KR20070028829A - Composition comprising the extract of dioscorea quinqueloba and acer tegmentosum showing anti-oxidative, anti-aging, anti-lipidperoxidative, anti-edema and expectorant activity - Google Patents

Abstract

A composition comprising the extracts of Dioscorea quinqueloba and Acer tegmentosum showing anti-oxidative, anti-aging, anti-lipidperoxidation, anti-edema and expectorant activity is provided to prevent and treat diseases associated with oxidation, aging and inflammation such as arteriosclerosis. The composition for preventing and treating diseases associated with oxidation, aging and inflammation comprises the extracts of Dioscorea quinqueloba and Acer tegmentosum as effective ingredients, wherein the mixing ratio of Dioscorea quinqueloba and Acer tegmentosum extracts is 1:1-5, and the amount of Dioscorea quinqueloba and Acer tegmentosum extracts is 0.1-50 wt.% based on the total weight of the composition. The extracts of Dioscorea quinqueloba and Acer tegmentosum are prepared with water or C1-C4 lower alcohol. The composition is a pharmaceutical or health food composition and the health food composition is formulated as powder, granule, tablet, capsule or drink.

Description

Composition comprising the extract of Dioscorea quinqueloba and Acer tegmentosum showing anti-oxidative, anti-aging, anti-lipidperoxidative, and antioxidative, anti-aging, lipid peroxidation inhibition, anti-edema and expectorant activity anti-edema and expectorant activity}

1 is a view schematically showing an antioxidant activity test by DPPH,

Figure 2 is a diagram schematically showing the inhibitory effect measurement test by the lipid peroxides (TBARS) in the LDL oxidation induced by the method Cu ^{+ 2,}

3 is a diagram schematically showing an experiment on the effect of suppressing edema in carrageenin-induced mice,

4 is a view schematically showing an experiment for measuring fluidity using mucin (mucin),

5 is a diagram showing the electron donating ability of the mixed extract of each part of maple and bee in the antioxidant activity experiment using DPPH,

Figure 6 is a diagram showing the IC ₅₀ value of the mixed extract of each region of maple and bee in the antioxidant activity experiment using DPPH,

7 is a diagram showing the effect of the mixed extract of each part of maple and bee on LDL lipid peroxidation using TBARS method,

8 is an IC ₅₀ value showing the inhibitory effect of LDL lipid peroxidation of the mixed extracts of maple and bees each part,

9 is a diagram showing the increase rate of the foot edema caused by the administration of the mixed extract of each part of maple and bee trees induced by carrageenin,

10 is a diagram showing the foot edema inhibition rate by the administration of the mixed extract of each part of maple and bee trees induced by carrageenan,

11 is a diagram showing the effect of the administration of the mixed extract of each part of the maple and bee trees using mucin (mucin) on the fluidity change.

The present invention prevents and treats inflammation-related diseases such as oxidation and arteriosclerosis, which contain a mixed extract of maple and bee trees as an active ingredient, which has an antioxidant effect, an anti-aging effect, a lipid peroxidation inhibitory effect, an anti-edema activity, and an expectorant effect. It relates to pharmaceutical compositions and food compositions.

Recently, economic development and cultural benefits have been enjoyed by the development of the industrial society, but our lives are threatened directly and indirectly due to environmental pollution and overnutrition. Therefore, modern people's interest in health is higher than ever, and among them, there is a high interest in functional foods that have the effect of maintaining health or regulating biorhythms, and thus the focus of recent research.

All aerobic organisms, including humans, use oxygen to metabolize energy and are fundamentally self-defense against free radical damage in the process, but the production of free radicals beyond the defenses of tissues is associated with aging and aging. Oxygen detriment has recently been recognized for its associated degenerative diseases and atherosclerosis, hypertension, diabetes, arthritis, circulatory disorders, as well as other causes of cancer.

Active oxygen, commonly called noxious oxygen, is a superoxide anion that is a singlet oxygen produced by reduction of triplet oxygen ( ³ O ₂ ), the safest form of oxygen, during oxidation and reduction. ¹ O ₂ ), free radicals such as hydrogen peroxide (H ₂ O ₂ ) and hydroxyl radicals (OH), which are factors of the immune system such as proteins, DNA, enzymes and T-cells Causes a variety of diseases, especially the problem is that the active oxygen attack the unsaturated fatty acid component of the cell membrane of the cell, causing lipid peroxidation reaction to accumulate lipid peroxide in the body, deteriorate the body function and at the same time cause aging and adult diseases It is known.

