WO2018008999A1 - Composition comprising aster koraiensis extract or fraction thereof as active ingredient for regulating blood glucose and preventing or treating diabetes - Google Patents

Abstract

The present invention relates to a composition comprising an Aster Koraiensis extract or a fraction thereof as an active ingredient for preventing or treating diabetes and, specifically, to a pharmaceutical composition comprising an Aster Koraiensis extract or a fraction thereof as an active ingredient for preventing or treating diabetes, to a food composition, and a method for prevention or treatment of diabetes by using the pharmaceutical composition. The Aster Koraiensis extract of the present invention exhibits an effect of reducing blood glucose by promoting glucose uptake of adipocytes, and thus can be advantageously used in the prevention or treatment of diabetes. In addition, the composition of the present invention is derived from a natural material, and thus can be safely used without side effects in the regulation of blood glucose and the prevention, treatment, and alleviation of diabetes.

Description

Composition for the control of blood sugar, prevention or treatment of diabetes mellitus, including bee odor extract or fractions thereof as an active ingredient

The present invention relates to a composition for the prevention or treatment of diabetes mellitus comprising a bee anesthetist extract or fractions thereof as an active ingredient, specifically, for the control of blood glucose, or a prevention or treatment of diabetes mellitus including a bee herbicide extract or a fraction thereof as an active ingredient A composition, a food composition, a feed composition and a method for preventing or treating diabetes using the pharmaceutical composition.

Diabetes is a disease in which abnormally high blood glucose levels are constantly increasing. Type 1 diabetes is caused by the destruction of the pancreatic insulin-producing beta cells due to autoimmune diseases. Type 2 diabetes is caused by insulin secretion of pancreatic beta cells and insulin target cells such as liver, muscle, and adipose tissue. Insulin resistance (impaired reactivity to insulin) and the like are related to the etiology. Diabetes is one of the world's most prevalent adult diseases. In 2009, the American Medical Association (JAMA) said that the number of diabetics in the world today is more than 240 million people and will increase to 380 million by 2025. It is announced. Recently, with the rapid economic growth in Korea, the prevalence of diabetes reaches 10%, and in particular, the onset of diabetes is being brought to the young, middle and old age.

Meanwhile, the overseas market and research progress of veterinary medicine targeting pets is exploding. In other words, the overseas veterinary medicine market is valued at $ 23 billion (about 25 trillion won) as of 2014, which is 40 times the size of the domestic market, growing about 4% annually. Pets are also suffering from the same diseases as humans, but there is no professional and appropriate countermeasure, so they are using medicines for humans.

In addition, pets live in closed spaces like apartments, and the incidence of diabetes in pets is increasing due to the consumption of high-sugar foods such as westernized eating habits, lack of exercise, obesity, and carbohydrates. Most of the products needed for pets are imported.

On the other hand, when diabetes continues to be chronic, advanced glycation end products (AGE), one of the leading triggers of diabetic complications, are irreversibly produced. That is, if time passes without treating diabetes, abnormally excessive final glycation products are produced and combined with proteins or lipids of specific tissues to progress to various diabetic complications. In addition, it has already been confirmed and reported that final glycosylation inhibitors cannot regulate blood glucose normally ( J. of Diabetes Mellitus , 2014, 4, 107-114; Diabetes Research and Clinical Practice , 1999, 43, 81-89; Arterioscler Thromb Vasc Biol , 2008, 28 (8), 1432-1438).

Therefore, steady management is essential for the prevention or treatment of diabetes in order to maintain blood sugar normally at the beginning of diabetes before the production of excessive final glycated products.

Insulin, now commonly prescribed for diabetics, can cause side effects of hypoglycemic shock, edema and congestive heart failure throughout the body. It also causes local allergic reactions of the skin due to continuous insulin injections. Accordingly, studies on natural materials are actively conducted as candidate materials for preventing, improving and treating diabetes, preventing diabetes including ginseng fruit extract (Korean Patent No. 2016-0000041) as a natural extract as an active ingredient, Improved or therapeutic compositions have been developed.

