KR20200113918A - Pharmaceutical composition comprising the extract of loranthus yadoriki as an effective component for prevention or treatment of diabetes and health functional food comprising the same - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and health functional food for preventing or treating/alleviating diabetes containing a Taxillus yadoriki extract as an active component. More specifically, the present invention relates to a pharmaceutical composition and health functional food for preventing or treating/alleviating diabetes through inhibition of beta-amylase and alpha-glucosidase, which contain a Taxillus yadoriki extract as an active component. The Taxillus yadoriki extract, as the active component of an antidiabetic composition, as demonstrated through embodiment of the present invention, has an excellent effect of being able to be used as medicines and health functional food for preventing or treating/alleviating diabetes. In addition, the Taxillus yadoriki extract of the present invention has excellent thermal stability, and does not exhibit loss of starch degrading enzyme inhibitory activity even in the acidic condition of pH 2 and plasma, so that the Taxillus yadoriki extract can be easily processed into various forms such as liquid, cream, powder, pills, tablets, etc., thereby being able to be very usefully used in the pharmaceutical industry, and food industry.

Description

Pharmaceutical composition and health functional food for prevention or treatment of diabetes containing oak mistletoe extract as an active ingredient {PHARMACEUTICAL COMPOSITION COMPRISING THE EXTRACT OF LORANTHUS YADORIKI AS AN EFFECTIVE COMPONENT FOR PREVENTION OR TREATMENT OF DIABETES AND HEALTH FUNCTIONAL FOOD COMPRISING THE SAME}

The present invention relates to a pharmaceutical composition and health functional food for the prevention or treatment/improvement of diabetes (diabetes) containing an oak mistletoe ( Loranthus yadoriki ) extract as an active ingredient, and more specifically, an oak mistletoe stem extract as an active ingredient It relates to a pharmaceutical composition for preventing or treating/improving diabetes through inhibition of beta-amylase (β-amylase) and alpha-glucosidase, and a health functional food.

Diabetes, one of the most common diseases in modern people, is a type of metabolic disease such as insufficient secretion of insulin or inability to function normally. It is characterized by a high blood glucose concentration and causes various symptoms and signs due to high blood sugar. Diabetes is divided into type 1 and type 2. Type 1 diabetes is caused by the inability to produce insulin at all due to genetic effects, and type 2 diabetes is caused by poor insulin function and insulin resistance. ) Is known as the cause. In particular, type 2 diabetes is known to be largely affected by environmental factors such as high calorie, high fat, high protein diet, lack of exercise, and stress due to westernization of diet. For the treatment of diabetes, insulin injection is essential in the case of type 1 diabetes, and lifestyle correction and drug treatment are necessary in the case of type 2 diabetes. Representative insulin secretion promoters (e.g., repaglinide, mitigl) Nide) and a delaying agent for the absorption of carbohydrates in the small intestine (glucobi: ingredient name-acarbose, bason: ingredient name-voglibose), and the like are used.

On the other hand, mistletoe is a parasitic plant distributed throughout Asia and Europe, including Korea, and is an evergreen shrub that grows on the branches of various host plants such as oak, chestnut, teal, hackberry, birch, pine, sakura, and apple trees. . In the private sector, the leaves and stems of mistletoe are known to be effective in preventing high blood pressure, pain, low back pain, frostbite, and miscarriage. In particular, oak mistletoe ( Loranthus yadoriki ) has been used as a valuable medicinal material in oriental medicine since ancient times because it has no toxicity and excellent medicinal properties.

The oak mistletoe is an evergreen parasitic shrub of the monocotyledonous mistletoe family, and is a semi-parasitic plant that performs photosynthesis while parasitic to plants. Leaves are opposite or alternate, wide egg-shaped, 3~6cm long, flat edge, and reddish brown spread hairs grow densely on the back side. Flowers are male and female, and flowers with 2-3 peduncles bloom in one leaf axil. The flower bud is bent and covered with a narrow egg-shaped tubular shape, with reddish-brown spread hairs on the outside, and the inside is black-purple, glossy, and the tip is split into 4 pieces and bends back, with 4 stamens, and the style of the stylus comes out of the perianth. The fruit is a berry (oval shape), ripens yellow after wintering, and has spread hairs, and the flesh is viscous. In Korea, it is usually collected from autumn to spring, but it is known that it is best to collect it mainly in midwinter. In private and oriental medicine, it is used for cancer cell death, high blood pressure, diabetes, cholesterol lowering, aging prevention, and bedsore removal.

