KR20030070232A - Composition containing an extract of evodiae fructus for the alleviation of alcohol-induced hangover and antioxidative activity - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing an extract of Evodia officinalis Dode having hangover-reducing effect and antioxidation effect is provided. Therefore, the composition can be used in a drug and health supplementary food showing hangover-reducing effect and antioxidant effect for protection of the liver from damage caused by alcohol consumption. CONSTITUTION: The composition for hangover and antioxidant action comprises 0.5 to 50% by weight of an Evodia officinalis Dode extract as an effective ingredient and a pharmaceutically acceptable carrier. The Evodia officinalis Dode is obtained by extracting in lower alcohol or lower alcohol acetate at 10 to 50deg.C for 1hr to 2 days, concentrating and then fractioning in an organic solvent selected from the group consisting of n-hexane, methanol, ethanol and ethyl acetate. The extract is administered to a patient in an amount of 0.1 to 500mg/kg from one time to several times on a daily basis.

Description

COMPOSITION CONTAINING AN EXTRACT OF EVODIAE FRUCTUS FOR THE ALLEVIATION OF ALCOHOL-INDUCED HANGOVER AND ANTIOXIDATIVE ACTIVITY}

The present invention relates to a composition for hangover relief and antioxidant activity, including sewage oil extract.

While new drug development has emerged as an important issue in modern medicine, natural products with a history of clinical use, especially for thousands of years, are emerging in the treatment of diseases.

Osuyu (吳 vo, Evodia officinalis ) is a deciduous arborescent from China. It grows mainly in Gyeongju province. It is 5 m high and has hairs on young branches. It is similar to a citrus tree, but has many leaflets, hairs on the back, and rounded fruit tips. In Oriental medicine, the fruit is called Evodiae Fructus , and it is used as dry stomach, old wind, detoxification and diuretic (李 昌 福. 大大 物 韓 植. 鄕 文 社. (1989) )).

As economic and cultural levels improve, alcohol consumption is on the rise with the introduction of westernized eating habits (Economic Bureau of Economic Planning and Statistics, Korean Social Indicators, pp.164, Seoul (1995)). In Korea, the average annual alcohol consumption per capita is the 23rd in the world, and 1 out of 10 adult males are regular drinkers who drink almost daily, and 68.4% of all adults over 20 years of age consumed alcohol (Kim In-sook, Ethanol intake). Effects of Sitau Reinforcement on Body Ethanol Concentration and Metabolism in Rats, MS Thesis, Department of Food and Nutrition, Yonsei University (1999). In addition, since 1989, the drinking rate has generally decreased, but the male drinking rate has been decreasing, while the drinking rate of women has increased, so that not only social problems but also health effects are emerging. (National Institute of Economic Planning and Statistics Bureau; National Health and Nutrition Survey, Ministry of Health and Welfare) (1998).

Chronic alcohol intake not only leads to hepatic cell disorders that play an important role in body metabolism, but can also have a devastating effect on the major organs of the body: the gastrointestinal tract, pancreas, brain, nerves, endocrine organs, hematopoietic organs, and immune system (Linder MC, Nutrition). and Metabolism of fats.In. Linder MC ed.Nutrical Biochemistry and Metabolism with Clinical Applications, 2nd ed. 79-83, Elsevier, New York, Amsterdam, Oxford, 1991), in particular for liver diseases such as fatty liver, alcoholic hepatitis, cirrhosis, etc. (Cha Chae-bok and Jung Hwan-kuk, Catholic University Medical School, 31 , 1 (1978)). The pathogenesis of ethanol liver disease remains unclear. However, in addition to the toxic effects of ethanol itself, it is known to be influenced by various factors such as direct or indirect nutritional disorder, gene effects and immunological mechanisms associated with drinking. (Sanders JB et al., J. Roy Soc. Med. 77, 204-216 (1984); and Srensen TIA., Liver 9, 489-197 (1989)).

Many people are interested in drugs that can get rid of hangovers because of heavy drinking and frequent drinking. The hangover that appears after drinking alcohol is not only toxic but also can be converted into a harmful substance to the human body during metabolism in the body, which is caused by harmful substances to the digestive system including the brain and liver. Hangovers appear as headaches or heartburn after ethanol intake, and many studies have been conducted to find a drug that can reduce it, and many drugs have already been introduced, but few have a remarkable effect (Lieber CS, Acta Scand.supple). , 703 , 11 (1985); and Blane HT, The personality of alcoholics: Guise of dependency, Harjper and Fow, New York, p 109 (1968).

