KR20080093663A - A composition comprising an extract of ailanthus altissima having antioxidative effect - Google Patents

Abstract

An extract of Ailanthus altissima is provided to show high polyphenol content, excellent electron donating capability, SOD like activity, nitrite scavenging capability, xanthine oxidase inhibiting capability and tyrosinase inhibiting capability, thereby being usefully used as a pharmaceutical composition and a health functional food such as tablet, capsule or pill for preventing and treating oxidation related diseases. A pharmaceutical composition for preventing and treating oxidation related diseases comprises 0.1-50 wt.% of an extract of Ailanthus altissima as an effective ingredient, wherein the extract is obtained by extracting roots, stems or leaves of the Ailanthus altissima with water, C1-4 alcohol, or a mixture solvent thereof.

Description

 A composition comprising an extract of Ailanthus altissima having antioxidative effect}

       1 is a diagram showing the total polyphenol content of the leather extract,

       Figure 2 is a diagram showing the electron donating ability of the leather extract,

       3 is a diagram showing the SOD-like activity of the leather extract,

      Figure 4 is a diagram showing the nitrite scavenging ability of the leather extract in the condition of PH 1.2,

      5 is a diagram showing the nitrite scavenging ability of the leather extract in the condition of PH 3.0,

      Figure 6 is a diagram showing the nitrite scavenging ability of the leather extract in the condition of PH 6.0,

      7 is a diagram showing the xanthine oxidase inhibitory activity of the leather extract,

8 is a diagram showing the tyrosinase inhibitory ability of the leather extract.

     The present invention relates to a composition for preventing and treating oxidation-related diseases containing leather extract as an active ingredient.

Recently, as interest in aging and adult diseases increases, studies are being actively conducted to prevent disease by eliminating excessively generated reactive oxygen in vivo. Free radicals occur in aerobic organisms where oxygen, which is absolutely necessary to maintain life, is an electron acceptor and a continuous biochemical reaction is carried out to supply energy. However, due to metabolic imbalances and physicochemical factors such as chemicals and pollution, hydrogen peroxide (H ₂ O ₂ ), superoxide anion (O _2- ), singlet oxygen ( ¹ O ₂ ) , is converted to the hydroxy radical (hydroxy radical, ^_ OH) a high active oxygen (reactive oxygen) such as reactive. These free radicals cause fatal oxygen toxicity due to strong oxidative force, causing cell membrane destruction, enzyme inactivation, lipid oxidation, DNA denaturation, and cell aging, leading to serious diseases such as cancer, brain diseases, stroke, arteriosclerosis, inflammation and autoimmune diseases. Cause physiological disorders (Hammond et al., J. Physion . Pharmacol . 63 , pp 173-187, 1985).

Currently, active research is being conducted to discover and develop useful substances for the treatment of antioxidants and antioxidants that remove free radicals. Various studies have been made to develop antioxidants from compounds such as In addition, the development of antioxidants and bioactive substances useful in herbal medicines used mainly in oriental medicine is being actively promoted (Nam and Kang, J. Food). Sci . Technol ., 36 , pp 338-338, 2000). Bioactive substances are high value-added substances that show remarkable activity in very small amounts, and many kinds are usefully used, and new substances are being actively researched (Kang, S. S et al., Natural product science . Seoul Univ . Publishers , p 71, 1988). Most of the bioactive substances present in natural products are phenolic compounds and are one of the secondary metabolites of various structures and molecular weights. These phenolic compounds include flavonoids, simple phenols, phenolic acids, and phenylprophanoids, and are antibacterial, anti-allergic, antioxidant, anticancer, and decayed. It has been reported to be effective in preventing heart disease and diabetes prevention (Azuma et al., J. Agric . Food Chem ., 47, pp 3963-3966, 1999). Phenolic antioxidants are also known to inhibit oxidation by removing hydrogen by donating hydrogen to alkyl radicals or alkylperoxy radicals during the chain reaction (Labuza, CRC). Rev. Food Technol ., 2, p335, 1973).

Natural antioxidants known to date include tocopherols, flavonoids, gossypol, sesamol, oryzanol and vitamin C (Pszcczola, DE, Food). Tech . , 55 , pp 51-59, 2001). Among them, tocopherol and L-ascorbic acid are preferred as natural antioxidants. Among them, tocopherol has high safety but low ability to inhibit oxidation reaction (Halliwell et al., FASEB J. , 2 , pp2867-2870, 1988) is expensive.