Studies related to oak mistletoe include the chemical composition of saccharides and tripterponoids (Soohee Lee et al., 1996, Wood Engineering, 24: 28-36), evaluation of antioxidant and antibacterial activity of stem leaf extract (Wonyoung Ahn, 1998, research report), and extraction conditions. (Hyejin Lee et al., 2011, Korean Journal of Food Storage and Distribution 18: 72-78) and cancer cell killing effect (Kim EJ et al., 2018, Biotechnology and Bioprocess Engineering 23: 201-207), related to antithrombotic activity In 2003, "Study on the analysis of components of the blood coagulation inhibitory effect of mistletoe" (Seon-kyung Lee, 2003, Master's thesis at Sungshin Women's University), however, studies on oak mistletoe are very insignificant, especially β-amylase and α-glucosidase. There are no studies related to antidiabetic by inhibitory activity against

In addition, as patents related to oak mistletoe, Korean Patent Registration No. 10-1329727 [composition containing oak mistletoe extract having monoamine oxidase inhibitory activity], [Mistletoe extract or separation therefrom] in No. 10-0923217 Composition for the prevention and treatment of neuropathic pain containing the active ingredient] and [Method for mass production of cultured cells and stems by culturing oak mistletoe] in No. 10-1386261 are disclosed. Japanese Patent JP-0099319 [HAIR TONIC] to promote hair growth in International Patent Publication No. WO-0081831, [COMPOSITION COMPRISING THE EXTRACT OF LORANTHUS YADORIKI SIEB HAVING MONOAMINE OXIDASE-INHIBITING ACTIVITY-Contains oak mistletoe extract having monoamine oxidase inhibitory activity One composition] has been disclosed, and [Pharmaceutical composition for thrombosis treatment] is disclosed in Korean Patent Registration No. 10-0247569 in relation to antithrombotic activity, but inhibition of β-amylase and α-glucosidase of oak mistletoe extract to date There are no known patents related to antidiabetic activity.

KR 10-0247569 B KR 10-1329727 B

Lee Seon-kyung, 2003, Master's thesis at Sungshin Women's University

The present invention has been conceived to solve the problems of the prior art as described above, and the problem to be solved in the present invention is to provide a pharmaceutical composition for preventing or treating diabetes containing oak mistletoe extract as an active ingredient and a health functional food I want to.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating diabetes containing an oak mistletoe ( Loranthus yadoriki ) extract as an active ingredient.

It is preferable that the oak mistletoe is an oak mistletoe stem.

In addition, the present invention provides a health functional food for preventing or improving diabetes containing an oak mistletoe ( Loranthus yadoriki ) extract as an active ingredient.

It is preferable that the oak mistletoe is an oak mistletoe stem.

The oak mistletoe extract as an active ingredient of the antidiabetic composition of the present invention has an excellent effect that can be used as a drug for preventing or treating/improving diabetes and a health functional food, as demonstrated through the examples of the present specification. In addition, the oak mistletoe extract of the present invention has excellent thermal stability, does not show a loss of starch degrading enzyme inhibitory activity even in the acidic condition of pH 2 and plasma, and can be easily processed into various forms such as liquid, powder, pills, tablets, etc. It can be very useful in the pharmaceutical industry and food industry.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention prepared an oak mistletoe stem extract by a certain method in order to test the antidiabetic effect on oak mistletoe, and recovered the extract as an excellent antidiabetic active ingredient, and the extract had hemolytic activity against human red blood cells. While not showing at all, by confirming that it has excellent characteristics of thermal stability and acid stability, the extract was intended to be utilized as a pharmaceutical composition and health functional food having antidiabetic activity.

