KR20120110947A - Composition for treating diabete-induced complication containing an extract from ligularia fischeri - Google Patents

Abstract

PURPOSE: A composition containing Ligularia fischeri extract or fraction thereof is provided to safely treat diabetic complication without toxicity and side effects. CONSTITUTION: A composition for preventing or treating diabetic complication contains Ligularia fischeri extract prepared by extracting Ligularia fischeri using water, straight or branched alcohol of C1-C4, or a mixture thereof, or Ligularia fischeri fraction prepared by fractioning the Ligularia fischeri extract with water, hexane, methylene chloride, ethyl acetate, or butanol. A composition for preventing or treating diabetes or diabetic complication contains the extract or fraction as an active ingredient.

Description

Composition for treating diabete-induced complication containing an extract from Ligularia fischeri}

The present invention bear ( Ligularia fischeri ) extract or a composition for the prevention and / or improvement of diabetic complications containing the extract as an active ingredient.

Diabetes, which is rapidly increasing as the awareness of health increases due to the improvement of living standards and the increase of human lifespan of modern people, is a chronic disease that is attracting worldwide attention.

Diabetes mellitus is a metabolic syndrome characterized by high blood sugar caused by a deficiency of insulin hormone or abnormal insulin resistance produced by the pancreatic beta cells and by both defects. These diabetes can be divided into insulin dependent diabetes mellitus (IDDM; Type I) and insulin independent diabetes mellitus (NIDDM; Type II) caused by insulin resistance and insulin secretion impairment.

Diabetes mellitus, with chronic hyperglycemia, causes not only glucose metabolism but also lipid and protein metabolic disorders. The conditions vary and are directly attributable to hyperglycemia and include diabetic peripheral neuropathy, diabetic retinopathy, diabetic nephropathy, diabetic cataracts, keratosis and diabetic arteriosclerosis in the retina, kidney, nerve and cardiovascular system. The number of diabetics is steadily increasing due to lifestyle changes such as overeating, lack of exercise, westernized diet, stress, drinking, and smoking.

In both type I and type II diabetes, various complications such as heart disease, intestinal disease, eye disease, neurological disease, stroke, etc. occur, which causes elevated levels of blood sugar and insulin for a long time, leading to chronic neurological and cardiovascular diseases. Short-term hypoglycemia and hyperglycemic reactions cause acute complications.

Diabetes treatment includes drug therapy, exercise therapy, and dietary therapy. Insulin medicines and various glycemic agents are used depending on the symptoms of the patient. However, diabetes is a complex disease that is characterized by excessive glucose production in the liver, insulin resistance, and decreased ability to process glucose in muscle and fat cells. Therefore, specific treatment alone cannot prevent the occurrence of various side effects. Among them, pharmacotherapy uses insulin and chemicals, which is a constant problem for side effects and patient resistance due to taking medications. Recently, it is possible to treat diabetes by using natural products with low side effects. There is a need for research to treat and further prevent and treat diabetic complications.

Thus, the present inventors have found that bear odor has an excellent effect on the treatment of diabetic complications in addition to the antioxidant effect has been completed the present invention.

Accordingly, one embodiment of the present invention provides a composition for preventing and / or improving diabetic complications containing at least one selected from the group consisting of extracts and fractions of bear odor as an active ingredient.

Another example provides a composition for preventing and / or improving diabetic complications comprising at least one selected from the group consisting of extracts and fractions of bear odor.

Another example provides an antioxidant composition containing at least one selected from the group consisting of extracts and fractions of bear odor as an active ingredient.

Hereinafter, the present invention will be described in more detail.

Provided is a composition for preventing and / or improving diabetic complications comprising an extract of bear and / or a fraction of the extract as an active ingredient.

