KR102266271B1 - Composition for preventing or treating diabetes containing extract of Ixeris strigosa - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of diabetes, containing an extract of Ixeris strigosa or a fraction thereof as an active component. More specifically, by using the extract of Ixeris strigosa or the fraction thereof, an effect of inhibiting alpha-amylase and alpha-glucosidase can be provided. Therefore, since the composition has an effect of preventing, treating or alleviating diabetes, the composition can be usefully used in various fields such as pharmaceuticals and health functional foods for preventing, treating or alleviating diabetes.

Description

BACKGROUND OF THE INVENTION Field of the Invention [0001] A composition for preventing or treating diabetes containing an extract or a fraction thereof as an active ingredient, the composition for preventing or treating diabetes containing extract of Ixeris strigosa

The present invention relates to a composition for the prevention or treatment of diabetes, comprising an extract of sagebrush or a fraction thereof as an active ingredient.

Diabetes is a multifactorial metabolic disorder characterized by chronic hyperglycemia caused by defective insulin secretion or insulin action. Various complications occur due to disability and chronic vascular damage.

Diabetes mellitus is a representative adult metabolic disease, affecting about 5% of the world's population, and the resulting human and economic loss is enormous. Currently, the number of diabetic patients in Korea is estimated at 2.4 million, about 5% of the total population of 48 million, and more than half of them do not know that they are diabetic, but are prospective patients who will soon develop diabetes. The number of diabetic patients is expected to increase even more due to changes in lifestyle and the pursuit of convenience in lifestyle. More than 90% of diabetes mellitus are adult type 2 diabetes, and the increase in the number of type 2 diabetes patients due to aging and lifestyle changes is a trend that accounts for most of the increase in the total number of diabetes patients.

Most of the diabetic patients are taking oral therapeutics, but a safe therapeutic agent has not been developed so far. Although insulin resistance is known as the most important mechanistic cause, the exact mechanism is not yet clear, but it has been found that a complex cause of genetic predisposition and environment is acting.

Blood sugar-lowering drugs used to treat diabetes control postprandial blood sugar and prevent hypoglycemia. However, when insulin preparations, sulfonylurea preparations, and biguanide preparations, which are mainly used as oral hypoglycemic agents, are administered, blood sugar level increases rapidly after eating, whereas insulin caused by the administered drug The appearance in the blood is relatively slow, resulting in postprandial hyperglycemia, hyperinsulinemia and hypoglycemia between meals. In addition, in the case of commonly used drugs, since they are chemical drugs made of chemically artificial compounds, they lack reducibility in the body, generate free radicals in the body, damage DNA, and cause resistance and various side effects. This is being provoked Therefore, there is a need for a composition for preventing and treating antidiabetic activity obtained from natural plants and a treatment method using the same.

On the other hand, Ixeris strigosa is a perennial plant belonging to the Asteraceae family and is distributed in Korea and Japan. It grows on the roadside or in dry grass, and several stems come out from the bottom and are white in color, and the height is 20-50cm. The leaves from the root are 8~24cm long and have an upside down lancet shape or an upside down lancet shape, long oval shape. The leaves on the stem are 1~2, the length is 1~4cm, and the lower part surrounds the stem. Flowers bloom from May to June, and the petals are white, but the edges are very light red. At the end of the stem, 20 head-shaped flowers (頭狀花: flowers that do not have a peduncle to form a head by gathering many small flowers without a peduncle at the end of the stem) run in coaxial inflorescences. The capillary flower is 2~2.5cm in diameter and has 23~27 small flowers of 12mm in length. The gun is 9~10mm long, and the pieces of the gun are arranged in two rows. The fruit is aqueous, 5.5-7mm long, with 10 hairy lines, and the tube hairs are white and 6mm long. Roots and tender plants are eaten as vegetables.

As a result of intensive research on natural products having excellent antidiabetic effects, the present inventors have confirmed that the sagebrush has an antidiabetic effect and have completed the present invention.

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of diabetes containing a line sseumbagwi (Ixeris strigosa) extracts or fractions thereof.

It is another object of the invention to provide a line sseumbagwi (Ixeris strigosa) extract or a health food composition for the prevention or amelioration of diabetes, including fractions thereof.

