JP6486022B2 - A method for measuring the degree of hangover, a method for inducing hangover in a test animal, a method for measuring the effect of hangover symptoms, a screening method for hangover inhibitors, and a method for selecting liquors that are difficult to hangover - Google Patents

Description

  The present invention relates to a method for measuring the degree of hangover, a method for inducing a hangover in a test animal, a method for measuring the effect of suppressing a hangover symptom, a screening method for a hangover inhibitor, and a method for selecting liquors that are difficult to hangover.

As a method for preventing or reducing the unpleasant symptoms of hangover, which are harmful effects of drinking, research on hangover inhibitors such as alcohol metabolism promoters has been conducted.
However, as a test method for evaluating the effect of a hangover inhibitor, a method capable of quantitatively measuring the effect of preventing or reducing hangover symptoms and a method of inducing a hangover in a test animal have been well established so far. Not.

  In Patent Document 1, as a method for evaluating the efficacy of a composition for preventing or reducing hangover or hangover, the state of spontaneous movement is visually confirmed 5 to 10 minutes after administration of the composition and ethanol to an SD rat. Discloses a method for evaluating “hangover” and a method for evaluating “hangover” by having humans answer about subjective symptoms when drinking the composition after ingestion or after ingestion. However, since these evaluations are based on the subjectivity of the person performing the evaluation, an objective and quantitative evaluation method is required instead of an objective evaluation.

  Further, Patent Document 2 discloses an agent that has an effect of suppressing and preventing discomfort caused by drinking including “hangover”. As a method for evaluating the efficacy of the agent, 30 minutes after oral administration of ethanol to ICR mice. There is disclosed a method of measuring a straight walking distance after a minute to 240 minutes (a distance walked on a stick installed at 45 degrees obliquely and walking for 10 minutes). However, this evaluation method does not evaluate the “spontaneous” activity of the animal because the tester intentionally places the animal on the “stick”. In addition, this evaluation method evaluates 30 minutes to 240 minutes after ethanol is administered, and also observes the stagger when walking on a bar installed obliquely. It is thought that it is not possible to say “hangover”.

  Furthermore, Non-Patent Document 1 includes an open field test, a tight rope test, a footprinting pattern, a hanging wire test, a T-shaped maze, and a Hole-board test 6 hours after ethanol was intraperitoneally administered to Swiss mice. A method of evaluating “hangover” is disclosed. However, this method is not a simple method because it requires the skill of the tester, and it is a system in which the tester intentionally moves the animal. It is not.

  In Non-Patent Document 2, as an evaluation method of “drunk” associated with oral ingestion of ethanol in mice, ethanol is administered to C57BL / 6N mice 1 hour before the test, or 2 hours and 1 hour before oral administration, then elevated cross Method of conducting a maze test, ethanol in C57BL / 6N mice immediately before the start of the test, or after 2 oral administrations 1 hour before the start of the test and immediately before the start of the test, the locomotor activity using the infrared sensor is 12 hours or 48 hours A method of measuring is disclosed. However, since these methods are measured immediately after ethanol administration, they are considered to be just “sickness” evaluation, and not only “hangover” evaluation, but are not sufficient as an evaluation method for “hangover”. It is not considered.

JP 2011-001348 A JP 2003-137788 A

Behavioral Brain Research 247,165-173 (2013) The Japanese Journal of Animal Psychology, 58, 1,1-13 (2008)

  In view of the above-described present situation, an object of the present invention is to provide a method for quantitatively (objectively) measuring the degree of hangover. Another object of the present invention is to provide a method for inducing a hangover in a test animal, a method for measuring the titer of a hangover inhibitor, a screening method for a hangover inhibitor, and a method for selecting liquors that are difficult to get a hangover. And

As a result of intensive studies to solve the above problems, the present inventors have surprisingly found that (A) after administration of ethanol to a test animal, the blood alcohol concentration of the test animal in the active phase is substantially reduced. And (B) inducing the hangover state of the test animal by substantially reducing the amount of behavior and / or locomotor activity of the test subject in the active period after the ethanol administration, and (C ) It has been found that the degree of hangover can be measured quantitatively (objectively) by using a measuring method including a step of measuring the amount of behavior and / or the amount of spontaneous movement of the test animal from the start of the active period.
In addition to the above, the present inventors have obtained various unexpected new findings as described below, and have further conducted intensive studies to complete the present invention.