Ligularia , a representative wild vegetable that grows a lot in Gangwon-do fischeri ) is a type of odor characterized by broad leaves. It is mainly consumed in the form of herbs, raw vegetables and Ssam in Korea (Cho SD and Kim GH 2005), pharmacological components such as chamomile, jacobine, ameleme, polyphenol compounds and flavonoids. As is confirmed (Cho SD and Kim GH 2005, Chang SK et al. 2008), a lot of researches on the biological activity of bear odor at the animal experiment and molecular biological level. Representatively, the cytotoxic effect of bear odor (Ham SS 1998b), the antimutagenic and genotoxic effect (Ham SS 1998a, Jeong SW et al. 1998), and the high polyphenols of bear odor Inhibition of antioxidant effects (Jeong SW et al. 1998, Park JA and Kim MK 1999, Kwon YJ et al. 2002, Kim EY et al. 2004, Chang SK et al. 2008) and blood pressure elevation-enhancing enzyme ACE (angiotensin converting enzyme) The effectiveness (Choi GP et al. 2002) is already well proven scientifically. In terms of nutrition, bear odor is particularly rich in vitamins A, B ₁ , B ₂ , B ₃ , C and β-carotene. Among them, vitamin A (780 RE / 100 g) and β-carotene (4681 μg / 100 g), calcium (241 mg / 100 g), fiber (1.7 g / 100 g) and iron (5.7 mg / 100 g) are known to be relatively high compared to other vegetables. It is estimated to be high.

In the present invention bear ( Ligularia) fischeri ) was used as a short-term screening method for the prevention of diabetic complications. Inhibition of the formation of advanced glycation endproducts and aldose reductase inhibitory activity was confirmed.

       The bear odor used in the present invention is not particularly limited in its form, and the composition of the present invention is characterized by including extracts and / or fractions obtained from the bear odor as an active ingredient.

The bear odor extract may be obtained by a common bear odor extraction method using at least one solvent selected from the group consisting of water and C ₁ to C ₄ straight or branched alcohol or mixture.

       In one embodiment of the present invention, in order to achieve a better anti-diabetic complication effect, the extract may be an ethanol extract obtained by extracting the bear odor with 60 to 80% (v / v) ethanol,

The fraction may be obtained by fractionating the extract into at least one selected from the group consisting of hexane, methylene chloride, ethyl acetate and butanol. More preferably, the fraction may be selected from the group consisting of fractions obtained for each application by applying one or more selected from the group consisting of the hexane, methylene chloride, ethyl acetate and butanol in the order of the obtained extract in polar order. . For example, the hexane fraction obtained by fractionating the ethanol extract with hexane The methylene chloride fraction obtained by separating the hexane fraction and the remaining aqueous layer with methylene chloride The ethyl acetate fraction obtained by separating the methylene chloride fraction and fractionating the remaining aqueous layer with ethyl acetate The ethyl acetate fraction may be at least one selected from the group consisting of a butanol fraction obtained by separating the remaining aqueous layer with butanol and a water fraction obtained by separating the butanol fraction.

 Bear extract may be prepared according to the conventional method for producing a plant extract, preferably by pressure extraction method, reflux extraction method, hot needle extraction method, ultrasonic grinding extraction method, etc., preferably by pressure extraction method, It is not limited to this.

In addition, the extract prepared as described above may then be concentrated or removed by performing a reduced pressure filtration process or further concentrated and / or lyophilization. Thus, the bear extract in the present invention is used in the sense including a dried product dried in a conventional manner and a concentrate concentrated in a conventional method.

The present invention provides a diabetic complication treatment effect of bear extract and fraction composition. Diabetic complication refers to a number of complications caused by hyperglycemia and protein reactions resulting from diabetes, resulting in the production of advanced glycation endproducts (AGEs) and cross-linking with other proteins in the blood or tissues. Specifically, the diabetic complication of the bear extract and / or fractions in the present invention may have a diabetic vascular disorder, diabetic neuropathy or diabetic infection, and the like, preferably diabetic retinopathy, diabetic Cataract, diabetic nephropathy, diabetic neuropathy, diabetic angiopathy, and the like, more preferably diabetic retinopathy, diabetic cataract or diabetic nephropathy.

Diabetic complication treatment of the present invention was tested through the ability to inhibit the formation of advanced glycation end products (AGEs) and aldose reductase (Aldose reductase).

The final glycosylated product is produced by the non-enzymatic glycosylation reaction of the sugar with the protein via the schiff base and the amadori product. However, in chronic hyperglycemic conditions, the amadori product is accelerated by an irreversible reaction that reversibly produces the final glycation end product, resulting in mass production and accumulation in tissues and serum. Therefore, the production of final glycation end products is one of the main causes of serious diabetic complications.