Specifically, the present inventors have used oak mistletoe known to be effective in various diseases such as vascular and circulatory system, digestive system, metabolic system, and aging in the private sector to develop a pharmaceutical composition and health functional food with antidiabetic activity, oak mistletoe Ethanol extracts were prepared from stems, and their inhibitory activity against β-amylase and α-glucosidase was evaluated. As a result, strong β-amylase and α-glucosidase enzyme inhibitory activities were confirmed in the extract.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating diabetes containing an oak mistletoe ( Loranthus yadoriki ) extract as an active ingredient.

It is preferable that the oak mistletoe is an oak mistletoe stem.

In addition, the present invention provides a health functional food for preventing or improving diabetes containing an oak mistletoe ( Loranthus yadoriki ) extract as an active ingredient.

It is preferable that the oak mistletoe is an oak mistletoe stem.

Hereinafter, a method for producing an oak mistletoe extract of the present invention and an experiment of efficacy will be described in more detail.

The present invention comprises the steps of preparing an extract from oak mistletoe stems; Evaluating the antidiabetic activity of the extract; And investigating the stability of the active extract.

The "oak mistletoe extract" included in the composition of the present invention can be obtained by extracting oak mistletoe with an organic solvent and filtering the extract using a filter net of 0.06 mm or less, and concentrating it under reduced pressure.

The organic solvent used in the present invention is water (cold water, hot water), alcohol, anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, butanol, etc.), a mixed solvent of the lower alcohol and water, etc. Can be used, hot water, or 70% ethanol extraction is most preferred.

As a preferred embodiment, the oak mistletoe extract of the present invention may be used by extracting the oak mistletoe stem with hot water or ethanol. Here, the hot water or ethanol extract may be sequentially or fractionated with an organic solvent of hexene, ethyl acetate, and butanol to additionally obtain a hexene fraction, an ethyl acetate fraction, a butanol fraction, and a water residue.

The oak mistletoe extract of the present invention may be prepared as a powder through a conventional powdering process such as vacuum drying, freeze drying, or spray drying. The oak mistletoe extract is not decomposed by various degrading enzymes in plasma, and maintains activity even at a heat treatment of 100°C and a pH of 2 in the stomach.

The oak mistletoe extract of the present invention exhibits potent β-amylase and α-glucosidase inhibitory activity against starch degrading enzymes, and thus prevents diabetes complications such as type 1 diabetes, type 2 diabetes or diabetic retinopathy, diabetic nephropathy, etc. It can be used as a material for pharmaceutical compositions and health functional foods related to treatment/improvement.

In a preferred embodiment, the antidiabetic composition of the present invention can be applied as a pharmaceutical composition.

The pharmaceutical composition is formulated in various forms such as oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and injections of sterile injectable solutions according to the usual method for each purpose of use. It may be used, and may be administered orally, or administered through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and topical administration.

Such pharmaceutical compositions may further include a carrier, excipient, or diluent, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil And the like. In addition, the pharmaceutical composition of the present invention may further contain a filler, an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, a preservative, and the like.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, drug activity, Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

The effective amount of the component having antidiabetic activity in the pharmaceutical composition of the present invention may vary depending on the age, sex, and weight of the patient, and is generally 1 to 5,000 mg, preferably 100 to 3,000 mg per body weight, daily or every other day. It can be administered or divided into 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, the severity of the disease, sex, weight, age, etc., the dosage amount is not limited by any method.

The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration can be expected and can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intra-cerebroventricular injection. In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the route of administration of the pharmaceutical composition of the present invention is oral or parenteral through all general routes as long as it can reach the target tissue. It can be administered orally. In addition, the composition of the present invention may be administered using any device capable of delivering an active ingredient to target cells.

In the present invention, the "object" is not particularly limited, but includes, for example, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, mouse, rabbit, or guinea pig And, preferably, a mammal, more preferably a human.

As a preferred embodiment, the antidiabetic composition of the present invention can be applied as a health functional food.

Foods containing an active ingredient having excellent antidiabetic activity of the present invention include, for example, various foods, beverages, gum, tea, vitamin complexes, health supplement foods, etc., and powders, granules, tablets, capsules or beverages Can be used in form.

In general, the active ingredient of the present invention may be added in an amount of 0.01 to 15% by weight of the total food weight, and the health beverage composition may be added in an amount of 0.02 to 10g, preferably 0.3 to 1g, based on 100 ml.

