WO2007120007A1 - Extract of echinosophora koreensis removing hangover and having anti-oxidant activity - Google Patents

Extract of echinosophora koreensis removing hangover and having anti-oxidant activity Download PDF

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Publication number
WO2007120007A1
WO2007120007A1 PCT/KR2007/001868 KR2007001868W WO2007120007A1 WO 2007120007 A1 WO2007120007 A1 WO 2007120007A1 KR 2007001868 W KR2007001868 W KR 2007001868W WO 2007120007 A1 WO2007120007 A1 WO 2007120007A1
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Prior art keywords
extract
hangover
echinosophora koreensis
alcohol
echinosophora
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PCT/KR2007/001868
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French (fr)
Inventor
Hyun Ok Yang
Kil Choo Moon
Saet Byoul Lee
Su-Nam Kim
Kwang-Hyun Cha
Chun Woo Nam
Ki-Oug Yoo
Won-Bae Kim
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Korea Institute Of Science And Technology
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Publication of WO2007120007A1 publication Critical patent/WO2007120007A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages

Definitions

  • the present invention relates to the extract of Echinosophora koreensis removing hangover and having antioxidant activity.
  • Hangover indicates headache, diarrhea, lack of appetite, nausea, vomiting, chill and cold sweat after drinking alcohol and its objective symptoms include lowering of recognition and capacity for locomotion, hematological change and change in hormone.
  • Reasons for hangover are known as dehydration, toxicity of alcohol and alcohol products (acetaldehyde, formaldehyde, acetone, etc.) and deficiency of nutritive substances (blood sugar, vitamin and mineral) by absorption disorder.
  • a degree of hangover greatly depends on genetic individuality and environmental conditions (nutritional conditions, physical conditions, dehydration degree and ' health condition) .
  • Alcohol taken in is metabolized through three routes; 1) metabolization by the actions of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) existing in the gastrointestinal tract or the liver when ethanol density is low; 2) metabolization into acetaldehyde and acetic acid by Microsomal ethanol oxidizing system (MEOS) existing in endoplasmic reticulum when ethanol density is high, and; 3) finally dicomposed into CO 2 and H 2 O via actions of catalase existing peroxisome.
  • ADH alcohol dehydrogenase
  • ADH acetaldehyde dehydrogenase
  • MEOS Microsomal ethanol oxidizing system
  • liver is an organ most influenced by this condition.
  • Various liver disorders such as fatty liver, liver cirrhosis, etc. take place in the long term while symptoms such as headache or heaviness of the head, a decline of concentration, a sour stomach, indigestion, etc. are caused in the short term.
  • Bean-sprout soup, dried Pollack soup, etc. have been widely used in order to remove hangover from old times.
  • bean sprout roots contain affluent aspartic acid known to protect the liver by promoting discomposition of alcohol through helping the functions of ADH. Therefore, hangover removing beverages in the first stage were manufactured and sold using aspartic acid as a main ingredient .
  • ALT, AST, ALP, etc liver-relating enzymes
  • ALT, AST, ALP liver-relating enzymes
  • the materials that are being studied include xylem of Hovenia dulcis Thunberg, fruit extract, persimmon extract, green tea extract, ginseng (red ginseng) extract, etc.
  • the existing patents for hangover comprise, beverage, health food, chewing gum, etc. as formulation and ingredients used include silk fibroin (Korean Patent No. 0528740) , Acanthopanax sessiliflorus, Phragmites communis Trinius, Chaeomeles sinensis KOEBHNE, Prunus tomentosa THUNBERG, rice bran of unpolishing Oryza Sativa (Korean Patent No.
  • Prunus mune Sieb. et Zucc Prunus Patent No. 04965264
  • Patent No. 0403720-22 Pueraria, thunbergiana Bentham, a seed of Cirsium pendulum Fischer, taurine, Curcuma longa
  • Organism is considered to have evolved while developing antioxidative mechanism from self-defense mechanism removing active oxygen species. But generation of active oxygen species exceeding organism's defense ability is the cause of various diseases by damaging immune system genes such as protein, DNA, enzyme and T cells. In addition, it is known that active oxygen species cause peroxidation by attacking unsaturated fatty acid, a component of cell plasma membrane, and peroxidized lipid accumulated in the body causes aging and various diseases.
  • active oxygen species cause peroxidation by attacking unsaturated fatty acid, a component of cell plasma membrane, and peroxidized lipid accumulated in the body causes aging and various diseases.
  • Echinosophora koreensis is a whole plant of Korea sophora, a member of Leguminosae, which is also called as “GaeMiPul", “GaeGoSam” , “NeuSamNaMu” and "The Cane of a Thief” .
  • Echinosophora koreensis is distributed in the north of Yanggu of Gangwon-do, Pyeongannam-do, Hamgyeongnam-do, etc., which are provinces in Korea. The fallen leaves bush of 1 m height grows at the foot of a mountain or the roadside and subterranean stems are well developed.
  • Echinosophora koreensis has pinnate compound leaves and long oval-shaped 13-25 leaves. The surface of a leaf is white and hairy, and the end of the hair is round and its edge is flat. Its thorn-shaped stipules do not fall apart. Yellow flowers bloom in a type of raceme in May. Its inflorescence is 3-5 cm long and buds from the end of a new branch, and the diameter of its flower is about 15 mm. A small bract is lancet-shaped and black, and has hair. Its calyxes are divided into five while two of those on the backside * are somewhat smaller.
  • Echinosophora koreensis has anti-bacterial effect (Sohn, H. -Y. et al., Phytomedicine. 11:666-672, 2004) and antibacterial effect against MRSA (Hironori T et al . , J. Ethnopharmacol . 5027-5034, 1996) .
  • the objective of the present invention is to provide new uses as an anti-hangover agent and an anti-oxidazing agent containing the extract of Echinosophora koreensis.
  • the inventors of the present invention identified that the extract of Echinosophora koreensis, one of alpine plants, presents excellent radical removal ability and reduce density of alcohol and acetaldehyde in blood by promoting in vivo alcohol metabolism through improving activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) . Also they completed the present invention by identifying that the extract of Echinosophora koreensis can remove hangover more efficiently without any side effects than hangover removing beverages currently sold in the market .
  • ADH alcohol dehydrogenase
  • ADH acetaldehyde dehydrogenase
  • the present invention provides the extract of Echinosophora koreensis that is extracted using water, alcohol or a mixture thereof, and removes hangover and has anti-oxidant ability.
  • the present invention further provides an anti- hangover agent including the extract of Echinosophora koreensis as an effective ingredient.
  • the present invention further provides health food for removal hangover including the extract of Echinosophora koreensis as an effective ingredient.
  • the present invention further provides an anti- oxidizing agent including the extract of Echinosophora koreensis as an effective ingredient.
  • the present invention further provides health food for anti -oxidation including the extract of Echinosophora koreensis as an effective ingredient.
  • the present invention further provides cosmetics for anti-oxidation including the extract of Echinosophora koreensis as an effective ingredie ' nt .
  • the present invention further provides anti-aging agent including the extract of Echinosophora koreensis as an effective ingredient.
  • the present invention provides an extract of Echinosophora koreensis removing hangover and having antioxidant ability.
  • Any extraction method known in the art can be used to extract Echinosophora koreensis but the inventors used the following extraction method comprising: 1) obtaining an extract from Echinosophora. koreensis using water, alcohol or a mixture thereof; 2) filtrating the extract from Step 1); and, 3) concentrating the filtrate from Step 2) under the reduced pressure.
  • the extraction time is preferably 2-24 hours, most preferably 3 hours.
  • An extract of Echinosophora koreensis presents excellent radical removal ability (See Table 1 and Figure 1) , improves activities of alcohol dehydrogenase (See Table 2 and Figure 2) and acetaldehyde dehydrogenase (See Table 3 and Figure 3) by dose-dependent manner, and shows better alcohol decomposition ability than hangover removing beverages in the market if it is administrated to animals to be experimented (See Table 4 and Figure 4) . Further, it reduced headache and unpleasantness of gastrointestinal tract at a hangover effect test for human bodies, which also implies that it has better effect than hangover removing beverages in the market (See Table 6) .
  • an extract of Echinosophora koreensis can be useful for hangover removing beverage and health food because it can reduce alcohol density and acetaldehyde density in blood by promoting in vivo alcohol metabolism and remove hangover more efficiently without any side effects than hangover removing beverages currently sold in the market.
  • the present invention further provides an anti- hangover agent including the extract of Echinosophora koreensis as an effective ingredient.
  • the extract of Echinosophora koreensis of the present invention removing hangover can be administered orally or parenterally, and can me produced in general forms of pharmaceutical formulation by mixing effective dose of the extract of Echinosophora koreensis and one or more than two kinds of pharmaceutically allowable carrier or one or more than two kinds of additive.
  • lactose monohydrate, cornstarch, soybean oil, microcrystalline cellulose or D-mannitorl is preferable as a dilluent
  • magnesium stearate or talc is good as a lubricant
  • polyvinyipyrolidone (PVP) or hydroxypropylcellulose (HPC) is preferable as a binder.