That is, the present invention relates to the following methods for measuring the degree of hangover.
[1] (A) The step of substantially zeroing the blood alcohol concentration of the test animal in the active period after ethanol administration to the test animal, and then (B) the test animal in the active period after the ethanol administration Including the step of inducing a hangover state in the test animal by substantially reducing the amount of behavior and / or the amount of spontaneous exercise, and the step of (C) measuring the amount of behavior and / or the amount of spontaneous exercise of the test animal from the start of the active period A method for measuring the degree of hangover, characterized by:
[2] (A) The step of substantially zeroing the blood alcohol concentration of the test animal in the active period after ethanol administration to the test animal, and then (B) the test animal in the active period after the ethanol administration A method for inducing a hangover in a test animal, comprising a step of substantially reducing a behavior amount and / or a spontaneous exercise amount to induce a hangover state in the test animal.
[3] The method according to [1] or [2], wherein the test animal is a rodent.
[4] The behavior amount and / or the spontaneous exercise amount in the steps (B) and (C) are the behavior amount measured using an infrared sensor and / or the spontaneous exercise amount measured using a rotating car. The method according to any one of [1] to [3].
[5] The method according to any one of [1] to [4], wherein in the step (A), ethanol is administered once or twice or more at a specific time.
[6] The method according to any one of [1] to [5], wherein in the step (A), the concentration of ethanol to be administered is 1 to 100% (w / v).
[7] A method for measuring an effect of suppressing a hangover symptom, comprising the method according to any one of [1] to [6].
[8] A screening method for a hangover inhibitor, comprising the method according to any one of [1] to [6].
[9] A method for selecting liquors that are less prone to hangovers, comprising the method according to any one of [1] to [6].

According to the present invention, the degree of hangover can be measured quantitatively (objectively), and the potency of a hangover inhibitor can be measured. Further, according to the present invention, it is possible to measure the effect of suppressing hangover symptoms instead of sickness, and it is possible to screen for hangover inhibitors. Furthermore, according to the present invention, it is possible to induce a “hangover” in which the test animal is completely or partially separated from “sickness” or different from “sickness”.
Furthermore, according to the present invention, it is possible to select liquors that are less likely to have a hangover.

FIG. 3 shows time-dependent changes in the amount of ethanol in mice after oral administration of ethanol in Test Example 1. FIG. FIG. 6 shows changes with time in the amount of ethanol in rat blood after oral administration of ethanol in Test Example 2. FIG. It is the schedule on the 13th day of breeding in the ethanol single administration test of Example 1. It is a schedule from the test start of a mouse | mouth to the completion | finish of a test (from the 1st day to the 21st day) in the ethanol twice administration test of Example 2 (1-week interval administration). It is the schedule of the breeding 13th day in the ethanol twice administration test (4 hour interval administration) of Example 3. FIG. It is the data of the time-dependent change of the action amount measured using the infrared sensor on the 12th day of breeding in the ethanol twice administration test (4 hour interval administration) of Example 3. It is the data of the time-dependent change of the spontaneous exercise amount measured using the rotary wheel of the breeding 12th day in the ethanol twice administration test (4 hour interval administration) of Example 3. In the ethanol double administration test of Example 3 (4 hour interval administration), the second dose of ethanol was 2.5 g / kg b. w. In this case, it is data of change over time of the amount of behavior measured using an infrared sensor on the 13th day of breeding. In the ethanol double administration test of Example 3 (4 hour interval administration), the second dose of ethanol was 2.5 g / kg b. w. In this case, it is data of the time course change of the spontaneous exercise amount measured using the rotating vehicle on the 13th day of breeding.

Hereinafter, the present invention will be described in detail.
The method for measuring the degree of hangover according to the present invention comprises (A) a step of substantially zeroing the blood alcohol concentration of an active test animal after ethanol administration to the test animal, and then (B) the ethanol. After administration, a step of inducing a hangover state in the test animal by substantially reducing the amount of behavior and / or locomotor activity of the test animal in the active phase, and (C) the amount of behavior of the test animal from the start of the active phase and / or Alternatively, the step of measuring the amount of spontaneous exercise is an essential constituent feature.
The measurement method of the present invention is usually carried out in the order of (A) to (C), and may include steps other than the steps (A) to (C).

  In the present invention, “hangover” refers to a state in which the blood alcohol concentration of a test animal is substantially zero after alcohol intake, and a behavioral influence is caused by alcohol intake.

