KR20160066643A - Anti-diabetic composition containing the extract of actinidia arguta leaves or fractions thereof - Google Patents

Abstract

The present invention relates to an anti-diabetic composition comprising an Actinidia argute extract or a fraction thereof as an active ingredient and, more specifically, to a pharmaceutical composition for preventing or treating diabetes, comprising an Actinidia argute extract or a fraction thereof as an active ingredient, which is extracted to be soluble by adding an organic solvent, and to a health functional food composition for preventing or alleviating diabetes. The anti-diabetic composition of the present invention comprising food grade young leaves of Actinidia argute as a main ingredient from Actinidia argute, which is a natural plant, has low risk of side effects, has excellent inhibitory activities of α-glucosidase, and has properties of preventing, alleviating, or treating diabetes.

Description

TECHNICAL FIELD The present invention relates to an antidiabetic composition comprising an extract of Quercus grandiflorum or extract thereof as an active ingredient.

The present invention relates to a composition for an anti-diabetic comprising an extract of Dulaya japonica or a fraction thereof as an active ingredient.

Diabetes is a disease caused by abnormal production, secretion or utilization of insulin, with high blood sugar levels due to the metabolic disorder of carbohydrates, and excretion of glucose into the urine. Most of the food we ingest is broken down by the digestive juices into sugar called glucose, which is transported to the blood vessels and enters our body cells for growth and energy. When glucose enters the cell, insulin is present and insulin is produced in the pancreas behind the stomach. When we eat food, the pancreas automatically produces an adequate amount of insulin to move the glucose from the blood to the cells. In diabetics, the pancreas produces little insulin or the cells do not respond to insulin, And it is discharged into the urine as it is left in the blood, and the blood contains many glucose, but the body can not use it, and various abnormal symptoms appear.

The ultimate goal in the treatment of diabetes is to prevent complications by managing to maintain normal blood glucose levels. There are three main types of health care for diabetics: diet, exercise, and medication. Diet therapy is based on low-salt, low-fat, and moderate abstinence. Exercise therapy can reduce insulin resistance, reduce blood sugar, and facilitate the metabolism of lipids to prevent atherosclerosis by managing the health of patients according to the condition can be. Drug therapy is largely an oral hypoglycemic agent and insulin therapy. Regular blood glucose testing should be done while changing the dosage, drug, and side effects and complications.

However, in the case of the drugs currently used for the treatment of diabetes, there is a problem that the continuous drug effect is insufficient and various side effects are caused in the body, and in particular, many therapeutic agents cause renal toxicity and hepatotoxicity.

Therefore, it is urgent to develop a natural substance having excellent anti-diabetic pharmacological effect while reducing such side effects.

Therefore, the present inventors have found that the natural substance Actinidia arguta ) showed excellent antidiabetic activity, and thus completed the present invention.

It is therefore an object of the present invention to provide a process for the production of Actinidia The present invention also provides a pharmaceutical composition for preventing, ameliorating or treating diabetes, and a health functional food composition comprising the extract or fraction thereof as an active ingredient.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

In order to achieve the object of the present invention as described above, the present invention is the order of Actinidia (Actinidia The present invention also provides a pharmaceutical composition for preventing or treating diabetes mellitus comprising, as an active ingredient, an extract or a fraction thereof.

The present invention also relates to a method for the treatment of Actinidia The present invention also provides a health functional food composition for preventing or ameliorating diabetes, comprising an extract of arguta or a fraction thereof as an active ingredient.

In one embodiment of the present invention, the composition may have an? -Glucosidase inhibitory activity.

In another embodiment of the present invention, the extract may be a solvent extract obtained by the extraction with water or an organic solvent.

In another embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, and methylene chloride.

In another embodiment of the present invention, the ethanol may be 50% or 100% ethanol.

In another embodiment of the present invention, the pharmaceutical composition may have a formulation selected from the group consisting of ointments, gels, creams, patches and sprays.