Recently, as the theory that aging and adult disease are caused by free radicals is gradually acknowledged, studies on antioxidants known as substances that can control free radicals have been actively conducted, so that the enzyme-based preventive antioxidants, superoxide dismutase (SOD) and catalase Many antioxidants have been developed, including catalase, glutathione peroxidase, and natural antioxidants such as tocopherol, ascorbic acid, carotenoid, glutathione, and synthetic antioxidants BHA and BHT. have.

Meanwhile, bronchial asthma is a disease that is accompanied by coughing, shortness of breath, tightness of the chest, and bronchial spasms, which is statistically common to account for 7 to 10% of the population. It is a disease that can not be eradicated. If the symptoms are severe, it is a condition that replaces all of the treatment by taking bronchodilators, etc. on a temporary basis. In recent years, the number of respiratory related diseases due to environmental pollution tends to increase. have.

Airway mucus has several important donations in prayer. That is, it protects the airway from particles entering the airway from outside, maintains the humidity of the intake air, and removes the inhaled chemicals and gases by the protein of the mucus, cilia action. Mucus also contains substances that play an important role in immune function, such as immunoglobulin A (IgA), lysozyme, lactoferrin, and the like. Most expectorants have been used as folk medicine or experience prescription for thousands of years using plants, and by 1989, Ziment et al. Collectively classified drugs that affect mucus.

Currently, there are two major mechanisms for using plants for seawater asthma. The first is a drug that acts by suppressing the central nervous system's remedy. Codeine may be used as a drug, but it is addictive as a drug. Second, it stimulates the common vagus nerve of the stomach and airways to promote the secretion of mucous membranes. Saponin, akaloids, mucus, and emetic agents are examples of mechanisms that promote excretion, and examples include Gilpyeong and Seoksan.

As described above, the mechanism of saponin-containing plants mainly stimulates the mucous membrane of the stomach, thereby stimulating the mucous membrane of the bronchus with the same function, promoting the secretion of mucus, and promoting the discharge of phlegm by the saponin's surfactant activity. It acts to suppress seawater.

The main material of this experiment, Dioscorea quinqueloba , is a perennial herbaceous herb that belongs to the family Dioscoreaceae.

Several branches are split from the stem, and the surrounding tree trunk grows while winding the rocks. The petiole is long and the leaf shape resembles a maple leaf. It is divided into 5-7 leaves in the shape of a palm. Male and female, flowering in June-July. Fruits with wings ripen in October. Indigenous plants in Korea are D. batatas , chrysanthemum, D. septemloba , D. tokoro , and D. japonica .

The taste of maple leaves is bitter and cold and returns to cirrhosis and menopause. It is known that maple leaves remove customs, make blood flow well, give meridians, break down walls, and stop coughing.

To date, there are no studies on the components and pharmacological activity of maple leaf, but many studies have been conducted on the same genus, Dioscorea, and steroid saponin, dioscin and diosgenin. ) And high amounts of mucus and starch.

In pharmacological experiments to date, Dioscore plants have been reported to lower cholesterol in blood, lower blood pressure, improve blood circulation in coronary vessels, stop coughing and eliminate breathing symptoms. It has been reported to be used for the prevention and treatment of dyskinesia, pain, bruises, goiter, hyperthyroidism, sputum, coughing and shortness of breath, chronic bronchitis and atherosclerosis.

As mentioned above, much research has been conducted on the related plants of the genus Sok, but there are no studies on the composition and activity of maples with high distribution as a resource in Korea.

Accordingly, the present inventors have completed the present invention by confirming that the maple and bee extracts have an antioxidant effect, a lipid peroxidation inhibitory effect, an anti-edema activity and an expectorant effect.

An object of the present invention is to provide a novel use of the maple and bee mixed extract having antioxidant, anti-aging, lipid peroxidation inhibition, anti-edema and expectorant activity, the maple and bee mixed extract of the present invention using DPPH In the experiment, it showed antioxidant effect, reduced TBARS production, confirmed mouse foot edema inhibition effect and expectorant effect to provide pharmaceutical composition and food composition for prevention and treatment of inflammation-related diseases such as various oxidation and arteriosclerosis. For the purpose of

In order to achieve the above object, the present invention provides a pharmaceutical composition for the treatment and prevention of oxidative, aging or inflammation-related diseases containing maple and bee wood mixture extract as an active ingredient.