  • Ca-CMC sodium starch glycolate, polacrylin potassium or cross-linked polyvinylpyrrolidone is preferable as a disintegrant
  • white sugar, fruit sugar, sorbitol or aspartame is preferable as a sweetener
  • carboxymethylcellulose sodium (Ma-CMC) is preferable as a sweetener
  • beta-cyclodextrin is preferable as a sweetener
  • white bee's wax or xanthan gum is selected as a stabilizer
  • one selected out of methyl p-hydroxy benzoate (methlparaben) , propyl p-hydroxy benzoate (propylparaben) or potassium sorbate is preferable as a preservative.
  • the extract of Echinosophora koreensis of the present invention removing hangover is used in order to efficiently remove hangover through reducing alcohol density and acetaldehyde density in blood by promoting in vivo alcohol metabolism.
  • its formulations are not limited specifically, so it can be used in the forms of beverages, powders, capsules, soft capsules, tablets, gums or adhesive type liquid composition. It further can be used in percutaneous formulations such as emulsions, ointments, gel, creams, patches or inhalers, and agents such as infusion, pills, tablets, powders, granules, etc.
  • the total weight of the composition for removing hangover containing the extract of Echinosophora koreensis is preferably 0.0001 ⁇ 50 weight %, and the most preferable total weight is 0.01 ⁇ 5 weight %.
  • the extract of Echinosophora koreensis of the present invention removing hangover can contain more than one effective ingredient presenting the same or similar functions.
  • Vitamin B, C or E, betacarotin, compounds containing minerals like Ca, Mg, Zn, etc., phosphatide like lecithin, or compounds of maltol, oligosaccharide, amino acid, etc. can be used as supplementary ingredients for the extract of Echinosophora koreensis removing hangover of the present invention.
  • Mixing two or three ingredients among Vitamin C, E, betacarotin, maltol, etc. is more preferable because it can enhance activation effect of an organism.
  • known additives such as natural flavor like Japanese apricot flavor, lemon flavor, pineapple flavor or herb flavor, natural coloring matter like natural fruit juice, Chlorophyllin, Flovonoid, etc., sweetener like fruit sugar, honey, sugar alcohol, sugar, etc., and firming agent like citric acid and citric acid sodium can be used by mixing.
  • the present invention further provides health food for removing hangover including the extract of Echinosophora koreensis removing hangover as an effective ingredient.
  • Health food containing the extract of Echinosophora koreensis of the present invention removing hangover includes traditional remedies for removing hangover such as tea, jelly, juice, extract, beverage, etc.
  • the health food processed in various formulations of the present invention can be useful to remove hangover before and after drinking because it not only has excellent detoxification effect with no side effects, but also allows long-term storage.
  • the extract of Echinosophora koreensis of the present invention removing hangover can be added to health food in order to remove hangover.
  • the extract of Echinosophora koreensis of the present invention can be added as it is, used together with other food or ingredients, or used appropriately in general methods.
  • the mixing volume of effective ingredient can be determined properly according to purposes (prevention, health or treatment) .
  • the extract of Echinosophora koreensis of the present invention is added less than 15 weight part, more preferably less than 10 weight part, for 100 weight part of total ingredients when manufacturing food or beverage.
  • the amount can be less than the above amount, but the effective ingredient can be used more than the above amount because there is no problem in terms of safety issue.
  • the extract of Echinosophora koreensis removing hangover can be added to dairy products including meat, sausages, bread, chocolates, candies, snacks, pizzas, instant noodles, noodles, gums and ice-cream, various soups, beverages, tea, alcoholic drink and vitamin compositions. Actually, all kinds of health food in general meaning are included.
  • Health beverage compositions of the present invention can contain various flavors or natural carbohydrate as additional ingredient just like general beverage.
  • Said natural carbohydrate includes monosaccharide such as glucose and fructose, disaccharide such as maltose and sucrose and polysaccharide such as dextrin, cyclodextrin, and sugar alcohol such as xylitol, sorbitol, erythritol, etc.
  • natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used.
  • the content of said natural carbohydrate is generally 0.01 ⁇ 0.04 g / 100 mi of the composition of the present invention, preferably about 0 . 02 ⁇ 0 . 03 g / 100 mi .
  • the composition of the present invention can contain various nutrients, vitamins, an electrolyte, flavors, coloring agents, pectic acid and its salt, alginic acid and its salt, organic acid, protective colloidal thickener, pH regulator, stabilizer, preservative, glycerin, alcohol, carbonation drink for carbonated beverages, etc.
  • the composition of the present invention can contain natural fruit juice and flesh for producing fruit juice and vegetable juice. These ingredients can be used independently or by combination. Although the content of these ' additives are not that important, they may have weight parts between 0.01 and 0.1 with respect to 100 parts of the composition of the present invention.
  • the present invention provides an anti-oxidizing agent including the extract of Echinosophora koreensis as an effective ingredient.
  • the extract of Echinosophora koreensis of the present invention presents excellent radical removal ability (See Table 1) , so that it can be useful for anti-aging by its excellent anti-oxidant activity.
  • the present invention provides health food for anti- oxidation including the extract of Echinosophora koreensis having anti-oxidant activity.
  • the extract of Echinosophora koreensis of the present invention can be added as it is, used together with other food or ingredients, or used appropriately in general methods.
  • the mixing volume of effective ingredient can be determined properly according to purposes (prevention, health or treatment) .
  • the extract of Echinosophora koreensis of the present invention is added 0.1 ⁇ 5 weight part, preferably 2 or 10 weight part, with respect to 100 parts of the composition of the present invention when manufacturing food or beverage.
  • the amount can be less than the above amount, but the effective ingredient can be used more than the above amount because there is no problem in terms of safety issue.
  • the extract of Echinosophora koreensis removing hangover can be added to dairy products including meat, sausages, bread, chocolates, candies, snacks, pizzas, instant noodles, noodles, gums and ice-cream, various soups, beverages, tea, alcoholic drink and vitamin compositions. Actually, all kinds of health food in general meaning are included.
  • the present invention provides cosmetics for anti- oxidation including the extract of Echinosophora koreensis having anti-oxidant activity.
  • cosmetics for anti-oxidation of the present invention include emulsions, ointments, gel, creams, patches or inhalers, there is no limitation in formulations.
  • the extract of Echinosophora koreensis of the present invention can be added to generally contained skin external substance composition at the ratio of 1 ⁇ 15 weight parts, preferably 2 ⁇ 10 parts, with respect to 100 parts.
  • external preparation for skin of the present invention can contain fat substance, organic solvent, solvent, concentrating agent, gelling agent, softener, anti- oxidizing agent, suspending agent, stabilizer, foaming agent, aromatic, surfactant, water, ionic or non-ionic emulsifier, filler, metal ion sequesterer, chelator, preservative, vitamin, screening agent, humectant, essential oil, dye, color, hydrophile or lipophile activator, lipid vesicle, or supplementaries that are commonly used for external substance for skin in dermatology. Further, the above ingredients can be applied at the volume generally used in dermatology.
  • the present invention provides anti -aging agent including the extract of Echinosophora koreensis as an effective ingredient.
  • the present invention can be useful for anti-aging agent because it secures excellent anti-oxidant activity (See Table 1) .
  • Figure 1 is a graph illustrating the radical removal ability of the extract of Echinosophora koreensis.
  • Figure 2 is a graph illustrating the activities of the extract of Echinosophora koreensis toward alcohol dehydrogenase (ADH) .
  • Figure 3 is a graph illustrating the activities of the extract of Echinosophora koreensis toward acetaldehyde dehydrogenase (ALDH) .
  • Figure 4 is a graph illustrating the alcohol metabolism promotion effect of the extract of Echinosophora koreensis.
  • Example 1 Preparation of the extract of Echinosophora koreensis using alcohol 1 kg whole plant of the dried Echinosophora koreensis was put into an extraction bag and immersed in 10 times volume of 100% methanol. The solution was extracted with reflux flask for three hours, then the extraction solution was filtered with a filter paper after cooling at room temperature. The above extraction solution was concentrated under the reduced pressure using a concentrator at 50°C until the solution is vaporized completely, and crystal products were obtained.
  • Example 2 Preparation of the extract of Echinosophora koreensis using water 1 kg whole plant of the dried Echinosophora koreensis was put into an extraction bag and immersed in 10 times of water. The extraction was performed at 100 ° C for 90 minutes by hot water extraction and the extraction solution was filtered with a filter paper after cooling at room temperature. The extraction solution was concentrated under the reduced pressure using a concentrator at 50 ° C until the solution is vaporized completely, and crystal products were obtained.
  • Anti-oxidant activity was measured using DPPH (1,1- diphenyl-2-picryl-hydrazyl) method in order to confirm anti-oxidant activity of the extract of Echinosophora koreensis in Examples 1-3 prepared by the above methods (Taco, T et al . , Biosci . Biotech. Biochem. 58:1780-1783, 1994; Na, M. K et al . , Nat Prod. Sic. 8:26-29, 2002).