In another embodiment of the present invention, the health functional food composition may be in the form of a powder, a granule, a tablet, a capsule or a drink.

The present invention also provides a method of treating diabetes comprising administering an extract of Actinidia arguta or a fraction thereof to a subject in need of treatment for diabetes.

In one embodiment of the present invention, the administration may be by oral administration, intravenous injection, intraperitoneal injection, intramuscular injection, intraarterial injection or subcutaneous injection.

In another embodiment of the present invention, the subject may be a mammal, including a human.

The composition for anti-diabetes according to the present invention is a natural substance, and particularly, it contains the juvenile leaf of Dulaya, which is an edible part, as a main ingredient, so that it has a low α-glucosidase inhibitory activity while preventing the risk of side effects, Or treatment.

FIG. 1 is a schematic diagram showing a process for identifying a fraction showing an α-glucosidase inhibitory activity by partially purifying a 50% ethanol extract of Dulalae according to the present invention.
FIG. 2 shows HPLC chromatograms of the extract of Quercus granda and its fractions analyzed by reverse phase chromatography.
Fig. 3 shows the inhibitory activity of? -Glucosidase in extracts of Quercus mongolica var. Elder extracts obtained by using various solvents in comparison with a positive control acarbose.
Figure 4 compares the alpha -glucosidase inhibitory activity of partially purified purple nettle fractions using liquid chromatography.

The present invention relates to a process for the production of Actinidia The present invention also provides a pharmaceutical composition for preventing, ameliorating or treating diabetes, and a health functional food composition, which comprises an extract of arguta or a fraction thereof as an active ingredient.

In the present invention, as a natural substance having an excellent antidiabetic effect, it was used in the order of Dalaea, which is a deciduous tree which grows in mountains of each place in Korea. Leaves are broad ovate and oval, with sharp and sharp sawtooth on edge, flower is white to male, dioecious, about 3 ~ 10 are pushed downward. Especially, young leaves, such as purple seeds, are widely used for edible purposes. In recent years, there has been a growing interest in securing safe food, as the interest in health and longevity has increased. Therefore, studies have been focused on finding active substances of natural products that have useful physiological activities and have no side effects. There is a safer advantage.

Therefore, the fruit extract of the present invention or its fractions according to the present invention have excellent antidiabetic activity and can be safely used for pharmaceutical and food applications as an edible part.

In the present invention, the net extract of Dulone may be used as a solvent extract, as long as it is a method for obtaining an extract from a natural source known in the art, preferably by adding water or an organic solvent and then extracting it by a soluble method .

The process for obtaining an extract having alpha -glucosidase inhibitory activity from the order of the present invention will be described in more detail as follows.

First, the plant extracts are dried or lyophilized by low-temperature drying to obtain a solution of the extract of the genus Xanthais by a solvent extraction method known in the art using about 10 to 300 times, preferably about 100 to 300 times, .

In one embodiment of the present invention, the plant was dried and then extracted with water, 50% ethanol or 100% ethanol at room temperature for 12 hours. The extracted extracts were then prepared by centrifugation, filtration and concentration. The extracts were lyophilized and stored at -80 ° C.

The dried fruit can be used before the drying or before extraction with a solvent, and the pulverization can be carried out manually or by using a mechanical device.

In the present invention, the term " freeze-drying " refers to a method of freezing an aqueous solution or a material containing a large amount of water and decompressing to sublimate the ice to remove moisture to obtain a dried material. That is, the sample is frozen and the frozen ice is sublimated at a lower temperature and pressure (triple point, 0.01 ° C., 0.6113 kPa) to obtain a dried product. The freeze-drier usable for freeze-drying employed in the present invention is not particularly limited as long as it is a drying plate constituted by a heating plate for supplying the sublimation heat for lyophilization, a condenser for condensing water vapor generated during drying to ice, a vacuum chamber and a vacuum pump Do not.

