Composition for preventing or alleviating hangover, containing <i>Longan arillus </i>extract

Abstract

Provided are a pharmaceutical composition for preventing an alcoholic hangover or alleviating a hangover, comprising a Longan arillus extract as an active ingredient, and a food composition for alleviating a hangover, comprising a Longan arillus extract.

Claims

1. A method of activating alcohol metabolism in liver tissue, or preventing or reducing an alcoholic hangover, the method comprising a step of administering an effective amount of a Longan arillus extract to a subject in need of activating alcohol metabolism in liver tissue, or preventing or reducing an alcoholic hangover.

2. The method of claim 1, wherein the extract further comprises an extract from at least one selected from the group consisting of Angelicae tenuissimae Radix and Polygalae Radix.

3. The method of claim 2, wherein the extract is a combined extract from a mixture of Longan arillus, Polygalae Radix, and Angelicae tenuissimae Radix at a weight ratio of 1:0.1 to 10:0.1 to 10.

4. The method of claim 2, wherein the extract further comprises an extract from at least one selected from the group consisting of Puerariae Radix, Scutellariae Radix, Platycodonis Radix, Angelica dahurica, Cimicifugae rhizoma, Raphani semen, and Acori gramineri rhizoma.

5. The method of claim 2, wherein the extract is obtained using water, at least one organic solvent selected from the group consisting of linear or branched lower alcohols of C.sub.1 to C.sub.4, or a mixture of water and the at least one organic solvent.

6. The method of claim 5, wherein the extract is obtained using water, ethanol, or a mixture of water and ethanol.

7. The method of claim 1, wherein the extract is obtained using water, at least one organic solvent selected from the group consisting of linear or branched lower alcohols of C.sub.1 to C.sub.4, or a mixture of water and the at least one organic solvent.

8. The method of claim 7, wherein the extract is obtained using water, ethanol, or a mixture of water and ethanol.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a graph of latency of fallen off(s) accounting for an effect on the motor impairment of mice intraperitoneally injected with alcohol after oral administration of an ethanol extract from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix, as measured by a Rotarod test.

(2) FIG. 2 is a graph of latency of fallen off(s) accounting for an effect on the motor impairment of mice intraperitoneally injected with alcohol after oral administration of a water extract (DG) from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix, as measured by a Rotarod test.

(3) FIG. 3 is a graph of LORR duration accounting for an effect of waking from a sleep induced by intraperitoneal injection of alcohol after oral administration of an ethanol extract from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix, as measured by a LORR test.

(4) FIG. 4 is a graph of LORR duraction accounting for an effect of waking from a sleep induced by intraperitoneal injection of alcohol after oral administration of a water extract from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix, as measured by a LORR test.

(5) FIG. 5 is a graph showing changes in the activity of alcohol dehydrogenase in liver tissues of mice intraperitoneally injected with alcohol after oral administration of a combined ethanol extract (WIN-1001X) from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix.

(6) FIG. 6 is a graph showing changes in the activity of alcohol dehydrogenase in liver tissues of mice intraperitoneally injected with alcohol after oral administration of a combined water extract (DG) from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix.

(7) FIG. 7 is a graph showing changes in the activity of acetaldehyde dehydrogenase in liver tissues of mice intraperitoneally injected with alcohol after oral administration of a combined ethanol extract (WIN-1001X) from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix.

(8) FIG. 8 is a graph showing changes in the activity of acetaldehyde dehydrogenase in liver tissues of mice intraperitoneally injected with alcohol after oral administration of a combined water extract (DG) from a mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix.

MODE FOR CARRYING OUT THE INVENTION

(9) Hereafter, the present disclosure will be described in detail by examples. The following examples are intended merely to illustrate the invention and are not construed to restrict the invention.

EXAMPLE 1

Preparation of Longan arillus Extract and Combined Extract Containing Longan arillus

(10) <1-1> Preparation of Longan arillus Extract

(11) After being cleansed and chopped, 40 g of Longan arillus was added with four volumes of water and heated at 95° C. for 4 hours in a herbal medicine pot to obtain a primary extract. The solid residue after filtration was added with the half volume of water for secondary extraction in the same condition.

(12) The two extracts thus obtained were pooled and filtered to remove solid residues, followed by centrifugation at 3200 rpm for 20 min. The supernatant was filtered through a 2.0 μm microfilter. The filtrate was collected to afford a Longan arillus extract.

(13) <1-2> Preparation of Combined Extracts From Mixture of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix According to Various Mix Ratios

(14) <1-2-1> Preparation of Water Extract

(15) Combined extracts were prepared from mixtures at various mix ratios of Longan arillus and Angelicae tenuissimae Radix, Longan arillus and Polygalae Radix, and Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix in the same manner as in Example 1-1. All the medicinal herbs were purchased from Cheonan Oriental Hospital of Daejeon University.

