KR20020088243A - A composition inhibiting hangover - Google Patents

Abstract

PURPOSE: A composition containing an effective active material induced from natural products capable of inhibiting activity of alcohol dehydrogenase is provided which reduces cytoxicity and inhibits hangover caused by consumption of alcoholic beverages by inhibition of the formation of aldehyde as a first metabolite in alcohol metabolism thereof. CONSTITUTION: The composition for reducing hanover contains as an effective ingredient one or more selected from Puerarin, mulberroside A, resveratrol, Rhaponticin, desoxyrhaponticin, rutin and quercetin. For an example, the Puerarin is separated from Pueraria thunbergiana(Sieb. et Zucc.) Benth., the mulberroside A is separated from Morus alba Linne, Rhaponticin and desoxyrhaponticin are separated from Rheum officinale Baillon.

Description

[0001] The present invention relates to a composition for inhibiting an alcohol dehydrogenase,

Resveratrol, rutin, quercetin with flavonoid structure, alcohol dehydrogenase inhibitory effect of puerarin, mulberroside A, rhaponticin, desoxyrhaponticin and stilbene chemical structure, which are active ingredients of methanol extract of Puerariae radix, . The present invention relates to a hangover inhibiting composition containing, as an active ingredient, puerarin, mulberroside A, resveratrol, rhaponticin, desoxyrhaponticin, rutin, quercetin derived from natural products as an active ingredient. The hangover inhibiting composition of the present invention is characterized in that, It inhibits the toxicity and hangover of aldehydes.

Accordingly, by using the hangover-inhibiting composition of the present invention before, during, or after mixing with alcohol, it is possible to expect a gastrointestinal or hepatic protection effect and an effect of inhibiting hangover.

Toxicity and hangover due to alcohol are known to be caused by aldehydes generated from alcohol by the action of alcohol dehydrogenase, which is present in gastrointestinal or hepatic cells during drinking.

In normal alcohol metabolism, when alcohol (ethyl alcohol, methyl alcohol, ethylene glycol, etc.) is ingested, it is absorbed in the gastrointestinal tract and is transported to the liver. Alcohol dehydrogenase, which is present in the stomach and liver cells, And then metabolized to acetic acid by aldehyde dehydrogenase. It is well known that the aldehyde produced during this degradation process is the main cause of cytotoxicity and hangover (Lieber CC, "Alcohol and the Liver, 1994 Update", Gastroenterology 1994; 106, 1085-1105). Therefore, inhibiting the reaction of aldehyde formation by alcohol dehydrogenase during drinking suppresses cytotoxicity and hangover phenomenon and maximizes the drinking effect. In addition, although the alcohol consumed is metabolized by the alcohol dehydrogenase and the aldehyde dehydrogenase in the liver, since the alcohol in the blood is also excreted from the kidney to the urine and from the lungs to the respiration, the present invention appropriately suppresses the metabolism from the alcohol to the aldehyde A significant portion of the alcohol consumed is excreted to the outside of the body in the same manner as described above before being metabolized to aldehyde. As a result, the total amount of the aldehyde, which is a toxic substance, is reduced, thereby exhibiting a hangover inhibition and hepatoprotective effect.

The use of 4-methylpyrazole (4-methylpyrazole, Formepizole) as an inhibitor of alcohol dehydrogenase in first-aid treatment instead of ethanol, which is used for methyl alcohol poisoning or ethylene glycol intoxication, 1999, 37 (5), 537-560, Intensive Care Med 1999, 25 (5), 528-531), which has been approved by the Agency for Medicinal Products and has been reported to be used in emergencies ( J. Toxicol .

Keung, WM: Biochemical studies of (a) and (b) showed that Pueraria japonica extract, or its components, daidzein, genistein, formononetin biochanin A and prunetin, 7-hydroxyflavone, apigenin, galangin and kaempferol new class of alcohol dehydrogenase inhibitors from Radix puerariae, alcohol Clin. Exp. Res. 17 (6), 1254 (1993)) is however, alcohols such as puerarin, mulberroside a, resveratrol, rhaponticin, desoxyrhaponticin, rutin, quercetin of the invention No dehydrogenase inhibiting effect has been reported, and a hangover inhibiting composition containing them as an active ingredient is not known.

SUMMARY OF THE INVENTION The present invention has been made in view of the foregoing circumstances, and it is an object of the present invention to provide a method for enhancing the drinking effect and to prevent toxicity of an aldehyde generated as a primary metabolite of alcohol metabolism in the body, And to provide toxicity and hangover suppression effect in drinking the gut in the gastrointestinal tract and the liver by using effective active ingredients derived from natural products inhibiting the production.