The type of final glycated glycosides in the blood of type 2 diabetic patients is correlated with the presence and severity of cardiovascular disease, and has been well correlated with several animal model studies. Final glycosylation products produced by nonenzymatic glycosylation of chronic hyperglycemia with biological proteins are one of the main causes of diabetic complications. Diabetic complications often occur despite the normal recovery of blood sugar. Final glycosylated products are protein glycosylated products that accumulate in tissues in proportion to the concentration of hyperglycemia. They exhibit specific absorbance and fluorescence. In particular, the amino groups of proteins have cross-link reactions. Once produced, glycosylated products accumulate in tissues without being degraded even though blood sugar is normally regulated, causing abnormal changes in the structure and function of tissues, and are known to be involved in atheroma atherosclerosis, DNA mutations, aging, Alzheimer's disease, and diabetic complications. have.

As the toxicity of aminoguanidine, which is known as an inhibitor of the final glycation endogenous product, has been shown to increase, there is a need for the development of a safe natural product with no toxicity to replace it.

As a result of testing the inhibitory activity on the final glycated product of the bear extract and fractions according to the present invention, it was found to have the effect of inhibiting the production of the final glycated product (see Table 1 and Table 2).

In addition, an increase in the polyol pathway, which is one of the trigger mechanisms of diabetic complications, is a mechanism causing cell damage. Aldose Reductase

reductase) is an enzyme involved in the process of converting glucose from sorbitol to fructose in the polyol pathway. In the hyperglycemic state, changes in intracellular glucose metabolism occur. When excess glucose enters the cell, it is converted into sorbitol by aldose reductase and is converted into fructose by sorbitol dehydrogenase. . In the normal blood glucose state, the Km value (70 mM) of aldose reductase to glucose is high and the intracellular sorbitol concentration is very low. However, if the intracellular glucose concentration is increased by hyperglycemia, not only the intracellular sorbitol concentration is increased but also the sorbitol dehydrogenase slows down the reaction so that sorbitol accumulates in the cell. Eventually, the high osmotic pressure of sorbitol accumulated in the cells expands the cells and damages the cells, resulting in complications such as cataracts, diabetic neurosis and retinopathy.

Therefore, in the present invention, in order to develop a new natural product that can effectively prevent diabetic complications, we tested the ability to inhibit the aldose reductase activity of the extract and fractions of bear odor. As a result, bear extract and fractions were found to have the ability to inhibit aldose reductase activity (see Table 3 and Table 4).

Bear extract and / or fractions according to the present invention have been shown to have good antioxidant activity. DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging effect was tested to test the antioxidant effect of the composition of the present invention. DPPH is a compound with chemically stable free cardial and is widely used to check the antioxidant capacity when it meets a substance with antioxidant activity. Electron-donating action using DPPH is used for the purpose of inhibiting fat oxidation in food by donating electrons to free radicals, and is used as an action for inhibiting aging by free radicals in the human body.

As a result of evaluating the antioxidant activity by testing DPPH radical scavenging ability of the bear extract and / or fractions, it was found that the bear extract and / or fractions had excellent antioxidant ability (see Table 5 and Table 6).

The composition of the present invention is a nutrient, vitamins, electrolytes, flavoring agents, coloring agents, neutralizing agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, in addition to the bear extract and / or fractions as an active ingredient , Stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The components can be added independently or in combination. The content of the additional component is preferably added in the range of 0.1 to 20 parts by weight or 100 to 10,000,000 parts by weight per 100 parts by weight of licoridine, but is not limited thereto. .

The content of the bear extract and / or fractions in the composition can be appropriately adjusted according to the symptoms of the disease, the progress of the symptoms, the condition of the patient, for example, 0.0001 to 99.9% by weight, preferably 0.001 based on the total weight of the composition It is preferably from 50% by weight, but is not limited thereto. The content ratio is a value based on the drying amount from which the solvent is removed.

The composition may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions, and oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like according to conventional methods It may be used in the form of a dosage form, an external preparation, a suppository, or a sterile injectable solution.

When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one or more excipients and / or lubricants. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like.

The preferred dosage of the composition depends on the condition and weight of the patient, the severity of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. For a more preferable effect, the dosage of the composition of the present invention is preferably 0.1 mg / kg to 20 mg / kg per day based on the active ingredient, but is not limited thereto. The administration may be carried out once a day or divided into several doses. The compositions of the present invention can be administered by various routes to an animal, preferably a mammal, including a human. All modes of administration can be envisaged, for example, by oral, intravenous, intramuscular, subcutaneous injection or the like. The pharmaceutical dosage form of the composition of the present invention may be used in the form of a pharmaceutically acceptable salt of the active ingredient, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

In another example, there is provided a composition for the prevention and / or improvement of diabetic complications comprising the bear odor extract and / or fraction as described above. In the present invention, the food means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through a certain processing process. , Dietary supplements, beverages, food additives and beverage additives. Examples of the foods include various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, the food in the present invention includes special nutritional products (e.g., prepared oils, infants, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasoned foods ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, ice cream, etc., natural seasonings (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages, and other health supplements. The health functional food, beverages, food additives or beverage additives may be prepared by a conventional manufacturing method.