In addition, the maple and bee are characterized in that the extract is mixed in a ratio of weight ratio (%) of 1: 1 to 5, preferably 1: 1 to 2.

The pharmaceutical composition exhibited antioxidant effects in experiments using DPPH, decreased TBARS production, suppressed mouse foot edema and expectorant effect, thereby exhibiting antioxidant, anti-aging, lipid peroxidation inhibition, anti-edema and expectorant activity. It is characterized by.

In addition, the maple and beech mixed extract includes an extract available in a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.

In addition, the oxidative and inflammation-related diseases include atherosclerosis, rheumatoid arthritis, asthma, bronchitis, acute pain, chronic pain, neuropathic pain, postoperative pain, pains such as migraine and joint pain, neuropathy, nerve damage, irritable bowel syndrome Shock, or inflammatory bowel disease caused by endotoxins.

Hereinafter, the present invention will be described in detail.

The maple and bee mixed extracts of the present invention are dried and dried each of the maple and bee trees, preferably the dried maple, preferably the root of the dried maple, root and root including the tubers and dried bees, respectively 1: 1 About 1 to 20 times, preferably about 10 to 15 times the amount of water, ethanol, methanol, etc., C1 to C4 lower alcohol or about 1: 0.1 to 1:10, preferably These mixed solvents having a mixing ratio (kg / L) of 1: 0.2 to 1: 5 are hydrothermal extraction, cold needle extraction, hot needle extraction, reflux cooling extraction at room temperature for about 1 to 24 hours, preferably 5 to 15 hours. Alternatively, by using an extraction method such as ultrasonic extraction, it is preferable to extract the hot needles, and then, by concentrating at low temperature, the maple and beech mixed extracts of the present invention can be obtained.

The pharmaceutical composition for the treatment and prevention of oxidative, aging or inflammation related diseases of the present invention comprises 0.1 to 50% by weight of the extract based on the total weight of the composition.

Pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical compositions comprising a mixture of maple and bee wood according to the present invention, powder, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, suppositories and sterilization, respectively, according to a conventional method It can be formulated and used in the form of injectable solutions. Carriers, excipients and diluents that may be included in the composition comprising maple leaf and beech extracts include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin , Calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the maple and beech mixed extracts of the present invention vary depending on the condition and weight of the patient, the extent 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. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

Maple and bee wood mixture extract of the present invention can be administered to various mammals such as mice, mice, livestock, humans. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The present invention provides a dietary supplement comprising the maple and bee wood mixture extract and food acceptable food supplements exhibiting the effect of preventing and treating oxidative, aging or inflammation-related diseases. Examples of the food to which the extract can be added include various foods, beverages, gums, teas, vitamin complexes, health functional foods, powders, granules, tablets, capsules or beverages.

In addition, suitability as a "food additive" as defined herein is determined according to the standards and standards for the relevant items in accordance with the General Regulations of the Food Additives Code and General Test Methods, etc., unless otherwise specified.

Items listed in the "Food Additives Code" include, for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as color pigments, licorice extract, crystalline cellulose, high color pigments, guar gum, Mixed preparations, such as a sodium L- glutamate preparation, a noodles addition agent, a preservative preparation, and a tar pigment preparation, etc. are mentioned.

It may also be added to foods or beverages for the purpose of preventing and treating oxidative, aging or inflammation related diseases. At this time, the amount of the extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml. have.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the above-mentioned maple and bee mixed extracts as essential ingredients in the indicated ratios, and various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. It may contain. 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 conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the maple and bee mixed extract of the present invention is a flavor, such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and its Salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. In addition, the extract of the present invention may contain natural fruit juice and fruit flesh for the production of fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Experimental Examples.

