KR102567159B1 - Extract of Ligularia fischeriof as α-glucosidase inhibitor - Google Patents

Abstract

The present invention relates to a composition for preventing and treating diabetes comprising a chloroform fraction of bear stink ( Ligularia fischeri ) as an active ingredient. In the present invention, as a result of comparing the α-glucosidase activity inhibitory effect of fractions obtained by extracting bear stink with 70% alcohol solvent and fractionating using hexane, chloroform, ethyl acetate, and butanol, the chloroform fraction was found to be alpha- Glucosidase (α-glucosidase) activity inhibitory effect is the most excellent, the present invention has an effect that can be usefully used for the treatment and prevention of diabetes. The bear odor fraction prepared by the method of the present invention can be usefully used as a pharmaceutical composition or food composition for the treatment and prevention of diabetes.

Description

Bear odor extract having inhibitory activity against alpha-glucosidase and its use {Extract of Ligularia fischeriof as α-glucosidase inhibitor}

The present invention relates to an extract of bear stink ( Ligularia fischeri ) having inhibitory activity against alpha-glucosidase and its use.

Diabetes is a disease caused by abnormally high plasma glucose levels in humans and mammals. This abnormal sugar level increases the level of hemoglobin in plasma, resulting in chronic hyperglycemia. ), atherosclerosis, microangiopathy, kidney disease, heart disease, diabetic retinopathy and other eye diseases.

Diabetes mellitus is characterized by two types. Insulin dependent diabetes (IDDM), which is an insulin-dependent type, is caused by a secretion deficiency of insulin, a glucose-regulating hormone in the blood, and mainly affects young people in their 10s to 20s It is also called juvenile diabetes because it develops in all age groups. Type Ⅱ diabetes (non-insulin dependent diabetes, NIDDM) mainly occurs after the age of 40, and accounts for most of the diabetic patients in Korea. Unlike type I, it is called adult type diabetes, and the cause of the onset is not yet clear, but it is known that genetic and environmental factors are involved together. As the pathogenesis of type II diabetes, both impaired insulin secretion in pancreatic beta cells and defects in insulin action (insulin resistance) in target cells are observed.

There are five groups of compounds currently used for complications such as type II diabetes and insulin resistance: biguanides, thiazolidinediones, sulfonylureas, and benzoic acid. Derivative compounds and α-glucosidase inhibitors are used, and among them, biguanide compounds such as metformin are known to prevent excessive blood gluconeogenesis. Thiazolidinedione compounds are thought to act to increase the rate of glucose consumption of peripheral nerves, sulfonylureas such as tolbutamide and glyburide, and benzoic acids such as repaglinide compounds. α-glucosidase inhibitors such as derivatives and acarbose act to lower plasma glucose by stimulating insulin secretion.

On the other hand, bear odor ( Ligularia fischeri ) is a perennial plant belonging to the genus Asteraceae and grows to about 1 meter in height. It grows wild in a slightly humid place in the hillside grass of a deep mountain, and its rhizome is short, thick, thin, and has many long beard roots. Heart-shaped radical leaves have long petioles and sawtooth edges. Usually, in summer, from July to September, yellow flowers are densely gathered on the upper part of the stem, and there are 5 to 9 tongue flowers on the edge. Recently, it has been found to have anti-cancer activity, so it is highly valued as a health food, and is used as an antitussive, expectorant, pain reliever, and blood circulation promoter.

In addition, it is stated in part A of 1020090047638 that bear stink is useful for suppressing, improving, and treating diabetes and diseases caused by it, but the difference from the present invention is that when the bear stink extract is fractionated, the alpha-glucosidase ( α-glucosidase) showed excellent efficacy in inhibitory activity.

Accordingly, the present inventors confirmed that the chloroform fraction of the bear stinger extract was effective for inhibiting α-glucosidase inhibitory activity and completed the present invention.

1020090047638 A

An object of the present invention is to provide a food composition for the prevention and improvement of diabetes comprising a chloroform fraction of Ligularia fischeri as an active ingredient.

An object of the present invention is to provide a pharmaceutical composition containing a chloroform fraction of Ligularia fischeri as an active ingredient for inhibiting α-glucosidase activity in preventing, improving or treating diabetes.

To provide a pharmaceutical composition for preventing or treating diabetes comprising a chloroform fraction of Ligularia fischeri as an active ingredient.

It is to provide a method for inhibiting alpha-glucosidase activity, including treating the sample with the chloroform fraction of the ethanol extract of bear stipea in vitro.

The present invention provides a food composition for the prevention and improvement of diabetes comprising a chloroform fraction of Ligularia fischeri as an active ingredient.

The present invention provides a pharmaceutical composition for inhibiting α-glucosidase activity in preventing, improving or treating diabetes, comprising a chloroform fraction of bear stink ( Ligularia fischeri ) as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating diabetes comprising a chloroform fraction of Ligularia fischeri as an active ingredient.

The present invention provides a method for inhibiting alpha-glucosidase activity, comprising treating a sample with the chloroform fraction of the ethanol extract of bear stipea in vitro.