On the other hand, Aster koraiensis is a Korean native plant belonging to the chrysanthemum family, which is planted for ornamental purposes. It grows mainly in marshes. Its height is 50 ~ 60cm, and the leaves attached to the roots are lost when flowering. The leaves on the stem are shifted and lanceolate, with fine jagged teeth on the edges, and become smaller and lined up. Flowers bloom in June-October. Duhwa is light purple, 4-5cm in diameter, hanging on one end of stem and branch. Fruits are aquatic, bassos, ripen in November. Young sprouts are used as salads and herbs, and they are used for the treatment of seawater, phlegm, asthma, etc., but there have been no reports on the effects of diabetes mellitus according to hypoglycemia or glycemic control.

The present inventors conducted research to develop a prophylactic or therapeutic agent for diabetes using natural products. As a result, it was confirmed that bee anesthetist extract or fractions thereof can treat diabetes by exhibiting a hypoglycemic or modulating effect, thereby completing the present invention.

One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of diabetes mellitus comprising a bee odor an extract or fractions thereof as an active ingredient.

Another object of the present invention to provide a food composition for the prevention or improvement of diabetes mellitus including a bee herb extract or fractions thereof as an active ingredient.

Yet another object of the present invention is to provide a method for preventing or treating diabetes, comprising administering the composition to a subject.

Another object of the present invention is to provide a feed additive for the prevention or improvement of diabetes mellitus including a bee odor and an extract thereof as an active ingredient.

Yet another object of the present invention is to provide the use of the bee anesthetist extract or fractions thereof for the manufacture of a medicament for the prevention or treatment of diabetes.

Beetle anesthetic extract of the present invention promotes glucose absorption of adipocytes and thus lowers blood sugar, and thus can be usefully used for the prevention or treatment of diabetes. In addition, since the composition of the present invention is derived from natural products, it can be safely used without side effects in the prevention, treatment and improvement of diabetes.

Figure 1 is a graph showing the glucose uptake promoting effect of the hummingbird odor 30% ethanol extract (ASKO-30) and hot water extract (ASKO)), *** means p <0.001 compared to the negative control (CON) ( N = 15).

Figure 2 is a graph showing the glucose uptake promoting effect of the bee odor ethanol extract (KIOM83A) and its fractions, ** means that p <0.001 compared to the negative control (CON) (N = 15).

3 is a graph comparing the effect of promoting the absorption of glucose of the honey bee extract (ASKO, ASKO-30, ASKO-50, ASKO-80) and Wanghaeguk extract, *** is p <0.001, * compared to the negative control (CON) * Means p <0.01 compared to negative control (CON), ### means p <0.001 compared to Wanghaeguk extract (N = 8).

Figure 4 is a graph comparing the effect of promoting the absorption of glucose from the honey bee odor extract (ASKO-30, ASKO-50, ASKO-80) and sesame extract, *** is p <0.001, ** is negative compared to the negative control (CON) P <0.01 compared to the control (CON), ### means p <0.001 compared to the scented extract (N = 8).

One aspect of the present invention for achieving the above object provides a pharmaceutical composition for the prevention or treatment of diabetes mellitus comprising a bee odor extract or fractions thereof as an active ingredient.

Another aspect of the present invention for achieving the above object provides the use of the bee anesthetized extract or fractions thereof for the manufacture of a medicament for the prevention or treatment of diabetes.

In the present invention, the honey bee anesthetized extract by promoting glucose absorption of adipocytes by confirming the effect of lowering blood sugar, it was confirmed that the extract can be used as an active ingredient of diabetes treatment.