Ailanthus , the material of this experiment altissima ) is a deciduous arborescent of the Simarou baceae, also called a weighted tree, and is wild throughout the country and planted for timber, roadside trees and ornamentals (Kim, TW The woody plants of korea in color 4 th . Kyo-Hak Publishing Co, p 485-486, 1996). In general, the names of leather and oak are often used in confusion. In early spring, young shoots of leaves are used for cooking such as reliefs and wild vegetables, and the plant called the leather is called the oak tree of Meliaceae. Cedrela sinensis A.) (Kim, TW The woody plants of korea in color 4 th . Kyo-Hak Publishing Co, p485-486, 1996; Choi ago How to use wild plants and cultivated. Oh Seong Publishing Co., Seoul, p206, 1992) The leaves of the leather tree are called low leaves, and the fruit is dried by removing Bongancho, the roots or stems, and the dried one is called Low Skin or Low Root Back Skin. It is used as a herbal medicine (Ko Bon-hong, Hangeul- bogam Hangeul Comp. (1456) , Public Library, p555, 1994). Herbal wood's herbal efficacy is bitter and bitter, cool and not poisonous, which lowers heat and removes moistness. are known to be used in treatment of peptic ulcer (gubonhong. Donguibogam wanyeokbon Hangul (Huh me, 1456), the public library, p555, 1994). In addition, the roots of the roots of leather trees are known to have astringent, antimicrobial, anti-inflammatory, anti-inflammatory and insecticidal effects (Jeong, YM, et al., Korean J. Ori . Physiol , Pathol ., 17 , pp914-922, 2003).

Leather wood contains flavonoid compounds such as 5,7-dihydroxychrom-7-neohesperidoside and naringin, and phenols such as 3,4,5-trimethoxyphenol, p-commeric acid, vanillin and vanillic acid. It contains a substance, merosine, tannin, probafen, and alantone, and Kim (Kim, KW, et al., J. Life Sci ., 15 , pp715-725, 2005) reported strong antimicrobial activity. In addition, hepatic function, anticancer activity against acute lymphocytic leukemia, and cell cycle regulation are shown. Chang et al. (Chang, YS, et al., Kor . J. Pharmacogn ., 34 , pp28-32, 2003) It has been reported that the okotolone component exhibits cytofusion inhibition activity of 70.76% of the virus. In addition, he and Zheng, etc. (Ko, YS and Chung, BS , Kor. Life Sci ., 2 , pp129-137, 1970) reported on the composition and properties of leather seed oil for use as a fragrance resource.

      Herbal herbal medicine has been prescribed and used for the total treatment or prevention of related diseases, but its utilization is relatively low because it is not known about the active ingredient, toxicity and its efficacy. However, in Korea, there are folk remedies that have ingested herbal medicines in various ways in daily life. Therefore, it is expected that the development and commercialization of functional foods or food additives with antioxidant function will be easy using herbal medicines. Therefore, it is necessary to provide scientific evidence of useful ingredients and efficacy for their proper use, and the exploration and development of functional materials, including herbal and natural products and active substances in their by-products, promotes efficient use of resources and public health. It is meaningful in that it can contribute to.

The present inventors have completed the present invention by confirming that the leather extract of the present invention exhibits a high polyphenol content, excellent electron donating ability, SOD-like activity, nitrite scavenging activity, xanthine oxidase inhibitory activity and tyrosinase inhibitory activity.

An object of the present invention is to prevent and treat oxidation-related diseases containing a leather extract having a high polyphenol content, excellent electron donating ability, SOD-like activity, nitrite scavenging activity, xanthine oxidase inhibitory activity and tyrosinase inhibitory activity as an active ingredient. To provide a pharmaceutical composition and dietary supplements.

      In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of oxidation-related diseases containing the leather extract as an active ingredient.

      In another aspect, the present invention provides a health functional food for the prevention and improvement of oxidation-related diseases containing the leather extract as an active ingredient.

The leather tree extract comprises an extract extracted from the stem, seed, leaf or root of the leather tree, preferably the root, stem or leaf.

The leather extract may be selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water or a mixed solvent of water and ethanol, more preferably water or 60 to 80% ethanol. Contains extracts.

      The oxidation-related diseases include obesity, hypertension, diabetes, arteriosclerosis, liver disease, lung disease, hyperthyroidism, hypothyroidism, chronic fatigue syndrome, heart disease, hyperlipidemia, stroke, kidney disease, gout, cancer, aging and neurodegenerative diseases It includes, and preferably includes obesity, diabetes, arteriosclerosis, hyperlipidemia, kidney disease and gout.

Hereinafter, the method for obtaining the extract of the present invention will be described in detail.