Therefore, by using the hangover-inhibiting composition of the present invention before, during, or after mixing with alcohol, it is possible to obtain a gastrointestinal or hepatic protection effect and an effect of inhibiting hangover.

Alcohol dehydrogenase is a primary enzyme that metabolizes alcohol in the stomach or liver. When the enzyme activity is inhibited, the production of aldehyde, which is an oxidizing substance of alcohol, is inhibited. Therefore, gastrointestinal or hepatoprotective effect and inhibition of hangover phenomenon can be expected.

It is effective to inhibit aldehyde dehydrogenase in alcohol metabolism and to increase aldehyde concentration 5-10 times in blood. When taking disulfiram which is used as an inducing agent, severe headache, nausea, vomiting, The symptoms of hangover are mainly caused by aldehyde in alcohol metabolism. In addition, aldehydes are highly reactive substances that react with body proteins and other biomaterials to form complexes, resulting in inactivation of various enzymes and undesired immune reactions. Physiological changes lead to increased lipid peroxidation, damage cell membrane and mitochondrial membrane, lead to depletion of vitamins and trace metals, resulting in necrosis and fibrosis of hepatocytes, lowering liver function, and increasing liver fibrosis and liver cancer incidence (Goodman &Amp; Gilman ' s The Pharmacological Basis of Therapeutics, 8th ed. Pp370-378, 1991).

Among the natural alcoholic extracts showing alcohol dehydrogenase inhibitory activity, active substances were examined by the active ingredients of Puerariae radix, rhubarb, and bark bark. As a result, puerarin having an isoflavonoid structure was identified as an active ingredient of Pueraria and mulberroside A having a stilbene structure As an ingredient, rhaponticin and desoxyrhaponticin were identified as active ingredients of the sulfa. In addition, the inhibitory effect of alcohol dehydrogenase on rutin and quercetin, which have flavonoid structure contained in resveratrol, collapsed bean curd, tangerine, etc. The inhibitory effect of these substances was tested at concentrations of 7.44-163.93 μM (0.0003% -0.01%) according to the experimental examples. Resveratrol, rutin, and quercetin were purchased from Sigma Chemical Co..

Hereinafter, the specific structure and activity of the present invention will be described in detail based on the following examples, but the present invention is not limited to the following examples.

Reference Example 1: Extraction and Separation of Puerarin from Puerariae

50 g of granules were extracted with 500 ml of 95% ethanol (10 times) for 3 hours or more by reflux cooling and concentrated under reduced pressure to prepare herbal extracts. The extracts were separated by silica gel column, TLC, Sephadex LH-20 column, etc., and the structure of the active substance was confirmed by using MS and NMR apparatus.

Reference Example 2: Extraction and Separation of Mulberoside A from Mulberry

50 g of bark extract was extracted with 500 ml of 95% ethanol (10 times) by reflux cooling for 3 hours and concentrated under reduced pressure to prepare a herbal extract. The extracts were separated by silica gel column, TLC, Sephadex LH-20 column, etc., and the structure of the active substance was confirmed by using MS and NMR apparatus.

Reference Example 3: Extraction and separation of rhaponticin and desoxyrhaponticin from rhubarb

50 g of rhubarb was extracted with 500 ml of 95% ethanol (10 times) by reflux cooling for 3 hours and concentrated under reduced pressure to prepare a herbal extract. The extracts were separated by silica gel column, TLC, Sephadex LH-20 column, etc., and the structure of the active substance was confirmed by using MS and NMR apparatus.

Measurement of alcohol dehydrogenase activity

After adding a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ , 5.0 mM ethanol and 1% methanol into a cuvette, 10 μl of the alcoholic dehydrogenase enzyme or 10 μl of the liver alcohol dehydrogenase The enzyme activity is measured (control group) by measuring the absorbance change at 340 nm by NADH produced when the cell extract (100 μl) is added (total volume of reaction solution: 1 ml).

In order to examine the inhibitory activity of the sample, the herbal active ingredients isolated in the Reference Example are dissolved in methanol, appropriately diluted, and 10 μl of the diluted solution is added to the enzyme reaction solution to determine the enzyme activity.

Experimental Example 1: Activity of puerarin by concentration in alcohol dehydrogenase

The puerarin solution was added to a cuvette such that the puerarin concentration was 20.55-102.76 μM in a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ , 5.0 mM ethanol. The puerarin concentration was measured by ultraviolet spectrophotometer The enzyme activity is measured by measuring the change in absorbance at 340 nm caused by NADH produced when 10 μl of alcohol dehydrogenase or rat liver cell extract (100 μl) is added (total volume of reaction solution: 1 ml). Calculate the IC ₅₀ (μM) by creating a calibration curve of the relative% activity and the log value of the puerarin concentration for the control experiment.