In the present invention, as a result of comparing the α-glucosidase activity inhibitory effect of fractions obtained by extracting bear stink with 70% alcohol solvent and fractionating using hexane, chloroform, ethyl acetate, and butanol, the chloroform fraction was found to be alpha- Since it has the best inhibitory effect on α-glucosidase activity, the chloroform fraction of bear stinger of the present invention can be usefully used for the treatment and prevention of diabetes.

1 is a diagram showing a method for preparing a bear odor fraction.
Figure 2 is a diagram showing the measurement of the α-glucosidase activity inhibitory effect of the fractions of the extract of bear chinensis alcohol.

The present invention provides a pharmaceutical composition for preventing and treating diabetes comprising a chloroform fraction of bear odor as an active ingredient.

The fraction may be extracted with a solvent selected from the group consisting of water, C1 to C4 lower alcohol and lower alcohol aqueous solution, more preferably ethanol extract, 60% to 80% ethanol extract or 70% chloroform of ethanol extract It's about fractions.

In the present invention, the chloroform fraction may be sequentially fractionated into hexane, chloroform, ethyl acetate, and n-butanol fractions after extracting bear odor with ethanol, and among them, the chloroform fraction may be used.

The extract may be extracted with bear stink and the solvent at a volume ratio of 1:10.

The extract may be extracted from a part selected from the group consisting of leaves, stems, roots, fruits and seeds of bear stink.

The composition can inhibit alpha-glucosidase activity, so it can be usefully used for the prevention, improvement or treatment of diabetes, and in the prevention and treatment of diabetes, alpha-glucosidase (α-glucosidase) ) can be used for activity inhibition.

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

The pharmaceutical composition according to the present invention may be prepared in the form of incorporating the active ingredient into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers usable in the pharmaceutical composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, 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 and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories or sterile injection solutions according to conventional methods, respectively. .

When formulated, it may be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations contain at least one or more excipients such as starch, calcium carbonate, sucrose, lactose, and gelatin in addition to active ingredients. It can be prepared by mixing etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions for oral administration, emulsions, syrups, etc. In addition to commonly used diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. can Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for suppositories, witepsol, tween 61, cacao paper, laurin paper, glycerogelatin, and the like may be used.

The pharmaceutical composition according to the present invention can be administered to a subject by various routes. All modes of administration are contemplated, eg oral, intravenous, intramuscular, subcutaneous, intraperitoneal injection.

The dosage of the pharmaceutical composition according to the present invention is selected in consideration of the age, weight, sex, and physical condition of the subject. It is obvious that the concentration of the active ingredient included in the pharmaceutical composition can be variously selected according to the subject, and is preferably included in the pharmaceutical composition at a concentration of 0.01 to 5,000 μg/ml. If the concentration is less than 0.01 μg/ml, pharmacological activity may not appear, and if the concentration exceeds 5,000 μg/ml, toxicity to the human body may be exhibited.

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

Formulations for oral administration include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, granules, etc. chlorose, mannitol, sorbitol, cellulose and/or glycine), lubricants such as silica, talc, stearic acid and magnesium or calcium salts thereof 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 in some cases starch, agar, alginic acid or a disintegrant or effervescent mixture, such as its sodium salt, and/or absorbents, colorants, flavors, and sweeteners. The formulation may be prepared by conventional mixing, granulating or coating methods.

In addition, a typical formulation for parenteral administration is an injection formulation, and water, Ringer's solution, isotonic physiological saline or suspension may be used as a solvent for the injection formulation. Sterile fixed oils of the above injectable preparations may be used as a solvent or suspending medium, and any bland fixed oil may be used for this purpose, including mono- and di-glycerides.

In addition, the formulation for injection may use a fatty acid such as oleic acid.

The present invention may provide a quasi-drug composition for preventing or improving diabetes comprising an extract of Ligularia fischeri or a fraction thereof as an active ingredient.

In addition, the present invention can provide a food composition for prevention or improvement of diabetes comprising a chloroform fraction of Ligularia fischeri as an active ingredient.

In addition to containing the active ingredient, the food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients like conventional food compositions.

Examples of the aforementioned 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 dextrins and cyclodextrins, and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agents described above, natural flavoring agents (thaumatin), stevia extracts (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used. The food composition of the present invention can be formulated in the same way 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, gum, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements, etc. there is

In addition, the food composition, in addition to the active ingredient extract, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like may be contained. In addition, the food composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages, and vegetable beverages.