  • DPPH relatively stable free radical, presents maximum absorbance at 517 nm when it exists in the form of radical while it looses absorbance when it is removed.
  • the method can measure anti-oxidant activity using this principle.
  • the extract of Echinosophora koreensis was diluted using 70% ethanol solution half and half in turns in 10 steps at the concentration of 500 ⁇ g/v ⁇ l on a 96 well plate, then, the total volume of the solution was made to 200 ⁇ l by adding DPPH (1, l-diphenyl-2-picryl-hydrazyl) prepared using 100 ⁇ M ethanol solution. After 30 minute reaction at 37 ° C absorbance was measured using a 520 nm ELISA reader and calculated according to [Expression 1] , having a group with no sample as a control group. The concentration of the sample removing 50% of DPPH radical was IC 50 (inhibitory Concentration) (See Table 1 and Figure D .
  • Vitamin C and Trolox were used as positive control groups.
  • the extract of Echinosophora koreensis has excellent radical removal ability. Therefore, the extract of Echinosophora koreensis of the present invention can be useful for anti- aging effect .
  • Zymogen was prepared by dissolving 26.4 mg of liver homogenate from a freeze-dried S9 rat (MOLTOX Co., USA) in 8 ml. of 0.1% BSA (bovine serum albumin) solution in order to make the final concentration 3.3 mg/mC, and filtering with a 0.45 ⁇ m syringe filter.
  • BSA bovine serum albumin
  • Reactive solution was prepared by putting a mixture of 1 mi of distilled water, 0.75 mi of 1 M tris-HCl (pH 8.8), 0.3 mi of 20 nM NAD + , 0.3 mi of ethanol and 0.1 mi of the sample and 0.15 mi of the zymogen into a cuvette, so that the total volume reached 3 mi. Then, the changes in absorbance were measured at 340 nm for 5 minutes after preliminary reaction at 30 ° Cfor 5 minutes. At this time, a group with no sample added was considered as a control group. The activity of alcohol dehydrogenase (ADH) of the sample was measured with relative activity (%) to the control group. (See Table 2 and Figure 2) .
  • ADH alcohol dehydrogenase
  • ADH alcohol dehydrogenase
  • acetaldehyde dehydrogenase For measurement of activity of acetaldehyde dehydrogenase (ALDH) , the generation speed of NADH at 340 nm was used as an index. Reactive solution was prepared by putting a mixture of 2 ml of distilled water, 0.3 vd of 1 M tris-HCl(pH 8.8), 0.1 ml of 20 nM NAD + , 0.1 ml of acetaldehyde and 0.1 ml of the sample and 0.1 ml of the zymogen into a cuvette, so that the total volume reached 3 ml. Then, the changes in absorbance were measured at 340 nm for 5 minutes after preliminary reaction at 30 ° C for 5 minutes. At this time, a group with no sample added was considered as a control group. The activity of acetaldehyde dehydrogenase (ALDH of the sample was measured with relative activity (%) to the control group (See Table 3 and Figure 3) .
  • Example 1 380 280 210
  • Example 3 360 250 190 As a result of the experiment, it was found that the extract of Echinosophora koreensis improves the activity of acetaldehyde dehydrogenase (ALDH) relatively more excellently than the control group and the activity of acetaldehyde dehydrogenase (ALDH) improves in a dose- dependent manner .
  • ADH acetaldehyde dehydrogenase
  • a Sprague Dawley rat of 200-25Og weight was used for the experiment after two-week preliminary breeding through letting it take compressed food and tap water freely. After 18-hour fasting, the test sample was orally administrated by mixing with 0.5% CMC (Carboxy methyl cellulose) solution at the concentration of 500 mg/Kg. After 30 minutes, 3 g/Kg of alcohol was orally administrated, then, blood was collected from the heart after 1, 3 and 5 hours respectively. Collected blood was mixed with the equal amount of perchloric acid and centrifugation was performed at 3,000 rpm for 10 minutes. Then, the above supernatant was mixed with 0.5 volume of buffer solution I and incubated at 4 ° Cfor 30 minutes. Then, centrifugation was performed at 3,000 rpm for 10 minutes again in order to remove protein, and the concentration of ethanol was measured using an ethanol measuring kit (Roche, USA) (See Table 4 and Figure 4) .
  • CMC Carboxy methyl cellulose
  • Preparation Examples 1-3 were implemented by quantifying 5g obtained from Examples 1-3 and 5g of CMC and making the total volume 1,000 ral using reagent grade water. Preparation Examples 1- 3 and Comparison Examples 1-3 were achieved as shown in Table 5. Then, hangover removal effect was measured after having 60 healthy adult men in their 30s divided into 6 groups take the composition.
  • the subjects were not allowed to eat but water from 12 hours prior to the experiment . And they were asked to take 100 ml of compositions obtained from Preparation Examples 1-3 and Comparison Examples 1-3 respectively. Then, they were asked to drink 100 m ⁇ - of 40% alcohol drink after 30 minutes.
  • CMC Carboxy methyl cellulose
  • a separate group administrated with only CMC solution was established as a control group because CMC solution was used for preparation of the sample to be administrated to the subjects.
  • a survey on headache and unpleasantness of gastrointestinal tract was conducted as shown in [Table 6] .
  • the above components are mixed and prepared powders by filling in a sealed package.
  • the above components are mixed and prepared in the form of pill according to the conventional pill preparation method.
  • the above components are mixed and prepared in the form of capsule by filling in gelatin capsules according to the conventional capsule preparation method.
  • Seasoning was prepared by adding 10-15 weight parts of the extract of Echinosophora koreensis of the present invention to 100 weight parts of seasoning to remove hangover .
  • Soup and gravies for meat products and noodles for health improvement were prepared by adding by adding 0.1-5 weight parts of the extract of Echinosophora koreensis of the present invention to 100 weight parts of soup and gravies .
  • Various dairy products such as butter and ice cream were prepared by adding by adding 5-10 weight parts of the extract of Echinosophora koreensis of the present invention to 100 weight parts of milk.
  • Black bean, black sesame and wild sesame were also steamed and dried with known method, and then ground into grain size of 60 mesh powder using a grinder.
  • the pressure of the extract of Echinosophora koreensis of the present invention was reduced and concentrated using a vacuum concentrator, then the atomized extract was dried using a hot-air dry machine. Then, the dried substance was ground into grain size of 60 mesh powder using a grinder
  • the grains, sesames and the extract of Echinosophora koreensis prepared above were mixed according to the following ratio.
  • Grains unpolished rice 30 weight parts, tear-grass 15 weight parts, barley 20 weight parts
  • Sesames wild sesame 7 weight parts, black bean 8 weight parts, black sesame 7 weight parts
  • Echinosophora koreensis of the present invention was prepared by mixing 5-10 weight parts of sugar, 0.05- 0.3 weight parts of citric acid, 0.005-0.02 weight parts of caramel, 0.1-1 weight parts of vitamin and 1-5 weight parts of the extract of Echinosophora koreensis, making syrup by adding 79-94 weight parts reagent grade water to it, sterilizing said syrup at the temperature of 85-98 ° C for 20- 180 minutes, mixing it with cooling water with the ratio of 1:4, and injecting 0.5-0.82 weight parts of carbon dioxide.
  • Health beverages were prepared by mixing supplementary materials such as 0.5 weight parts of liquid fruit sugar, 2 weight parts of oligosaccharide, 2 weight parts of sugar, 0.5 weight parts of salt and 75 weight parts of water, and 10 weight parts of the extract of Echinosophora koreensis, and packing it into glass bottles or PET bottles after sterilization.
  • supplementary materials such as 0.5 weight parts of liquid fruit sugar, 2 weight parts of oligosaccharide, 2 weight parts of sugar, 0.5 weight parts of salt and 75 weight parts of water, and 10 weight parts of the extract of Echinosophora koreensis
  • Vegetable juice for health improvement was prepared by adding 5 g of the extract of Echinosophora koreensis of the present invention to 1 I of tomato or carrot juice.
  • Fruit juice for health improvement was prepared by adding 1 g of the extract of Echinosophora koreensis of the present invention to 1 ( of apple or grape juice.
  • the extract of Echinosophora koreensis that is extracted using water, alcohol or a mixture thereof can be used for an anti-hangover agent because it can reduce alcohol density and acetaldehyde density in blood by promoting in vivo alcohol metabolism through improving activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) .
  • ADH alcohol dehydrogenase
  • ADH acetaldehyde dehydrogenase
  • ADH acetaldehyde dehydrogenase
  • the extract of Echinosophora koreensis of the present invention can get rid of hangover more efficiently than hangover removing beverages in the market, and has no side effects as an anti-oxidant activator due to its effective radical removal.