Experimental Example 2: Activity of mulberoside A by concentration of alcohol dehydrogenase

10 μl of the mulberoside A solution was added to a cuvette such that the concentration of mulberoside A was 8.80-66.19 μM in a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ and 5.0 mM ethanol, and the resultant was analyzed by ultraviolet spectrophotometer ), The enzyme activity is measured by measuring the change of absorbance at 340 nm by NADH produced when 10 μl of a horse liver alcohol dehydrogenase or a mouse cell extract (100 μl) is added (total volume of reaction solution: 1 ml). Calculate the IC ₅₀ (μM) by creating a calibration curve of the relative% activity for the control experiment and the log value of the mulberoside A concentration.

Experimental Example 3: Activation experiments of resveratrol on alcohol dehydrogenase

10 μl of resveratrol solution was added to a cuvette such that resveratrol concentration was 13.69-82.16 μM in a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ and 5.0 mM ethanol, and the resultant was analyzed by ultraviolet spectrophotometer The enzymatic activity is measured by measuring the change in absorbance at 340 nm caused by NADH produced when 10 μl of horse liver alcohol dehydrogenase or rat liver cell extract (100 μl) is added (total volume of reaction solution: 1 ml). Calculate the IC ₅₀ (μM) by creating a calibration curve of the relative% activity and the log value of resveratrol concentration for the control experiment.

Experimental Example 4: Activity test of rhaponticin on alcohol dehydrogenase by concentration

10 μl of rhaponticin solution was added to a cuvette such that rhaponticin concentration was 7.44-59.52 μM in a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ and 5.0 mM ethanol, and the resultant was analyzed by ultraviolet spectrophotometer The enzymatic activity is measured by measuring the change in absorbance at 340 nm caused by NADH produced when 10 μl of horse liver alcohol dehydrogenase or rat liver cell extract (100 μl) is added (total volume of reaction solution: 1 ml). Calculate the IC ₅₀ (μM) by creating a calibration curve of the relative% activity and the log value of rhaponticin concentration for the control experiment.

Experimental Example 5: Activity test of desoxyrhaponticin on alcohol dehydrogenase by concentration

10 μl of desoxyrhaponticin solution was added to a cuvette such that desoxyrhaponticin concentration was adjusted to 12.07-48.27 μM in a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ and 5.0 mM ethanol, and the resultant was subjected to ultraviolet spectrophotometer The enzymatic activity is measured by measuring the change in absorbance at 340 nm caused by NADH produced when 10 μl of horse liver alcohol dehydrogenase or rat liver cell extract (100 μl) is added (total volume of reaction solution: 1 ml). Calculate the IC ₅₀ (μM) by creating a calibration curve of relative activity and the log value of desoxyrhaponticin concentration for the control experiment.

Experimental Example 6: Activity test of rutin on alcohol dehydrogenase by concentration

10 μl of a rutin solution was added to a cuvette such that the rutin concentration was 81.97-163.93 μM in a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ and 5.0 mM ethanol, and the resultant was analyzed by ultraviolet spectrophotometer The enzymatic activity is measured by measuring the change in absorbance at 340 nm caused by NADH produced when 10 μl of horse liver alcohol dehydrogenase or rat liver cell extract (100 μl) is added (total volume of reaction solution: 1 ml). Calculate the IC ₅₀ (μM) by creating a calibration curve of the relative% activity and the log value of the rutin concentration for the control experiment.

Experimental Example 7: Activity of quercetin on alcohol dehydrogenase activity by concentration

10 μl of quercetin solution was added to a cuvette such that quercetin concentration was 73.90-147.80 μM in a 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ and 5.0 mM ethanol, and the resultant was analyzed by ultraviolet spectrophotometer The enzymatic activity is measured by measuring the change in absorbance at 340 nm caused by NADH produced when 10 μl of horse liver alcohol dehydrogenase or rat liver cell extract (100 μl) is added (total volume of reaction solution: 1 ml). Calculate the IC ₅₀ (μM) by creating a calibration curve of relative activity and quercetin concentration log value for the control experiment.

The inhibitory activity of each substance in the above experimental examples is shown in Table 1.