The food composition may include food acceptable food additives, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of functional foods.

In addition, in the food, the amount of the bear extract and / or fraction may be included in 0.00001% to 50% by weight of the total weight of the food, if the food is a beverage 0.001 g based on 100 ml of the total volume of food To 50 g, preferably 0.01 g to 10 g, but is not limited thereto.

Since the composition comprising bear extract and / or fractions according to the present invention is a composition derived from natural products which is excellent in treating diabetic complications but does not have toxicity and side effects, it can be more safely applied to the treatment of diabetic complications.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are only intended to illustrate the present invention, but the scope of the present invention is not limited by these examples.

Material preparation

Ligularia fischeri ) was purchased from Gangwon-do, Yanggu-gun pharmacist, DPPH (1,1-diphenyl-2-picrylhydrazyl), DMSO (dimethyl sulfoxide), BSA (Albumin from bovin serum), methylglyoxal, amino Guanidine (Aminoguanidine), NADPH, DL-glyceraldehyde, Quercetin (Quercetin), NaCO ₃ was purchased from Sigma (USA), and all other reagents used in the analysis were express.

sample Preparation Example 1

Bear extract and fractions of the present invention is added to 10 times 70% ethanol of 1000 g of dried bear odor weight and extracted at 65 ℃ in an aqueous solution for 3 hours and then filtered to obtain an extract. Repeat the extraction process three times. The obtained extract was concentrated under reduced pressure at 40 ° C. or lower, and then freeze-dried to obtain a powdery extract.

sample Preparation Example 2

Bear odor obtained in Sample Preparation Example 1 500 L of distilled water was added to the extract and suspended. 500 L of hexane was added to the mixture, and the mixture was separated into a hexane-soluble fraction and a water-soluble fraction. The same process was repeated three times to collect only hexane-soluble fractions and dried under reduced pressure to obtain hexane soluble extract powder. Similarly, methylene chloride, ethyl acetate, butanol-soluble fractions and water-soluble fractions were collected separately and dried under reduced pressure. A powder was obtained.

Example  One. Of final glycation end products (AGEs)  Inhibitory effect test

In this example, to examine the therapeutic effect of diabetic complications, the 24-hour short-term screening method using methylglyoxal was used to determine the effect of suppressing the final glycosylated product (AGEs) of the extracts and fractions of the bear odor on the basis of the blank and the control. Measured.

A sample dissolved in DMSO (dimethyl sulfoxide) in BSA (5 mg / mL) and methylglyoxal (2 mg / mL) (extract and fraction of the odor obtained in Example 1) (11 mg / mL) was added thereto, and 50 mM phosphate buffer ( pH 7.4) was used to adjust the total volume to 5 mL. After reacting at 36 ° C. for 24 hours, the absorbance (Excitation: 355 nm, Emission: 460 nm) of the sample before and after the reaction was measured using a spectrofluorometer (Fluoroskan Ascent FL, Thermo Scientific, USA). Inhibition rate of absorbance was calculated by comparing the absorbance increase value of the sample based on the absorbance increase value of the control group.

Calculation = {1- (S ₁ -S ₀ ) / (C ₁ -C ₀ )} * 100

(S ₀ : sample 0hr absorbance value, S ₁ : sample 24hr absorbance value, C ₀ : control 0hr absorbance value, C ₁ : control 24hr absorbance value) were used. In addition, the fluorescence inhibition rate was compared using the method described above using aminoguanidine (aminoguanidine, Sigma Chemical Co. (St Louis, MO, USA)) as a standard substance known to have an inhibitory effect on the final glycation product. The concentrations (IC ₅₀ ) of the samples show a 50% reduction in absorbance relative to the control, and each sample was run three times and averaged.

The obtained results are shown in Table 1 and Table 2. As a result of measuring the increase of fluorescence based on the blank and the control group, the 70% ethanol extract was 71.9% at the concentration of 5.5 mg / mL for the bear extract, showing the highest fluorescence inhibition among the extracts.