The process of extracting water or an organic solvent by adding water or an organic solvent may be carried out by stirring or standing. The solvent extraction method includes, but is not limited to, hot water extraction, cold extraction, Etc. can be extracted once or repeatedly several times. The organic solvent that can be added at this time is not limited thereto but may be methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, , And most preferably 50% ethanol.

The present inventors measured alpha -glucosidase inhibitory activity to induce blood glucose elevation in order to investigate whether or not the antler extract of the present invention has anti-diabetic activity. As a result, all of the extracts of alpha-glucosidase were found to have activity In particular, it was found that, in the case of the extract of P. multocida using 50% ethanol as a solvent, the α-glucosidase inhibitory activity was as excellent as the positive control group acarbose at the concentration of 0.23 mg / mL (See Example 2).

In addition, the present inventors fractionated the extract through partial purification of the extract of the genus Darla, and measured the activity of the α-glucosidase of the fractions. As a result, it was found that the fraction (fraction 3-1) had excellent α-glucosidase Inhibitory activity (see Example 3).

Therefore, the inventors of the present invention have confirmed that the extract of Dulone or its fractions has anti-diabetic activity by inhibiting the activity of? -Glucosidase.

Therefore, the present invention can provide an antidiabetic composition comprising as an active ingredient a pure extract of Dulone or a fraction thereof, and the composition can be used as a pharmaceutical composition for the prevention and treatment of diabetes, ≪ RTI ID = 0.0 > and / or < / RTI >

In the pharmaceutical composition according to the present invention, the net fruit extract or fraction thereof may be contained in an amount of 0.1 to 50% by weight based on the total weight of the composition, and the diabetes may be type 2 diabetes mellitus.

The pharmaceutical composition according to the present invention may comprise a pharmacologically effective amount of a nettle extract or fraction thereof, alone or in combination with one or more pharmaceutically acceptable carriers, excipients or diluents. As used herein, the term " pharmaceutically effective amount " refers to an amount sufficient to prevent or treat diabetic symptoms.

The pharmacologically effective amount of the extract of Dulone or its fractions according to the present invention is 0.5 to 100 mg / day / kg body weight, preferably 0.5 to 5 mg / day / kg body weight. However, the pharmaceutically effective amount may be appropriately changed depending on the degree of diabetic symptoms, the age, body weight, health condition, sex, administration route and treatment period of the patient.

The term " pharmaceutically acceptable " as used herein refers to a composition that is physiologically acceptable and does not normally cause an allergic reaction such as gastrointestinal disorder, dizziness, or the like when administered to humans. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

In addition, the composition of the present invention can be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to a subject in need of diabetes treatment, including human . Formulations may be powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatine capsules, sterile injectable solutions, sterile powders.

The pharmaceutical composition according to the present invention may be administered through various routes including oral, transdermal, subcutaneous, intravenous or muscular, and the dose of the active ingredient may be appropriately determined depending on the administration route, age, sex, And can be appropriately selected according to various factors. In addition, it can be administered in combination with a known compound having an effect of preventing or ameliorating diabetic symptoms.

Furthermore, the fruit extract of the present invention or its fractions according to the present invention not only provides an effect of alleviating the symptoms of diabetes through an excellent antidiabetic effect, but also is a natural plant which can be used as a raw material for the extract of the present invention, It has very stable characteristics in the body and has the advantage that it can be used safely even when taken for a long time without toxicity and side effects.

Therefore, the present invention can be used as a food composition for preventing or ameliorating the symptoms of diabetes by adding to the food for the purpose of preventing or improving diabetes.

Therefore, the present invention can be applied to the production of foodstuffs, additives, food additives, functional foods or beverages which are effective for prevention and improvement of diabetes mellitus according to the present invention.

The term " food " as used herein means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be directly eaten through a certain degree of processing, , Food additives, functional foods and beverages.