The health functional food of the present invention may contain the compound as an essential component in the indicated ratio as an additional component, as well as food supplementary additives acceptable for food, such as natural carbohydrates and various flavoring agents. Examples of the natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and conventional sugars such as polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, taumatin, rebaudioside A, glycyrrhizin, saccharin, aspartame, and the like may be used. The proportion of the flavoring agent is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention.

In addition to the above, the health functional food of the present invention includes a variety of nutrients, vitamins, minerals, synthetic flavors, and flavoring agents such as natural flavors, colorants and thickeners, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloids. It may contain thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.

In addition, the health functional food of the present invention may contain pulp for the production of natural fruit juices, fruit juice drinks, and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is generally selected from 0.01 to about 20 parts by weight per 100 parts by weight of the active fraction of the present invention.

Hereinafter, the present invention will be described in more detail through examples. The following examples are only one preferred specific example of the present invention, and the scope of the present invention is not limited to the scope of the following examples.

[Example]

Example 1: Preparation of oak mistletoe extract and analysis of useful ingredients

Oak mistletoe harvested in Pyeongchang, Gangwon-do in 2018 was purchased and used to prepare ethanol extract. Specifically, 10 times the amount of ethanol (70% ethanol) was added to the stem sample of oak mistletoe, extracted twice at room temperature, collected and filtered, and concentrated under reduced pressure to prepare an ethanol extract.

The content of total polyphenols, total flavonoids, total sugars and reducing sugars was measured by component analysis of the oak mistletoe extract. For the total polyphenol content, 50 μl of Folin-ciocalteau and 100 μl of Na ₂ CO ₃ saturated solution were added to 400 μl of the extraction sample, and allowed to stand at room temperature for 1 hour, and the absorbance was measured at 725 nm. Tannic acid was used as a standard reagent. For the total flavonoid content, each sample was extracted with methanol for 18 hours, and then 4 ml of 90% diethylene glycol was added to 400 μl of the filtered extraction sample solution, 40 μl of 1 N NaOH was added, and the absorbance was measured at 420 nm after reaction at 37° C. for 1 hour. Rutin was used as a standard reagent. Reducing sugar was quantified by DNS method and total sugar was quantified by phenol-sulfuric acid method.

[Table 1] Analysis of extraction efficiency and useful components of oak mistletoe extract

As shown in Table 1, the ethanol extraction efficiency of oak mistletoe stem was 15-20%, and as a result of the analysis of useful components of the extract, the total polyphenol content and total flavonoid content were very high, 172.7mg/g and 25.16mg/g. appear. This is a lower value compared to the general berry extract, but the content was higher than that of conventional herbal medicines. On the other hand, as a result of analysis of total sugar and reducing sugar, 836mg/g and 436mg/g were very high.

Example 2: Evaluation of antidiabetic activity of oak mistletoe extract

The antidiabetic activity of the oak mistletoe extract prepared in Example 1 was evaluated, and in-vitro β-amylase inhibitory activity and α-glucosidase inhibitory activity were evaluated.

First, β-amylase (4-alpha-D-glucan maltohydrolase) inhibitory activity was obtained by mixing 2.5 μl of a sample and 25 μl of α-amylase (0.25 U/ml) diluted with 50 mM phosphate buffer (pH 6.8) at 37°C for 10 minutes. After the first reaction, 25 μl of 0.5% soluble starch (Samchun Chemicals Co., Korea) was added and the second reaction was performed at 37°C for 10 minutes, and then the reaction was stopped by heating at 100°C for 5 minutes. DNS (3,5-dinitrosalicylic acid, Sigma Co., st. Louis, USA) solution was added, heated at 100° C. for 5 minutes to develop color, and then allowed to cool at room temperature. The color developing solution was calculated by measuring the absorbance at 540 nm.

Inhibition rate (%) = [1-(Enzyme activity after addition of sample/Enzyme activity after addition of control)] x 100

α-glucosidase inhibitory activity was evaluated using pNPG (p-nitrophenol glucoside; Sigma Co., USA), and α-glucosidase (0.25 U/ml) 25 μl diluted with 2.5 μl of sample and 50 mM sodium acetate buffer (pH 5.6) The mixture was first reacted at 37° C. for 10 minutes, and 25 μl of 1 mM pNPG solution was added, followed by second reaction at 60° C. for 10 minutes. Thereafter, 25 μl of 1M NaOH was added to stop the reaction, and the inhibition rate was calculated by measuring the absorbance at 405 nm.