The leather extract of the present invention is finely ground the roots, stems or leaves of the dry leather trees, such as water, ethanol and methanol in volumes of about 1 to 40 times its weight, preferably about 1 to 30 times its weight. Hot water extraction, ultrasonic extraction, reflux cooling extraction for about 1 to 10 hours, preferably about 2 to 5 hours with C ₁ to C ₄ lower alcohols or mixed solvents thereof, preferably water or 60 to 80% ethanol The extract obtained by extraction under reduced pressure, preferably by reflux cooling extraction, is concentrated under reduced pressure at about 40 to 80 ℃, preferably about 60 to 70 ℃, and then suspended by distilled water. The leather extract of the present invention can be obtained by vacuum lyophilization, hot air drying or drying by spraying, preferably lyophilization.

The present invention provides a pharmaceutical composition for the prevention and treatment of oxidation-related diseases containing the leather wood extract obtained by the above manufacturing process as an active ingredient.

In addition, the leather is a herbal medicine that has been used for a long time as a herbal medicine and edible extracts of the present invention extracted from them can also be used safely in long-term use because there is no problem such as toxicity and side effects.

The pharmaceutical composition for preventing and treating oxidation-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 extracts according to the invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract 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 such solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. Or lactose, gelatin and the like. 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 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 10 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.

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

      In addition, the present invention provides a health functional food containing the leather wood extract obtained by the above manufacturing process as an active ingredient.

      Examples of the food to which the extract of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

      It may also be added to food or beverages for the purpose of preventing and ameliorating oxidation-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 5g, preferably 0.3 to 1g based on 100ml. .

Health functional food of the present invention includes the form of tablets, capsules, pills, liquids and the like.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for having the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. 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 mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said 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 extract of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, colorants and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the extracts of the present invention may contain flesh for the production of natural fruit juices and 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.

Below, The invention is illustrated in detail by the following examples and experimental examples.

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

Reference Example  1. Preparation of Sample

Leather wood used in the present invention After collecting and identifying at Yasan, Namyang-ri, Seolcheon-myeon, Namhae-gun, Gyeongnam, Korea, the stems and leaves were separated, washed with distilled water, and used after drying.The roots were used at the Herbal Medicine Ingredients (Daegu Hannam Pharmaceutical Co.) The roots, stems and leaves of leather trees were prepared by purchasing the low root skins sold.

Reference Example  2. Statistical Processing

The following experimental example shows the results of experiments repeated independently three times as the average ± standard deviation. In order to test the significance between the experimental groups, the ANOVA test was performed using SPSS 12.0, and when there was a significance, Duncan's multiple range test was performed at the p <0.05 level.

Example  1. Water extract of leather roots ( WE -1) Manufacture

      In a round flask attached with a reflux condenser, 1 L of distilled water was added to 100 g of the root sample prepared in Reference Example 1, and the extract obtained by repeating extraction three times for three hours in an 80 ° C water bath was filtered through a filter paper. The mixture was concentrated under reduced pressure using a rotary vacuum evacuator (Eyela 400 series, Japan) and freeze-dried (FD 5510 SPT, Ilshin Korea) to make 13.77 g of powder (hereinafter referred to as WE-1). It used as the sample of an experiment example.

Example  2. 70% Ethanol Extract of Leather Tree Root EE -1) manufacturing

1 L of 70% ethanol was added to a 100 g root sample prepared in Reference Example 1 in a round flask to which a reflux cooling tube was attached, and the extract obtained by repeating extraction three times for 3 hours in a 60 ° C water bath was filtered through a filter paper. It was then concentrated under reduced pressure with a rotary vacuum evaporator (Eyela 400 series, Japan) and lyophilized (FD 5510 SPT, Ilshin Korea) to make 13.38 g of powder (hereinafter referred to as EE-1). It used as the sample of the following experiment example.

Example  3. of leather roots Hydrothermal  extract( HWE -1) manufacturing

3L of distilled water was added to the 100 g seed sample prepared in Example 1 in a round flask to which a reflux cooling tube was attached, and the extract obtained by extracting for 3 hours at 110 ° C. and 1.5 atm using a pressure extractor was filtered through a filter paper, followed by rotary decompression. The mixture was concentrated under reduced pressure with a rotatory vacuum evaporator (Eyela 400 series, Japan) and lyophilized (FD 5510 SPT, Ilshin Korea) to prepare 19.94 g of powder (hereinafter referred to as HWE-1). Used as.

Example  4. Water Extract of Leather Tree Trunk WE -2) Manufacture

Except for changing the root sample of Example 1 to the stem sample was carried out the same process to prepare a water extract of the leather tree stem 3.80 g (hereinafter referred to as WE-2), was used as a sample of the following experimental example.