Examples of the food to which the composition containing the extract of the genus Dianthus according to the present invention or the fraction thereof as the active ingredient can be added include various foods, beverages, gums, tea, vitamin complex, and functional foods. In addition, the above foods may include special nutritional foods (eg, crude oil, spirit, baby food, etc.), meat products, fish products, tofu, jelly, noodles (eg, ramen, noodles, etc.) (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce, etc.), sauces, confectionery (eg snacks), candy, chocolate, gums, ice cream, milk products (E.g., various kinds of kimchi, pickles, etc.), beverages (e.g. fruit drinks, vegetable beverages, beverages, fermented beverages, etc.) The additive can be produced by a conventional production method.

The above-mentioned " functional food " refers to a food group which is imparted with added value to function and express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, Refers to a food prepared by processing a body so as to sufficiently express the body's control function with respect to the body, and preferably, it may be a health functional food. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

In the present invention, the term " beverage " means a general term for drinking or enjoying a taste, and includes a functional beverage. The beverage includes, as an essential ingredient, a composition comprising a nettle extract or its fraction for preventing or ameliorating the symptoms of diabetes, and there is no particular limitation on other ingredients, and various flavors or natural carbohydrates, such as ordinary beverages, And may be contained as an additional component.

In addition to the above, the food containing the composition of the present invention may be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers (cheese, chocolate, etc.) An organic acid, a protective colloid thickener, a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in a carbonated beverage, and the like, Can be used in combination.

In the food containing the food composition according to the present invention, the amount of the composition comprising the net fruit extract or fraction thereof as an active ingredient may be 0.001% by weight to 90% by weight, 0.1% by weight to 40% by weight, and in case of beverage, it may be contained in a proportion of 0.001 g to 2 g, preferably 0.01 g to 0.1 g, based on 100 ml, and in the case of long- However, since the active ingredient has no problem in terms of safety, it can be used in an amount of more than the above-mentioned range, so it is not limited to the above range.

Therefore, the present invention relates to a method for producing < RTI ID = The composition of the present invention may be in the form of a powder, a granule, a tablet, a capsule or a drink. The composition may be in the form of a powder, a granule, a tablet, a capsule or a drink. have.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

Dalae  Preparation of extract

Actinidia arguta ) was extracted with 200 mL of solvent. The mixture was centrifuged (Hanil combi-514R, Korea) at 7,000 g for 5 minutes, and then washed with water (distilled water), 50% ethanol or 100% And only the supernatant (solution) was taken. The supernatant was then filtered through a filter (Whatman No. 1 paper, GE Healthcare, UK), and the filtrate was concentrated using a rotary evaporator (Eyela, Japan) The extracts were stored at -80 ℃ until freeze-drying.

Dalae  The α- Glucosidase (? glucosidase ) Inhibition activity measurement

As in Example 1, lyophilized pure extracts were fractionated with three solvents and dissolved in distilled water to a concentration of 0.23 and 0.92 mg / mL, and the extracts from Watanabe et al. (1997) method was modified to evaluate the? -Glucosidase inhibitory activity in vitro. More specifically, 5 mM of yeast α-glucosidase 0.7 unit / mL and p-nitrophenyl-α-D-glucopyranoside (Sigma Co., USA) were dissolved in potassium phosphate buffer (50 mM, pH 6.8) ) And substrate solution. After mixing 50 μL of the enzyme solution and 10 μL of the diluted nettle extract, the mixture was pre-incubated at 25 ° C for 5 minutes. Subsequently, 50 μL of the substrate solution was added, reacted again for 5 minutes, and the absorbance was measured at 405 nm using a microplate reader (SpectraMax 190, Molecular Devices, USA). The inhibitory activity was calculated as a percentage of the blank control.

As a result, among the extracts of three solvents (water, 50% ethanol and 100% ethanol), the highest inhibitory activity against α-glucosidase was shown at 61.6 ± 5.51% at a concentration of 0.92 mg / mL of 50% The water extracts showed 9.5 ± 1.90% and the 100% ethanol extract had 17.97 ± 3.16% inhibitory activity, and 50% ethanol was the most effective extraction solvent.