How to measure the degree of hangover
Step (A)
The test animal used in the present invention is not particularly limited, and examples thereof include rodents, and preferably, an inbred strain, a mutant strain, a hybrid strain, a closed colony, a diabetes model, or an immunodeficiency model mouse (AKR, BALB / cA, BALB / cB, CBA, C3H / HeN, C57BL, C57BL / cd, C57L, C58, DBA, KK, NZW, RF, SJL, SWR, FVB / N, 129 ^{+ Ter} / Sv, C57BL / 6-Fv ^S , DDD-Fv ^S , CDF ₁ , BDF ₁ , B6C3F2, CD2F1, B6D2F1, B6C3F1, dd, ICR, CFW, CF # 1, Swiss mouse, kk / Ta, KK / A ^y BKS.Cg-m + ^{/ + Lepr db /, GK,} SDT, NOD / Shi, BALB / AJcl-mu / mu, F344 / NJcl-mu / mu, CB-17 / Icr-scid / scid, NOD / ShiJic-scid, ARY / NscJcl-ally / ally), inbred or closed colony or diabetes model Rats (ACI, BN, F344, LEW, M520, SHR, WAG, SD, Wister, Long-Evans, Danryu, GK, SDT) and the like, more preferably inbred mice (A, AKR, BALB / cA) , BALB / cB, CBA, C3H / HeN, C57BL, C57BL / cd, C57L, C58, DBA, KK, NZW, RF, SJL, SWR, FVB / N, 129 ^{+ Ter} / Sv), inbred rats (ACI, BN, F344, LEW, M520, SHR, WAG), etc. The preferred are mouse (BALB / cA, C3H / HeN, C57BL).

  In the present invention, the term “active period” refers to a period of time during which a living organism performs daily activities from when it wakes up until it goes to bed. For example, in the case of nocturnal animals, it is during the dark period. Say.

  In the present invention, the “dark period” usually means an environment without lighting, and corresponds to an active period of nocturnal animals. The “light period” usually means an environment with lighting, which corresponds to a dormant period of nocturnal animals. The time corresponding to the dark period and the time corresponding to the light period can be changed as appropriate according to the facility for performing the measurement method of the present invention. For example, as a first embodiment when using a mouse as a test animal, the dark period can be a period from 19:00 to 7 o'clock, the light period can be a period from 7 o'clock to 19 o'clock, As an embodiment, the dark period may be a period from 7 o'clock to 19 o'clock, and the light period may be a period from 19 o'clock to 7 o'clock. The term “nocturnal animal” refers to an animal that mainly performs activities such as feeding and reproduction at night, or an animal that increases its activity at night or in the dark period of a light-dark cycle, such as rodents. Genus, bat, owl, etc.

  The ethanol used in the present invention is not particularly limited, and a commercially available product can be used, or a commercially available product purified by a conventional method can also be used. Ethanol may be used alone, a mixture of ethanol and water (aqueous solution), a beverage containing ethanol, or the like may be used. These aqueous solutions and beverages can be prepared using conventionally known methods, and may contain substances other than ethanol. The beverage containing ethanol (ethanol-containing beverage) may be an alcoholic beverage.

  Although it does not specifically limit as said alcoholic beverage, For example, wine, beer, sake, whiskey, brandy, shochu, gin, rum, vodka, tequila etc. are mentioned.

  The concentration of ethanol in the aqueous ethanol solution and the ethanol-containing beverage is not particularly limited, but is usually about 1 to 100% (w / v), preferably about 3% to 50% with respect to the total amount of the aqueous solution or beverage. % (W / v), more preferably about 10 to 40% (w / v).

In the present invention, the method of administering ethanol to the test animal is not particularly limited. For example, a method of adding ethanol to food or water of the test animal to ingest the test animal, a method of artificially administering, etc. Is mentioned.
The dose of ethanol is not particularly limited, but is usually about 0.5 to 50 g / kg b.w., preferably about 2 to 30 g / kg b.w. per administration. As such, it is administered to test animals.

  As a method for causing a test animal to ingest ethanol into food or water, for example, a method of allowing a test animal to freely ingest food or water containing ethanol for a certain period of time can be used. The time for ingesting the food or water is not particularly limited, but the blood alcohol concentration of the test animal in the active period needs to be substantially zero, and the intake end time is determined by the ethanol concentration of the food or water. To do.

  The method of artificially administering ethanol is not particularly limited, but is preferably oral intragastric administration, intraperitoneal administration, intravenous administration, enteral administration, etc., more preferably intragastric absorption of ethanol from the intestine. Oral administration, enteral administration, and the like, more preferably intragastric oral administration.