In addition, the inhibitory rate and IC ₅₀ of each concentration of the bear odor fraction are shown in Table 2 below.

Table 1. Inhibitory Effect of Bear Extract on AGEs Formation

Table 2. Effect of Inhibitory Formation on AGEs Formation

¹⁾ IC ₅₀ values represent concentrations showing 50% inhibition of AGEs formation. The calculated value is the least-square regression equation for a log curve of three concentrations.

²⁾ Positive Control (Typical AGEs Formation Inhibitors).

As can be seen in Table 1, 2, 70% ethanol extract of Goji odor was shown to have high activity, and the ethyl acetate and butanol fractions had a inhibition rate of 50% at concentrations of 0.795 and 0.621 μg / mL, respectively. appear.

Example  5. Aldos  Reductase ( Aldose reductase Inhibitory efficacy test

The effect of bear extract on aldose reductase activity, a catalytic enzyme of the polyol pathway, a glucose metabolism known to play an important role in the development of diabetic complications, was confirmed. An aldose reductase inhibitor is an enzyme inhibitor that inhibits aldose reductase, which converts glucose into sorbitol, and aims to reduce the production of sorbitol. Since many studies have shown that polyol metabolic abnormalities cause diabetic complications, aldose reductase inhibitors may be useful as treatments for diabetic complications.

Thus, in this embodiment, in order to prove the effect of treating diabetic complications, the ability of inhibiting the aldose reductase activity of the extract and fractions of bear odor.

Lenses weighing more than 200 g (Sprague-Dawly (SD) rats; 250-280 g) were extracted and 0.05 μL sodium buffer (pH 6.8) was added to the lens, depending on the wet weight of the lens therein. Was added to homogenize in ice. After centrifugation (centrifuge UNION 55 R Hanil, Korea) for 30 minutes at 4 ℃, 7500rpm, the supernatant was taken and used as the enzyme source of the aldose reductase test.

The ability to inhibit aldose reductase activity was tested by modifying the method used by Haymanh and Kinoshita (Hayman S, Kinoshita JH. 1965. Isolation and properties of lens aldose reductase. J. Biol. Chem. 240 (2) ): 877-882). Into 530 μL of 0.05M potassium phosphate buffer (pH 7.0), put 160 μL of the above prepared enzyme source, 100 μL of 1.6 mM NADPH (Applichem, Germany), and 10 μL of the sample (Gear extract or fraction obtained in Sample Preparation Example 1, 1 mg / mL). Finally, 100 μL of 0.1M DL-glyceraldehyde (DL-Glyceraldehyde, Sigma, USA) was added. Reduction of NADPH absorbance was measured at 340 nm by reacting for 4.4 minutes in the cuvette. The reaction was carried out for 4.4 minutes in the cuvette, and the absorbance values of the samples were calculated with absorbance values of 0s and 280s. Formula 1 = (absorbance value at 0 s-absorbance value at 280 s) / time of reaction} was used. Inhibition rate was used as the following formula.

Calculation = {(control-sample)-(control-blank)}

In addition, using a quercetin (Quercetin, Sigma Chemical Co. (St Louis, MO, USA)) as a control substance known to have an aldose reductase inhibitory effect was measured in the same manner as described above to measure the rate of decrease in absorbance sample Preparation Example 1 It was compared with the results in the samples prepared in, 2. The concentration of the sample (IC ₅₀ ), indicating a 50% reduction in absorbance for the control material, was expressed, and each sample was repeated three times and averaged.

Table 3. Inhibitory Effects of Bear Extracts on Aldose Reductase Activity

Table 4. Inhibitory Effect of Aldose Reductase Activity on the Bear Fraction

¹⁾ IC ₅₀ value represents the concentration showing 50% inhibition of aldose reductase activity,% inhibition vs. The calculated value is the least-square regression equation for a log curve of three concentrations.

²⁾ positive control substance (typical aldose reductase activity inhibitor).

As can be seen in Tables 3 and 4, high activity was observed in the 70% ethanol extract of Goji odor, and accordingly, the ethyl acetate and butanol fractions had 50% inhibition rate at concentrations of 0.003 and 0.013 μg / mL, respectively. appear. The fractions of the odor ethyl acetate and butanol fractions showed much lower concentrations than the IC ₅₀ values of quercetin.