Inhibition rate (%) = [1-(Enzyme activity after addition of sample/Enzyme activity after addition of control)] x 100

[Table 2] Antidiabetic activity of oak mistletoe extract

As a result, as shown in Table 2, acarbose, which has been used clinically as a therapeutic agent for diabetes, showed strong β-amylase inhibitory activity and α-glucosidase inhibitory activity in a concentration-dependent manner. In the case of oak mistletoe stem extract, 44.9% of potent β-amylase inhibitory activity and 5.2% of α-glucosidase inhibitory activity were found at 0.5mg/ml concentration. The antidiabetic activity of this oak mistletoe stem extract was more potent when treated with high concentration, and showed a concentration-dependent inhibitory activity. Therefore, it was confirmed that the stem extract of oak mistletoe can be used for the prevention and treatment of type 2 diabetes.

Example 3: Human Red Blood Cell Hemolytic Activity of Oak Mistletoe Extract

Oak mistletoe has been drinking in the private sector for a long time, and as an oriental medicine mentioned in Donguibogam, it has been used for cancer cell death, high blood pressure, diabetes, cholesterol lowering, aging prevention, and bedsore removal. Therefore, it is judged that the extract of oak mistletoe does not cause a separate acute toxicity. In order to evaluate the possibility of acute toxicity of the oak mistletoe extract, the hemolytic activity of human red blood cells was evaluated, and the results are shown in Table 3. At this time, the hemolytic activity was evaluated according to the previous report (Ho-Yong Son, 2014 ㆍKorean J. Microbiol. Biotechnol. 42: 285-292), and simply 100 μl of human red blood cells washed three times with PBS was placed in a 96-well microplate. Then, 100 μl of sample solution of various concentrations was added and reacted for 30 minutes at 37°C. After that, the reaction solution was centrifuged for 10 minutes (1,500 rpm) to transfer 100 μl of the supernatant to a new microtiter plate, and the degree of hemoglobin outflow due to hemolysis was 414 nm. It was measured at. DMSO (2%) was used as a solvent control for the sample, and Triton X-100 (1 mg/ml) was used as an experimental control for hemolysis of red blood cells. Hemolytic activity was calculated using the following formula.

[Table 3] Hemolytic activity of oak mistletoe extract in human red blood cells

First, it was confirmed that DMSO and water used as control cells had no hemolytic activity, and triton X-100 hemolytic 100% red blood cells at a concentration of 1 mg/ml. In addition, it was confirmed that amphotericin B, which is used as an anticancer agent and antifungal agent, hemolyzes more than 50% of red blood cells at a concentration of 0.025mg/ml. It was confirmed that the oak mistletoe extract of the present invention did not show any red blood cell hemolysis up to a concentration of 1 mg/ml, and thus had no acute toxicity and red blood cell hemolytic activity.

Example 4: Plasma, acid and heat stability evaluation of oak mistletoe extract

Plasma stability, thermal stability, and acid stability for antidiabetic activity were confirmed for the oak mistletoe stem extract obtained in Example 1 above. Oak mistletoe stem extract showed no decrease in β-amylase and α-glucosidase inhibitory activities even after 1 hour heat treatment at 100°C, 1 hour treatment at pH 2 (0.01M HCl), and 1 hour treatment in plasma. Therefore, it could be expected that the oak mistletoe stem extract contained various components of antidiabetic active substances having acid resistance and heat resistance.

Claims (4)

Oak mistletoe ( Loranthus yadoriki ) A pharmaceutical composition for the prevention or treatment of diabetes containing an extract as an active ingredient. The pharmaceutical composition according to claim 1, wherein the oak mistletoe is a stem of oak mistletoe. A health functional food for preventing or improving diabetes containing an oak mistletoe ( Loranthus yadoriki ) extract as an active ingredient. The health functional food according to claim 3, wherein the oak mistletoe is an oak mistletoe stem.