As used herein, the term "beetle odor" refers to a domestic native plant belonging to the chrysanthemum family having the scientific name of Aster koraiensis . Young sprouts are used as salads and herbs, and are used for the treatment of seawater, phlegm and asthma. However, the effect of bee anesthesia on the hypoglycemic or glycemic control effect of the diabetes is not known at all, and was first identified by the present inventors.

In the present invention, the roots, stems, leaves, fruits, flowers, etc. of the hummingbird can be used for the prevention or treatment of diabetes mellitus, more specifically, leaves and / or stems can be used, as long as it has a prevention or treatment effect of diabetes Is not particularly limited thereto.

The hummingbird odor can be purchased commercially, or may be used collected or cultivated in nature.

As used herein, the term "extract" refers to extracts obtained by extracting and processing the bee odor, diluents and concentrates of the extracts, dried products obtained by drying the extracts, modifiers and purified products of the extracts, and mixtures thereof. And extracts of all formulations that can be formed using extracts.

The method of extracting the hummingbird odor is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method, hot water extraction method, ultrasonic extraction method, filtration method, reflux extraction method, and the like, these may be performed alone or in combination of two or more methods.

In the present invention, the type of extraction solvent used to extract the bee odor is not particularly limited, and any solvent known in the art may be used.

Non-limiting examples of the extraction solvent may include water, alcohols having 1 to 4 carbon atoms or mixed solvents thereof, and these may be used alone or in combination of one or more thereof. Specifically, ethanol may be used, but is not limited thereto. The ethanol may be 0.1 to 99.99% (v / v), specifically 30 to 90% (v / v), and more specifically 80% (v / v) ethanol. May be, but is not limited to such.

In the present invention, the bee odorous extract may be a bee odorous 80% (v / v) ethanol extract or water extract.

As an example of preparing the bee odorless ethanol extract, after extracting the bee odorous leaf alone, or the bee odorous leaf and stem pulverized with ethanol, it can be concentrated under reduced pressure and lyophilized to prepare a bee odor ethanol extract.

As used herein, the term "fraction" refers to the result obtained by performing fractionation to separate a specific component or a specific group of components from a mixture comprising several various components.

The fractionation method of obtaining the fraction in the present invention is not particularly limited, and may be performed according to a method commonly used in the art. Non-limiting examples of the fractionation method include a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed through an ultrafiltration membrane having a constant molecular weight cut-off value, and various chromatography (size, charge, hydrophobicity). Or chromatographed fractions), and combinations thereof, which are prepared for separation according to affinity. Specifically, a method of obtaining a fraction from the extract by treating a predetermined solvent with an extract obtained by extracting the bee sting odor of the present invention.

The kind of the fractionation solvent used to obtain the fraction in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvents include polar solvents such as water and alcohols having 1 to 4 carbon atoms; Nonpolar solvents such as hexane, ethyl acetate, chloroform and dichloromethane; Or a mixed solvent thereof. These may be used alone or in combination of one or more thereof, but is not limited thereto. Specifically, hexane, ethyl acetate, butanol or water may be used alone or in combination of one or more thereof, but is not limited thereto. It doesn't happen.

In addition, the extract or fraction may be prepared and used in the form of a dry powder after extraction, but is not limited thereto.

As used herein, the term "diabetes" refers to metabolic diseases such as insufficient insulin secretion or normal function. It is characterized by high blood glucose, which increases the concentration of glucose in the blood, and causes various symptoms and signs due to high blood sugar and releases glucose into the urine.

As used herein, the term "prevention" refers to any action that inhibits or delays diabetes by administration of a pharmaceutical composition of the present invention comprising a cane odor extract or a fraction thereof.

As used herein, the term "treatment" refers to any action in which diabetes is improved or beneficially altered by administration of the pharmaceutical composition.

In a specific embodiment of the present invention, as a result of the treatment of bee anesthetic extract on adipocytes, it was confirmed that glucose uptake of adipocytes was increased in a concentration-dependent manner (FIGS. 1 and 2). This suggests that bee anesthetized extract of the present invention promotes glucose absorption of adipocytes and thus regulates blood sugar normally, so that it can be used as an active ingredient in diabetes treatment.