Therefore, in the present invention, using the extract of sewage oil used as a detoxification and diuretic in civilian, to compare the hangover removal effect in the animal model with the efficacy of a commercial hangover removal drink. In addition, we intended to measure the antioxidant effect of the consumption of sesame oil extract on the oxidative stress induced by alcohol intake. Also, based on this, we want to investigate the functionality as an effective hangover drink.

Unlike other foods, alcohol that can't be stored in tissues is oxidized to acetaldehyde by the catalytic action of alcohol dehydrogenase (ADH), and then to acetic acid and hydrogen by the catalytic action of aldehyde dehydrogenase (ALDH). (Jae-Yeol Cho et al., Korean Society of Food Science and Technology, 29 (1), 167-172 (1997)). 90% of the acetic acid formed is directly CO ₂ and H ₂ O and about 10% is completely degraded through the TCA cycle (Cho Jae-Hyeol et al .; Lieber CS and Leo MA, In Progress in liver diseases, Popper, H. and Schaffner, F (Ed), Grune amd Stration, New York, p. 253 (1986); and Linder MC). However, when excessive alcohol intake, in addition to ADH, microsomal ethanol oxidizing system (MEOS) is oxidized and converted to aldehyde and converted to the final CO ₂ and H ₂ O (Fig. 1) (Linder MC). The mechanism by which chronic ethanol intake causes hyperlipidemia and fatty liver results in metabolism of ethanol by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) resulting in increased liver and intracellular NADH / NAD ⁺ ratios. Theorem has been shown to increase fatty acid oxidation and TCA cycle activity in hepatocytes and result in fatty liver (Murray R. K. et al., Haper's Biochemistry, Appleton & Lange, Connechcut, USA. 3rd ed, p260 (1993)). In addition, acetaldehyde, known as a cause of hangover, has a high affinity with sulfur-containing substances such as cysteine and glutathione in the liver microsomes, causing lipid changes in the plasma membrane and causing liver damage such as fatty liver and hepatic necrosis. Kaufman N. et al . , AM A. Arch.Path 70 , 331 (1960).

Alcohol affects the body depending on the amount of intake, and excessive alcohol consumption in a short time causes vomiting, headache, lowering blood pressure, tachycardia and shock with accumulation of aldehydes, especially in liver lipid oxidation or endoplasmic reticulum It interferes with metabolism and chronic adaptation leads to adaptability, which increases the metabolism of alcohol and drugs and accelerates the production of lipoproteins, resulting in damage to liver cells (Rosser BG and Gores GJ, Gastroenterology, 108, 252 (1995)). These toxic phenomena are due to the peroxidation reactions occurring during the production of acetaldehyde and the destruction of hepatocytes by excess NADH formed by the catalyst of MEOS, the chemical equilibrium of hepatocytes, metabolic abnormalities and the inhibition of mitochondrial function (Bunsel RG and Lehmann AG, Behav.Vrain Res . 1, 351 (1980); and State Chungno, Chemical World, 34 , 767 (1994). These harmful reactions and harmful substances cause hepatic cell damage, leading to the accumulation of fatigue substances such as lactic acid, reducing the activity of ALDH, inhibiting the activation of vitamins, reducing the amount of vitamins in the blood and inhibiting the synthesis of muscle protein in the heart. (Note Chung). Furthermore, the increased hydrogen is directly or indirectly involved in the synthesis of fatty acids to form fats and the like, resulting in the pathological phenomenon of fatty liver (Lieber CS and Leo MA; Tkabe M. and Itokawa Y., J. Nutr. Sci. Viaminol). ., 29, 509 (1983)).