In order to achieve the above object,

The present invention provides a pharmaceutical composition for preventing or treating diabetes containing a line sseumbagwi (Ixeris strigosa) extracts or fractions thereof.

The present invention also provides a line sseumbagwi (Ixeris strigosa) extract or a health food composition for the prevention or amelioration of diabetes, including fractions thereof.

Since the extract of the present invention has excellent α-glucosidase and α-amylase inhibitory effects, it can be widely used as a pharmaceutical composition for preventing or treating diabetes or a food composition for preventing or improving diabetes.

1 is a diagram showing a process for preparing a fraction from an extract of sagebrush.
Figure 2 is a diagram showing the inhibitory effect of α-glucosidase of the extract of Sunflowers.
3 is a diagram showing the α-glucosidase inhibitory effect of a fraction fractionated from an extract of S. serrata.
Figure 4 is a diagram showing the α-amylase inhibitory effect of the extract of sagebrush.
5 is a diagram showing the α-amylase inhibitory effect of a fraction fractionated from an extract of S. serrata.

Hereinafter, the present invention will be described in detail as an embodiment of the present invention with reference to the accompanying drawings. However, the following embodiments are presented as examples of the present invention, and when it is determined that detailed descriptions of well-known techniques or configurations known to those skilled in the art may unnecessarily obscure the gist of the present invention, the detailed description may be omitted, and , the present invention is not limited thereby. Various modifications and applications of the present invention are possible within the scope of equivalents interpreted therefrom and the description of the claims to be described later.

In addition, the terms used in the present invention (terminology) are terms used to properly express the preferred embodiment of the present invention, which may vary according to the intention of the user or operator, or customs in the field to which the present invention belongs. Accordingly, definitions of these terms should be made based on the content throughout this specification. Throughout the specification, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

Unless otherwise defined, all technical terms used in the present invention have the meanings commonly understood by those skilled in the art in the relevant field of the present invention. In addition, although preferred methods and samples are described herein, similar or equivalent ones are also included in the scope of the present invention. The contents of all publications herein incorporated by reference are incorporated herein by reference.

The present invention provides a pharmaceutical composition for preventing or treating diabetes containing a line sseumbagwi (Ixeris strigosa) extracts or fractions thereof.

As used herein, the term "prevention" may refer to any action of inhibiting or delaying the onset of diabetes by administering the pharmaceutical composition for preventing or treating diabetes according to the present invention to an individual.

As used in the present invention, the term "treatment" may refer to any act of administering the composition of the present invention to an individual suspected of having diabetes so that symptoms of diabetes mellitus are improved or beneficial.

As used herein, as used herein, the term "Snowfly extract" is a generic term for preparations concentrated by squeezing sagebrush into an appropriate leaching solution and evaporating the leachate. It can be obtained by an extraction process using a , and may include an extract, a diluted or concentrated solution thereof, or a dry powder of the extract, or any form formulated using the same. In addition, in the extraction method, methods such as hot water extraction, hot water extraction, cold-chim extraction, warm-chim extraction, pressure extraction, reflux cooling extraction, or ultrasonic extraction may be used, but are not limited thereto.

In obtaining the E. sagebrush extract, it may be preferably extracted using water, an organic solvent, or a mixed solvent thereof. In case of extraction using an organic solvent, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or an organic solvent that is a mixed solvent thereof can be used, and extraction can be performed at room temperature or by heating under conditions in which the active ingredient of the herbal medicine is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and loss of the active ingredient of the drug may be different, so an appropriate organic solvent should be selected and used.

The solvent extract may further comprise filtering the extract to remove suspended solid particles. It is possible to filter the particles using cotton, nylon, or the like, or use ultrafiltration, cryofiltration, centrifugation, and the like.

Methods such as reduced pressure concentration, reverse osmosis concentration, etc. may be used for concentration of the extract.

The drying step after concentration includes freeze drying, vacuum drying, hot air drying, spray drying, reduced pressure drying, foam drying, high frequency drying, or infrared drying. In some cases, it may further include a process of pulverizing the final dried extract.