In addition, in another embodiment of the present invention, the results of comparing the efficacy of the bee-flavored odor with the same chrysanthemum Wanghae-guk or chamshu, proved that the effect of the bee-flavored odor is significantly superior to the Wanghae-guk and chamcheuk (Fig. 3; compared to Wanghaeguk extract ### P <0.001 / FIG. 4; compared to scent extract ### P <0.001).

The pharmaceutical composition of the present invention may include 0.0001 to 80% by weight of the bee odor extract relative to the weight of the total composition, specifically may include 0.01 to 40% by weight, but is not limited thereto.

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent commonly used in the manufacture of the pharmaceutical composition, the carrier comprising a non-naturally occuring carrier. can do.

The pharmaceutical composition may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. .

"Pharmaceutically acceptable" means to exhibit properties that are not toxic to cells or humans exposed to the composition.

Specifically, the type of the carrier is not particularly limited, and any carrier can be used as long as it is a conventionally used and pharmaceutically acceptable carrier in the art. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and the like. These may be used alone or in combination of two or more thereof. In addition, if necessary, other conventional additives such as antioxidants, buffers and / or bacteriostatic agents may be added and used, and diluents, dispersants, surfactants, binders, lubricants, and the like may be additionally added to provide a solution such as an aqueous solution, a suspension, an emulsion, or the like. It may be formulated into a use formulation, pills, capsules, granules or tablets.

The mode of administration of the pharmaceutical composition for preventing or treating diabetes mellitus according to the present invention is not particularly limited, and may be in accordance with methods commonly used in the art. As a non-limiting example of the mode of administration, the composition may be administered by oral or parenteral administration. In addition, the composition for preventing, ameliorating or treating diabetes of the present invention may be prepared in various formulations according to the desired mode of administration.

Another aspect provides a method of preventing or treating diabetes comprising administering the pharmaceutical composition to a subject.

At this time, the definition of diabetes, prevention and treatment is as described above.

As used herein, the term "administration" means the introduction of certain substances into an individual in a suitable manner.

As used herein, the term "individual" means all animals, such as rats, mice, and livestock, including humans who may or may have diabetes. As a specific example, it may be a mammal including a human.

More specifically, the "individual" may include a companion animal. The companion animal refers to an animal that lives together with a human, and includes, but is not limited to, a mammal such as a dog, a cat, a hamster, and a guinea pig, a bird such as a parrot, a canary, and the like.

Specifically, the method for preventing or treating diabetes of the present invention may include administering to the individual a pharmaceutical composition comprising a bee-smelling extract or a fraction thereof in a pharmaceutically effective amount.

The "pharmaceutically effective amount" means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to the medical treatment and not causing side effects, and the effective dose level refers to the sex, age, weight, Well-known in the field and other medical fields, including health status, type of disease, severity, drug activity, drug sensitivity, method of administration, time of administration, route of administration, and rate of release, duration of treatment, combination or drug used simultaneously Depending on the factor, it can be readily determined by one skilled in the art.

The pharmaceutical composition of the present invention may be administered at 0.0001 to 500 mg / kg body weight per day, more specifically at 0.01 to 500 mg / kg body weight, based on solids. Dosing may include administering the recommended dose once a day or in several divided doses.

The pharmaceutical compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. It may also be single or multiple doses. Taking all of these factors into consideration it is important to administer an amount that will achieve the maximum effect in a minimum amount without causing side effects, which can be readily determined by one skilled in the art.

In the method for preventing or treating diabetes of the present invention, the route of administration and the mode of administration of the composition are not particularly limited, and any route of administration and administration can be achieved as long as the composition comprising the composition can reach the desired site. You can follow the way. Specifically, the composition may be administered through various routes, oral or parenteral, and non-limiting examples of the route of administration include oral, rectal, topical, intravenous, intraperitoneal, intramuscular, intraarterial, transdermal, nasal What is administered through intralateral or inhalation etc. are mentioned.