In addition, ethanol intake has been suggested to be toxic in another way than to increase the synthesis of triglycerides in the liver can damage the liver cells. In other words, alcohol consumption increased with the report that oxygen consumption in the liver is increased by the oxidation of alcohol, leading to oxygen deficiency and necrosis in liver cells (Mezey E., Amerian Journal of Clinical Nutrition, 33, 2709-2718 (1980)). The theory that it promotes lipid peroxidation in hepatocytes causes damage to liver tissue (Valenzuela A. et al . , REBS Letters. 111, 11-13 (1980)), and acetaldehyde and fatty acid ethyl, intermediate metabolites from ethanol oxidation It has been reported that the ester itself is toxic and damages major organs (Laposata EA and Lange LG, Science. 231, 497-499 (1986)). The production of these intracellular free radicals is formed from enzymes in the cytoplasm and various oxidases. These free radicals cause damage to DNA and mitochondria and promote fatty oxidation in the plasma membrane to cause fatty liver (Figure 2) (Said EM et al., European Jounal of obsterics and gynecology and reproductive biology, 89 , 1-6 (2000) ). On the other hand, the generated free radicals are converted to H ₂ O ₂ by Cu-Zn superoxide dismutase (Cu-Zn SOD), which in turn is glutathione reductase 5 (GPX 5) or antioxidant catalase. Is separated into H ₂ O and O ₂ . Therefore, Cu-Zn SOD and GPX 5 and the like can be said to play a secondary role in restoring the oxidation of cells (Said EM, etc.) (Fig. 2).

Therefore, an effective hangover drink can protect the cells from the lowering of blood alcohol concentration, the rapid elimination of fatigue substances and radical damage caused by alcohol metabolism, the first step in the reduction of substances that are the main cause of pathological phenomena such as acetaldehyde. The antioxidants present should be used together.

Prior research on sewage oil showed that Xu et al. In China found that dehydroevodiamine isolated from Evodiae Fructus had an effect on lowering blood pressure (Xu SB et al . , Am. J. Chin). Med. 10 , 1-4 (1982)), and Yamaha et al. Reported that oxygen deficiency symptoms in mice induced by anoxia with KCN for the development of new drugs using natural products (Yamahara J. et al. , Chem Pharm Bull (Tokyo) 37 , 7 (1989)), EVO diamine in the ohsuyu (evodia) Chinese medicinal plant (evodiamine), capric (rutaecarpine) and indole in rutile -. alkaloids (indole-alkaloids) are It was found to contain a large amount and this was identified as the main material for this effect. In addition, methanol extract of sewage oil has been reported to be effective in inhibiting the activity of acetylcholinesterase. Dehydroevodiamine has a specific effect on acetylcholinesterase, and thus acetylcholinesterase Has been reported to reversibly and non-competitively inhibit the activity of acetylcholinesterase from sorghum milk and its mechanism of action, Seoul National University, Seoul, Korea (1996). In addition, Yu et al. Reported that water-soluble extracts of Evodiae fructus play a role of antidiarrheal agents in diarrhea induced by castor oil (Yu LL et al . , J Ethnopharmacol. 73 , 1-2 (2000)).

It is an object of the present invention to provide a pharmaceutical composition exhibiting hangover relief and antioxidant effects.

Still another object of the present invention is to provide a dietary supplement having a hangover relief and antioxidant effect.

1 is a schematic of ethanol metabolic pathways.

2 is a schematic of oxidative stress.

3 is a process diagram of sewage oil extraction of the present invention.

4A and 4B are graphs showing blood alcohol concentrations at 1 hour after 20% and 40% alcohol administration, respectively.

5A and 5B are graphs showing blood alcohol concentrations at 2 hours after 20% and 40% alcohol administration, respectively.

6A and 6B are graphs showing blood alcohol concentrations at 3 hours after 20% and 40% alcohol administration, respectively.

7A and 7B are graphs showing blood alcohol concentrations at 4 hours after 20% and 40% alcohol administration, respectively.

8A and 8B are graphs showing changes in blood alcohol concentrations for 4 hours after 20% and 40% alcohol administration, respectively.

Figure 9 is a graph and electrophoresis picture confirmed by RT-PCR change in mRNA amount of ADH and ALDH 1 hour after 20% alcohol administration.

Figure 10 is a graph and electrophoresis picture confirmed by RT-PCR changes in mRNA amount of ADH and ALDH 1 hour after 40% alcohol administration.

Figure 11 is a graph and electrophoresis picture confirmed by RT-PCR changes in the mRNA amount of ADH and ALDH 2 hours after 20% alcohol administration.

Figure 12 is a graph and electrophoresis picture confirmed by RT-PCR change in mRNA amount of ADH and ALDH 2 hours after 40% alcohol administration.

Figure 13 is a graph and electrophoresis picture confirmed by RT-PCR changes in mRNA amount of ADH and ALDH 3 hours after 20% alcohol administration.

Figure 14 is a graph and electrophoresis picture confirmed by RT-PCR change in mRNA amount of ADH and ALDH 3 hours after 40% alcohol administration.

Figure 15 is a graph and electrophoresis picture confirmed by RT-PCR change in mRNA amount of ADH and ALDH 4 hours after 20% alcohol administration.