In addition, the extract extracted with the solvent is then subjected to a fractionation process with a solvent selected from the group consisting of n-butanol, hexane, dimethyl chloride, methylene chloride, acetone, ethyl acetate, ethyl ether, chloroform, water, and mixtures thereof. and preferably selected from the group consisting of hexane, chloroform, ethyl acetate, n-butanol, and mixtures thereof, but is not limited thereto.

The treatment method of the present invention may include administering a pharmaceutical composition comprising an extract of Pseudomonas albacorea in a pharmaceutically effective amount.

As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is dependent on the subject's type and severity, age, sex, and disease. It can be determined according to the type, drug activity, drug sensitivity, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. and may be administered single or multiple. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, and can be easily determined by those skilled in the art.

The preferred dosage of the composition of the present invention varies depending on the patient's condition and weight, the degree of disease, the drug form, the route and duration of administration, and a suitable total daily amount can be determined by the treating physician within the scope of sound medical judgment.

The pharmaceutical composition of the present invention may further include an adjuvant in addition to the active ingredient. The adjuvant may be used without limitation as long as it is known in the art, and for example, Freund's complete adjuvant or incomplete adjuvant may be further included to increase the immunity thereof.

The pharmaceutical composition according to the present invention may be prepared in a form in which the active ingredient is incorporated into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers that can be used in the pharmaceutical composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories or sterile injection solutions according to conventional methods, respectively. .

In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the active ingredient, for example, starch, calcium carbonate, sucrose, lactose, gelatin. It can be prepared by mixing and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. can Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As the base of the suppository, witepsol, tween 61, cacao butter, laurin, glycerogelatin, etc. may be used.

The pharmaceutical composition according to the present invention may be administered to an individual by various routes. Any mode of administration can be envisaged, for example, by oral, intravenous, intramuscular, subcutaneous, intraperitoneal injection.

The pharmaceutical composition may be formulated in various oral or parenteral dosage forms.

Formulations for oral administration include, for example, tablets, pills, hard, soft capsules, solutions, suspensions, emulsifiers, syrups, granules, etc., and these formulations include diluents (eg, lactose, dextrose, water crose, mannitol, sorbitol, cellulose and/or glycine), lubricants (eg silica, talc, stearic acid and its magnesium or calcium salts and/or polyethylene glycol). In addition, the tablet may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, and optionally starch, agar, alginic acid. or disintegrants such as sodium salts thereof or effervescent mixtures and/or absorbents, coloring, flavoring and sweetening agents. The formulation may be prepared by conventional mixing, granulating or coating methods.

In addition, representative formulations for parenteral administration are injection formulations, and water, Ringer's solution, isotonic physiological saline, or suspension can be exemplified as a solvent for the injection formulation. The sterile, fixed oil of the injectable preparation may be used as a solvent or suspending medium, and any non-irritating fixed oil including mono- and di-glycerides may be used for this purpose. The injection preparation may use a fatty acid such as oleic acid.

In addition, the present invention provides a health functional food for the prevention or improvement of diabetes, comprising the extract or a fraction thereof as an active ingredient.

As used herein, the term “improvement” refers to any action that at least reduces a parameter related to the condition being treated, for example, the severity of symptoms. In this case, the functional food composition may be used simultaneously with or separately from a drug for treatment before or after the onset of the disease in order to prevent or improve diabetes.

The food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, like a conventional food composition, in addition to containing the compound as an active ingredient.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The above-mentioned flavoring agents can advantageously use natural flavoring agents (Taumatin), stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). The food composition of the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, and health supplements. There is this.

In addition, the food composition includes various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, colorants and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages.