Yet another aspect of the present invention provides a food composition for preventing or improving diabetes comprising a bee anesthetist extract or a fraction thereof as an active ingredient.

At this time, the definition of bee odor, extract, fraction, diabetes and prevention is as described above.

As used herein, the term " improvement " means any action that at least reduces the parameters associated with the condition to be treated with the administration of the compositions of the present invention, eg the extent of symptoms.

The term "food" of the present invention, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages , Vitamin complexes, nutraceuticals and health foods, and includes all foods in the usual sense.

Since the food composition of the present invention can be consumed on a daily basis, a high diabetic improvement effect can be expected, and thus can be very useful for health promotion purposes.

The functional food (functional food) is the same term as a food for special health use (Food for special health use, FoSHU), in addition to the nutritional supply, the processed food, so that the bioregulatory function appears efficiently, high medical effect Means food. Here, the term 'function (sex)' refers to obtaining a useful effect for health purposes such as nutrient control or physiological action on the structure and function of the human body. The food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding the raw materials and ingredients commonly added in the art. In addition, the formulation of the food may be prepared without limitation as long as the formulation is recognized as a food. Food composition of the present invention can be prepared in a variety of dosage forms, unlike the general medicine has the advantage that there is no side effect that can occur when taking a long-term use of the drug as a natural product, and because the portability is excellent, The food of the present invention can be taken as an adjuvant for enhancing the diabetic improvement effect.

The health food refers to foods having active health maintenance or promotion effect as compared to general foods, and the health supplement food refers to foods for health supplementation purposes. In some cases, the terms nutraceutical, health food, dietary supplement are used.

Specifically, the health functional food is a food prepared by adding the compound of the present invention to food materials such as beverages, teas, spices, gums, confections, or the like, encapsulated, powdered, suspensions, etc. Means to bring effect, but unlike the general medicine has the advantage that there is no side effect that can occur when taking long-term use of the drug as a raw material.

The food composition may further include a physiologically acceptable carrier, and the type of carrier is not particularly limited and may be any carrier that is commonly used in the art.

In addition, the food composition may include additional ingredients that are commonly used in food compositions to improve the smell, taste, time and the like. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, and the like. In addition, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu) and chromium (Cr); And amino acids such as lysine, tryptophan, cysteine, valine and the like.

In addition, the food composition is a preservative (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetic acid, etc.), fungicides (bleaching powder and highly bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisol (BHA), butylhydride) Oxytoluene (BHT), etc.), colorant (such as tar pigment), colorant (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasoning (MSG glutamate, etc.), sweetener (ducin, cyclate, saccharin Foods such as sodium, etc.), fragrances (vanillin, lactones, etc.), swelling agents (alum, potassium D-tartrate, etc.), reinforcing agents, emulsifiers, thickeners (foils), coatings, gum herbicides, foam inhibitors, solvents, modifiers, etc. It may include food additives. The additive may be selected according to the type of food and used in an appropriate amount.

An example of the food composition of the present invention may be used as a health beverage composition, in which case it may contain various flavors or natural carbohydrates and the like as additional ingredients, such as a general beverage. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin, cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumartin, stevia extract; Synthetic sweeteners such as saccharin and aspartame; The ratio of the natural carbohydrate may be generally about 0.001 to 0.4 g, specifically about 0.002 to 0.3 g per 100 mL of the health beverage composition of the present invention.

In addition to the above, the health beverage composition includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, Alcohol or carbonation agent and the like. Others may contain fruit flesh for the production of natural fruit juices, fruit juice drinks, or vegetable drinks. These components can be used independently or in combination. Although the ratio of such an additive is not critical, it is generally selected in the range of 0.001 to 0.10 parts by weight per 100 parts by weight of the health beverage composition of the present invention.