Example  3. DPPH Antioxidant Effect Test

DPPH is a compound with chemically stable free cardial and is widely used to check the antioxidant capacity when it meets a substance with antioxidant activity. Thus, in this embodiment, the antioxidant effect was evaluated by evaluating the free radical scavenging effect (hydrogen donating ability to DPPH) of the extract and fractions of the bear odor extract using DPPH.

Bear extract and each fraction were prepared by diluting with DMSO (dimethyl sulfoxide). 2 mL of 5Χ 10 ^-4 M DPPH solution (dissolved in 99% ethanol) was added to 1 mL of the prepared sample at a constant concentration, mixed well, and left at room temperature for 30 minutes. The obtained reaction solution was measured by electron donating ability at 517 nm absorbance. The inhibitory capacity of each fraction was calculated from the radical scavenging rate from the reduced absorbance compared to the positive control.

Calculation = {1- (Control / Sample)} * 100

At this time, ascorbic acid (Showa Chemicals Inc., Japan), which is known to have an antioxidant effect, was used as a positive control group, and the results were described by measuring DPPH quenching ability by treating the same method as described above. And compared with the content required for 50% inhibition of DPPH free radicals (IC ₅₀ ), and the results obtained by the three repeated experiments the sample is shown as the average value.

The results of comparing DPPH scavenging ability of the extracts of Goji odor and fractions are shown in Tables 5 and 6, respectively.

 Table 5. DPPH Radical Scavenging Effects of Bear Extract

Table 6. DPPH radical scavenging effect of goji odor fraction

In Table 5 and Table 6,

¹⁾ IC ₅₀ value represents the concentration showing 50% inhibition of DPPH radical,% inhibition vs. The calculated value is the least-square regression equation for a log curve of three concentrations.

²⁾ Positive Control (Typical DPPH Radical Scavengers).

As confirmed in Table 5 and Table 6, it was confirmed that the 70% ethanol extract in the extract of Goji odor has excellent oxidation inhibitory ability, fraction based on this to compare the DP ₅₀ radical inhibition activity of the Goji odor fraction by comparing the IC ₅₀ value Ethyl acetate fraction and butanol fraction showed 0.039 and 0.663 ug / mL, respectively. That is, it was confirmed that Goji odor ethyl acetate fraction and methylene chloride fractions have excellent oxidation inhibitory ability.

Statistical processing

Statistical analysis of all data was carried out using SAS statistical program (SAS institute, 1987), and the results of analysis were expressed as mean and standard error per experimental group. Significance test was performed at the p <0.05 level by Duncan's multiple range test.

Claims (7)

An extract obtained by extracting bear odor from water and at least one selected from the group consisting of C ₁ to C ₄ linear or branched alcohols or mixtures and the extract selected from the group consisting of water, hexane, methylene chloride, ethyl acetate and butanol A composition for the prevention or improvement of diabetic complications containing at least one selected from the group consisting of fractions obtained by fractionating at least one as an active ingredient. The extract obtained by extracting the bear odor with 60 to 80% (v / v) ethanol and the extract obtained by fractionating the extract into at least one selected from the group consisting of hexane, methylene chloride, ethyl acetate, butanol and the like. A composition for preventing or improving diabetes or diabetic complications, containing at least one selected from the group consisting of fractions as an active ingredient. The methylene obtained by separating the hexane fraction obtained by fractionating the extract with hexane and the remaining aqueous layer from methylene chloride, according to claim 2, wherein the extract is obtained by extracting an odor from 60 to 80% (v / v) ethanol. The ethyl acetate fraction obtained by separating the chloride fraction and the methylene chloride fraction and then fractionating the remaining aqueous layer with ethyl acetate, and the ethyl acetate fraction and the ethyl acetate fraction are separated from the remaining water fraction. Prevention or improvement composition. According to any one of claims 1 to 3, wherein the bear odor is a composition for preventing or improving diabetic complications due to diabetes. The composition for preventing or ameliorating diabetic complications according to any one of claims 1 to 3, wherein the diabetic complications are diabetic vascular disorders, diabetic neuropathy or diabetic infections. The method of claim 5, wherein the diabetic complication is characterized by diabetic retinopathy, diabetic cataract, diabetic nephropathy, diabetic neuropathy, diabetic angiopathy A composition for preventing or improving diabetic complications. A composition for preventing or improving diabetes or diabetic complications, comprising the composition according to any one of claims 1 to 3.