is shown in FIG.

As shown in FIG. 16, it was clarified that the alcohol-ingested Kuro-moji diet group had less vacuole formation in hepatocytes than the alcohol-intake group, and was comparable to the control diet group not ingesting alcohol. rice field. This indicates that the fatty liver formation caused by alcohol is suppressed.

The GOT value and GPT value, which are indicators of liver function, were measured in the control diet group, the alcohol intake group, and the alcohol intake Kuro-moji diet group by the same method as in Test Example 3-1. The results of GOT value and GPT value in serum are shown in FIG. 17 or FIG. 18, respectively. In FIG. 17 or 18, the vertical axis of the graph indicates the unit (U).

As shown in FIG. 17 or FIG. 18, as a result of measuring the GOT value and the GPT value, which are indicators of liver function, both the GOT value and the GPT value are significantly decreased in the alcohol intake Kuro-moji diet group as compared with the alcohol intake group. Was recognized.

(Prescription example)
The unit of the numerical value in each of the following prescription examples is shown as "% by weight" with the total amount as 100.

(Prescription Example 1) Composition (tablet) for prevention and / or improvement of lifestyle-related diseases
Using the extract obtained in the above example, tablets having the following composition for prevention and / or improvement of lifestyle-related diseases can be produced by a conventional method.

(Prescription Example 2) Composition (granule) for prevention and / or improvement of lifestyle-related diseases
Using the extract obtained in the above example, the following composition can be mixed and stirred to uniformly prepare, and granules can be obtained by a fluidized bed granulator. Granules can be taken with beverages.

(Prescription Example 3) Composition (capsule) for prevention and / or improvement of lifestyle-related diseases
Using the extract obtained in the above example, the following composition can be mixed and stirred to make a uniform preparation to obtain a powder for capsules to be used as a capsule.

(Prescription Example 4) Beverage for Prevention and / or Improvement of Lifestyle-related Disease A beverage for prevention and / or improvement of lifestyle-related disease having the following composition by a conventional method using the extract obtained in the above example. Can be manufactured.

(Prescription Example 5) Chewing gum for prevention and / or improvement of lifestyle-related diseases Using the extract obtained in the above example, chewing gum for prevention and / or improvement of lifestyle-related diseases having the following composition by a conventional method is used. Can be manufactured.

Claims (4)

Contains an extract of the genus Lindera,
The genus Lindera is derived from the stem and leaves of Kuro-moji.
A composition for promoting GLUT4 gene expression and / or UCP-3 gene expression for those who are on a high-fat diet. The composition according to claim 1, wherein the extract is a water extract, an organic solvent extract, or a water-containing organic solvent extract. The composition according to claim 1 or 2 , which is a food and drink product, a quasi-drug, or a pharmaceutical product. The composition according to claim 3 , wherein the food and drink is a tea beverage.

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