Moreover, although the timing which administers ethanol is not specifically limited, It is preferable to administer at a specific time. The specific time is not particularly limited, but the blood alcohol concentration of the test animal in the active phase needs to be substantially zero. For example, when ethanol is administered orally at 3 g / kg b.w. 4 hours before, 8 hours before, etc. from the activity period of animal testing are preferable.
Furthermore, the administration frequency of ethanol is not particularly limited, and can be administered once or twice or more depending on the ethanol concentration of feed or water. For example, when orally administering ethanol 3 g / kg b.w. 2 times.

In the present invention, the test animal may be ingested with a hangover inhibitor.
In the present invention, the timing at which the test animal ingests the hangover inhibitor is not particularly limited, but it can be ingested before or simultaneously with or after ethanol administration, depending on the mechanism of action of the agent. For example, in the step (A), the test animal may be ingested with a hangover inhibitor before ethanol administration. The ingestion method is not particularly limited, and examples thereof include a method of causing a test animal to contain a hangover inhibitor in food or water of the test animal, a method of artificially administering the method, and the like.
The intake of the hangover inhibitor is not particularly limited, and can be appropriately changed according to the type of the agent.

  As a method of causing a test animal to contain a hangover inhibitor in food or water, for example, a method of allowing a test animal to freely ingest food or water containing a hangover inhibitor for a certain period of time is used. Can do.

  The method for artificially administering a hangover inhibitor to a test animal is not particularly limited, but preferably includes oral intragastric administration, intraperitoneal administration, intravenous administration, enteral administration, and the like.

  In the present invention, the hangover inhibitor to be ingested by the test animal is not particularly limited. For example, an alcohol metabolism promoter, an acetaldehyde degradation promoter, or a hangover inhibitor is dissolved or dispersed in an aqueous methylcellulose solution, water, food, etc. Etc. are preferred. These dissolution and dispersion methods can use conventional methods. In addition, as a hangover inhibitor, for example, a hangover symptom recovery agent described in JP 2012-31080 A can be used.

  The hangover suppressing component is not particularly limited, and is any food, medicine, compound, composition or the like generally used as a hangover suppressing component, and examples thereof include turmeric extract.

In a test animal in which the blood alcohol concentration is substantially zero after ethanol intake, if the behavioral effect of ethanol intake occurs, it indicates “hangover” instead of just “drunk”. In addition, the measuring method of the blood alcohol concentration of a test animal is not specifically limited, A conventionally well-known method can be used and the method as described in the below-mentioned Example can be used.
That is, the present invention includes a step (B) of inducing a hangover state in a test animal by substantially reducing the amount of behavior and / or the amount of spontaneous movement of the test animal in the active period after the ethanol administration, and from the start of the active period Before the step (C) of measuring the amount of behavior and / or the amount of spontaneous movement of the subject animal, it is confirmed that the blood alcohol concentration of the subject animal in the active phase is substantially zero.

  The blood alcohol concentration of the test animal is determined by the type of animal, the intake of food and water (such as satiety or fasting), the dose and concentration of alcohol, and the like. Examples of a method for substantially reducing the blood alcohol concentration of a test animal to zero include, for example, about 10 to 40% (w / v) ethanol 3 g / kg b. w. Can be made substantially zero by allowing about 4 hours or more, more preferably about 6 hours or more, from the last administration to the start of the active phase.

Process (B)
The measurement method of the present invention comprises the step (B) of inducing a hangover state of the test animal by substantially reducing the amount of behavior and / or the amount of spontaneous movement of the test animal in the active phase following the step (A). contains. As a method for measuring the amount of behavior and / or the amount of spontaneous exercise, the method described in the step (C) described later can be used.
In the step (B), the degree of decrease in the activity amount and / or the spontaneous exercise amount of the test animal in the active period is not particularly limited. Specifically, in the test animal not taking the hangover inhibitor, ethanol administration The amount of behavior using the infrared sensor on the day of ethanol administration with respect to the previous day is preferably about 0 to 90%, more preferably about 20 to 90%, still more preferably about 20 to 70%, and / or Alternatively, the amount of spontaneous exercise using a rotating wheel on the day of ethanol administration relative to the day before ethanol administration is preferably about 0 to 90%, more preferably about 10 to 90%, and more preferably about 10 to 70%. State.

Process (C)
In addition to the steps (A) and (B), the measurement method of the present invention includes a step (C) of measuring the amount of behavior and / or the amount of spontaneous movement of the test animal from the start of the active phase.
In the present invention, the behavior amount and / or the spontaneous exercise amount of the test animal is preferably the behavior amount measured using an infrared sensor and / or the spontaneous exercise amount measured using a rotating wheel.