Abstract

The present invention relates to use of the extract of Echinosophora koreensis that is extracted using water, alcohol or a mixture thereof. Particularly, the extract of Echinosophora koreensis can be used for an ant i -hangover agent because it can reduce alcohol density and acetaldehyde density in blood by promoting in vivo alcohol metabolism through improving activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). In addition, it can be used as an ant i -oxidizing agent because it presents radical removal ability. The extract of Echinosophora koreensis of the present invention can get rid of hangover more efficiently than hangover removing beverages in the market, and has no side effects as an anti-oxidant activator due to its effective radical removal.

Description

EXTRACT OF ECHINOSOPHORA KOREENSIS REMOVING HANGOVER AND HAVING ANTI-OXIDANT ACTIVITY
[Technical Field]
The present invention relates to the extract of Echinosophora koreensis removing hangover and having antioxidant activity.
[Background Art]
Hangover indicates headache, diarrhea, lack of appetite, nausea, vomiting, chill and cold sweat after drinking alcohol and its objective symptoms include lowering of recognition and capacity for locomotion, hematological change and change in hormone. Reasons for hangover are known as dehydration, toxicity of alcohol and alcohol products (acetaldehyde, formaldehyde, acetone, etc.) and deficiency of nutritive substances (blood sugar, vitamin and mineral) by absorption disorder. A degree of hangover greatly depends on genetic individuality and environmental conditions (nutritional conditions, physical conditions, dehydration degree and' health condition) .
In Korea, alcohol consumption has increased dramatically along with economic growth after 1980s. In the year of 1996, average drinking per one adult a day was 6 bottles of beer (Chosun Ilbo, Change in Drinking Culture) . Alcohol was used as a basic excipient for all types of drug in ancient times and has played a great role in social life since it makes people feel good and forget worries by affecting the central nervous system. In the Korean drinking culture where excessive drinking and frequent drinking habits are prevailing, many people are taking much interest in drugs or beverages for hangover. Alcohol itself not only has toxicity, but also changes into harmful substances to human bodies during in vivo metabolism. The substances function in the digestive organs including a brain and a liver harmfully, which creates hangover after drinking alcohol .
A small quantity of alcohol is absorbed at the gullet and the oral mucosa, and approximately 10% is absorbed at the stomach and 90% at the small intestines. Alcohol taken in is metabolized through three routes; 1) metabolization by the actions of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) existing in the gastrointestinal tract or the liver when ethanol density is low; 2) metabolization into acetaldehyde and acetic acid by Microsomal ethanol oxidizing system (MEOS) existing in endoplasmic reticulum when ethanol density is high, and; 3) finally dicomposed into CO2 and H2O via actions of catalase existing peroxisome. In case that proper amount of alcohol is taken in, all symptoms caused from alcohol are not generated because said metabolism system becomes harmoniously, but if a great deal of alcohol is taken in, balance of metabolism is destroyed and homeostasis in the body cannot be maintained. The liver is an organ most influenced by this condition. Various liver disorders such as fatty liver, liver cirrhosis, etc. take place in the long term while symptoms such as headache or heaviness of the head, a decline of concentration, a sour stomach, indigestion, etc. are caused in the short term.
Bean-sprout soup, dried Pollack soup, etc. have been widely used in order to remove hangover from old times. Particularly, bean sprout roots contain affluent aspartic acid known to protect the liver by promoting discomposition of alcohol through helping the functions of ADH. Therefore, hangover removing beverages in the first stage were manufactured and sold using aspartic acid as a main ingredient .
There are currently dozens of hangover removing beverages in the Korean market such as "Condition" (CJ) using Jari (a leaf of Juglandaceae) , Astragalus membranaceus Bunge, Lotus seed, embryo bud of rice, etc., "Thank You" (ChongKunDang) using chitosan with high purity as a main ingredient, Dawn 808 (Glami) using Alnus japonica Thunberg Steudel and Sorbus commixta extract, etc. In addition, relevant studies have been announced and related patent applications are filed. Particulary, companies are executing clinical researches into changes in ethanol density and acetaldehyde density in blood, changes in activity of liver-relating enzymes (ALT, AST, ALP, etc) and asking patients detailed questions about hangover as function comparison data among products are allowed to be used for advertisement these days. The materials that are being studied include xylem of Hovenia dulcis Thunberg, fruit extract, persimmon extract, green tea extract, ginseng (red ginseng) extract, etc.
In Korea, most studies on hangover are being performed through animal test while human body tests are commonly performed overseas. Researchers in Japan are actively studying focusing on development of hangover removal ingredients, and researchers in the US and Europe is focusing on the symptoms of ethanol (methanol) addiction, causal substances of toxicity and the mechanism of toxicity. Particularly, structures of tetrahydroisoquinoline or tetrahydro-β-carbolines produced by in vivo metabolism of ethanol are similar to morphine, which generates withdrawal symptoms causing ethanol dependence. It is generally known that reasons for ethanol dependence are a decrease in enzyme metabolizing ethanol and partially lowered reaction of brain nerve cells. Recently, metabolization of a very small amount of methanol contained in alcohol (ethanol) and formaldehyde have been pointed as reasons for hangover. Studies are entering new stage as many studies claims that hangover by alcohol is because of alcohol by-products such as acetaldehyde, acetone, etc. (Moser J, Bagchi D, Akubue PI, Stohs SJ.. Alcohol Alcohol. 28:287-295, 1993). In Japan, a patent teaches that fermented products of Oryza sativa extract contributes to lowering the density of acetaldehyde, not that of alcohol in blood. Also a study revealed that one with severe hangover showed higher density of acetaldehyde than one with light hangover while there was little difference in the densities of ethanol (Wall TL, Horn SM, Johnson ML, Smith TL, Carr LG., J Stud. Alcohol. 61:13-17, 2000), which implies that the causal substance of hangover by alcohol is acetaldehyde.
The existing patents for hangover comprise, beverage, health food, chewing gum, etc. as formulation and ingredients used include silk fibroin (Korean Patent No. 0528740) , Acanthopanax sessiliflorus, Phragmites communis Trinius, Chaeomeles sinensis KOEBHNE, Prunus tomentosa THUNBERG, rice bran of unpolishing Oryza Sativa (Korean Patent No. 0528388), Artemisia asiatica, Lycium chinense Mill, Zingiber officinale Rose, Fraxini Cortex, Lotus seed, Hovenia dulcis Thunberg, Morus alba Linn, Schizandra chinensis Baillon (Korean Patent No. 0512912), Astaxanthin, aspartic acid (Korean Patent No. 0520985), persimmon leaves, persimmon (Korean Patent No. 0504351) , Fraxini Cortex, Poria cocos Wolf, Artemisiae Scopariae Herba,
Pueraria thunbergiana Bentham, Pueraria thunbergiana Benth
(Korean Patent No. 0500373), asparagus, green tea, onion,
Prunus mune Sieb. et Zucc (Korean Patent No. 0496524),
Aralia elata seem (Korean Patent No. 0462329), bean, bean sprout, grape seeds (Korean Patent No. 0455075) , cricket
(Korean Patent No. 0453475), Hovenodulinol - an active ingredient of Hovenia dulcis Thunberg fruit (Korean Patent
No. 0452128), radish, Artemisia capillaris Thunb, Pueraria lobata, persimmon leaves, Saururus chinensis Baill (Korean Patent No. 0451013), decursinol (Korean ' Patent No. 0448680), Hovenia dulcis Thunberg, Jigumock (stream skin of Hovenia dulcis) , Hovenia dulcis Thunberg mixture, Pueraria thunbergiana Benth, Cynanchum wilfordii Hemsley, Dioscorea japonica Thunberg, Paeonia aliflora Pall (Korean Patent No. 0446061) , Imperata cylindrica Beauvois, Morus alba Linn, Cuscuta chinensis Lamark, Pueraria thunbergiana Bentham,
Atractylodes lancea D. C, Rubus coreanus Miquel, Duchesnea indica Focke, Sonchus oleraceus L., Citrus unshiu Marcor
(Korean Patent No. 0442771) , pine needle (Korean Patent No. 0412425) , the xylem of Hovenia dulcis Thunberg (Korean
Patent No. 0403720-22), Pueraria, thunbergiana Bentham, a seed of Cirsium pendulum Fischer, taurine, Curcuma longa
Linn (Korean Patent No. 0392876) , Naringin, Naringenin
(Korean Patent No. 0375047) , Alnus japonica Thunberg Steudel, Sorbus commixta (Korean Patent No. 0181168) , etc. Other ingredients used in the existing patents include Fomes fomentarius, green tea catechin, Acanthopanax sessiliflorus, Opuntia ficus-indica, Gastrodiae Elata BL, Hericium erinaceum, Amomum globosum Loureiro, propolis, magnetized hexagonal water, Capsoεfiphon fulvescens, Evodiae Fructus, etc. However, although hangover removing beverage, food, chewing gum, etc. have been developed using the above ingredients so far, none of them presents outstanding effects .