(16) Particularly, a water extract from a mixture of 1:1:1 of Longan arillus, Angelicae tenuissimae Radix, and Polygalae Radix was named DG.

(17) TABLE-US-00001 TABLE 1 Double Combined Extract Triple Combined Extract Name Mix Ratio Name Mix Ratio Y1W1 Longan arillus 30 g:Polygalae Y1W2G1 Longan arillus 15 g:Polygalae Radix 30 g Radix 30 g:Angelicae tenuissimae Radix 15 g Y1G1 Longan arillus 30 g:Angelicae tenuissimae Y1W1G2 Longan arillus 15 g:Polygalae Radix 30 g Radix 15:Angelicae tenuissimae Radix 30 g Y1W2 Longan arillus 20 g:Polygalae Y1W2G2 Longan arillus 12 g:Polygalae Radix 40 g Radix 24 g:Angelicae tenuissimae Radix 24 g Y1G2 Longan arillus 20 g:Angelicae tenuissimae Y1W1G1 (DG) Longan arillus 20 g:Polygalae Radix 40 g Radix 20 g:Angelicae tenuissimae Radix 20 g Y2W1 Longan arillus 40 g:Polygalae Y2W1G1 Longan arillus 30 g:Polygalae Radix 20 g Radix 15 g:Angelicae tenuissimae Radix 15 g Y2G1 Longan arillus 40 g:Angelicae tenuissimae Y2W2G1 Longan arillus 24 g:Polygalae Radix 20 g Radix 24 g:Angelicae tenuissimae Radix 12 g Y2W1G2 Longan arillus 24 g:Polygalae Radix 12 g:Angelicae tenuissimae Radix 24 g

(18) <1-2-2> Preparation of Ethanol Extract

(19) After being cleansed and chopped, the mixture of 20 g of Longan arillus, 20 g of Polygalae Radix, and 20 g of Angelicae tenuissimae Radix was added with four-fold volumes of 20% (v/v) ethanol/water solvent (aqueous ethanol solution) and heated at 95° C. for 4 hours in an earthenware pot to obtain a primary extract. The solid residue after filtration was added with the half volume of the 20% (v/v) ethanol/water solvent for secondary extraction in the same condition.

(20) The two extracts thus obtained were pooled and filtered to remove solid residues, followed by centrifugation at 3200 rpm for 20 min. The supernatant was filtered through a 2.0 μm microfilter. The filtrate was collected to afford an ethanol extract of Longan arillus, Polygalae Radix, and Angelicae tenuissimae Radix extract. The extract was named WIN-1001X.

(21) <1-3> Preparation of Combined Extracts From Longan arillus, Puerariae Radix, Angelicae tenuissimae Radix, Scutellariae Radix, Platycodonis Radix, Angelicae dahuricae Radix, Cimicifugae rhizoma, Raphani semen, Polygalae Radix, and Acori gramineri rhizoma

(22) A mixture of 12 g of Puerariae Radix, 8 g of Angelicae tenuissimae Radix, 4 g of Scutellariae Radix, 2 g of Platycodonis Radix, 4 g of Angelicae dahuricae Radix, 4 g of Cimicifugae rhizome, 4 g of Raphani semen, 8 g of Polygalae Radix, 12 g of Acori gramineri rhizome, and 12 g of Longan arillus was prepared and the mixture was subjected to extraction in the same manner as in Example 1-1 to obtain a combined extract.

(23) TABLE-US-00002 TABLE 2 Medicinal Herb Weight (g) and Composition (%) Longan arillus 12 g (17.14 Wt %) Angelicae tenuissimae Radix 8 g (11.43 Wt %) Polygalae Radix 8 g (11.43 Wt %) Puerariae Radix 12 g (17.14 Wt %) Scutellariae Radix 4 g (5.71 Wt %) Platycodonis Radix 2 g (2.86 Wt %) Cimicifugae rhizoma 4 g (5.71 Wt %) Angelicae dahuricae Radix 4 g (5.71 Wt %) Raphani semen 4 g (5.71 Wt %) Acori Gramineri rhizoma 12 g (17.14 Wt %) Total 70 g

EXPERIMENTAL EXAMPLE 1

Test for Suppressive Effect on Motor Impairment in Ethanol-Administered Mice (Alcohol-Induced Motor Impairment-Rotarod Test)

(24) The Longan arillus extract or combined extracts from Longan arillus plus other medicinal herbs, obtained in Example 1, were tested for suppressive effects on ethanol-induced motor impairment. To this end, male C57BL/6J mice (Japan SLC Inc., Shizuoka, Japan) 6 weeks old were used as experimental animals and acclimated for one week with free access to solid feeds and tap water before experiments. Two days before experiment, the mice were trained on a Rotarod machine (2-16 rpm) every day four times, each for 3 min.

