WO2022188762A1 - Pharmaceutical composition for treating or alleviating hangover from alcohol, and use thereof - Google Patents

Abstract

A pharmaceutical composition for treating or alleviating a hangover from alcohol, which comprises radix codonopsis pilosulae, poria, and pericarpium citri reticulatae, or further comprises rhizoma zingiberis and rhizoma atractylodis macrocephalae. Further provided is a use in preparing a pharmaceutical for treating or alleviating a hangover from alcohol and liver, kidney, and gastrointestinal tract injury by means of using an effective amount of the pharmaceutical composition.

Description

[Title of invention made by ISA pursuant to Rule 37.2] Pharmaceutical composition for the treatment or reduction of alcohol hangover and use thereof technical field

The present invention relates to a novel pharmaceutical composition for treating or reducing alcohol hangover, liver, kidney and gastrointestinal damage.

Background technique

Alcohol-induced hangovers are the most commonly reported adverse effect of excessive drinking and occur when blood alcohol concentration (BAC) returns to almost zero. Since ethanol is miscible with water, it will target water-rich tissues, such as the brain, where it will cause familiar symptoms such as general distress, headache, tiredness, concentration problems, thirst, dizziness , nausea, cognitive impairment, and mood changes. An alcohol hangover can lead to absenteeism in the workplace, impaired job performance, reduced productivity, poor academic performance, and can jeopardize potentially dangerous daily activities such as driving a car or operating heavy machinery. These socioeconomic consequences and health risks of alcohol hangovers are much higher when compared to a variety of common diseases and other health risk factors.

The search for a cure for alcohol-induced damage and side effects, including hangovers, is as old as alcohol itself. There are many cures and preventives available, but scientific evidence about their effectiveness is generally lacking.

Accordingly, there is a need for methods and compositions capable of rapidly reducing symptoms of elevated blood alcohol concentration (BAC) or regaining sobriety after drinking alcohol, or reducing or preventing symptoms of drinking. The present invention addresses this need and others.

SUMMARY OF THE INVENTION

In one embodiment, the present invention discloses a pharmaceutical composition comprising (1) an herbal mixture comprising at least two herbs selected from the group consisting of dried ginger; Codonopsis; Poria; Chenpi and Atractylodes; and (2) A pharmaceutically acceptable carrier.

In another embodiment, there is provided a pharmaceutical composition comprising: (1) a herbal mixture comprising Codonopsis pilosula; Poria cocos and tangerine peel; and (2) a pharmaceutically acceptable carrier.

In another embodiment, there is provided a pharmaceutical composition comprising: (1) an herbal mixture comprising dried ginger; Codonopsis; Poria; tangerine peel and Atractylodes; and (2) a pharmaceutically acceptable carrier.

Also included is the use of a pharmaceutical composition described herein for the manufacture of a medicament for treating or reducing the effects of alcohol consumption.

Also included is the use of the pharmaceutical composition described herein for the manufacture of a medicament for the treatment or reduction of hepatocyte inflammation and fat accumulation.

Also included is the use of the pharmaceutical compositions described herein for the manufacture of a medicament for the treatment or reduction of impairment of the mucosal or intestinal function of the small intestine.

Also included is the use of the pharmaceutical compositions described herein for the manufacture of a medicament for the treatment or reduction of glomerular damage or renal function damage.

Also provided are methods of treating or reducing the effects of alcohol consumption in an individual in need thereof by administering the pharmaceutical compositions described herein.

The terms "invention" and "present invention" as used in this patent are intended to refer broadly to all of the subject matter within the scope of this patent and the following patent claims. Statements containing these terms should be understood not to limit the subject matter described herein or to limit the meaning or scope of the following patent claims. Embodiments of the invention covered by this patent are defined by the following claims rather than this Summary. This summary is a high-level overview of various aspects of the invention, and introduces some concepts that are further described in the following detailed description section. This Summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended solely for use in determining the scope of the claimed subject matter. The subject matter should be understood by reference to appropriate portions of the entire specification, any or all drawings, and the scope of each claim.

The present invention will become clearer when reading the following figures and embodiments.

Description of drawings

Figure 1 is a line graph illustrating the effect of the pharmaceutical composition of the present invention on the body weight of mice.

2 and 3 are histograms illustrating the effects of the pharmaceutical composition of the present invention on the liver/body weight ratio and the kidney/body weight ratio in mice.

4 and 5 are histograms illustrating the effects of the pharmaceutical composition of the present invention on the levels of serum liver function (AST and ALT) in mice.