All kinds of aerobic organisms including human survive through energy metabolism using oxygen (O2) . However, in vivo oxygen changes to harmful active oxygen species such as superoxide anion radical (O2-) , hydrogen peroxide (H2O2) , hydroxy radical ( • OH), etc. if it gaining various physical, chemical and biological stress, causing fatal physiological disorder and diseases, and further taking life.
Organism is considered to have evolved while developing antioxidative mechanism from self-defense mechanism removing active oxygen species. But generation of active oxygen species exceeding organism's defense ability is the cause of various diseases by damaging immune system genes such as protein, DNA, enzyme and T cells. In addition, it is known that active oxygen species cause peroxidation by attacking unsaturated fatty acid, a component of cell plasma membrane, and peroxidized lipid accumulated in the body causes aging and various diseases. (Halliwell, B., Drugs 42:569, 1999; Fukuzawa, K and Takaishi, Y., J. Act. Dxyg. Free Rad. 1:55, 1990; Neuzil, J., Gebicki, J. M. and Stocker, R., Biochem. J. 293:601, 1993; Sies, H., ed., Oxidative stress, 1985; Steinberg. D., et al . , N. Engl. J. Med. 320:915. 1989) .
As a theory that active oxygen species cause aging and geriatric diseases has been approved recently, investigation of anti-oxidizing agents regulating active oxygen species is being conducted actively. Specifically, a lot of studies on anti-oxidant enzymes such as superoxide dismutase, peroxidase, catalase, glutathione peroxidase, etc. and low molecular anti-oxidants from natural ingredients such as tocopherol, ascorbate, carotenoid, glutathione, etc. are being implemented (Chang, S. S., et al., J. Food Sic. 42:1102, 1977; Hammerschmidt , P. A. and Pratt, D. E., J". Food Sic. 43:556, 1964). Also many of synthetic anti-oxidizing agents such as 2 , 6-di-tert-butyl- 4-hydroxyanisole (BHA) were developed and are widely used in the pharmaceutical and food industry (Kitahara, K. , et al., Chem Pharm. Bull. 40:2208, 1992; Hatano, T., Natural Medicines 49:357, 1995; Masaki, H., et al . , Biol. Pharm. Bull. 18:162, 1995) .
However, use of synthetic anti-oxidizing agents is gradually restricted because of consumers' avoidance of synthetic anti-oxidizing agents and its carcinogenicity reported through animal test using a large amount of administration of synthetic anti-oxidizing agents (Branen, A. L., JAOCS 52:59, 1975). Therefore, development of new safer natural anti-oxidizing agents with excellent effect is desperately needed.
Echinosophora koreensis is a whole plant of Korea sophora, a member of Leguminosae, which is also called as "GaeMiPul", "GaeGoSam" , "NeuSamNaMu" and "The Cane of a Thief" . Echinosophora koreensis is distributed in the north of Yanggu of Gangwon-do, Pyeongannam-do, Hamgyeongnam-do, etc., which are provinces in Korea. The fallen leaves bush of 1 m height grows at the foot of a mountain or the roadside and subterranean stems are well developed. The plant proliferates with subterranean stems and its dark brown branches have hair. Winter buds are not visible because the hair covers them. Echinosophora koreensis has pinnate compound leaves and long oval-shaped 13-25 leaves. The surface of a leaf is white and hairy, and the end of the hair is round and its edge is flat. Its thorn-shaped stipules do not fall apart. Yellow flowers bloom in a type of raceme in May. Its inflorescence is 3-5 cm long and buds from the end of a new branch, and the diameter of its flower is about 15 mm. A small bract is lancet-shaped and black, and has hair. Its calyxes are divided into five while two of those on the backside* are somewhat smaller. It contains alkaloid and flavonoid. It has been reported that Echinosophora koreensis has anti-bacterial effect (Sohn, H. -Y. et al., Phytomedicine. 11:666-672, 2004) and antibacterial effect against MRSA (Hironori T et al . , J. Ethnopharmacol . 5027-5034, 1996) .
[Disclosure] [Technical Problem]
The objective of the present invention is to provide new uses as an anti-hangover agent and an anti-oxidazing agent containing the extract of Echinosophora koreensis.
[Technical Solution]
The inventors of the present invention identified that the extract of Echinosophora koreensis, one of alpine plants, presents excellent radical removal ability and reduce density of alcohol and acetaldehyde in blood by promoting in vivo alcohol metabolism through improving activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) . Also they completed the present invention by identifying that the extract of Echinosophora koreensis can remove hangover more efficiently without any side effects than hangover removing beverages currently sold in the market .
The present invention provides the extract of Echinosophora koreensis that is extracted using water, alcohol or a mixture thereof, and removes hangover and has anti-oxidant ability.
The present invention further provides an anti- hangover agent including the extract of Echinosophora koreensis as an effective ingredient. The present invention further provides health food for removal hangover including the extract of Echinosophora koreensis as an effective ingredient.
The present invention further provides an anti- oxidizing agent including the extract of Echinosophora koreensis as an effective ingredient.
The present invention further provides health food for anti -oxidation including the extract of Echinosophora koreensis as an effective ingredient. The present invention further provides cosmetics for anti-oxidation including the extract of Echinosophora koreensis as an effective ingredie'nt .
The present invention further provides anti-aging agent including the extract of Echinosophora koreensis as an effective ingredient.
Hereinafter, the present invention is described in detail.
The present invention provides an extract of Echinosophora koreensis removing hangover and having antioxidant ability.
The inventors extracted using the whole plant of dried Echinosophora koreensis in order to study its effect on hangover. Any extraction method known in the art can be used to extract Echinosophora koreensis but the inventors used the following extraction method comprising: 1) obtaining an extract from Echinosophora. koreensis using water, alcohol or a mixture thereof; 2) filtrating the extract from Step 1); and, 3) concentrating the filtrate from Step 2) under the reduced pressure. It is preferable to use ethanol or methanol for alcohol in Step 1) and its volume is preferably 5~10 times. The extraction time is preferably 2-24 hours, most preferably 3 hours.
An extract of Echinosophora koreensis presents excellent radical removal ability (See Table 1 and Figure 1) , improves activities of alcohol dehydrogenase (See Table 2 and Figure 2) and acetaldehyde dehydrogenase (See Table 3 and Figure 3) by dose-dependent manner, and shows better alcohol decomposition ability than hangover removing beverages in the market if it is administrated to animals to be experimented (See Table 4 and Figure 4) . Further, it reduced headache and unpleasantness of gastrointestinal tract at a hangover effect test for human bodies, which also implies that it has better effect than hangover removing beverages in the market (See Table 6) . Therefore, an extract of Echinosophora koreensis can be useful for hangover removing beverage and health food because it can reduce alcohol density and acetaldehyde density in blood by promoting in vivo alcohol metabolism and remove hangover more efficiently without any side effects than hangover removing beverages currently sold in the market.
The present invention further provides an anti- hangover agent including the extract of Echinosophora koreensis as an effective ingredient.
The extract of Echinosophora koreensis of the present invention removing hangover can be administered orally or parenterally, and can me produced in general forms of pharmaceutical formulation by mixing effective dose of the extract of Echinosophora koreensis and one or more than two kinds of pharmaceutically allowable carrier or one or more than two kinds of additive.
In the present invention, all kinds of pharmaceutically allowable carrier and additive are available. Specifically, lactose monohydrate, cornstarch, soybean oil, microcrystalline cellulose or D-mannitorl is preferable as a dilluent, magnesium stearate or talc is good as a lubricant, and polyvinyipyrolidone (PVP) or hydroxypropylcellulose (HPC) is preferable as a binder.
Also one selected out of carboxymethylcellulose calcium
(Ca-CMC) , sodium starch glycolate, polacrylin potassium or cross-linked polyvinylpyrrolidone is preferable as a disintegrant , white sugar, fruit sugar, sorbitol or aspartame is preferable as a sweetener, carboxymethylcellulose sodium (Ma-CMC) , beta-cyclodextrin, white bee's wax or xanthan gum is selected as a stabilizer, and one selected out of methyl p-hydroxy benzoate (methlparaben) , propyl p-hydroxy benzoate (propylparaben) or potassium sorbate is preferable as a preservative.
The extract of Echinosophora koreensis of the present invention removing hangover is used in order to efficiently remove hangover through reducing alcohol density and acetaldehyde density in blood by promoting in vivo alcohol metabolism. And its formulations are not limited specifically, so it can be used in the forms of beverages, powders, capsules, soft capsules, tablets, gums or adhesive type liquid composition. It further can be used in percutaneous formulations such as emulsions, ointments, gel, creams, patches or inhalers, and agents such as infusion, pills, tablets, powders, granules, etc.
Based on in vitro and in vivo experiments, the total weight of the composition for removing hangover containing the extract of Echinosophora koreensis is preferably 0.0001 ~ 50 weight %, and the most preferable total weight is 0.01 ~ 5 weight %. The extract of Echinosophora koreensis of the present invention removing hangover can contain more than one effective ingredient presenting the same or similar functions.