Figure 16 is a graph and electrophoresis picture confirmed by RT-PCR changes in the mRNA amount of ADH and ALDH 4 hours after 40% alcohol administration.

Figure 17 is a graph and electrophoresis picture confirmed by RT-PCR changes in the mRNA amount of Cu-Zn SOD, GPX 5 and catalase 1 hour after 20% alcohol administration.

Figure 18 is a graph and electrophoresis picture confirmed by RT-PCR changes in the mRNA amount of Cu-Zn SOD, GPX 5 and catalase 1 hour after 40% alcohol administration.

Figure 19 is a graph and electrophoresis picture confirmed by RT-PCR changes in mRNA amount of Cu-Zn SOD, GPX 5 and catalase 2 hours after 20% alcohol administration.

Figure 20 is a graph and electrophoresis picture confirmed by RT-PCR changes in the mRNA amount of Cu-Zn SOD, GPX 5 and catalase 2 hours after 40% alcohol administration.

Figure 21 is a graph and electrophoresis picture confirmed by RT-PCR changes in mRNA amount of Cu-Zn SOD, GPX 5 and catalase 3 hours after 20% alcohol administration.

Figure 22 is a graph and electrophoresis picture confirmed by RT-PCR changes in mRNA amount of Cu-Zn SOD, GPX 5 and catalase 3 hours after 40% alcohol administration.

Figure 23 is a graph and electrophoresis picture confirmed by RT-PCR changes in mRNA amount of Cu-Zn SOD, GPX 5 and catalase 4 hours after 20% alcohol administration.

Figure 24 is a graph and electrophoresis picture confirmed by RT-PCR changes in the mRNA amount of Cu-Zn SOD, GPX 5 and catalase 4 hours after 40% alcohol administration.

In accordance with the above object, the present invention provides a composition for the hangover and antioxidant action comprising a sewage extract.

The composition for the hangover elimination and antioxidant action of the present invention, it is effective to contain the sewage extract 0.5 to 50% by weight based on the total weight of the composition.

The sewage oil extract of the present invention may be prepared according to a conventional method, for example, dried sewage oil may be lower alcohol such as methanol, ethanol, or lower alcohol acetate at an extraction temperature of 10 ° C. to 50 ° C., preferably room temperature. Preferably, extraction with 100% ethanol or ethyl acetate for about 1 hour to 2 days, preferably about 2 hours to 1 day can be obtained by repeating 1 to 5 times, preferably 3 times.

In addition, the sewage oil extract of the present invention may be further fractionated by a conventional fractionation method, for example, concentrated under reduced pressure at 10 ° C to 50 ° C, preferably 30 to 40 ° C and lower alcohols such as methanol and ethanol. After dilution in 90% ethanol, preferably, an organic solvent, preferably a lower alcohol such as lower alkyl and methanol, ethanol, more preferably a mixture of n -hexane and 90% ethanol (preferably volume ratio of 1: Extract fractions 1 to 5 times, preferably 3 times in 1), concentrate the lower alcohol fraction phase at 10 ° C to 50 ° C, preferably 30 to 40 ° C, and concentrate this concentrate with distilled water and lower alcohol acetate, preferably The fraction may be extracted once to five times, preferably three times, in a mixed solution of ethyl acetate (preferably in a volume ratio of 1: 1) to finally obtain an ethyl acetate fraction (FIG. 3).

The present invention provides a composition for hangover and antioxidant action comprising lower alcohol extract and acetate extracts obtained in the extraction and fractionation process.

The composition comprising the sewage oil extract of the present invention may further comprise a suitable carrier, excipient and diluent according to conventional methods.

Carriers, excipients and diluents that may be included in the composition comprising the sewage oil extract of the present invention 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.

Compositions comprising sewage oil extracts according to the present invention are formulated in the form of powders, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be used.

The amount of sewage extract may vary depending on the age, sex, and weight of the patient, but the amount of 0.1 to 500 mg / kg may be administered once to several times daily. The dosage of sewage extract and fractions may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition comprising the sewage oil extract of the present invention may be used in various forms, such as drugs, foods and beverages for the hangover removal and antioxidant activity. Examples of the food to which the sesame oil extract can be added include various foods, beverages, gums, teas, vitamin complexes, and health supplements.

Fructus extract itself of the present invention has little toxicity and side effects, so it is a drug that can be used safely even for long-term administration for the purpose of prevention.