Example  5. 70% Ethanol Extract of Leather Tree Trunk EE -2) manufacturing

Except for changing the root sample of Example 2 to the stem sample was carried out the same process to prepare 3.37 g of 70% ethanol extract of the leather trunk (hereinafter referred to as EE-2), used as a sample of the following experimental example It was.

Example  6. of leather tree trunk Hydrothermal  extract( HWE -2) manufacturing

Except for changing the root sample of Example 3 to the stem sample was carried out the same process to prepare a 4.69 g of hydrothermal extract of the leather tree trunk (hereinafter referred to as HWE-2), was used as a sample of the following experimental example.

Example  7. Water Extract of Leather Leaves ( WE -3) Manufacture

Except for changing the root sample of Example 1 to the leaf sample was carried out in the same process to prepare a water extract of 12.50 g of leather leaves (hereinafter referred to as WE-3), was used as a sample of the following experimental example.

Example  8. 70% Ethanol Extract of Leather Tree Leaf EE -3) manufacturing

Except for changing the root sample of Example 2 to the leaf sample was carried out the same process to produce 18.75 g of 70% ethanol extract of the leather leaves (hereinafter referred to as EE-3), used as a sample of the following experimental example It was.

Example  9. of leather leaf Hydrothermal  extract( HWE -3) manufacturing

Except for changing the root sample of Example 3 to the leaf sample was carried out the same process to prepare 31.33 g of hydrothermal extract of leather leaves (hereinafter referred to as HWE-3), was used as a sample of the following experimental example.

Experimental Example  1. Measurement of polyphenol content

In order to determine the polyphenol content of the leather extract, experiments were performed using the method described in the literature (AOAC; Association of official analytical chemists, Washington DC USA , 45 , pp 21-22, 2005).

Leather tree extracts prepared in Examples 1 to 9 (WE-1, EE-1, HWE-1, WE-2, EE-2, HWE-2, WE-3, EE-3, HWE-3) Was diluted in distilled water at a concentration of 1 mg / mL, 0.2 mL of Folin-ciocalteu's phenol reagent was added thereto, mixed, and the mixture was allowed to stand at room temperature for 3 minutes, followed by sodium carbonate, Na ₂ CO ₃ ) 0.4 mL saturated solution was added and mixed. 1.4 mL of distilled water was added thereto, reacted at room temperature for 1 hour, and the absorbance was measured at 725 nm using a UV / VIS spectrophotometer. Take the tannic acid so that the final concentration of the total polyphenolic compound is 0, 37.5, 75, 150, 300 ug / mL, and from the standard curve of absorbance measured above, the Association of official analytical chemists, Washington DC USA , 45 , pp21-22, 2005) method to determine the total polyphenol compound content of the leather extract, the results are shown in Table 1 and FIG.

extract Total Polyphenols (mg / g) extract Total Polyphenols (mg / g) extract Total Polyphenols (mg / g)  WE-1  60.14 ± 1.54 WE-2 68.12 ± 0.92 WE-3 80.40 ± 1.06  EE-1 50.67 ± 1.10 EE-2 62.86 ± 1.33 EE-3 78.56 ± 1.75  HWE-1 52.16 ± 1.32 HWE-2 87.51 ± 2.24 HWE-3 64.26 ± 1.47

       As a result, as shown in Table 1, HWE-2 contained 87.51 mg / g of the most polyphenols, and WE-3 and EE-3 contained 80.40 mg / g and 78.56 mg / g, respectively. Indicated. Root extract (WE-1, EE-1, HWE-1) was 52.16 to 60.14 mg / g showed the lowest content among the nine extracts. Therefore, the roots, stems and leaves of the leather tree have a great effect as a natural antioxidant because they contain a large amount of polyphenol, which is known as a major substance that exhibits antioxidant activity, and the stem and leaves have a greater effect than the root of the leather tree. Could know.

Experimental Example 2. Measurement of electron donating ability

To find out the antioxidant activity of the leather extracts, blots using DPPH ( 1,1-diphenyl-2-picryl hydrazyl), which can identify the antioxidant activity according to the degree of reduction of the bioactive substance and the color fading to purple. (Blois, M. S, Nature, 181 , pp1199-1200, 1958) was measured the electron donating ability of the leather extract. Each extract of leather trees prepared in Examples 1 to 9 at concentrations of 0.1, 0.3, 0.5 or 1.0 mg / mL (WE-1, EE-1, HWE-1, WE-2, EE-2, HWE-2, WE-3, EE-3, HWE-3) 2 mL of 0.2 mM DPPH solution dissolved in 99% ethanol was added to 2 mL, and the mixture was reacted at 37 ° C. for 30 minutes. The reaction solution was measured for absorbance at 517 nm, and the electron donating ability was obtained by expressing the difference in absorbance before and after sample addition as a percentage (%), and the results are shown in Table 2 and FIG. 2.