The amount of action measured using the infrared sensor can be obtained from the number of counts using the infrared sensor, as described in Examples below.
Further, the spontaneous momentum measured using the rotating wheel can be obtained from the number of rotations (counting number) using the rotating wheel, as described in the examples described later.

  The time for measuring the amount of behavior and / or the amount of spontaneous movement in step (C) is not particularly limited, but a fixed time from the start of the active period of the test animal is preferable. For example, in nocturnal animals such as rodents, Since the amount of action and the amount of spontaneous exercise increase immediately after the start of the period, 3 or 4 hours from the start of the dark period is more preferable.

  The amount of behavior and / or the amount of spontaneous exercise of the subject animal on the day before the steps (A) to (C) (ie, the day before the subject animal ingests ethanol) Alternatively, it is measured at the same time as the time for measuring the spontaneous exercise amount, and the behavior amount and / or the spontaneous exercise amount of the test animal on the previous day is compared with the behavior amount and / or the spontaneous exercise amount measured in the step (C). By doing so, the degree of hangover can be measured.

Method for inducing hangover in test animal After ethanol is administered in the step (A), the blood alcohol concentration becomes substantially zero, and in the step (B), the amount of behavior and / or the amount of spontaneous exercise is substantially reduced. A test animal that has decreased to a hangover condition. That is, the present invention also includes a method for inducing a hangover in a test animal, comprising the steps (A) and (B). In the method for inducing hangover of the test animal of the present invention, the aspects of steps (A) and (B) are the same as the aspects of steps (A) and (B) in the above-described method for measuring the degree of hangover of the present invention. Are the same.

Method for measuring hangover symptom suppression effect, method for screening for hangover inhibitor In the method for measuring the degree of hangover of the present invention described above, an animal to which a hangover inhibitor is administered before or simultaneously with or after administration of ethanol as a test animal And a non-administered animal, and comparing the behavioral amount and / or the amount of spontaneous movement of the test animal from the start of the active phase measured in step (C) between them, the hangover of the hangover inhibitor The symptom suppression effect can be measured and evaluated. Further, by using a test animal to which a different type of hangover inhibitor is administered, the strong agent can be selected. Moreover, the screening of a hangover inhibitor can be performed by using a test animal administered with any kind of different foods, pharmaceuticals, compounds or compositions. To a test animal not administered with a hangover inhibitor, only a solvent (dispersion medium) may be administered at the same timing as when the agent is administered to a test animal administered with a hangover inhibitor as a comparison target. The solvent is preferably the same as the solvent used for preparing the hangover inhibitor to be compared.

  In addition, the administration of the hangover inhibitor is preferably in the same manner as described in the above paragraphs [0026] to [0030].

Method for selecting liquors that are less likely to have hangovers The present invention uses different types of alcoholic beverages (alcohol) to be administered as ethanol in the step (A), and implements a method comprising the steps (A) to (C). Thus, it is possible to measure the degree of hangover depending on the type of liquor, and to select liquors that are difficult to hangover.

  The present invention will be specifically described with reference to the following examples and comparative examples, but the present invention is not limited thereto.

<Measurement of activity using an infrared sensor>
A mouse is placed under a behavior measuring device (Muromachi Kikai, Supermex PYS-001) using an infrared sensor, and the body temperature (far infrared) of the mouse is sensed by the lens of the device using the device. The number of times per hour was counted, and the counted number was used as the amount of action. Moreover, the value which integrated | accumulated the measured action amount per hour was made into the integrated action amount.

<Measurement of spontaneous momentum using a rotating car>
A rotating wheel (mouse rotating momentum measuring device, made by Muromachi Kikai Co., Ltd., Igloo Fast Track FT-713) is installed in the breeding environment of the mouse, and the number of times the mouse turns the rotating wheel is counted. The number was the amount of spontaneous exercise. Moreover, the value which integrated | accumulated the measured spontaneous exercise amount per hour was made into the integrated spontaneous exercise amount.

<Measurement of blood alcohol concentration>
Blood was collected from the test animals, and F-kit ethanol (JK International, 176290) was used for the measurement of blood alcohol concentration.