Besides the above main ingredients, Vitamin B, C or E, betacarotin, compounds containing minerals like Ca, Mg, Zn, etc., phosphatide like lecithin, or compounds of maltol, oligosaccharide, amino acid, etc. can be used as supplementary ingredients for the extract of Echinosophora koreensis removing hangover of the present invention. Mixing two or three ingredients among Vitamin C, E, betacarotin, maltol, etc. is more preferable because it can enhance activation effect of an organism.
In addition to the above ingredients, known additives such as natural flavor like Japanese apricot flavor, lemon flavor, pineapple flavor or herb flavor, natural coloring matter like natural fruit juice, Chlorophyllin, Flovonoid, etc., sweetener like fruit sugar, honey, sugar alcohol, sugar, etc., and firming agent like citric acid and citric acid sodium can be used by mixing.
The present invention further provides health food for removing hangover including the extract of Echinosophora koreensis removing hangover as an effective ingredient.
Health food containing the extract of Echinosophora koreensis of the present invention removing hangover includes traditional remedies for removing hangover such as tea, jelly, juice, extract, beverage, etc. The health food processed in various formulations of the present invention can be useful to remove hangover before and after drinking because it not only has excellent detoxification effect with no side effects, but also allows long-term storage.
The extract of Echinosophora koreensis of the present invention removing hangover can be added to health food in order to remove hangover. When it is used as food additive, the extract of Echinosophora koreensis of the present invention can be added as it is, used together with other food or ingredients, or used appropriately in general methods. The mixing volume of effective ingredient can be determined properly according to purposes (prevention, health or treatment) . Generally, the extract of Echinosophora koreensis of the present invention is added less than 15 weight part, more preferably less than 10 weight part, for 100 weight part of total ingredients when manufacturing food or beverage. However, for health and hygiene or long-term intake for health control, the amount can be less than the above amount, but the effective ingredient can be used more than the above amount because there is no problem in terms of safety issue.
There is no specific limitation in kinds of the food. The extract of Echinosophora koreensis removing hangover can be added to dairy products including meat, sausages, bread, chocolates, candies, snacks, pizzas, instant noodles, noodles, gums and ice-cream, various soups, beverages, tea, alcoholic drink and vitamin compositions. Actually, all kinds of health food in general meaning are included.
Health beverage compositions of the present invention can contain various flavors or natural carbohydrate as additional ingredient just like general beverage. Said natural carbohydrate includes monosaccharide such as glucose and fructose, disaccharide such as maltose and sucrose and polysaccharide such as dextrin, cyclodextrin, and sugar alcohol such as xylitol, sorbitol, erythritol, etc. For sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used. The content of said natural carbohydrate is generally 0.01 ~ 0.04 g / 100 mi of the composition of the present invention, preferably about 0 . 02 ~ 0 . 03 g / 100 mi .
In addition to the above, the composition of the present invention can contain various nutrients, vitamins, an electrolyte, flavors, coloring agents, pectic acid and its salt, alginic acid and its salt, organic acid, protective colloidal thickener, pH regulator, stabilizer, preservative, glycerin, alcohol, carbonation drink for carbonated beverages, etc. Besides, the composition of the present invention can contain natural fruit juice and flesh for producing fruit juice and vegetable juice. These ingredients can be used independently or by combination. Although the content of these ' additives are not that important, they may have weight parts between 0.01 and 0.1 with respect to 100 parts of the composition of the present invention.
The present invention provides an anti-oxidizing agent including the extract of Echinosophora koreensis as an effective ingredient.
Radical generated in the body causes fatal physiological disorder and diseases, and further taking life. The extract of Echinosophora koreensis of the present invention presents excellent radical removal ability (See Table 1) , so that it can be useful for anti-aging by its excellent anti-oxidant activity.
The present invention provides health food for anti- oxidation including the extract of Echinosophora koreensis having anti-oxidant activity.
When it is used as food additive, the extract of Echinosophora koreensis of the present invention can be added as it is, used together with other food or ingredients, or used appropriately in general methods. The mixing volume of effective ingredient can be determined properly according to purposes (prevention, health or treatment) . Generally, the extract of Echinosophora koreensis of the present invention is added 0.1~5 weight part, preferably 2 or 10 weight part, with respect to 100 parts of the composition of the present invention when manufacturing food or beverage. However, for health and hygiene or long-term intake for health control, the amount can be less than the above amount, but the effective ingredient can be used more than the above amount because there is no problem in terms of safety issue.
There is no specific limitation in kinds of the food. The extract of Echinosophora koreensis removing hangover can be added to dairy products including meat, sausages, bread, chocolates, candies, snacks, pizzas, instant noodles, noodles, gums and ice-cream, various soups, beverages, tea, alcoholic drink and vitamin compositions. Actually, all kinds of health food in general meaning are included.
The present invention provides cosmetics for anti- oxidation including the extract of Echinosophora koreensis having anti-oxidant activity.
Although cosmetics for anti-oxidation of the present invention include emulsions, ointments, gel, creams, patches or inhalers, there is no limitation in formulations. When preparing cosmetics for anti-oxidation including the extract of Echinosophora koreensis, the extract of Echinosophora koreensis of the present invention can be added to generally contained skin external substance composition at the ratio of 1 ~ 15 weight parts, preferably 2 ~ 10 parts, with respect to 100 parts.
Besides the extract of Echinosophora koreensis, external preparation for skin of the present invention can contain fat substance, organic solvent, solvent, concentrating agent, gelling agent, softener, anti- oxidizing agent, suspending agent, stabilizer, foaming agent, aromatic, surfactant, water, ionic or non-ionic emulsifier, filler, metal ion sequesterer, chelator, preservative, vitamin, screening agent, humectant, essential oil, dye, color, hydrophile or lipophile activator, lipid vesicle, or supplementaries that are commonly used for external substance for skin in dermatology. Further, the above ingredients can be applied at the volume generally used in dermatology.
The present invention provides anti -aging agent including the extract of Echinosophora koreensis as an effective ingredient.
Active oxygen generated during metabolism is the cause of aging (Harman, D., Free radical theoryof aging: Role of free radicals in the origination and evolution of life,
Aging and Disease Processes, p.3-49, Alan R Liss, New York,
1986) . The present invention can be useful for anti-aging agent because it secures excellent anti-oxidant activity (See Table 1) .
[Description of Drawings]
The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which:
Figure 1 is a graph illustrating the radical removal ability of the extract of Echinosophora koreensis.
Figure 2 is a graph illustrating the activities of the extract of Echinosophora koreensis toward alcohol dehydrogenase (ADH) .
Figure 3 is a graph illustrating the activities of the extract of Echinosophora koreensis toward acetaldehyde dehydrogenase (ALDH) .
Figure 4 is a graph illustrating the alcohol metabolism promotion effect of the extract of Echinosophora koreensis.
[Mode for Invention]
The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless otherwise specified.
<Example 1> Preparation of the extract of Echinosophora koreensis using alcohol 1 kg whole plant of the dried Echinosophora koreensis was put into an extraction bag and immersed in 10 times volume of 100% methanol. The solution was extracted with reflux flask for three hours, then the extraction solution was filtered with a filter paper after cooling at room temperature. The above extraction solution was concentrated under the reduced pressure using a concentrator at 50°C until the solution is vaporized completely, and crystal products were obtained.
As a result of the above method, 68 g of Echinosophora koreensis extract was prepared out of 1 kg of the dried Echinosophora koreensis .
< Example 2> Preparation of the extract of Echinosophora koreensis using water 1 kg whole plant of the dried Echinosophora koreensis was put into an extraction bag and immersed in 10 times of water. The extraction was performed at 100 °C for 90 minutes by hot water extraction and the extraction solution was filtered with a filter paper after cooling at room temperature. The extraction solution was concentrated under the reduced pressure using a concentrator at 50 °C until the solution is vaporized completely, and crystal products were obtained.
As a result of the above method, 46 g of Echinosophora koreensis extract was prepared out of 1 kg whole plant of the dried Echinosophora koreensis .
< Example 3> Preparation of the extract of Echinosophora koreensis using a mixture of water and alcohol
1 kg whole plant of the dried Echinosophora. koreensis was put into an extraction bag and immersed in 10 times of 80% methanol (water: methanol = 2 : 8) . The solution was extracted with reflux flask for three hours, then the extraction solution was filtered with a filter paper after cooling at room temperature. The extraction solution was concentrated under the reduced pressure using a concentrator at 50 °C until the solution is vaporized completely, and crystal products were obtained. As a result of the above method, 59 g of Echinosophora koreensis extract was prepared out of 1 kg whole plant of the dried Echinosophora koreensis .