Concentration (mg / mL) 0.1 0.3 0.5 1.0 WE-1 33.49 ± 1.76 62.77 ± 3.65 77.33 ± 0.46 74.83 ± 0.40 EE-1 24.71 ± 1.41 48.21 ± 1.50 66.25 ± 0.95 64.04 ± 1.82 HWE-1 51.61 ± 2.17 80.29 ± 0.51 90.42 ± 0.96 91.25 ± 0.25 WE-2 48.04 ± 2.79 68.86 ± 0.33 69.91 ± 0.25 70.01 ± 0.79 EE-2 46.43 ± 1.76 61.45 ± 0.74 62.78 ± 0.26 63.27 ± 0.37 HWE-2 50.39 ± 3.25 60.86 ± 0.55  63.33 ± 0.67 67.03 ± 0.96 WE-3 27.14 ± 0.76 28.29 ± 0.29 28.38 ± 0.82 29.24 ± 0.17 EE-3 6.92 ± 1.38 10.49 ± 1.44 11.07 ± 1.73 17.47 ± 1.71 HWE-3 39.90 ± 3.00 46.17 ± 1.09 47.64 ± 0.26 47.94 ± 0.13

       As a result, as shown in Table 2, HWE-1 showed the highest activity of 91.25 at the concentration of 1.0 mg / mL, and the highest electron donating ability was found in the order of root> stem> leaf. Compared with water and ethanol extract showed higher electron donating ability. Therefore, the root, stem and leaf extract of the leather tree was found to be a herbal herbal resource with a very high electron donating ability.

Experimental Example  3. SOD Pseudo-activity  Measure

      In order to determine the antioxidant activity of the leather extract, the experiment was performed using the SOD-like activity measurement method related to the antioxidant and anti-aging activity.

Pyrogallol, an oxidase that catalyzes the conversion of hydrogen peroxide (H ₂ O ₂ ) according to the method of Marklund and Marklund, Eur . J. Biochem . , 47 , pp468-474, 1975. ) SOD-like activity was measured by measuring the amount of production. Each extract of leather trees prepared in Examples 1 to 9 at concentrations of 0.1, 0.3, 0.5 or 1.0 mg / mL (WE-1, EE-1, HWE-1, WE-2, EE-2, HWE-2, WE-3, EE-3, HWE-3) 2.6 mL of tris-HCl buffer, 50 mM tris [hydroxymethyl] amino-methane + 10 mM EDTA, calibrated to pH 8.5 in 0.2 mL and pyrogallol ) 0.2 mL was added and reaction was carried out at 25 ° C. for 10 minutes, and then 0.1 mL of 1N hydrochloric acid (HCl) was added to stop the reaction. The amount of oxidized pyrogallol in the reaction solution measured the absorbance at 420 nm, and the difference in absorbance between the addition and non-addition groups of leather extract was expressed as a percentage (%), and the results are shown in Table 3 and FIG. 3. It was.

Concentration (mg / mL) 0.1 0.3 0.5 1.0 WE-1 1.12 ± 0.56 1.30 ± 0.58 2.14 ± 0.85 3.82 ± 0.58 EE-1 1.21 ± 0.91 2.62 ± 1.06 4.73 ± 0.76 10.07 ± 1.52 HWE-1 8.84 ± 0.91 10.94 ± 0.61 12.54 ± 1.02 14.31 ± 0.39 WE-2 0 0 0 0 EE-2 1.36 ± 0.04 4.69 ± 0.43 6.67 ± 1.92 7.66 ± 1.54 HWE-2 5.52 ± 0.97 6.24 ± 0.00 7.84 ± 0.21 10.78 ± 0.85 WE-3 3.21 ± 1.45 5.89 ± 0.47 15.93 ± 1.81 26.77 ± 2.45 EE-3 45.22 ± 1.10 47.79 ± 1.84 49.14 ± 1.49 50.00 ± 0.97 HWE-3 7.81 ± 0.37 18.34 ± 1.50 27.51 ± 0.00 49.07 ± 0.74

       As shown in Table 3, at the concentration of 1.0 mg / mL, EE-3 (50.00%)> HWE-3 (49.07%)> WE-3 (26.77%)> HWE-1 (14.31%)> HWE-2 (10.78%)> EE-1 (10.07%)> EE-2 (7.66%)> WE-1 (3.82%)> WE-2 (0%). Compared to the stem and stem showed the highest SOD activity, and as the concentration of the sample increased SOD-like activity significantly increased. Therefore, the leather extract was found to have an excellent effect on antioxidant and anti-aging by having a high SOD activity.