In addition, as shown in Fig. 3, the inhibitory activity of the 50% ethanol extract was 32.6 ± 5.77% at 0.23 mg / mL, and was similar to that of the positive control group, Acabose (30.0 ± 1.10%).

This result implies that the extract of Dulaya extract obtained by using 50% ethanol has a remarkable effect as a diabetic drug on the market.

Using preparative liquid chromatography Dalae  Partial purification of the extract

As shown in the schematic diagram of FIG. 1, the 50% ethanol extract of Dalaea was confirmed to exhibit the most excellent inhibitory activity through Example 2, and was partially purified by an in vitro enzyme analysis method. For this, fractions were separated using preparative liquid chromatography (LC-Forte / R, YMC, Japan) with a double plunger pump and a UV detector (200-400 nm). The fractionation process was carried out by reverse phase liquid chromatography (YMC-DispoPack AT C18, 50 μm; C18 column, 20 μm, 15 × 250 mm, YMC, Japan) using acetonitrile with different gradient concentrations.

The 50% ethanol extract suspended in water by reversed phase liquid chromatography was divided into fractions of 1 to 4 and the inhibitory activity of? -Glucosidase was measured. As shown in FIG. 4, fraction (1) at a concentration of 0.09 mg / The activity of inhibiting the? -Glucosidase activity of the fractions 2 to 4 was 10.6 ± 4.1%, 80.8 ± 3.4%, 29.7 ± 2.9% and 16.6 ± 0.7%, respectively, and fraction 2 as the active fraction was further purified. That is, fraction 2 was again divided into 2-1 to 2-4 fractions using 32-50% acetonitrile gradient solvent, and α-glucosidase inhibitory activity was measured. As a result, the fractions were 4.4 ± 2.1%, 59.1 ± (23-28% of acetonitrile elution) and 2-3 (28-37%) of the active fraction, respectively, as the? -Glucosidase inhibitory activity was 3.9%, 63.2 ± 1.0% and 26.5 ± 4.1% % Acetonitrile) were mixed and further purified to 1 to 5 subfractions. As a result of measuring α-glucosidase inhibitory activity of the small fraction, the small fraction 3 showed the most excellent inhibitory activity as 58.0 ± 3.1%, so that the small fraction 3 was again divided into 3-1 to 3-4 small fractions . Glucosidase inhibitory activities of the 3-1 to 3-4 subfractions were confirmed to be 82.5 ± 4.6%, 73.9 ± 5.9%, 34.3 ± 9.0% and 0.1 ± 1.7%, respectively, and the final α-glucosidase inhibitory activity The 3-1 small fractions were collected as active fractions.

As shown in FIG. 2, the number of peaks was decreased and the intensity was increased according to the purification step. As a result, the final fractions (3-1 small fractions ) Showed a relatively high intensity at 5 to 6 minutes.

From the above results, it can be seen that the 3-1 small fraction is a fraction containing an active ingredient capable of obtaining an anti-diabetic effect.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

Claims (8)

Actinidia The present invention relates to a pharmaceutical composition for preventing or treating diabetes mellitus comprising an extract or fraction thereof as an active ingredient.
Actinidia arguta ) extract or a fraction thereof as an active ingredient for preventing or ameliorating diabetes.
3. The method according to claim 1 or 2,
Wherein the composition has an? -Glucosidase inhibitory activity.
3. The method according to claim 1 or 2,
Wherein the extract is a solvent extract obtained by extracting with water or an organic solvent.
5. The method of claim 4,
Wherein the organic solvent is selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, and methylene chloride.
6. The method of claim 5,
Wherein the ethanol is 50% or 100% ethanol.
The method according to claim 1,
Wherein the composition has a formulation selected from the group consisting of ointments, gels, creams, patches, and sprays.
3. The method of claim 2,
Wherein the composition is in the form of a powder, a granule, a tablet, a capsule or a drink.

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