Another aspect of the present invention provides a feed composition for the prevention or improvement of diabetes mellitus including a bee herb extract or fractions thereof as an active ingredient.

The bee odor, extract, fraction, prevention and improvement are as described above.

The composition comprising the bee anesthetized extract of the present invention or a fraction thereof may be used for preventing or treating diabetes onset in a subject other than humans, such as livestock or a companion animal, and may be utilized as a functional feed additive or a feed composition.

The content of the bee-flavored extract or fractions thereof in the feed composition according to the present invention can be appropriately adjusted according to the type and age of the livestock, the type of application, the desired effect, and the like, for example, 1 to 99% by weight, preferably 10 to 90% by weight. %, More preferably 20 to 80% by weight, but is not limited thereto.

Feed composition of the present invention for administration, in addition to the bee anesthetic extract or fractions thereof, organic acids such as citric acid, fumaric acid, adipic acid, lactic acid; Phosphates such as potassium phosphate, sodium phosphate and polymerized phosphate; One or more of natural antioxidants such as polyphenol, catechin, tocopherol, vitamin C, green tea extract, chitosan and tannic acid can be mixed and used. In addition, diluents, dispersants, surfactants, binders or lubricants may be additionally added to formulate injectable formulations, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

In addition, the feed composition of the present invention is a supplementary ingredient such as amino acids, inorganic salts, vitamins, antioxidants, antifungal agents, antibacterial agents, and vegetable protein feed such as milled or crushed wheat, barley, corn, blood meal, meat meal, fish meal as auxiliary ingredients. In addition to the main components such as animal protein feed, animal fat and vegetable fat, such as can be used with nutritional supplements, growth promoters, digestive absorption accelerators, disease prevention agents.

When the feed composition of the present invention is used as a feed additive, the feed composition may be added as it is or used together with other ingredients, and may be suitably used according to a conventional method. Dosage forms of the feed composition may be prepared in immediate release or sustained release formulation in combination with a nontoxic pharmaceutically acceptable carrier. Such edible carriers may be corn starch, lactose, sucrose, propylene glycol. In the case of a solid carrier, it may be a dosage form such as a tablet, a powder, a torokee agent, or the like, and in the case of a liquid carrier, it may be a dosage form of a syrup, a liquid suspension, an emulsion, or a solution. In addition, the dosage may contain preservatives, lubricants, solution promoters, stabilizers, and may also contain other inflammatory disease ameliorating agents and substances useful for virus prevention.

The feed composition of the present invention is applicable to a number of animal diets, ie feeds, including mammals, poultry, fish and shellfish. Commercially important mammals such as pigs, cows and goats, zoo animals such as elephants and camels, and livestock such as dogs and cats. Commercially important poultry includes chickens, ducks, geese and the like, and may include commercially raised fish and crustaceans such as trout and shrimp.

The feed composition according to the present invention may be mixed in a livestock feed in an amount of about 10 to 500 g, preferably 10 to 100 g per kg, on a dry weight basis, and then thoroughly mixed and then fed into a mesh or further processing. Palletization, expansion, extrusion through the process is preferred.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example  1. Preparation of Beetle Odor Boiled Extract

After cutting the leaves and stems of the hummingbird odor, hot water was extracted at 100 ° C with 100% water. The extract was filtered, concentrated and lyophilized in a dark condition to obtain a bee-flavored water extract (ASKO).

Example  2. Preparation of Beetle-odorous Water-soluble Ethanol Extracts

After cutting the leaves and stems of the bee-flavored odor, they were extracted with 30% ethanol, 50% ethanol, and 80% ethanol at room temperature, filtered, concentrated, dried, and then dried by the bee-flavored water-soluble ethanol extract (ASKO-30, ASKO-50, ASKO- 80) was obtained respectively.