Example  One. Maple leaves  And Bee  Mixed Extract

1-1. Maple leaves  Underground and Bee  Preparation of Leaf Blend Extract

Maple was supplied with wild products near Jinju Co., Ltd., and the bee trees were dried using the wild products to dry the underground and beech leaves including maple roots, rhizomes, and tubers, and then dried maple underground and beech leaves. Was mixed at a weight ratio of 1 to 1, and then sufficiently weighed, and 70% ethanol of 10 to 15 times the dry weight was added thereto, and the resultant was concentrated by low temperature by immersion for 5 hours at a total of 5 hours and freeze-dried to obtain ethanol extracts of maple leaf and leaves of maple. (Yield: 64.5%)

1-2. Maple leaves  Underground and Bee  Preparation of Stem Blend Extract

Using the same method as in Example 1-1, the ethanol extract of the maple leaf and the trunk of the maple was obtained by using a 1: 1 ratio of the maple leaf and the trunk of the maple. (Yield: 68.2%)

1-3. Maple leaves  Underground and Bee  Preparation of Blood Extract

Using the same method as in Example 1-1, the ethanol extract of the maple leaf and the beech blood was obtained using the 1: 1 weight ratio of the maple leaf and the beech blood. (Yield: 60.5%)

Reference Example  1. Preparation of experimental animals

Experimental animals were purchased from Hallim Experimental Animal Research Institute (Hwaseong, Gyeonggi-do, Korea) with male white mice with an average weight of 230 ± 10 g of Sprague-Dawley system. 12 hour contrast cycle and constant temperature (21-23 ° C). It was used for experiments after adapting for at least 4 days with free feeding of water and water under constant humidity (55%).

Reference Example  2. Statistical Processing

The analytical results of the experiments were expressed as mean ± standard error (SE), and the significance was the Duncan's multiple test using the one-way ANOVA of the SPSS / PC + package. Analyzed.

Experimental Example  1. Antioxidant Test

1-1. DPPH Using Antioxidant activity  Experiment

Maple underground and bee leaf extract (hereinafter referred to as ATHM), maple underground and bee stem extract (hereinafter referred to as ATSM), maple underground and beech blood extract (hereinafter referred to as ATCM) are referred to as Hatano, etc. 0.1mM DPPH solution (99.5% ethanol) in 0.1 ml (control: 99.5% ethanol) of each fraction prepared in five concentrations of 3000, 2000, 1000, 500 and 250 ppm (99.5% ethanol) 1.9 ml were added. After shaking for 10 seconds with a vortex mixer (Vortex mixer) and incubated for 30 minutes at 37 ℃, the absorbance was measured at 515nm using a spectrophotometer. As a positive control drug, L-ascorbic acid was measured by preparing five concentrations of 500, 250, 100, 50, and 25 ppm (99.5% ethanol). The approximate experimental method is shown in FIG. 1. Antioxidant activity of each sample was represented by the electron donating ability (EDA%) and IC ₅₀ value (μl concentration required to inhibit DPPH radical formation by 50%) for DPPH, and the results are calculated by the following equation (1). 1, Table 2, FIG. 6 and FIG.

As a result, the antioxidant activity by DPPH was significant (p <0.05) compared with the control drug ascorbic acid.

IC ₅₀ also showed significant results (p <0.05).

Sample O.D.value = absorbance of the test solution to which the sample was added

Control O.D.value = 0.1 ml ethanol (instead of sample) + 0.1 ml DPPH 1.9 ml

sample EDA (%) 250 (ppm) 500 (ppm) 1000 (ppm) 2000 (ppm) 3000 (ppm) ATSM 20.58 ± 0.01 ^* 34.69 ± 0.02 ^** 60.84 ± 0.04 ^* 90.81 ± 0.01 ^** 94.08 ± 0.01 ^** ATHM 31.93 ± 0.03 49.03 ± 0.05 81.97 ± 0.07 93.05 ± 0.01 ^** 93.36 ± 0.01 ^** ATCM 31.08 ± 0.01 ^** 55.82 ± 0.02 ^* 87.99 ± 0.01 ^** 93.29 ± 0.02 ^* 94.22 ± 0.01 ^** L-ascorbic acid 25 (ppm) 50 (ppm) 100 (ppm) 200 (ppm) 300 (ppm) 35.41 ± 0.04 ^** 50.28 ± 0.01 ^** 88.89 ± 0.01 ^** 93.26 ± 0.01 ^** 95.75 ± 0.01 ^**