The functional food composition of the present invention may be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills, and the like. In the present invention, 'health functional food composition' refers to a food manufactured and processed using raw materials or ingredients having useful functionality for the human body according to Health Functional Food Act No. 6727, and the structure and function of the human body It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions. The health functional food of the present invention may contain ordinary food additives, and the suitability as a food additive is determined according to the general rules of the Food Additive Code and General Test Methods approved by the Food and Drug Administration, unless otherwise specified. It is judged according to standards and standards. Examples of the items listed in the 'Food Additive Code' 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, kaoliang pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations. For example, a health functional food in the form of a tablet is obtained by granulating a mixture obtained by mixing the active ingredient of the present invention with an excipient, a binder, a disintegrant, and other additives in a conventional manner, and then adding a lubricant or the like to compression molding, or as described above. The mixture can be directly compression molded. In addition, the health functional food in the form of a tablet may contain a flavoring agent and the like as needed. Among health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture in which the active ingredient of the present invention is mixed with additives such as excipients in a normal hard capsule. It can be prepared by filling the mixture mixed with gelatin in a capsule base. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary. The health functional food in the form of a pill can be prepared by molding a mixture of the active ingredient of the present invention mixed with an excipient, a binder, a disintegrant, etc. by a conventionally known method, and can be coated with sucrose or other coating agent if necessary, Alternatively, the surface may be coated with a material such as starch or talc. Health functional food in the form of granules can be prepared in granular form by a conventionally known method of mixing the active ingredient of the present invention with excipients, binders, disintegrants, etc., and, if necessary, flavoring agents, flavoring agents, etc. can

Hereinafter, the present invention will be described in more detail by examples. These examples are merely for explaining the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

<Example 1>

Preparation of bear odor extract of the present invention

The above-ground parts of bear sting (harvested in Pyeongchang-gun, Gangwon-do in 2018) were dried in a freeze dryer (PVTFD 50R, Ilshinbio, Seoul, KOREA) and powdered. For extraction, 50 g of dried aerial parts of bear stinger were repeatedly extracted with 70% alcohol (v/v) (1:10 ratio of sample to solvent) at room temperature for 24 hours for 3 times. The extract solution was concentrated by filtration and freeze-dried. Each extract was dissolved in dimethylsulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) at a concentration of 100 mg/mL and diluted before use.

<Example 2>

Method for producing bear odor fraction of the present invention

For the sequential solvent fractionation of bear stinger extract, 1 g of 70% alcohol extract was re-dissolved in 200 mL of water, put into a separatory funnel, and hexane, chloroform, ethyl acetate, and water standard n- Butanol (water standard n-butanol) was added in order to fractionate. Five fractions, including the water layer remaining after fractionation, were concentrated under reduced pressure, re-dissolved in dimethyl sulfoxide (DMSO), and used as samples. The yield of each solvent was confirmed as 16.0% of n-butanol, 2.0% of hexane, 3.0% of chloroform, 4.0% of ethyl acetate, and 64.0% of water (see FIG. 1). )

<Example 3>

Confirmation of Alpha-Glucosidase Inhibitory Effect of the Chloroform Fraction of the Ethanol Extract of Gomchidae of the Present Invention

In order to evaluate the α-glucosidase inhibitory effect of the bear odor fraction of the present invention, an α-glucosidase activity colorimetric assay kit (biovision, Milpitas, CA, USA) was used. 10ul of the sample diluted by 1/10 was added to each well, 60ul of the assay buffer was added, and 10ul of alpha-glucosidase was added, followed by reaction at room temperature by blocking light for 30 minutes. Then, 20ul alpha-glucosidase (α-glucosidase) substrate mixture was added to each well, and then the absorbance was measured in a kinetic mode at 410 nm for 40 minutes with a microplate reader (Biotek, Winooski, VT, USA). .

As a result, when the α-glucosidase activity inhibitory effect of the fractions fractionated using hexane, chloroform, ethyl acetate, and butanol of the bear odor fraction was measured, the activity of the bear odor chloroform fraction was the highest and the positive control group It was confirmed that the inhibition rate was the highest than that of acarbose (see FIG. 2).

So far, the present invention has been looked at with respect to its preferred embodiments. Those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent scope will be construed as being included in the present invention.

Claims (12)

A food composition for the prevention and improvement of diabetes comprising a chloroform fraction of the aerial part of bear odor as an active ingredient,
The chloroform fraction of the aerial part of the bear stink is obtained by extracting the aerial part of the bear stink with 70% ethanol and then fractionating it with chloroform. delete According to claim 1,
Wherein the extraction is to extract bear odor and the solvent at a volume ratio of 1:10. delete According to claim 1,
Wherein the fraction inhibits alpha-glucosidase activity, composition delete delete A pharmaceutical composition for inhibiting α-glucosidase activity in the prevention, improvement or treatment of diabetes, comprising a chloroform fraction of the aerial parts of bear stink bug as an active ingredient,
The chloroform fraction of the aerial part of the bear stink is obtained by extracting the aerial part of the bear stink with 70% ethanol and then fractionating it with chloroform. delete delete A pharmaceutical composition for the prevention or treatment of diabetes comprising a chloroform fraction of the aerial part of bear odor as an active ingredient,
The chloroform fraction of the aerial part of the bear stink is obtained by extracting the aerial part of the bear stink with 70% ethanol and then fractionating it with chloroform. A method for inhibiting alpha-glucosidase activity, comprising treating a sample with a chloroform fraction of the aerial part of bear stinger in a test tube,
The chloroform fraction of the aerial part of the bear stink is obtained by extracting the aerial part of the bear stink with 70% ethanol and then fractionating it with chloroform.