The sewage oil extract of the present invention may be added to food or beverage for the purpose of relieving hangovers and antioxidant activity. At this time, the amount of the sewage oil extract in the food or beverage is generally added to the health food composition of the present invention 0.1 to 15% by weight, preferably 1 to 10% by weight of the total food weight, the food health beverage composition is 100m1 It can be added in the ratio of 1-30g, Preferably it is 3-10g on the basis of.

The health beverage composition of the present invention has no particular limitation on the liquid component except for containing the sewage oil extract as an essential ingredient in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of one natural carbohydrate 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 (e.g., rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of said natural carbohydrate is generally about 1-20g, preferably about 5-12g per 100m1 of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp 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 critical but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Hangover-relieving effect of the sewage oil extract of the present invention is a first step for reducing the main causative agent of pathological phenomena such as acetaldehyde, lowering blood alcohol concentration, and alcohol dehydrogenase (ADH) and acetaldehyde for oxidizing acetaldehyde. It can be confirmed by the expression amount of aldehyde dehydrogenase (ALDH) which decomposes into acetic acid and hydrogen, the antioxidant activity of the sewage oil extract of the present invention is the rapid removal of fatigue substances and radicals generated during alcohol metabolism to H ₂ O ₂ Peroxide dismutase (Cu-Zn SOD) to be converted and glutathione reductase (GPX 5) to decompose H ₂ O ₂ into H ₂ O and O ₂ and the expression of catalase are identified.

Fructus extract of the present invention promotes alcohol degradation during alcohol administration by increasing hepatic ADH and ALDH activity, and lowers blood and hepatic alcohol concentrations, and also has high antioxidant activity by increasing expression of genes that restore oxidative stress. Indicates.

The present invention is described in more detail based on the following examples, but the present invention is not limited thereto.

Reference Example 1. Measurement of Blood Alcohol Concentration

Absorbance at 340 nm was measured using a spectrophotometer (Jasco, V -550, Japan) using an ethanol quantitative kit (332-UV, Sigma Chemical Co, St. Lowis, Mo, USA) based on enzyme colorimetric method. By calculation, the alcohol concentration in the serum was measured.

Reference Example 2 Measurement of Enzyme mRNA Amount by RT-PCR

RNA is extracted from tissue according to the methods described in Chomczynski P., BioTechniques, 15, 532-537 (1993) and Chomczynski P. and Sacchi N., Anal. Biochem. 162 , 156-159 (1987). MRNA amount of each enzyme was measured.

RT-PCR was performed using the RT-PCR kit (Promega), and the amplification cycle was determined to be 40 cycles through preliminary experiments.

Primers used for PCR are described in ADH: Zhang K. et al., Gene , 57 , 27-36 (1987); ALDH: Bond SL and Singh SM, Biochem. Med. Metab. Biol. , 52 (2), 155-159 (1994); Cu-Zn SOD: Bewley GC, Nucleic Acids Res. , 16 (6), 2728 (1988); GPX 5: Ghyselinck NB et al . , Mol. Endocrinol., 7 , 258-272 (1993); And catalase: Srausburg R., Direct submission. National Institutes of Health. Mammalian gene collection. Cancer genomics office. National cancer Institute} was used to synthesize a sequence (Bionia, Korea) and the base sequence is shown in Table 1.

RT-PCR reactants include 1 x AMV / T f1 reaction buffer, 0.2 mM dNTP mixture, 1M downstream primer, 1M upstream primer, 1 mM MgSO ₄ , 0.1 u / μL AMV reverse transcriptase, 0.1 u / μl T f1 DNA polymerase and 1 μg RNA sample were mixed to a total volume of 50 μl. The first cDNA strand by reverse transcription was synthesized at 48 ° C. for 45 minutes and RNA / cDNA / primers were denatured at 94 ° C. for 2 minutes. Synthesis and PCR amplification of the second cDNA strand were carried out for 30 seconds of denaturation at 94 ° C., 1 min of annealing, extension at 68 ° C. for 2 minutes, and a total amplification cycle of 40 cycles. The last extension was reacted for 7 minutes at 68 ° C. (Robocycler gradient 96, STRATAGENE, USA). PCR results were confirmed by electrophoresis with 2% agarose gel added with ethidium bromide. In order to confirm the efficiency of amplification, the mRNA expression of β-actin was expressed together.