The functional food composition of the present invention may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, and the like. In the present invention, the term 'health functional food composition' refers to a food manufactured and processed using raw materials or ingredients useful for the human body according to Act No. 6727 of the Health Functional Food Act, and It refers to ingestion for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological effects. The health functional food of the present invention may include normal food additives, and unless otherwise specified, whether it is suitable as a food additive is related to the item according to the general rules and general test method of food additives approved by the Food and Drug Administration. It is judged according to standards and standards. The items listed in the 'Food Additives Codex' include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high pigment, and guar gum; Mixed preparations, such as a sodium L-glutamate preparation, a noodle-added alkali agent, a preservative preparation, and a tar dye preparation, etc. are mentioned. For example, a health functional food in tablet form is granulated by a conventional method by mixing the active ingredient of the present invention with an excipient, a binder, a disintegrant and other additives, followed by compression molding with a lubricant, etc. The mixture can be compression molded directly. In addition, the health functional food in the form of tablets may contain a corrosive agent and the like, if necessary. Among health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture of the active ingredient of the present invention with additives such as excipients in ordinary hard capsules, and soft capsules include the active ingredient of the present invention with additives such as excipients. It can be prepared by filling a mixture mixed with a capsule base such as gelatin. The soft capsules may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary. A health functional food in the form of a ring can be prepared by molding a mixture of the active ingredient of the present invention with an excipient, a binder, a disintegrant, etc. by a conventionally known method, and can be coated with sucrose or other peeling agent if necessary, Alternatively, the surface may be coated with a material such as starch or talc. The health functional food in the form of granules can be prepared in granular form by a conventionally known method by mixing a mixture of the active ingredient excipients, binders, disintegrants, etc. of the present invention, and may contain flavoring agents, flavoring agents, etc. as needed. can

Throughout this specification, '%' used to indicate the concentration of a specific substance is (w/w) % for solid/solid, (w/v) % for solid/liquid, and Liquid/liquid is (v/v) %.

Hereinafter, the present invention will be described in more detail by way of Examples. However, the following examples are only examples for easily explaining the content and scope of the technical idea of the present invention, and thereby the technical scope of the present invention is not limited or changed. In addition, it can be easily determined by those skilled in the art that various modifications and changes are possible within the scope of the technical spirit of the present invention based on these examples.

<Example 1>

Manufacture of extract of sagebrush

The sample of Ixeris strigosa was harvested from the ginseng special production department in Eumseong-gun, Chungcheongbuk-do in 2018, and the above-ground part of the ginseng plant was freeze-dried and powdered. For extraction, 50 g of the dried above-ground part of the dried sagebrush was extracted three times for 24 hours at room temperature with 70% alcohol and water (sample/solvent ratio, 1:10). After filtration, all extracts were concentrated in vacuo and freeze-dried. The extract was dissolved in DMSO (Sigma-Aldrich, St.Louis, MO, USA) at a concentration of 100 mg/mL and used for the experiment.

The extraction yield is shown in Table 1 below.

Sample name
(Scientific name) Growing area Solvent
(Alcohol %) Extraction weight (g) extract
Weight (g) Yields
(%) good word
( Ixeris strigosa ) Chungbuk Eumseong 0 50 17.69 35.38 70 50 17.15 34.3

<Example 2>

Preparation of fractions fractionated from the extract of sagebrush

Solvent fractions of sagebrush were prepared by sequential solvent fractionation (Jeon et al., 2012).

First, after dissolving 2 g of the 70% alcohol extract of the sagebrush of Example 1 in 200 mL of water, put it in a separatory funnel, and hexane, chloroform, ethyl acetate, and water saturated n -Butanol (water saturated n-butanol) was added in the order of polarity from non-polarity to fractionate. The five fractions, including the water layer remaining after fractionation, were re-dissolved in DMSO after concentration under reduced pressure and used as samples.

The yield of the fraction obtained as described above is shown in Table 2 below.

<Experimental Example 1>

The α-glucosidase inhibitory effect of the P. serrata extract and the fractions fractionated therefrom

<1-1> Confirmation of α-glucosidase inhibitory effect of P. serrata extract

The α-glucosidase inhibitory activity was measured using the α-glucosidase Inhibitor Screening Kit (BioVision K938, Milpitas, USA).

After mixing and reacting algae extract (Samples, S) and assay buffer (Enzyme Control, EC) with a solution containing α-glucosidase and substrate, absorbance was measured at 410 nm for 60 minutes at 5-minute intervals, The enzyme inhibitory activity was calculated from the change in absorbance. As a control (Inhibitor Control, IC), acarbose was used, and it was prepared and measured at the same concentration as the sample.

Relative Inhibition(%) =

As a result, as shown in FIG. 2 , the 70% ethanol extract of S. serrata of the present invention has an α-glucosidase inhibitory effect of about 60%, which has superior α-glucosidase inhibitory effect compared to the water extract of S. confirmed that.