Example  3. Beetle anesthetic extract and Fraction  Produce

After cutting the leaves and stems of the hummingbird odor, they were extracted at room temperature in 80% (v / v) ethanol. The extract was filtered, concentrated and dried in the dark, to obtain a bee-flavored alcohol extract (KIOM83A). Thereafter, the KIOM83A was separated into hexane, ethyl acetate, butanol, and water, and the hexane layer (KIOM-83H), the ethyl acetate layer (KIOM-83EA), the butanol layer (KIOM-83Bu), and the water layer (KIOM-) of the bee odor extract were extracted. 83W) was obtained.

Example  4. Booze  Preparation of Extract

Aster scaber Thunb. (Wing leaf squirrel) was purchased and purchased in July 2016 from Sanju Botanical Garden of Chungju, Chungju, Korea and stored in a low temperature room for sample storage at Korea Institute of Oriental Medicine (Daejeon, Korea).

As a specific sesame extract manufacturing process, 5 g of sesame seed was added to 50 mL of primary distilled water corresponding to 10 times the weight to prepare hot water extract, filtration, and concentrated freeze-dried extract at 100 ° C.

Example  5. Wanghae Guk  Preparation of Extract

Wang Hae- guk ( Aster spathulifolius var. Pharai ) was purchased and used in Chungcheongju National Petrochemical Botanical Garden in July 2016, and was stored in the low temperature room for sample storage of Korea Institute of Oriental Medicine (Daejeon, Korea). Wanghaeguk is a plant belonging to Aster spathulifolius .

As a specific Wanghaeguk extract manufacturing process, 5g of Wanghaeguk was added to 50 mL of primary distilled water corresponding to 10 times the weight, and hot water extraction, filtration and concentrated freeze-dried extracts were prepared at 100 ° C.

Example  6. Of the preadipocyte  culture

Adipose precursor cells 3T3-L1 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), and all the media used for cell culture were purchased from WELGENE. The progenitor cells were cultured at 37 ° C. and 5% CO ₂ using Dulbecco's modified Eagle's medium (DMEM) containing 1% P / S (penicillin-streptomycin) and 10% FBS. Cultivation was carried out at 3 days intervals, when cells were grown to about 80% at the bottom of a 100 mm plate, washed with PBS (phosphate buffered saline), 0.25% trypsin-EDTA solution was added at 37 ℃, 5% CO ₂ conditions 3 Cells were detached and passaged and maintained by treatment for a minute.

Example  7. Beetle anesthetic extracts ( ASKO , ASKO -30, ASKO -50, ASKO -80, KIOM83A , KIOM-83H, KIOM - 83EA , KIOM - 83Bu , KIOM -83W) hypoglycemic effect

To analyze the hypoglycemic activity of ASKO, ASKO-30, ASKO-50, ASKO-80, KIOM83A, KIOM-83H, KIOM-83EA, KIOM-83Bu, KIOM-83W It was.

First, adipocytes were prepared by differentiating 3T3-L1, an adipocyte. To differentiate 3T3-L1, an adipocyte, into adipocytes, the 3T3-L1 cells were dispensed in 96 well plates at 1 × 10 ⁴ cells / well and incubated for 5 days in a 5% CO ₂ incubator. The differentiated adipocytes were cultured in low concentration glucose medium without FBS, and were treated with ASKO, ASKO-30, ASKO-50, ASKO-80, KIOM83A, KIOM-83H, KIOM-83EA, KIOM-83Bu, KIOM-83W. Each concentration was treated and glucose uptake was analyzed using Glucose Uptake Cell-Based Assay Kit (Cayman Chemical Company, MI, USA). At this time, the result was measured in the fluorescence analyzer 485/535 nm.

As a result, as shown in Figures 1 and 2, ASKO, ASKO-30, KIOM83A, KIOM-83H, KIOM-83Bu was confirmed to increase the concentration of glucose uptake of adipocytes. Through the above results, it was found that ASKO, ASKO-30, KIOM83A, KIOM-83H, and KIOM-83Bu have excellent glycemic control activity to promote glucose absorption of adipocytes during blood sugar rise to maintain blood sugar normally.