(25) To begin with, WIN-1001X extract, prepared in Example 1-2-2, was tested for a suppressive effect on ethanol-induced motor impairment: After starvation for 18 hours, a saline solution was orally administered to a normal group and a control group and a solution of WIN-1001X extract in saline was administered at a dose of 300 mg/kg or 600 mg/kg to an experimental group. For a comparative group, one of the commercially available hangover-reducing beverages (Morning Care (Dong-A PHARM.)) was provided. For use in oral administration, a single recommended amount of intake of the beverage was lyophilized and then dissolved at a concentration corresponding to an amount for the adult standard weight (70 kg) in a saline solution (administration concentration : lyophilized Morning Care 3 g/10 g). One hour after oral administration, alcohol (20% (v/v) ethanol) was intraperitoneally injected at a dose of 4 g/kg to the normal, the control, the comparative, and the experimental group. 5, 30, 60, 120, 180, and 300 min later, the mice were put on a Rotarod machine (2-16 rpm, constant acceleration for 3 min) and measured for the time (seconds) taken to fall to the ground due to the loss of motor ability (Latency of Fallen off (s)).

(26) The results (Latency of Fallen off (s)) are depicted in FIG. 1. As shown in FIG. 1, WIN-1001X according to the present disclosure was observed to significantly counteract against alcohol-induced motor impairment at all of the concentrations tested and measured to have a hangover reducing effect higher than that of the commercially available beverage.

(27) In addition, the DG extract (Longan arillus:Polygalae Radix:Angelicae tenuissimae Radix=1:1:1 weight ratio) prepared in Example 1-2-1 was subjected to the same test. In this regard, a control was orally administered Morning Care at a dose of 0.17 ml/10 g while experimental groups were orally administered DG extract at a dose of 0.085 m1/10 g or 0.17 ml/10 g. The administration of Morning Care or DG extract was conducted twice in total 1.5 and 0.5 hours after alcohol injection.

(28) The result (Latency of Falling off (s)) thus obtained are summarized in Table 3 and depicted in FIG. 2:

(29) TABLE-US-00003 TABLE 3 Latency of Fallen Off (s) Time 5 30 60 90 180 300 min min min min min min Normal Control 160.8 143.8 150.0 152.3 146.5 146.8 Alcohol Control 2.8 2.1 13.2 27.5 59.3 87.7 Morning Care (3 g/10 g) 3.3 11.3 21.9 37.3 90.6 136.0 DG (0.085 ml/10 g) 4.3 4.7 20.0 36.0 72.3 128.0 DG (0.17 ml/10 g) 5.8 14.3 27.2 49.2 105.5 141.8

(30) In FIG. 2, Morning (-.box-tangle-solidup.-) stands for a Morning Care-administered group, DG (½) (-.circle-solid.-) for a DG (0.085 ml/10 g)-administered group, DG (-.square-solid.-) for a DG (0.17 ml/10 g)-administered group, and control (-□-) for a group injected with alcohol only (alcohol control).

(31) As is understood in FIG. 2 and Table 3, the DG-administered groups restored motor ability to a significant level after the alcohol-induced motor impairment, compared to the alcohol control. The restoration was found to be in a DG dose-dependent pattern.

<EXPERIMENTAL EXAMPLE 2>

Test for Awakening Effect on Ethanol-Administered Mice (Alcohol-Induced Sedation: LORR Test)

(32) 2.1. WIN-1001X Test

(33) For use as experimental animals in testing the Longan arillus extract or combined extracts from Longan arillus plus other medicinal herbs, obtained in Example 1, for awakening effects on ethanol-induced sedation, male C57BL/6J mice (Japan SLC Inc., Shizuoka, Japan) 10 weeks old were acclimated for one week with free access to solid feeds and tap water before experiments.

(34) After starvation for 18 hours, a saline solution was orally administered to a control and a solution of WIN-1001X extract, prepared in Example 1-2-2, in saline was administered at a dose of 300 mg/kg or 600 mg/kg to an experimental group. For a comparative group, one of the commercially available hangover-reducing beverages (Morning Care (Dong-A PHARM.)) was provided. A single recommended amount of intake of the beverage was lyophilized and then dissolved at a concentration corresponding to an amount for the adult standard weight (70 kg) in a saline solution (administration concentration : lyophilized Morning Care 3 g/kg).