Experimental Example  4. Nitrite Scavenging power  Measure

To determine whether leather extracts are involved in antioxidant activity by removing nitrite (NaNO ₂ ), which is an easy carcinogen to produce nitrosamines, Kato et al. al ., Agric . Biol . Chem ., 51 , pp1333-1338, 1987) and the nitrite (NaNO ₂ ) removal effect of the pH was measured according to the method.

Each extract of leather wood (WE-1, EE-1, HWE-1, WE) prepared in Examples 1 to 9 at a concentration of 0.1, 0.3, 0.5 or 1.0 mg / mL in 2 mL of 1 mM nitrite (NaNO ₂ ) solution -2, EE-2, HWE-2, WE-3, EE-3, HWE-3), and 0.1N hydrochloric acid (HCI, pH 1.2) and 0.2M citric acid buffer After adjusting the pH of the reaction solution to 1.2, 3.0 or 6.0, respectively, the reaction solution was made up to 10 mL and reacted at 37 ° C. for 1 hour, followed by 1 mL each. To this was added 5 mL of 2% acetic acid and the grease reagent A: B = 1: 1, A; 1% sulfanilic acid in 30% acetic acid, B; 1% naphthylamine in 30% acetic acid) 0.4 mL was added and mixed, followed by reaction at room temperature for 15 minutes. The absorbed samples were measured at 520 nm, and the control group was measured in the same manner as above by adding 0.4 mL of distilled water instead of the grease reagent to determine the percentage difference in absorbance between the addition and non-addition groups of the leather extract. %), And the results are shown in Tables 4 to 6 and FIGS. 4 to 6.

Concentration (mg / mL) (PH 1.2) 0.1 0.3 0.5 1.0 WE-1 20.64 ± 2.75 21.69 ± 2.43 28.04 ± 0.92 33.33 ± 0.00 EE-1 6.31 ± 1.41 11.95 ± 0.42 26.43 ± 1.09 45.77 ± 0.90 HWE-1 4.28 ± 0.93 15.49 ± 1.43 24.13 ± 0.87 45.97 ± 0.87 WE-2 10.61 ± 0.65 16.61 ± 1.39 35.79 ± 0.48 55.17 ± 0.78 EE-2 15.18 ± 1.34 16.17 ± 2.64 33.18 ± 0.79 56.25 ± 0.24 HWE-2 14.43 ± 0.92 19.32 ± 0.24 36.39 ± 1.12 54.07 ± 0.99 WE-3 46.89 ± 0.49 65.83 ± 0.22 93.64 ± 0.19 95.18 ± 0.00 EE-3 55.64 ± 0.11 75.22 ± 0.11 97.66 ± 0.11 98.58 ± 0.18 HWE-3 15.00 ± 0.00 48.95 ± 0.28 74.26 ± 0.19 93.33 ± 0.00

Concentration (mg / mL) (pH 3.0) 0.1 0.3 0.5 1.0 WE-1 4.17 ± 0.00 11.11 ± 2.41 15.28 ± 2.41 29.17 ± 0.00 EE-1 0.21 ± 0.18 1.17 ± 0.46 3.20 ± 1.42 4.58 ± 0.88 HWE-1 5.60 ± 1.00 10.54 ± 1.34 11.59 ± 1.02 17.94 ± 0.38 WE-2 2.12 ± 0.37 5.42 ± 0.23 22.21 ± 0.73 33.04 ± 0.31 EE-2 9.15 ± 0.92 12.44 ± 1.41 22.51 ± 0.37 34.41 ± 0.43 HWE-2 12.70 ± 0.43 16.69 ± 0.36 24.58 ± 0.36 34.34 ± 0.14 WE-3 37.40 ± 0.86 50.88 ± 1.59 80.10 ± 0.11 89.19 ± 0.11 EE-3 41.05 ± 0.10 56.13 ± 0.37 92.99 ± 0.20 98.94 ± 0.78 HWE-3 9.67 ± 0.83 37.77 ± 0.54 58.23 ± 0.40 85.40 ± 0.42

Concentration (mg / mL) (pH 6.0) 0.1 0.3 0.5 1.0 WE-1 0 0 0 0 EE-1 0 0 0 0 HWE-1 0 0 0 5.24 ± 0.65 WE-2 0.67 ± 0.35 1.38 ± 0.08 1.79 ± 0.16 2.54 ± 0.13 EE-2 0 0 1.09 ± 0.17 2.01 ± 0.17 HWE-2 0.67 ± 0.35 1.38 ± 0.08 1.78 ± 0.15 2.54 ± 1.33 WE-3 0 0 0 1.78 ± 1.41 EE-3 1.09 ± 0.89 1.03 ± 0.25 2.72 ± 0.76 4.41 ± 0.57 HWE-3 0 0 0 1.05 ± 0.09