Example  8. Beetle Anesthetic Extracts Wanghae Guk  Verification of extract and hypoglycemic

The blood glucose lowering effects of the bee-flavored extracts prepared in Examples 1, 2 and 5 and the extract of Wanghaeguk were compared. Specific hypoglycemic effect measurement is the same as in Example 7.

As a result, as can be seen in Figure 3, the honey bee extracts (ASKO, ASKO-30, ASKO-50, ASKO-80) It was confirmed that the glucose absorption efficiency of fat cells was significantly better than that of Wanghaeguk extract. That is, when compared with the negative control group (CON), Wanghaeguk extract did not show a significant effect, but all of the bee-flavored extracts were very good (** P <0.01, *** P <0.001).

In addition, as a result of comparing the hypoglycemic effect of the Wanghaeguk and the bee-flavored extracts, it can be seen that the honeysuckle extracts exhibited a better blood sugar-lowering effect than the extract of Wanghaeguk (### P <0.001).

Example  9. Beetle Anesthetic Extracts Booze  Verification of extract and hypoglycemic

The blood sugar lowering effects of the bee-flavored extracts prepared in Examples 1, 2, and 4 and sesame extract were compared. Specific hypoglycemic effect measurement is the same as in Example 7.

As a result, as shown in Figure 4, the bee sting odor extract (ASKO-30, ASKO-50, ASKO-80) It was confirmed that glucose absorption efficiency of adipocytes was significantly better than that of scent extract. In other words, compared to the negative control group (CON), the extract of the scavenging did not show a significant effect, but all of the bee-flavored extracts were very good (** P <0.01, *** P <0.001).

In addition, as a result of comparing the blood glucose lowering effect of the scavenging extract and the bee sting odor extracts, it can be seen that the bee sting odor extracts show a better blood sugar lowering effect than the scavenging extract (### P <0.001).

Through the results of Examples 7 and 8, the bee anesthetized extract such as ASKO, ASKO-30, KIOM83A, KIOM-83H, KIOM-83Bu promotes glucose absorption of adipocytes during blood sugar rise, thereby maintaining excellent blood sugar. Since it has modulating activity, it can be seen that it can be usefully used as a composition for preventing and treating diabetes. In addition, it can be seen that the effect is significantly superior to the plants of the same genus Hwanghaeguk and chamcheuk.

Claims (9)

Pharmaceutical composition for the prevention or treatment of diabetes mellitus including a bee odor extract or fractions thereof as an active ingredient. The pharmaceutical composition of claim 1, wherein the beetail odor extract is prepared by extraction with a solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof. The pharmaceutical composition of claim 1, wherein the prevention or treatment of diabetes is by glycemic control. The method according to claim 1, wherein the fraction comprises a bee odor extract, water, alcohol having 1 to 4 carbon atoms, hexane (Hexane), ethyl acetate (Ethyl acetate), chloroform (Chloroform), dichloromethane and mixed solvents thereof It is prepared by fractionation with a solvent selected from the group consisting of, pharmaceutical compositions. The pharmaceutical composition of claim 1, wherein the bee odor extract is 0.5 to 80% (v / v) ethanol extract. A method for the prevention or treatment of diabetes comprising administering to a subject a pharmaceutical composition according to any one of claims 1 to 5. Food composition for glycemic control, prevention or improvement of diabetes mellitus comprising bee odor an extract or fractions thereof as an active ingredient. A pharmaceutical composition or a food composition or feed additive for controlling blood sugar, preventing or improving diabetes, comprising a bee anesthetist extract or a fraction thereof administered to an individual except a human as an active ingredient. Use of said beetail extract or fractions thereof for the manufacture of a medicament for the prevention or treatment of diabetes.

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