Experimental Example 1> Measurement of radical removal ability
Anti-oxidant activity was measured using DPPH (1,1- diphenyl-2-picryl-hydrazyl) method in order to confirm anti-oxidant activity of the extract of Echinosophora koreensis in Examples 1-3 prepared by the above methods (Taco, T et al . , Biosci . Biotech. Biochem. 58:1780-1783, 1994; Na, M. K et al . , Nat Prod. Sic. 8:26-29, 2002). DPPH, relatively stable free radical, presents maximum absorbance at 517 nm when it exists in the form of radical while it looses absorbance when it is removed. The method can measure anti-oxidant activity using this principle. Particularly, the extract of Echinosophora koreensis was diluted using 70% ethanol solution half and half in turns in 10 steps at the concentration of 500 βg/vλl on a 96 well plate, then, the total volume of the solution was made to 200 μl by adding DPPH (1, l-diphenyl-2-picryl-hydrazyl) prepared using 100 μM ethanol solution. After 30 minute reaction at 37°C absorbance was measured using a 520 nm ELISA reader and calculated according to [Expression 1] , having a group with no sample as a control group. The concentration of the sample removing 50% of DPPH radical was IC50 (inhibitory Concentration) (See Table 1 and Figure D .
[Expression l]
Control Group Absorbance - Sample Absorbance
DPPH Radical Removal Activity(%) = X 100
Control Group Absorbance
[Table l]
Radical Removal Ability of the extract of Echinosophora koreensis (IC50, Unit: μg/m4)
Figure imgf000029_0001
)
Example 10 . 62
1
Example 9 . 97
2
Example 10 . 27
3
Vitamin 2 . 51
C
Trolox 3 . 14
In the experiment, Vitamin C and Trolox were used as positive control groups. As a result, it was found that the extract of Echinosophora koreensis has excellent radical removal ability. Therefore, the extract of Echinosophora koreensis of the present invention can be useful for anti- aging effect .
< Experimental Example 2> Measurement of Activity of Alcohol Dehydrogenase (ADH) (in vitro)
Zymogen was prepared by dissolving 26.4 mg of liver homogenate from a freeze-dried S9 rat (MOLTOX Co., USA) in 8 ml. of 0.1% BSA (bovine serum albumin) solution in order to make the final concentration 3.3 mg/mC, and filtering with a 0.45 μm syringe filter. For measurement of activity of alcohol dehydrogenase (ADH) , the generation speed of NADH at 340 nm was used as an index. Reactive solution was prepared by putting a mixture of 1 mi of distilled water, 0.75 mi of 1 M tris-HCl (pH 8.8), 0.3 mi of 20 nM NAD+, 0.3 mi of ethanol and 0.1 mi of the sample and 0.15 mi of the zymogen into a cuvette, so that the total volume reached 3 mi. Then, the changes in absorbance were measured at 340 nm for 5 minutes after preliminary reaction at 30°Cfor 5 minutes. At this time, a group with no sample added was considered as a control group. The activity of alcohol dehydrogenase (ADH) of the sample was measured with relative activity (%) to the control group. (See Table 2 and Figure 2) .
[Table 2]
Activity of alcohol dehydrogenase (ADH) of the extract of Echinosophora koreensis (Unit: %, Concentration βg/mi)
Figure imgf000030_0001
As a result of the experiment, it was found that the extract of Echinosophora koreensis. improves the activity of alcohol dehydrogenase (ADH) relatively more excellently than the control group and the activity of alcohol dehydrogenase (ADH) improves in a dose-dependent manner. < Experimental Example 3> Measurement of Activity of Acetaldehyde Dehydrogenase (ALDH) (in vitro)
For measurement of activity of acetaldehyde dehydrogenase (ALDH) , the generation speed of NADH at 340 nm was used as an index. Reactive solution was prepared by putting a mixture of 2 ml of distilled water, 0.3 vd of 1 M tris-HCl(pH 8.8), 0.1 ml of 20 nM NAD+, 0.1 ml of acetaldehyde and 0.1 ml of the sample and 0.1 ml of the zymogen into a cuvette, so that the total volume reached 3 ml. Then, the changes in absorbance were measured at 340 nm for 5 minutes after preliminary reaction at 30 °C for 5 minutes. At this time, a group with no sample added was considered as a control group. The activity of acetaldehyde dehydrogenase (ALDH of the sample was measured with relative activity (%) to the control group (See Table 3 and Figure 3) .
[Table 3] Activity of acetaldehyde dehydrogenase (ALDH) of the extract of Echinosophora koreensis (Unit: %, Concentration
100 μg/ml 50 μg/ml 10 μg/ml
Example 1 380 280 210
Example 2 350 250 180
Example 3 360 250 190 As a result of the experiment, it was found that the extract of Echinosophora koreensis improves the activity of acetaldehyde dehydrogenase (ALDH) relatively more excellently than the control group and the activity of acetaldehyde dehydrogenase (ALDH) improves in a dose- dependent manner .
< Experimental Example 4> Measurement of Ethanol Decomposition Ability (in vitro)
A Sprague Dawley rat of 200-25Og weight was used for the experiment after two-week preliminary breeding through letting it take compressed food and tap water freely. After 18-hour fasting, the test sample was orally administrated by mixing with 0.5% CMC (Carboxy methyl cellulose) solution at the concentration of 500 mg/Kg. After 30 minutes, 3 g/Kg of alcohol was orally administrated, then, blood was collected from the heart after 1, 3 and 5 hours respectively. Collected blood was mixed with the equal amount of perchloric acid and centrifugation was performed at 3,000 rpm for 10 minutes. Then, the above supernatant was mixed with 0.5 volume of buffer solution I and incubated at 4°Cfor 30 minutes. Then, centrifugation was performed at 3,000 rpm for 10 minutes again in order to remove protein, and the concentration of ethanol was measured using an ethanol measuring kit (Roche, USA) (See Table 4 and Figure 4) .
[Table 4]
Alcohol decomposition ability of the extract of Echinosophora koreensis (Unit: q/i)
Figure imgf000033_0001
As a result, it was found that the extract of Echinosophora koreensis reduces the amount of alcohol by promoting alcohol metabolism in blood very excellently.
< Experimental Example 5> Measurement of Hangover Removal Effect
In order to check hangover removal effect of the composition of the present invention, Preparation Examples 1-3 were implemented by quantifying 5g obtained from Examples 1-3 and 5g of CMC and making the total volume 1,000 ral using reagent grade water. Preparation Examples 1- 3 and Comparison Examples 1-3 were achieved as shown in Table 5. Then, hangover removal effect was measured after having 60 healthy adult men in their 30s divided into 6 groups take the composition.
[Table 5] Composition
Figure imgf000034_0001
The subjects were not allowed to eat but water from 12 hours prior to the experiment . And they were asked to take 100 ml of compositions obtained from Preparation Examples 1-3 and Comparison Examples 1-3 respectively. Then, they were asked to drink 100 m^- of 40% alcohol drink after 30 minutes. CMC (Carboxy methyl cellulose) is widely used for preparation of beverages because it functions as thickener and stabilizer when it is used for beverages. And a separate group administrated with only CMC solution was established as a control group because CMC solution was used for preparation of the sample to be administrated to the subjects. After 4 hours of taking alcohol, a survey on headache and unpleasantness of gastrointestinal tract was conducted as shown in [Table 6] .
[Table 6] A survey on hangover removal
Figure imgf000035_0001
Figure imgf000036_0001
As shown in the above results, it was found that the extract of Echinosophora koreensis has an effect on removing hangover and the extract can reduce major symptoms of hangover such as headache and unpleasantness of gastrointestinal tract more efficiently than the existing C Company's product.
The following are manufacturing examples for composition removing hangover of the present invention.
Manufacturing Example 1> Preparation of Pharmaceutical Agent
1. Preparation of powders Extract of Echinosophora koreensis 2 g Lactose 10 g
The above components are mixed and prepared powders by filling in a sealed package.
2. Preparation of tablets
Extract of Echinosophora koreensis 100 mg
Corn starch 100 mg
Lactose 100 mg Magnesium stearate 2 mg
The above components are mixed and prepared in the form of pill according to the conventional pill preparation method.
3. Preparation of capsules
Extract of Echinosophora koreensis 100 mg Corn starch 100 mg
Lactose 100 mg Magnesium stearate 2 mg
The above components are mixed and prepared in the form of capsule by filling in gelatin capsules according to the conventional capsule preparation method.
< Manufacturing Example 2> Preparation of Food
The following are food preparations including.
1. Preparation of seasoning
Seasoning was prepared by adding 10-15 weight parts of the extract of Echinosophora koreensis of the present invention to 100 weight parts of seasoning to remove hangover .
2. Preparation of flour food Food for health improvement was prepared by adding 0.5-5 weight parts of the extract of Echinosophora koreensis of the present invention to 100 weight parts of flour and making bread, cakes, cookies, crackers and noodles using said mixture.
3. Preparation of soup and gravies
Soup and gravies for meat products and noodles for health improvement were prepared by adding by adding 0.1-5 weight parts of the extract of Echinosophora koreensis of the present invention to 100 weight parts of soup and gravies .
4. Preparation of dairy products
Various dairy products such as butter and ice cream were prepared by adding by adding 5-10 weight parts of the extract of Echinosophora koreensis of the present invention to 100 weight parts of milk.