      As a result, as shown in Tables 4 to 6, EE-3 showed the highest nitrite scavenging effect at 98.58% at pH 1.2, and EE-3 had the highest scavenging effect at 98.94% at pH 3.0. At pH 6.0, HWE-1 showed the highest scavenging effect of 5.24%. In general, the lower the pH concentration, the higher the sample concentration. The nitrite scavenging effect increased in the order of leaf> stem> root. Therefore, the root (low root skin), stem and leaf extracts of leather trees, which are used as herbal medicines, have an excellent nitrite scavenging effect, and inhibit the formation and antioxidant activity of nitrosamines when ingested or processed together with products containing nitrite and amine. It was found that there is an excellent effect.

Experimental Example  5. Xanthine Oxidase Inhibition  Measure

Xanthine oxidase oxidizes xanthine into uric acid, which is involved in purine metabolism in vivo and causes gout and kidney disease (Kramer and Curhan, American). Journal of Kidney diseases, 40 , pp37-42, 2002), Stirpe, Corte, Biol . Chem . , 244 , pp3855-3861, to determine whether leather extracts are involved in inhibiting these xanthine oxidases. 1969) xanthine oxidase inhibitory activity measurement experiment was performed.

      Leather tree extracts prepared in Examples 1 to 9 diluted to a concentration of 0.1, 0.5, 1.0 or 2.0 mg / mL (WE-1, EE-1, HWE-1, WE-2, EE-2, HWE- 2, WE-3, EE-3, HWE-3) 0.6 mL of 0.1 M potassium phosphate buffer (pH 7.5) and 0.2 mL of substrate solution containing 2 mM xanthine were added. 0.1 mL of xanthine oxidase (0.2 U / mL) was added thereto and reacted at 37 DEG C for 5 minutes, and then 1 mL of 1N hydrochloric acid (HCl) was added to stop the reaction. The absorbance of the uric acid produced in the reaction solution was absorbed. Measured at 292 nm. The xanthine oxidase inhibitory activity of the leather extract was determined by expressing the percentage decrease in absorbance of the sample solution addition group and the no addition group as a percentage (%), and the results are shown in Table 7 and FIG. 7.

Concentration (mg / mL) 0.1 0.5 1.0 2.0 WE-1 44.24 ± 1.05 91.52 ± 1.05 93.94 ± 1.05 97.58 ± 2.10 EE-1 41.95 ± 1.99 81.61 ± 3.59 91.95 ± 0.99 95.40 ± 1.99 HWE-1 32.05 ± 1.11 86.54 ± 0.00 96.80 ± 1.38 97.44 ± 1.28 WE-2 40.60 ± 0.00 89.60 ± 2.33 93.62 ± 3.08 98.32 ± 1.16 EE-2 32.98 ± 2.13 87.23 ± 2.13 90.07 ± 2.46 93.62 ± 3.69 HWE-2 49.52 ± 0.00 84.47 ± 1.94 87.70 ± 2.97 97.41 ± 1.12 WE-3 74.25 ± 2.32 90.30 ± 1.16 92.64 ± 1.16 94.65 ± 1.16 EE-3 75.71 ± 0.98 89.27 ± 2.59 93.22 ± 1.70 94.62 ± 0.00 HWE-3 78.25 ± 3.53 89.27 ± 3.05 90.11 ± 2.59 92.09 ± 1.96

      As a result of the experiment, as shown in Table 7, all the extracts of the leatherwood showed a very high inhibition rate of 90% or more at a concentration of 2.0 mg / mL, and among them, WE-2 was particularly high at 98.32%. Inhibitory activity was shown. Therefore, the leather extract was effective in preventing goutine from being oxidized to uric acid and preventing gout and kidney disease, and it was found to be excellent as an antioxidant.

Experimental Example  6. Tyrosinase Inhibition  Measure

Synthesis of melanin (melanin) due to tyrosinase has the problem of forming blemishes, freckles, blotch, etc., promoting skin aging and causing skin cancer, and degrading the quality due to browning of vegetables, fruits and fish (Sanchez- Ferrer et al ., Biochem . biophys . Acta ., 1247 , pp 1-11, 1995), to determine whether leather extracts are involved in inhibiting tyrosinase (Yagi et al ., Planta Medica 53 , pp517-519, 1987) and the degree of tyrosinase inhibitory activity was measured according to the method.