Table 1. Primer Sequences and PCR Reaction Conditions

Example 1. Preparation of Sewage Extract

Thinly sliced 1 Kg of dried sewage oil purchased from Gyeongdong market in Seoul was extracted three times with 10 L of 100% ethanol at room temperature for 1 day. The ethanol extract was concentrated under reduced pressure at 30 to 40 ° C. The concentrate was diluted with 1 L of water to form a 90% ethanol solution, and then partitioned three times with n -hexane and 90% ethanol (1: 1), and the 90% ethanol fractions were concentrated at 30 to 40 ° C. The concentrated ethanol fraction was partitioned and concentrated three times with distilled water and ethyl acetate (1: 1), and ethyl acetate extract was used as a sample.

Example 2. Hangover Removal Effect of Sewage Extract

1) Treatment of experimental animals

About 20 g to 25 g of ICR male mice were purchased from Seoul National University Experimental Animal Breeding Center, and then adapted to a general solid feed (Korean experimental animal, Korea) for one week, and treated with alcohol control (C, n = 5) as a control. As the group, the alcohol / sewage extract administration group (T, n = 5) and the positive control group alcohol / commercial drug (condition, CheilJedang) administration group (P, n = 5) were set as follows. After fasting overnight before the experiment, the drug was orally administered using sonde for mice. In the control group (C), 10% / kg of alcohol at a concentration of 40% or 20% was administered, and in the case of the treatment group (T), 10 mg / kg, 5 mg / kg, or 2.5 mg / kg of sewage extract were administered. Positive control group (P) was administered with alcohol 10ml / kg, 5ml / kg, or 2.5ml / kg drug having a commercial hangover effect at the same time.

C20A: 20% alcohol control

2. P20A: 20% alcohol and commercial drug high concentration (10ml / kg) positive control

3. P20B: 20% alcohol and commercial drug concentration (5ml / kg) positive control

4. P20C: 20% alcohol and commercial drug low concentration (2.5ml / kg) positive control

5. T20A: 20% alcohol and sewage extract high concentration (10mg / kg) administration group

6. T20B: 20% alcohol and sewage extract medium concentration (5mg / kg) administration group

7. T20C: 20% alcohol and sewage extract low concentration (2.5mg / kg) administration group

8.C40A: 40% alcohol control control

9.P40A: 40% alcohol and commercial drug high concentration (10ml / kg) positive control

10.P40B: 40% alcohol and commercial drug concentration (5ml / kg) positive control

11.P40C: 40% alcohol and commercial drug low concentration (2.5ml / kg) positive control

12.T40A: 40% alcohol and sewage extract high concentration (10mg / kg) administration group

13. T40B: 40% alcohol and sewage extract medium concentration (5 mg / kg) administration group

14. T40C: 40% alcohol and sewage extract low concentration (2.5mg / kg) treatment group

2) Sample Collection and Pretreatment

The blood and liver of the mice were collected at 1, 2, 3 and 4 hours after alcohol administration the day after fasting for 12 hours or more. Blood was collected from the orbital vein of the mouse using a tube treated with heparin, and the blood was centrifuged at 4 ° C. and 10000 g for 15 minutes to separate serum, and then divided into tubes until freezing and stored at -70 ° C .. The liver of the mouse was immediately removed, snap-freeze in liquid nitrogen, stored in a -70 ° C freezer (Operon, Korea) and used for the experiment.

3) Statistical processing

All experimental results of the present invention were statistically processed using a SAS (Statistical Analysis System) PC package (ver. 6.12, 1998, USA), and all the analysis values were expressed as mean ± SD. Significance of reinforcement of sewage extract during ethanol administration was determined by using a paired t-test of ethanol / positive control group (P) and ethanol / sewage extract group (T) based on the ethanol control group (C). It was verified by.

4) blood alcohol concentration

According to the method of Reference Example 1, blood alcohol concentration was measured, and the results are shown in FIGS. 4 to 8.

In general, blood alcohol concentrations were lower in the positive control group and sewage extract group than in the control group. The blood alcohol concentrations of the positive control group and the sewage group were significantly decreased at the high concentration of 10 mg / kg.

Specifically, blood alcohol concentration at 1 hour after 20% or 40% alcohol administration was lower in the positive control group and the sewage treatment group than the control group. Especially, in the sewage oil treated group treated with sesame oil extract, both the 20% and 40% alcohol treated groups showed significantly lower blood alcohol levels in the high, medium and low concentration groups than the control group. (FIGS. 4A and 4B). However, no significant difference was found between the positive control group and the sewage treatment group in the 20% and 40% alcohol groups.