<Time of disappearance of ethanol in blood of mice administered with oral ethanol>
(Test Example 1)
As a mouse breeding condition, the mouse breeding room was set to a 12-hour light / dark cycle (light period: 7:00 to 19:00).
ICR mice were orally administered with a 30% (w / v) aqueous ethanol solution at 3 g / kg bw or a 40% (w / v) aqueous ethanol solution at 4 g / kg bw. The amount of ethanol in blood was measured before ethanol administration (0 hour) and 0.5, 2, 4, and 6 hours after ethanol administration. The measurement results are shown in FIG. FIG. 1 shows the value of Mean ± SD (mean ± standard deviation) for N = 4.
From the results shown in FIG. 1, it was found that approximately 3 hours were required when a 30% (w / v) ethanol aqueous solution was administered to mice at 3 g / kg bw, and a 40% (w / v) ethanol aqueous solution was administered to mice at 4 g / kg. In about 6 hours of administration, the blood ethanol concentration becomes substantially zero.
That is, when a 30% (w / v) ethanol aqueous solution is administered to a mouse at 3 g / kg bw, the blood ethanol concentration is preferably substantially reduced during the mouse active phase (19:00 to 7:00). At 4 hours before the active phase, and more preferably at 4 hours before the start of the dark period so that the blood ethanol concentration is substantially zero at the start of the dark period. . In addition, when a 40% (w / v) ethanol aqueous solution is administered to a mouse at 4 g / kg b.w., it is preferably administered 6 hours before the active phase of the mouse, and more preferably at the start of the dark phase. Administer 6 hours before.

<Ethanol disappearance time in rats after oral administration of ethanol>
(Test Example 2)
The SD rat was orally administered with a 40% (w / v) aqueous ethanol solution at 2 g / kg bw. The amount of ethanol in blood was measured before ethanol administration (0 hour) and 1, 2, 3, and 4 hours after ethanol administration.
The measurement results are shown in FIG. FIG. 2 shows the value of Mean ± SD (mean ± standard deviation) for N = 5.
From the results of FIG. 2, the blood ethanol concentration becomes substantially zero in about 4 hours when a 40% (w / v) ethanol aqueous solution is administered to rats at 2 g / kg bw.
That is, when a 40% (w / v) ethanol aqueous solution is administered to a rat at 2 g / kg bw, the blood ethanol concentration is preferably substantially reduced during the rat active period (19: 00 to 7:00). At 4 hours before the active phase, and more preferably at 4 hours before the start of the dark period so that the blood ethanol concentration is substantially zero at the start of the dark period. .

<Selection of mouse used for measurement>
(Test Example 3)
The following types of mice were placed in a plastic cage with a rotating wheel and raised for 14 days. From the start of the breeding until the 12th day, the mice were allowed to freely ingest food and water. On the 13th day of breeding, treatment was performed according to the following schedule.
Test animals: Mice (inbred lines: C57BL / 6N (B6N), BALB / cA (BALB / c), C3H / HeN (C3H), closed colony: ICR) were used.
Schedule: Mice were fasted and dehydrated from 12:30 to 15:00, and at 15:00, 30% (w / v) aqueous ethanol solution was orally administered to mice at 3 g / kg bw.
The amount of behavior and the amount of spontaneous exercise on the 12th, 13th, and 14th days of breeding were measured.

  Table 1 shows the results of the accumulated action amount and the accumulated spontaneous exercise amount for 3 hours (19: 00 to 22:00) from the start of the dark period on the 12th day of breeding (the day before ethanol administration). From the results in Table 1, B6N, BALB / c, and C3H inbred mice have low CV and are suitable for the method of the present invention.

In addition, on the 13th day of breeding (on the day of ethanol administration), the accumulated action amount on the 12th day in Table 1 above for the accumulated action amount and the accumulated spontaneous exercise amount for 3 hours (19: 00 to 22:00) from the start of the dark period, Table 2 shows values obtained by calculating the integrated action amount and the ratio (%) of the integrated spontaneous exercise amount on the 13th day with respect to the integrated spontaneous exercise amount. In Tables 1 and 2, Mean represents an average value, SD represents a standard deviation, CV represents a coefficient of variation, and A. U. Indicates an arbitrary unit.
From the results shown in Table 2, the B6N mouse has the most reduced amount of behavior and spontaneous movement, and there is little variation. Therefore, the B6N mouse is optimal as the mouse used in the method of the present invention.