Numeric = mean ± standard error (n = 2), significance from negative control: ^* p <0.05, ^** p <0.01

sample IC ₅₀ (μg / ml) ATSM 50.85 ± 0.25 ^* ATHM 33.02 ± 0.82 ^** ATCM 29.83 ± 0.02 L-ascorbic acid 3.20 ± 0.04

Value = mean ± standard error (n = 2), significance from negative control: ^* p <0.05

1-2. Lipid peroxide ( TBARS Inhibition Effect of Lipid Peroxidation Using

16 mL of human plasma LDL (low-density lipoprotein, 400 μg), 1 mM CuSO ₄ , and 100 μl of each sample (2000, 1000, 500, 250, and 100 ppm) prepared at different concentrations were mixed with PBS (pH 7.4). Was made. The mixture was mixed with a vortex mixer and oxidized by shaking culture for 4 hours in a 37 ° C. water bath, and then 20 µl of 1 mM EDTA was added to stop the oxidation.

1 ml of 25% trichloroacetic acid was added to the oxidized LDL solution to precipitate the protein, and 1 ml of 1% thiobarbituric acid was added to the supernatant, followed by color development at 95 ° C., followed by cooling. . The amount of malondialdehyde (MDA) produced was measured using a spectrophotometer at 532 nm. The approximate experimental method is shown in FIG. 2.

As the MDA standard sample, 10 nM 1,1,3,3-tetraethoxypropane solution was prepared and used, and it was calculated by the following Equation 2, and also to compare the LDL lipid peroxidation inhibitory effect of each sample. the generation of lipid peroxides induced by Cu ^{2 +} concentration of the sample required to inhibit _50% (IC 50) the following Table for each result calculated in equation (3) 3, Table 4, is shown in Figs. 8 and 9 .

MDA concentration (nM / mL) = (f / F) × 10

F: absorbance of standard sample (532 nm)

f: absorbance of the sample (532 nm)

[Ox-LDL]: of oxidized LDL by Cu ^{2 +} MDA

[Sample LDL]: MDA of LDL oxidized by sample addition

[LDL]: MDA of non-oxidized LDL

 sample MDA (nmol / mg protein) 100 ppm 200 ppm 500 ppm 1000 ppm 2000 ppm 3000 ppm LDL 0.618 ± 0.147 Ox-LDL 12.119 ± 2.87 ATSM - 7.31 ± 0.13 ^* 5.56 ± 0.19 ^* 4.10 ± 0.08 ^* 3.60 ± 0.11 ^* 2.52 ± 0.05 ^* ATHM - 7.77 ± 0.23 ^* 5.73 ± 0.26 ^* 3.35 ± 0.20 2.36 ± 0.09 ^* 2.91 ± 0.04 ^* ATCM - 6.68 ± 0.15 ^* 3.44 ± 0.12 ^* 3.02 ± 0.04 ^* 2.01 ± 0.05 ^* 1.45 ± 0.04 ^* L-ascorbic acid 7.55 ± 0.15 ^* 3.95 ± 0.39 2.62 ± 0.25 0.67 ± 0.38 - -

LDL: non-oxidized LDL, Ox-LDL: LDL oxidized by Cu ^{2 +,} value = mean ± SE (n = 2), significant from the negative control ^{group: * p <0.05, ** p} <0.01

sample IC ₅₀ (μg / ml) ATSM 7.07 ± 0.16 ^* ATHM 9.42 ± 0.72 ATCM 2.92 ± 0.73 L-ascorbic acid 0.96 ± 0.08

Value = mean ± standard error (n = 2), significance from negative control: ^* p <0.05, ^** p <0.01

As a result, as shown in Table 3, the results showed significant results when compared with the control drug ascorbic acid, and the concentration-dependent TBARS production was reduced, so that the treatment of maple and bee extracts inhibited lipid peroxidation. Could confirm. IC ₅₀ also showed significant results.

Experimental Example  2. Mouse Carrageenan ( carrageenin Foot Edema Test in Model

Eight white mice of 230 ± 10 g prepared in Reference Example 1 were used as a group, and the weight of each rat and the volume of the foot accurately indicated were measured. The control group was 200 mg / kg of saline, the positive control group was 1 mg / kg of indomethacin (named SMC), and the rest of the experimental groups prepared the extracts of the subterranean and beech parts of maple leaf. Through the experiment, oral administration of 200 mg / kg having the highest discrimination of efficacy for each sample. Each sample and drug were prepared by dissolving in sarin. After 30 minutes of oral administration, 25 mg / kg of carrageenin (Sigma) was injected subcutaneously into the soles of each group. Each group measured the volume of feet accurately indicated from 1 hour to 4 hours later. All experiments were conducted twice. The approximate experimental method is shown in FIG. 3.