After 2 hours of administration, the blood alcohol concentration in the 40% alcohol-treated group dropped sharply in the positive control group and the sewage-treated group compared to the control group, especially in the high control group and in the positive control group and the sewage-treated group at 294.93 ± 28.47 mg / dl. Compared to the levels of 119.90 ± 19.73 mg / dl and 95.73 ± 15.13 mg / dl, respectively (FIG. 5B). On the other hand, there was no significant difference between the positive control group and the sewage-treated group in the blood alcohol concentration of 2 hours after administration (FIG. 5A).

The blood alcohol concentration at 3 hours after alcohol administration was lower in the positive control group and sewage group than in the control group in the 20% alcohol group, and the low level group was treated with the low concentration group in the sewage group treated with low concentration of 2.5 mg / kg. It was lower than that, but there was no significant difference (Fig. 6a), and there was no statistically significant difference between the groups treated with 40% alcohol (Fig. 6b).

The blood alcohol concentration at 4 hours after alcohol administration did not show a difference between the control group, the positive control group and the sewage treatment group in the 20% alcohol administration group (FIG. 7A). However, in the 40% alcohol-treated group, the sewage-treated group in the high concentration group of 10 mg / kg and the medium concentration group of 5 mg / kg was lower in blood alcohol concentration to a statistically significant level than the positive control group (FIG. 7B).

On the other hand, the level of blood alcohol in the 20% alcohol group was found to be only about 30% of the 40% alcohol group (Figs. 8A and 8B). The value of alcohol concentration tended to drop most rapidly over time (FIG. 8B).

5) Expression level of ADH and ALDH

According to the method of Reference Example 2, mRNA amounts of ADH and ALDH were measured, and the results are shown in FIGS. 9 to 16.

RT-PCR using hepatic mRNA showed that ADH and ALDH activities were inversely proportional to blood alcohol levels.

Specifically, 1 hour after alcohol administration, ADH and ALDH expression was higher in the positive control group and the sewage-treated group than the control group, and the activity was higher in the high concentration group. On the other hand, in the group treated with medium and low concentrations, there was no significant difference in the expression of ALDH between the groups (FIG. 9).

As with the decrease in blood alcohol concentration, the RT-PCR result after 2 hours of alcohol administration showed higher expression of ADH and ALDH in the positive control group and the sewage treatment group than the control group, especially in the 20% alcohol group. The expression of ALDH was more expressed in the group treated with high concentration, indicating that the activity of ALDH was higher (FIG. 11). In the group treated with 40% alcohol, the expression of ADH was higher in the positive control group and the sewage treatment group than the control group. It was higher in the group, but there was no significant difference, the expression of ALDH was very high in the positive control group and sewage treatment group, compared to the control group (Fig. 12).

Three hours after alcohol administration, when 20% alcohol was administered, the expression of ADH and ALDH in the sewage-treated group was higher than that of the control group and the positive control group (FIG. 13). When 40% alcohol was administered, the expression of ADH and ALDH was higher in the positive control group and the sewage treatment group than the control group, but there was no difference between the positive control group and the sewage treatment group (FIG. 14).

At 4 hours after administration of 20% alcohol, there was little expression of ADH and only slightly ADH was expressed in the high sewage treatment group (FIG. 15). At 4 hours after the 40% alcohol administration, ADH and ALDH expression was higher in the positive control group and the sewage treatment group than in the control group, but there was no difference between the positive control group and the sewage treatment group (FIG. 16).

Example 3. Antioxidant Effect of Sewage Extract

Statistical treatment of the experimental animal treatment, liver sample collection and experimental results was carried out in the same manner as in Example 2, according to the method of Reference Example 2, the mRNA amount of Cu-Zn SOD, GPX 5 and catalase was measured. 17 to 24 are shown.

One hour after 20% alcohol administration, Cu-Zn SOD expression was higher in the sewage-treated group than in the control group and the positive control group, but there was no significant difference in the expression levels of GPX 5 and catalase (FIG. 17). At the same time, the group treated with 40% alcohol showed higher expression of Cu-Zn SOD in the positive control group and the sewage treatment group than the control group, but there was no difference between the positive control group and the sewage treatment group. There was no difference (FIG. 18).