(35) One hour after oral administration, alcohol (20% (v/v) ethanol) was intraperitoneally injected at a dose of 4 g/kg to the normal, the control, the comparative, and the experimental group (Longan arillus extract administered at doses of 150 and 300 mg/kg/10 ml). In order to determine whether mice in an alcohol-induced sleep phase were awakened, a loss of righting reflex (LORR) test was conducted. Sleeping mice were laid in a breeding cage with straw littered on the bottom thereof so that the limbs faced upward. Measurement was made of a time (LORR duration) taken for the mice to awake from the sleep and take an upright posture with the limbs standing on the bottom) (whether the upright posture was taken three times or more within 60 seconds).

(36) The results (LORR duration) were depicted in FIG. 3. As shown in FIG. 3, WIN-1001X exhibited a significant effect of waking from alcohol-induced sleeps and an improved performance at all doses tested, compared to the commercially available hangover-reducing beverage.

(37) 2.1. DG Extract Test

(38) The DG extract prepared in Example 1-2-1 was subjected to the same test. In this regard, a control was orally administered Morning Care at a dose of 0.17 ml/10 g while experimental groups were orally administered DG extract at a dose of 0.085 ml/10 g or 0.17 ml/10 g. The administration of Morning Care or DG extract was conducted twice in total 1.5 and 0.5 hours after alcohol injection and the LORR test was conducted 30 min after alcohol injection.

(39) The results (LORR duration) were depicted in FIG. 4. As shown in FIG. 4, DG extract exhibited a significant effect of waking from alcohol-induced sleeps, compared to the alcohol control, and an improved performance particularly at a dose of 0.17 ml/10 g, compared to the commercially available beverage.

EXPERIMENTAL EXAMPLE 3

Test for Activation of Alcohol Dehydrogenase (ADH) and Acetaldehyde Dehydrogenase (ALDH)

(40) In order to examine the activation effect of the Longan arillus extract and combined extracts from Longan arillus plus other medicinal herbs, obtained in Example 1, on alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH), liver tissues were taken from the mice at an LORR duration of 2 hours in Experimental Example 2.1 (LORR test for WIN-1001X-administered group) and 2.2 (LORR test for DG-administered group). The liver tissue was homogenized with Protein Extraction Solution (PRO-PREP™, iNtRON Biotechnology, Korea) at a concentration of 600 μl per 10 mg of liver tissue and stored on ice for 20 min, followed by centrifugation (13,000 g, 20 min, 4° C.). The supernatant was used as a measurement sample.

(41) 3.1. Measurement of Activity of Alcohol Dehydrogenase (ADH)

(42) The activity of alcohol dehydrogenase was measured using a reaction buffer containing 5.214 ml of distilled water, 2.642 ml of 1.0 M Tris-HCl buffer (pH 8.8), 1.072 ml of 20 mM NAD+, and 1.072 ml of 99.9% absolute ethanol per 10 ml. Briefly, 285 μl of the reaction buffer and 15 μl of the measurement sample were incubated together at 30° C. for 10 mM in each well of 96-well plates. Thereafter, absorbance at 340 nm was measured 0, 10, 20, and 30 mM after the incubation. Absorption deviations in the individual times with reference to 0 mM were calculated, and the results are depicted in FIG. 5 (test results for WIN-1001X-administerd groups) and FIG. 6 (test results for DG-administered groups).

(43) As can be seen in FIGS. 5 and 6, the Longan arillus extract (WIN-1001X or DG) invoked a remarkable increase of alcohol dehydrogenase activity in a dose-dependent manner.

(44) 3.2. Measurement of Activity of Acetaldehyde Dehydrogenase (ALDH)

(45) The activity of alcohol dehydrogenase was measured using a reaction buffer containing 7.492 ml of distilled water, 1.072 ml of 1.0 M Tris-HC1 buffer (pH 8.8), 0.359 ml of 20 mM NAD+, 0.359 ml of 1.0 M acetaldehyde, 0.359 ml of 3.0 M KCl, and 0.359 ml of 0.33 M 2-mercaptoethanol per 10 ml. Briefly, 285 μl of the reaction buffer and 15 μl of the measurement sample were incubated together at 30° C. for 10 mM in each well of 96-well plates. Thereafter, absorbance at 340 nm was measured 0, 10, 20, and 30 mM after the incubation. Absorption deviations in the individual times with reference to 0 mM were calculated.

(46) The results are depicted in FIG. 7 (test results for WIN-1001X-administerd groups) and FIG. 8 (test results for DG-administered groups).

(47) As can be seen in FIGS. 7 and 8, the Longan arillus extract (WIN-1001X or DG) invoked an increase of alcohol dehydrogenase activity in a dose-dependent manner.