Observing the edema increase and decrease of each time, the edema increase rate (calculated by Equation 4) and edema inhibition rate for each time was calculated by comparing with the control group and the results are shown in Table 5, Table 6, FIG. 10 and FIG.

sample Edema growth rate% 1 hour later 2 hours later 3 hours later 4 hours later Control 11.60 ± 0.76 ^** 16.53 ± 1.17 ^** 18.18 ± 1.66 ^** 9.63 ± 1.30 ATSM 6.58 ± 0.47 8.59 ± 0.75 * 7.11 ± 1.05 2.86 ± 0.27 ATSM 7.07 ± 0.57 11.73 ± 0.73 12.41 ± 0.59 4.44 ± 0.96 ATCM 9.03 ± 0.36 14.00 ± 0.138 12.82 ± 0.88 6.35 ± 0.67 IDMC 6.01 ± 0.42 4.18 ± 0.32 3.42 ± 1.03 2.03 ± 0.53

Value = mean ± standard error (n = 2), significance from negative control: ^* p <0.05, ^** p <0.01

sample Edema inhibition rate% 1 hour later 2 hours later 3 hours later 4 hours later ATSM 43.28 ± 2.32 48.03 ± 3.75 60.89 ± 5.24 70.30 ± 1.36 ATSM 39.05 ± 2.87 29.04 ± 3.66 31.74 ± 2.93 53.89 ± 4.79 ATCM 22.16 ± 1.78 15.31 ± 6.90 29.48 ± 4.43 34.06 ± 3.34 IDMC 48.19 ± 2.06 74.71 ± 1.62 81.19 ± 5.09 78.92 ± 2.64

Value = mean ± standard error (n = 2), significance from negative control: ^* p <0.05, ^** p <0.01

As a result, when compared with the reference drug IDMC (Indometacin), it was confirmed that the foot edema suppression effect of the group administered with the extract of each part of the basement of maple and beeswax.

Experimental Example  3. expectoration

3-1. Mucin  Liquidity Measurement

To 10 ml of 0.1% mucin solution, each sample and positive control group (N-Acetyl-L-cysteine) of each part of maple leaf and bee extracts were added dropwise in concentrations of 5, 10, 15, and 20 mg, respectively. Incubation was for 10 minutes, and the control group was treated with only 10 ml of saline. The approximate experimental method is shown in FIG. 4.

The test group treated as described above was measured using a Oswald capillary viscometer (Ostwald viscometer) at 37 ℃ to measure the fall time according to the viscosity to calculate the flow rate (Flow rate) values are shown in Table 7 and FIG.

sample Flow rate St (1St = 1㎠ / second) 5mg 10mg 15mg 20mg ATSM 248 ± 2.83 231 ± 2.48 225 ± 0.71 204 ± 0.71 ATHM 209 ± 3.18 182 ± 0.71 162 ± 1.06 160 ± 1.41 ATCM 228 ± 2.83 220 ± 4.95 225 ± 0.71 223 ± 1.77 NaLc 173 ± 2.12 138 ± 2.83 124 ± 2.83 112 ± 0.71 Saline 278 ± 1.77 269 ± 3.54 275 ± 4.24 276 ± 0.71

Value = mean ± standard error (n = 2), significance from negative control: ^* p <0.05, ^** p <0.01

As a result of the experiment, the saline treated as a control group showed a little activity in the maple leaf and each part of the beech extract administration group, and the maple leaf and beech leaf extract showed the best effect.

Experimental Example  4. Acute Toxicity  exam

Acute toxicity experiments were performed using 6-week-old specific pathogen-free (SPF) SD rats. Two animals of each group were orally administered with maple leaf and bee extract of the present invention at a dose of 100 mg / kg. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed, and hematological and hematological examinations were performed.

As a result, no significant clinical symptoms or dead animals were noted in all animals treated with the test substance, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings. As a result, the extract of the present invention did not show a toxicity change even in rats up to 100 mg / kg, respectively, the minimum lethal dose (LD ₅₀ ) was determined to be a safe substance of more than 100 mg / kg.