5. Preparation of grain powder Alpha unpolished rice, barley, glutinous rice and tear-grass dried with known method were ground into grain size of 60 mesh powder using a grinder.
Black bean, black sesame and wild sesame were also steamed and dried with known method, and then ground into grain size of 60 mesh powder using a grinder. The pressure of the extract of Echinosophora koreensis of the present invention was reduced and concentrated using a vacuum concentrator, then the atomized extract was dried using a hot-air dry machine. Then, the dried substance was ground into grain size of 60 mesh powder using a grinder
The grains, sesames and the extract of Echinosophora koreensis prepared above were mixed according to the following ratio.
Grains (unpolished rice 30 weight parts, tear-grass 15 weight parts, barley 20 weight parts) ,
Sesames (wild sesame 7 weight parts, black bean 8 weight parts, black sesame 7 weight parts) ,
Dried powder of the extract of Echinosophora koreensis (3 weight parts) ,
Ganoderma lucidum Karst (0.5 weight parts), Rehmannia glutinosa (0.5 weight part) .
< Manufacturing Example 3> Preparation of Beverage
1. Preparation of carbonated beverage
Carbonated beverage containing the extract of
Echinosophora koreensis of the present invention was prepared by mixing 5-10 weight parts of sugar, 0.05- 0.3 weight parts of citric acid, 0.005-0.02 weight parts of caramel, 0.1-1 weight parts of vitamin and 1-5 weight parts of the extract of Echinosophora koreensis, making syrup by adding 79-94 weight parts reagent grade water to it, sterilizing said syrup at the temperature of 85-98 °C for 20- 180 minutes, mixing it with cooling water with the ratio of 1:4, and injecting 0.5-0.82 weight parts of carbon dioxide.
2. Preparation of functional beverage to remove hangover A mixture of 0.1 weight parts of Vitamin, 5.8 weight parts of fruit sugar, 3.8 weight parts of white sugar, 0.12 weight parts of citric acid, 0.03 weight parts of malic acid, 0.04 weight parts of citric acid sodium, 0.02 weight parts of gardenia colors and 3 weight parts of the extract of Echinosophora koreensis of the present invention was completely dissolved in measured water. The above beverage was made to 100 weight parts using measured water.
3. Preparation of health beverages Health beverages were prepared by mixing supplementary materials such as 0.5 weight parts of liquid fruit sugar, 2 weight parts of oligosaccharide, 2 weight parts of sugar, 0.5 weight parts of salt and 75 weight parts of water, and 10 weight parts of the extract of Echinosophora koreensis, and packing it into glass bottles or PET bottles after sterilization.
4. Preparation of vegetable juice
Vegetable juice for health improvement was prepared by adding 5 g of the extract of Echinosophora koreensis of the present invention to 1 I of tomato or carrot juice.
5. Preparation of fruit juice
Fruit juice for health improvement was prepared by adding 1 g of the extract of Echinosophora koreensis of the present invention to 1 ( of apple or grape juice.
The following are manufacturing examples for antioxidant composition of the present invention.
< Manufacturing Example 4> Preparation of external substance for skin
1. Preparation of cream cetostearyl alcohol 2.8 weight parts beeswax 2.6 weight parts stearic acid 1.4 weight parts lipophile glyceryl monostearate 2 weight parts peg-100 stearate 1 weight part sorbitan sesquioleate 1.4 weight parts jojoba oil 4 weight parts squalane 3.8 weight parts polysorbate 60 1.1 weight parts macadamia oil 2 weight parts tocopherol acetate 0.2 weight parts methyl polysiloxane 0.4 weight parts ethyl paraben 0.1 weight parts propyl paraben 0.1 weight parts
Euxyl K-400 0.1 weight parts
1,3-butylene glycol 7 weight parts methyl paraben 0.05 weight parts glycerine 6 weight parts d-panthenol 0.2 weight parts extract of Echinosophora koreensis 4.6 weight parts triethanolamine 0.2 weight parts pt 41891 0.2 weight parts
P-H2O 46.05 weight parts
2. Preparation of emulsion cetostearyl alcohol 1.6 weight parts stearic acid 1.4 weight parts lipophile glyceryl monostearate 1.8 weight parts peg-100 stearate 2.6 weight parts sorbitan sesquioleate 0.6 weight parts squalane 4.8 weight parts macadamia oil 2 weight parts jojoba oil 2 weight parts tocopherol acetate 0.4 weight parts methyl polysiloxane 0.2 weight parts ethyl paraben 0.1 weight parts propyl paraben 0.1 weight parts
1,3- butylene glycol 4 weight parts methyl paraben 0.1 weight parts xanthan gum 0.1 weight parts glycerine 4 weight parts d- panthenol 0.15 weight parts allantoin 0.1 weight parts extract of Echinosophora koreensis 3.5 weight parts carvone (2% aq. Sol) 4 weight parts triethanolamine 0.15 weight parts ethanol 3 weight parts pt 41891 0.1 weight parts
P-H2O 48.3 weight parts
[industrial Applicability]
As studied in the above, the extract of Echinosophora koreensis that is extracted using water, alcohol or a mixture thereof can be used for an anti-hangover agent because it can reduce alcohol density and acetaldehyde density in blood by promoting in vivo alcohol metabolism through improving activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) . In addition, it can be used as an anti-oxidant activator because it presents radical removal ability. The extract of Echinosophora koreensis of the present invention can get rid of hangover more efficiently than hangover removing beverages in the market, and has no side effects as an anti-oxidant activator due to its effective radical removal.
Those skilled in the art will appreciate that the conceptions and specific embodiments disclosed in the foregoing description may be readily utilized as a basis for modifying or designing other embodiments for carrying out the same purposes of the present invention. Those skilled in the art will also appreciate that such equivalent embodiments do not depart from the spirit and scope of the invention as set forth in the appended claims.

Claims

What is Claimed is: [Claim l]
An extract of Echinosophora koreensis which is extracted using water, alcohol or a mixture of the two, and removes hangover and has anti-oxidant activity.
[Claim 2]
The extract of Echinosophora koreensis as set forth in Claim 1, wherein the alcohol is of Ci or C3 lower alcohol.
[Claim 3]
The extract of Echinosophora koreensis as in any one of Claims 1 and 2, wherein the alcohol is ethanol or methanol, and the extract is extracted with reflux flask.
[Claim 4]
The extract of Echinosophora koreensis as set forth in Claim 1, wherein hangover removal activity is accomplished through improving alcohol dehydrogenase (ADH) activity.
[Claim 5]
The extract of Echinosophora koreensis as set forth in Claim 1, wherein hangover removal activity is accomplished through improving Acetaldehyde dehydrogenase (ALDH) activity.
[Claim 6]
An anti-hangover agent comprising the extract of Echinosophora koreensis of Claim 1 as an effective ingredient .
[Claim 7]
An anti-oxidizing agent comprising the extract of Echinosophora koreensis of Claim 1 as an effective ingredient.
[Claim 8]
A cosmetic composition for anti-oxidant comprising the extract of Echinosophora koreensis of Claim 1 as an effective ingredient.
[Claim 9]
An anti-aging agent comprising the extract of Echinosophora koreensis of Claim 1 as an effective ingredient .
[Claim lθ]
A health food for removing hangover comprising the extract of Echinosophora koreensis of Claim 1 as an effective ingredient. [Claim ll]
An anti-oxidant health food comprising the extract of Echinosophora koreensis of Claim 1 as an effective ingredient .
PCT/KR2007/001868 2006-04-17 2007-04-17 Extract of echinosophora koreensis removing hangover and having anti-oxidant activity WO2007120007A1 (en)

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RU2625963C1 (en) * 2016-03-25 2017-07-20 Игорь Владимирович Кривопалов-Москвин Anti-alcoholic beverage "api"
KR20220105767A (en) * 2021-01-21 2022-07-28 충북대학교 산학협력단 Saururus chinensis extract using oriental processing and its use for external skin

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KR100912290B1 (en) 2007-11-02 2009-08-17 한국과학기술연구원 Novel 6,8-di?,?-demethylallyl-3,5,7,2',4',6'-hexahydroxy- flavanone or pharmaceutically acceptable salt thereof, preparation method thereof and composition for removing hangover containing the same as an active ingredient

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CN103083391A (en) * 2011-10-31 2013-05-08 韩国科学技术研究院 Composition for preventing and/or alleviating hangover comprising extracts of sophora flavescens
US9132160B2 (en) 2011-10-31 2015-09-15 Korea Institute Of Science And Technology Composition for preventing and/or alleviating hangover comprising extracts of sophora flavescens
RU2625963C1 (en) * 2016-03-25 2017-07-20 Игорь Владимирович Кривопалов-Москвин Anti-alcoholic beverage "api"
KR20220105767A (en) * 2021-01-21 2022-07-28 충북대학교 산학협력단 Saururus chinensis extract using oriental processing and its use for external skin
KR102478898B1 (en) 2021-01-21 2022-12-19 충북대학교 산학협력단 Saururus chinensis extract using oriental processing and its use for external skin

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