<1-2> Confirmation of the α-glucosidase inhibitory effect of the fractions fractionated from the extract

The α-glucosidase inhibitory activity was measured using the α-glucosidase Inhibitor Screening Kit (BioVision K938, Milpitas, USA).

After mixing and reacting the algae fraction (Samples, S) or Assay buffer (Enzyme Control, EC) with a solution containing α-Glucosidase and Substrate, the absorbance was measured at 410 nm for 60 minutes at 5 minute intervals. The enzyme inhibitory activity was calculated from the change. As a control (Inhibitor Control, IC), acarbose was used, and it was prepared and measured at the same concentration as the sample.

Relative Inhibition(%) =

As a result, as shown in FIG. 3 , it was confirmed that the ethyl acetate fraction and the butanol fraction among the fractions fractionated from the serrata extract of the present invention had a superior α-glucosidase inhibitory effect than Acarbose used as a treatment for diabetes.

<Experimental Example 2>

The α-amylase inhibitory effect of the sagebrush extract and the fractions fractionated therefrom

<2-1> Confirmation of the α-amylase inhibitory effect of P. serrata extract

The α-amylase inhibitory activity was measured using the α-amylase Inhibitor Screening Kit (BioVision K482, Milpitas, USA).

After mixing and reacting algae extract (Samples, S) or assay buffer (Enzyme Control, EC) with a solution containing α-Amylase and substrate, absorbance was measured at 405 nm for 25 minutes at 5-minute intervals. The enzyme inhibitory activity was calculated from the change. As a control (Inhibitor Control, IC), acarbose was used, and it was prepared and measured at the same concentration as the sample.

Relative Inhibition(%) =

As a result, as shown in FIG. 4 , the 70% ethanol extract of S. oleracea of the present invention showed an α-amylase inhibitory effect of about 80%, confirming that it had a superior α-amylase inhibitory effect compared to the water extract of S.

<2-2> Confirmation of the α-amylase inhibitory effect of the fractions fractionated from the P. serrata extract

The α-amylase inhibitory activity was measured using the α-amylase Inhibitor Screening Kit (BioVision K482, Milpitas, USA).

After mixing and reacting the antler fraction (Samples, S) or assay buffer (Enzyme Control, EC) with a solution containing α-Amylase and substrate, absorbance was measured at 405 nm for 25 minutes at 5 minute intervals. The enzyme inhibitory activity was calculated from the change. As a control (Inhibitor Control, IC), acarbose was used, and it was prepared and measured at the same concentration as the sample.

Relative Inhibition(%) =

As a result, as shown in FIG. 5 , it was confirmed that the ethyl acetate fraction and the butanol fraction among the fractions fractionated from the serrata extract of the present invention had an excellent α-amylase inhibitory effect comparable to that of Acarbose used as a treatment for diabetes.

Claims (8)

Sseumbagwi line (Ixeris strigosa) as above-ground ethanol extract or fraction A pharmaceutical composition for the prevention or treatment of diabetes, including thereof,
The composition, characterized in that the sagebrush fraction is the ethyl acetate and butanol fractions fractionated from the ethanol extract of the above sagebrush. delete delete delete According to claim 1,
A pharmaceutical composition for the prevention or treatment of diabetes mellitus, wherein the extract or a fraction thereof is excellent in an alpha-glucosidase inhibitory effect. According to claim 1,
A pharmaceutical composition for the prevention or treatment of diabetes mellitus, wherein the extract or a fraction thereof is excellent in the alpha-amylase inhibitory effect. Sseumbagwi line (Ixeris strigosa) as the ethanol extract of aerial parts or fractions prevention or health food composition for the improvement of diabetes, including thereof,
The composition, characterized in that the sagebrush fraction is the ethyl acetate and butanol fractions fractionated from the ethanol extract of the above sagebrush. A method for inhibiting the activities of alpha-glucosidase and alpha-amylase by treating cells with an ethanol extract of an above-ground part of the sagebrush or a fraction thereof in vitro, the method comprising:
The method according to claim 1, wherein the sagebrush fraction is an ethyl acetate and butanol fraction fractionated from an ethanol extract of an aerial part of sagebrush.

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