(Example 1: One administration)
B6N mice were acclimated in a wire mesh cage for 3 days, then transferred to a plastic cage with a rotating wheel and reared for 14 days. On the 13th day of breeding, treatment was performed according to the following schedule. In FIG. 3, the schedule of the breeding 13th day of the present Example 1 is shown. In FIG. 3, “measurement” indicates measurement of the amount of behavior and / or the amount of spontaneous exercise.
Schedule: Mice were fasted and watered from 12:30 to 15:00, and at 14:30, turmeric extract as a hangover inhibitor was 20 mg / kg b. w. Orally administered so that The administration liquid used was a 0.2% (w / v) turmeric extract dispersion. Only the solvent (0.5 w / v% methylcellulose 400 solution) was orally administered to the control group of mice. At 15:00, 4 hours before the start of the dark period, a 15% (w / v) ethanol solution was given to the mice at 3 g / kg b. w. Orally administered so that The amount of behavior and the amount of spontaneous exercise on the 12th and 13th day of breeding were measured.

(Example 2: Twice administration, 1 week interval)
B6N mice were bred for 14 days in the same manner as in Example 1, and further bred for 1 week. The animals were treated according to the following schedule on the 13th and 20th day of breeding. Except for the 13th and 20th days of breeding, the mice were allowed to freely ingest food and water. FIG. 4 shows a mouse schedule of the second embodiment. In FIG. 4, “measurement” indicates measurement of the amount of behavior and / or the amount of spontaneous exercise.
Schedule: The mice were fasted and watered from 12:30 to 15:30 on the 13th day of breeding, and at 15:00, turmeric extract as a hangover suppressing component was 20 mg / kg b. w. Orally administered so that The administration liquid used was a 0.2% (w / v) turmeric extract dispersion. Only the solvent (0.5 w / v% methylcellulose 400 solution) was orally administered to the control group of mice. Then, at 15:30, a 30% (w / v) ethanol solution was given to the mice at 2 g / kg b. w. Was orally administered (first administration).
Further, the mice were fasted and dehydrated from 12:30 to 16:00 on the 20th day of breeding, and at 15:30, turmeric extract as a hangover suppressing ingredient was 20 mg / kg b. w. Orally administered so that The administration liquid used was a 0.2% (w / v) turmeric extract dispersion. A control group of mice was orally administered with solvent alone. At 16:00, a 30% (w / v) ethanol solution was given to mice and 3 g / kg b. w. Was orally administered (second administration). The amount of behavior and the amount of spontaneous exercise on the 12th and 13th days of breeding, and the amount of activity and the amount of spontaneous exercise on the 19th and 20th days were measured.

(Example 3: 2 doses, 4 hour intervals)
B6N mice were raised for 14 days in the same manner as in Example 1. On the 13th day of breeding, treatment was performed according to the following schedule. In FIG. 5, the schedule of the breeding 13th day of the present Example 3 is shown. In FIG. 5, “measurement” indicates measurement of the amount of behavior and / or the amount of spontaneous exercise.
Schedule: Water from 8:30 to 11:00 and fast from 15:00, and at 10:30, turmeric extract is 20 mg / kg b. w. Orally administered so that The administration liquid used was a 0.2% (w / v) turmeric extract dispersion. A control group of mice was orally administered with solvent alone. At 11:00 (first dose) and 15:00 (second dose), a 15% (w / v) ethanol solution was given to the mice at 2.5 g / kg b. w. Or 3 g / kg b. w. Orally administered so that The amount of behavior and the amount of spontaneous exercise on the 12th and 13th day of breeding were measured.

About the said Examples 1-3, about the integrated action amount of 3 hours (19: 0 to 22:00) and the integrated spontaneous exercise amount from the dark period start, the ratio of the integrated action amount on the ethanol administration day to the day before ethanol administration, and the integrated spontaneous exercise amount Table 3 and Table 4 show the values obtained by calculating (%) and the results of a t test comparing the turmeric extract administration group (with turmeric extract) and the control group (without turmeric extract).
For Examples 1 and 3, the value on the 12th day was used as the day before ethanol administration, the value on the 13th day was used as the day of ethanol administration, and for Example 2, the value on the 19th day as the day before ethanol administration. The value on the 20th day of breeding was used as the day of ethanol administration. Moreover, in Table 3 and Table 4, P value shows a significant probability.

  As is clear from the results of Tables 3 and 4, since the cumulative amount of behavior and the amount of spontaneous locomotor activity on the day of administration were reduced compared to the day before ethanol administration, the amount of behavior and / or locomotor activity of the mice was reduced by ethanol administration. It turns out that it fell substantially. The amount of behavior and the amount of spontaneous exercise were further decreased by administration at 4 hour intervals (Example 3) among the two administrations. In addition, from the results of Test Example 1, when ethanol 3 g / kg bw was administered to mice, the blood ethanol concentration became substantially zero in about 4 hours. It can be seen that the amount of medium ethanol is substantially zero. Therefore, the method of the present invention can induce a hangover in a test animal.