Inhibitor enzyme IC ₅₀ ([mu] M) Chemical Structure Series Material name Concentration (μM) Isoflavonoid Puerarin 20.55-82.21 HLADH 59.21 41.11-102.76 RADH 120.48 stilbene Mulberrosidea 14.61-44.01 HLADH 50.97 8.80-66.19 RADH 210.62 Resveratrol 13.69-41.08 HLADH 39.08 13.69-82.16 RADH 196.92 Rhaponticin 7.44-59.52 HLADH 62.32 Desoxyrhaponticin 12.07-48.27 HLADH 58.44 Flavonoid Routine 81.97-163.93 HLADH 141.58 RADH 95.94 Quercetin 73.90-147.80 HLADH 137.40 RADH 101.39

HLADH; Horse liver alcohol dehydrogenase

RADH; Rat liver cytosolic fraction

IC ₅₀ ; 50% inhibitory concentration

Examples 1 to 9: Preparation of a hangover inhibiting composition

The hangover inhibitory health compositions of Examples 1 to 9 were prepared and the alcohol dehydrogenase activity inhibitory effect was tested with the compositions shown in Table 2 below (% concentration is the final concentration of each active substance in the composition).

Although the hangover-restraining health composition of this embodiment is prepared as a liquid agent according to a conventional method, it can also be prepared as a solid agent such as a candy.

Ingredients Puerarin Mulberrosidea Resveratrol Desoxy-rhaponticin Routine Quercetin Inhibitory effect Example 1 0.00046% 0.00631% 0.00286% 74.3% Example 2 0.00091% 0.00033% 0.00539% 0.00224% 58.1% Example 3 0.00065% 0.00062% 0.00023% 0.00026% 0.00460% 0.00176% 52.1% Examples 4-1 to 3 0.00091% 15.9% 0.00247% 35.3% 0.00669% 78.7% Examples 5-1 to 3 0.00091% 19.8% 0.00290% 50.0% 0.00920% 82.5% Examples 6-1 to 3 0.00033% 14.3% 0.00089% 39.2% 0.00243% 79.3% Examples 7-1 to 2 0.00039% 22.1% 0.00236% 55.8% Examples 8-1 to 2 0.00539% 20.8% 0.00865% 50.4% Examples 9-1 to 3 0.00224% 16.1% 0.00465% 40.8% 0.00962% 81.1%

As shown in Table 2, the single or mixed composition of the present invention shows an inhibitory effect on enzyme activity of 50% or more at a low concentration of 0.01% or less, thereby effectively inhibiting the production of aldehyde.

Experimental Example 8: Experiment for inhibiting hangover

To evaluate the preference of the hangover inhibiting compositions prepared in Examples 1 to 9, a 5-point scaling method was used. In addition, the composition of the inhibitory composition prepared in Examples 1, 4-3, 6-3, and 8-2 was evaluated in five healthy adults who usually drink 1 bottle of shochu (about 90 g of ethanol) Within 5 hours after the administration of 5% concentration of alcoholic beverage (1 bottle of shochu) corresponding to 90g ethanol within 3 hours, whether or not hangover symptoms such as headache, nausea, dizziness, The test was performed according to the scale method.

Sensory test results of hangover inhibition health drinks Example Test item One 2 3 4-3 5-3 6-3 7-2 8-2 9-3 color 4.0 3.9 3.8 5.0 5.0 5.0 4.5 4.2 4.0 flavor 3.5 3.7 3.4 4.2 4.0 4.0 3.5 4.1 3.6 Hangover inhibition effect 4.5 4.7 4.8 4.0

* Sensory test number: 5

* Test method: 5 point scale method

3: normal (as usual), 4: good (better than usual), 5: very good (much better than usual), 1: very disliked (much worse than usual);

As can be seen from the above description, the hangover inhibiting composition of the present invention inhibits the activity of the alcohol dehydrogenase from the active ingredient derived from the natural product contained in the composition, It is possible to inhibit the production of aldehyde and to reduce the cytotoxicity due to the gastrointestinal or hepatic protection effect and the drinking and to stop the hangover by taking the hangover inhibiting composition of the present invention or by mixing it with alcohol after drinking, It is a useful invention.

Claims (4)

A hangover inhibiting composition comprising at least one member selected from the group consisting of puerarin, mulberroside A, resveratrol, rhaponticin, desoxyrhaponticin, rutin and quercetin as active ingredients for inhibiting the activity of an alcohol dehydrogenase. The hangover inhibiting composition according to claim 1, wherein the hangover inhibiting composition contains puerarin having an isoflavonoid structure inhibiting the activity of an alcohol dehydrogenase as an effective active ingredient in an amount of 0.001 to 2%. The hangover inhibiting composition according to claim 1, wherein the hangover inhibiting composition comprises at least one selected from among mulberoside A, resveratrol, rhaponticin and desoxyrhaponticin having a stilbene structure inhibiting the activity of an alcohol dehydrogenase as an active ingredient . The hangover inhibiting composition according to claim 1, wherein the hangover inhibiting composition contains at least one selected from rutin and quercetin having a flavonoid structure inhibiting the activity of an alcohol dehydrogenase as an active active ingredient in an amount of 0.002 to 2%.