2 hours after alcohol administration, the expression of Cu-Zn SOD, GPX 5 and catalase was higher in the 20% alcohol-treated group than in the control group and the sewage-treated group (FIG. 19), but in the 40% alcohol-treated group There was little difference in the expression levels of Cu-Zn SOD and catalase, whereas GPX 5 was expressed more in the high concentration control group and the ewes group compared to the control group (FIG. 20).

At 3 hours after 20% alcohol administration, the expression of Cu-Zn SOD, GPX 5 and catalase was significantly higher in the sewage-treated group than in the control and the positive control group, especially in the high-treated group. Figure 21). On the other hand, in the group treated with 40% alcohol, the expression levels of the antioxidant-related genes were higher in the positive control group and the sewage-treated group than the control group, but there was no difference between the positive control group and the sewage-treated group. There was no change according to the concentration difference (FIG. 22).

At 4 hours after the 20% alcohol administration, the amount of expression of the antioxidant control genes was significantly higher in the positive control group and the sewage-treated group than in the control group, especially in the high concentration group (FIG. 23). In addition, the expression of each gene was higher in the sewage treatment group than in the positive control group (FIG. 23). Similarly, in the 40% alcohol-treated group, the expression was higher in the positive control group and the sewage-treated group than in the control group, but there was no difference between the positive control group and the sewage-treated group. Was absent (FIG. 24).

Formulation Example 1 Preparation of Powder

According to the preparation method of powder in the pharmacopeia formulation, it is prepared in the following component content per packet.

Sewage Oil Extract ㆍ ··········· 300 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Talc 10 mg

Formulation Example 2 Preparation of Tablet

According to the preparation method of tablets in the pharmacopeia formulation, it is prepared in the following component content per tablet.

Sewage Oil Extract ㆍ ············ 300 mg

Corn starch ㆍ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Magnesium Stearate 2 mg

Formulation Example 3 Preparation of Capsule

According to the preparation method of capsules in the pharmacopeia formulation, it is prepared in the following component content per capsule.

Sewage oil extract ·············· 300 mg

Corn starch ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose ····················· 100 mg

Magnesium stearate 2 mg

Formulation Example 4 Preparation of Injection

According to the preparation method of injection in the pharmacopeia formulation, it is prepared in the following component content per ampoules (2 ml).

Sewage Oil Extract ㆍ ············· 300 mg

Sterile distilled water for injection ㆍ ·············

pH regulator ㆍ ··················

Formulation Example 5 Preparation of Liquid

According to the preparation method of the liquid formulation in the Pharmacopoeia General Formulation, it is prepared in the following component content per 100 ml of the liquid formulation.

Sewage Oil Extract ㆍ ············· 300 mg

Heterosaccharides ················· 10 g

Mannitol ···················· 5 g

Purified water ㆍ ···················

In addition, the health beverage is prepared as follows.

Sewage oil extract 0.1-15%, sugar 5-10%, citric acid 0.05-0.3%, caramel 0.005-0.02%, vitamin C 0.1-1% of the additives were mixed and 79-94% purified water was mixed to make a syrup, The syrup was sterilized at 85 to 98 ° C. for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and then a carbonated beverage containing sewage oil extract was prepared by injecting carbon dioxide gas at 0.5 to 0.82%.

Instant sterilization by homogeneous mixing of subsidiary ingredients such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%) and sewage oil extract Health drinks were prepared by packaging in small packaging containers such as plastic bottles.

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

Composition containing sewage oil extract of the present invention, due to the hangover and antioxidant action of the sewage oil extract, can be usefully used as a drug or dietary supplement for the hangover and antioxidant action.

Claims (7)

Hangover oil ( Evodia officinalis ) as an active ingredient and a pharmaceutical composition for hangover resolution made by mixing with a conventional pharmaceutically acceptable carrier. A pharmaceutical composition for antioxidant activity, comprising an extract of Evodia officinalis as an active ingredient and mixing with a conventional pharmaceutically acceptable carrier. The composition according to claim 1 or 2, wherein the sewage extract is obtained by extraction with lower alcohol or lower alcohol acetate. The composition according to claim 3, wherein the sewage oil extract is a fraction obtained by distilling a lower alcohol or a lower alcohol acetate extract under reduced pressure and then dividing with an organic solvent. The composition of claim 4, wherein the organic solvent is selected from the group consisting of n-hexane, methanol, ethanol and ethyl acetate. A dietary supplement exhibiting a hangover effect, comprising a sesame oil extract and a food acceptable additive. A dietary supplement exhibiting antioxidant activity, comprising a sesame oil extract and a food acceptable additive.