  Also, in Tables 3 and 4, by comparing the cases with and without the turmeric extract, the decrease in the accumulated action amount and the accumulated spontaneous exercise amount on the day of administration relative to the day before ethanol administration is suppressed by the administration of the turmeric extract. I found out. Therefore, the method of the present invention can measure the degree of hangover and can measure the hangover symptom suppression effect. In addition, the method of the present invention can select a strong agent by using a test animal administered with a different type of hangover inhibitor, and any food, drug, compound or composition of different types Screening for a hangover inhibitor can also be performed by using a test animal to which an object or the like is administered.

In particular, of the two doses of Example 3 (4 hour intervals), the second dose of ethanol was 2.5 g / kg b. w. In this case, the cumulative action amount and the cumulative spontaneous exercise amount are significantly higher with the turmeric extract than with the turmeric extract, and are suitable as the method of the present invention.
Moreover, the time-dependent change of the action amount and the spontaneous exercise amount on the said conditions is shown to FIGS. The values in FIGS. 6 to 9 indicate the value of Mean (average value) where N = 7. 6 and 7 show changes over time in the amount of behavior and spontaneous exercise amount on the 12th day of breeding, and FIGS. 8 and 9 show changes over time in the amount of behavior and spontaneous exercise amount on the 13th day of breeding, respectively. Also from these results, the effect of the hangover inhibitor can be measured by comparing the amount of action in FIGS. 6 and 8 and / or the amount of spontaneous movement in FIGS. 7 and 9.

  Furthermore, from the results of Tables 3 and 4, the evaluation of the hangover inhibitor is based on the ratio of the cumulative action amount and the amount of spontaneous exercise on the day of ethanol administration relative to the day before ethanol administration without turmeric extract (no administration of hangover inhibitor), It can be particularly preferably performed in the range of 25 to 62% in cumulative action amount and 13 to 33% in cumulative spontaneous exercise amount.

  The method of the present invention can induce a hangover in a test animal, can measure the degree of hangover quantitatively (objectively), and can measure the titer of a hangover inhibitor. In addition, the method of the present invention can screen for hangover inhibitors. Furthermore, the method of the present invention can select liquors that are difficult to hangover.

Claims (9)

(A) The step of substantially zeroing the blood alcohol concentration of the test animal in the active period after ethanol administration to the test animal, and (B) the behavior of the test animal in the active period after the ethanol administration Including a step of substantially reducing the amount and / or the amount of spontaneous exercise to induce a hangover state of the test animal, and (C) a step of measuring the amount of behavior and / or the amount of spontaneous exercise of the test animal from the start of the active period A method for measuring the degree of hangover, characterized by: (A) The step of substantially zeroing the blood alcohol concentration of the test animal in the active period after ethanol administration to the test animal, and (B) the behavior of the test animal in the active period after the ethanol administration A method for inducing a hangover in a test animal, comprising a step of inducing a hangover state in the test animal by substantially reducing the amount and / or the amount of spontaneous exercise. The method according to claim 1 or 2, wherein the test animal is a rodent. The action amount and / or spontaneous exercise amount in the step (B) and / or (C) is an action amount measured using an infrared sensor and / or a spontaneous exercise amount measured using a rotating car. Item 4. The method according to any one of Items 1 to 3.   The method according to any one of claims 1 to 4, wherein in the step (A), ethanol is administered once or twice or more at a specific time.   The method according to any one of claims 1 to 5, wherein in the step (A), the concentration of ethanol to be administered is 1 to 100% (w / v). A method for measuring the effect of suppressing hangover symptoms, comprising the method according to any one of claims 1 and 3 to 6, wherein the test animal is a hangover before or simultaneously with or after administration of ethanol. Using the animal administered with the inhibitor and the animal not administered with the agent, the behavioral amount and / or the amount of spontaneous exercise of the test animal from the start of the active phase measured in the step (C) is compared between them. Method. A screening method for a hangover inhibitor, comprising the method according to any one of claims 1 and 3 to 6, wherein a food, a pharmaceutical, a compound or a composition administered with any different type of food, pharmaceutical, compound or composition is used. A method in which a test animal is used and a behavioral amount and / or a spontaneous exercise amount of the test animal from the start of the active phase measured in the step (C) is compared between the two. A method for selecting liquor that is less prone to hangover, comprising the method according to any one of claims 1 and 3 to 6, wherein different types of alcoholic beverages administered as ethanol are used, and the step ( The method of implementing the method containing A)-(C).