The formulation examples of the maple subterranean and bee extract are described, but this is not intended to limit the present invention only intended to be described in detail.

Formulation example  1. Preparation of Injection

Extract of Example 1 ............ 100 mg

Sodium Metabisulfite ... 3.0 mg

Methylparaben .................. 0.8 mg

Propylparaben ....................... 0.1 mg

Sterile Distilled Water for Injection ...

The above ingredients are mixed and made into 2 ml by a conventional method, and then filled into 2 ml ampoules and sterilized to prepare an injection.

Formulation example  2. Preparation of Tablets

Extract of Example 1 ... 200 mg

Lactose ................................... 100 mg

Starch ......................................... 100 mg

Magnesium stearate proper amount

The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.

Formulation example  3. Preparation of Capsule

Extract of Example 1 ..... 100 mg

Lactose ................................... 50 mg

Starch ... 50 mg

Talc ........................................ 2 mg

Magnesium stearate .....

Capsules are prepared by mixing the above ingredients and filling into gelatin capsules according to a conventional method for preparing capsules.

Formulation example  4. Liquid  Produce

Extract of Example 1 ..... 1000 mg

Sugar ... 20 g

Isomerized sugar ...................... 20 g

Lemon flavor ........................

Purified water was added to adjust the total volume to 1000 ml. According to the conventional method for preparing a liquid, the above components are mixed, and then filled into a brown bottle and sterilized to prepare a liquid.

Formulation example  5. Preparation of Ointment

Extract of Example 1 ..... 1000 mg

Purified Lanolin ......................................... 1000 mg

Vaseline ............... 8000 mg

The extract of Example 1 and purified lanolin were completely mixed, and then mixed with petrolatum to prepare an ointment.

Formulation example  6. Manufacture of healthy food

Extract of Example 1 ............................................ 1000 mg

Vitamin Blend ...

Vitamin A Acetate ......... 70 μg

Vitamin E .................................. 1.0 mg

Vitamin B1 ............ 0.13 mg

Vitamin B2 ............... 0.15 mg

Vitamin B6 ...................... 0.5 mg

Vitamin B12 ............... 0.2 μg

Vitamin C ................................. 10 mg

Biotin ......................................... 10 μg

Nicotinic Acid Amide ... 1.7 mg

Folic acid ......................................... 50 ㎍

Calcium Pantothenate ......................................... 0.5 mg

Inorganic Mixture ...

Ferrous Sulfate ............... 1.75 mg

Zinc Oxide ............... 0.82 mg

Magnesium Carbonate ........................... 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dicalcium Phosphate Dibasic ............... 55 mg

Potassium Citrate ... 90 mg

Calcium Carbonate ... 100 mg

Magnesium Chloride ............... 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

Extract of Example 1 ...................................... 1000 mg

Citric acid ..................... 1000 mg

Oligosaccharide ......................... 100 g

Plum concentrate ........................... 2 g

Taurine ..................................... 1 g

Purified water is added .............. 900 ml

After mixing the above components according to a conventional healthy beverage production method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a relatively suitable component for a preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As described above, the maple and bee wood mixture extract of the present invention shows an antioxidant effect, peroxidation inhibitory effect, anti-edema activity and expectorant effect, pharmaceutical composition and food for the prevention and treatment of oxidative, aging or inflammation-related diseases It can be used as a composition.

Claims (4)

Maple horse ( Dioscorea quinqueloba and bee ( Acer) tegmentosum ) A pharmaceutical composition for the treatment and prevention of oxidative, aging or inflammation-related diseases containing a mixed extract as an active ingredient. According to claim 1, wherein the oxidative and inflammation-related diseases include atherosclerosis, rheumatoid arthritis, asthma, bronchitis, acute pain, chronic pain, neuropathic pain, pain after surgery, migraine and joint pain, neuropathy, nerve damage Pharmaceutical composition, which is irritable bowel syndrome, shock or inflammatory bowel disease caused by endotoxin. Health supplements comprising maple and bee wood mixture extract and food acceptable food additives that have an effect of preventing and improving oxidation, aging or inflammation-related diseases. 4. The dietary supplement of claim 3 which is a powder, granule, tablet, capsule or beverage.

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