      In Examples 1 to 6, diluted with 0.2 mL of a substrate solution of 10 mM L-DOPA dissolved in 0.5 mL of 0.175M sodium phosphate buffer (pH 6.8) and a concentration of 0.1, 0.5, 1.0, or 2.0 mg / mL. To each solution of 0.1 mL of the leather extract (E-1, EE-1, HWE-1, WE-2, EE-2, HWE-2, WE-3, EE-3, HWE-3) 0.2 mL of mushroom tyrosinase (110 U / mL) was added thereto and reacted at 25 ° C. for 2 minutes, and the resulting DOPA chrome was measured at an absorbance of 475 nm. The tyrosinase inhibitory activity of the extract of the leather tree was determined by expressing the percentage decrease in absorbance of the sample solution addition group and no addition group as a percentage (%), and the results are shown in Table 8 and FIG. 8.

Concentration (mg / mL) 0.1 0.5 1.0 2.0 WE-1 2.84 ± 1.77 4.97 ± 1.88 6.62 ± 0.54 7.09 ± 0.36 EE-1 57.70 ± 0.35 60.32 ± 0.20 61.34 ± 0.51 62.01 ± 0.51 HWE-1 63.97 ± 0.15 64.80 ± 0.55 66.46 ± 0.20 67.38 ± 0.76 WE-2 1.41 ± 1.27 3.26 ± 1.30 4.45 ± 1.36 5.21 ± 0.65 EE-2 2.10 ± 0.92 2.55 ± 0.62 3.70 ± 0.86 4.05 ± 1.05 HWE-2 4.27 ± 1.29 5.87 ± 0.81 6.45 ± 1.09 7.73 ± 1.29 WE-3 9.31 ± 0.84 12.99 ± 0.70 14.72 ± 2.09 16.33 ± 1.27 EE-3 2.87 ± 0.45 3.67 ± 0.30 6.74 ± 1.69 11.94 ± 1.34 HWE-3 9.09 ± 1.15 11.36 ± 0.86 12.88 ± 0.67 15.04 ± 1.90

       As a result, as shown in Table 8, at the concentration of 2.0 mg / mL EE-1 and HWE-1 showed the highest inhibitory effect of 67.38%, 62.01%, respectively, in particular HWE-1 0.1 mg / mL In the concentration of 63.97% showed high tyrosinase inhibitory activity. Therefore, leather extract was found to have an excellent effect on the prevention of browning of food by inhibiting tyrosinase.

      Examples of the formulation of the composition are described below, but are not intended to limit the present invention but to explain in detail only.

Formulation Example 1 Preparation of Powder

20 mg of WE-1 of Example 1

Lactose 100 mg

Talc 10 mg

       The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

10 mg of HWE-1 of Example 3

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

       After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

10 mg of HWE-2 from Example 6

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

       According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation example  4. Preparation of Injectables

10 mg of EE-3 from Example 8

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

_{Na 2 HPO 4 · 12H 2 O} 26 mg

       According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

20 mg of HWE-3 from Example 9

10 g of isomerized sugar

5 g of mannitol

Purified water

       According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve it, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation example  6. Manufacture of healthy food

EE-2 1000 mg of Example 5

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 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 Preparation of Healthy Drink

100 mg of WE-3 of Example 7

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Water quantification

      After mixing the above components in accordance with the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated Used to prepare the healthy beverage composition of the invention.

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

      Leather extract of the present invention has a high polyphenol content, excellent electron donating ability, SOD-like activity, nitrite scavenging activity, xanthine oxidase inhibitory activity and tyrosinase inhibitory activity, and thus the pharmaceutical composition for the prevention and treatment of oxidation-related diseases and health It can be usefully used as a functional food.

Claims (7)

Leather Tree ( Ailanthus altissima ) A pharmaceutical composition for the prevention and treatment of diseases related to oxidation, containing the extract as an active ingredient. The pharmaceutical composition according to claim 1, wherein the extract is extracted from roots, stems or leaves of leather trees. The pharmaceutical composition according to claim 1, wherein the extract is an extract available in a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.        According to claim 1, wherein the oxidation-related diseases are obesity, hypertension, diabetes mellitus, arteriosclerosis, liver disease, lung disease, hyperthyroidism, hypothyroidism, chronic fatigue syndrome, heart disease, hyperlipidemia, stroke, kidney disease, gout, cancer Pharmaceutical composition, which is an aging and neurodegenerative disorder.        The pharmaceutical composition according to claim 1, which comprises 0.1 to 50% by weight of the extract based on the total weight of the composition. Health functional food for the prevention and improvement of oxidation-related diseases containing the leather extract as an active ingredient.        The dietary supplement of claim 6 which is a tablet, capsule, pill or liquid.

Images