6 and 7 are histograms illustrating the effects of the pharmaceutical composition of the present invention on the levels of serum total cholesterol (T-CHO) and triglyceride (TG) in mice.

Figure 8 is a histogram illustrating the effect of the pharmaceutical composition of the present invention on serum glucose levels in mice.

9 and 10 are histograms illustrating the effects of the pharmaceutical composition of the present invention on the levels of total cholesterol (T-CHO) and triglyceride (TG) in the liver of mice.

Figure 11 is a histogram illustrating the steatosis score of the pharmaceutical composition of the present invention in treating alcoholic liver disease mice.

Figure 12 is a histogram illustrating the effect of one embodiment of a pharmaceutical composition of the present invention on field sobriety test performance.

Figure 13 is a bar graph illustrating the effect of one embodiment of a pharmaceutical composition of the present invention on alcohol-induced symptoms 1 hour after drinking alcohol.

Figure 14 is a bar graph illustrating the effect of one embodiment of a pharmaceutical composition of the present invention on alcohol-induced symptoms 2 hours after drinking alcohol.

Figure 15 is a bar graph illustrating the effect of one embodiment of a pharmaceutical composition of the present invention on alcohol-induced symptoms 24 hours after drinking alcohol.

Figures 16A-16C are histograms illustrating the effect of one embodiment of a pharmaceutical composition of the present invention on liver function tests (AST, ALT and γ-GT).

Figures 17A and 17B are bar graphs illustrating the effect of one embodiment of a pharmaceutical composition of the invention on renal function tests (aldosterone and creatinine).

Figure 18 is a bar graph illustrating the effect of one embodiment of the pharmaceutical composition of the present invention on glucose-6-phosphate dehydrogenase (G6PD).

Figure 19 is a collection of hematoxylin and eosin stained (H and E) images showing alcohol-induced damage to the jejunal villi (panel B) and an embodiment of a pharmaceutical composition of the invention reduces alcohol-induced damage to the small intestinal capsule damage (C to E panels).

Figure 20 is a collection of H and E images showing alcohol-induced damage to glomeruli (Panel B) and an embodiment of a pharmaceutical composition of the invention reduces alcohol-induced damage to glomeruli (Panels C to E) .

Detailed description of the invention

definition

As employed above and throughout this disclosure, unless otherwise indicated, the following terms shall be understood to have the following meanings.

As used herein, the singular terms "a/an" and "the" include plural referents unless the context clearly dictates otherwise.

As used herein, an "effective amount" includes a dose of the pharmaceutical composition sufficient to treat or ameliorate at least one symptom due to alcohol consumption or BAC elevation.

As used herein, the term "treatment" refers to palliative use or outcome, and/or slowing or inhibiting the development of alcohol-induced symptoms or signs.

As used herein, the term "individual" generally refers to a human or animal having or suspected of having symptoms due to alcohol consumption or elevated BAC. Individuals who are about to consume alcohol, whether or not with symptoms or signs of alcohol consumption or elevated BAC, are also included within the scope of the term "individual".

All numbers herein are understood to be modified by "about". As used herein, the term "about" is meant to encompass a variation of ±10%.

In one embodiment, the present invention discloses a pharmaceutical composition comprising (1) an herbal mixture comprising at least two herbs selected from the group consisting of dried ginger; Codonopsis; Poria; Chenpi and Atractylodes; and (2) A pharmaceutically acceptable carrier.

In an exemplary embodiment, the herbal mixture includes dried ginger and Codonopsis. In an exemplary embodiment, the herbal mixture comprises Codonopsis and Poria. In an exemplary embodiment, the herbal mixture includes Poria cocos and tangerine peel. In an exemplary embodiment, the herbal mixture comprises Chenpi and Atractylodes. In an exemplary embodiment, the herbal mixture includes dried ginger and Poria cocos. In an exemplary embodiment, the herbal mixture includes dried ginger and dried tangerine peel. In an exemplary embodiment, the herbal mixture comprises dried ginger and Atractylodes Rhizoma. In an exemplary embodiment, the herbal mixture comprises Codonopsis and tangerine peel. In an exemplary embodiment, the herbal mixture comprises Codonopsis and Atractylodes. In an exemplary embodiment, the herbal mixture comprises Poria and Atractylodes.

In another embodiment, the present invention discloses a pharmaceutical composition comprising (1) an herbal mixture comprising at least three herbs selected from the group consisting of dried ginger; Codonopsis; Poria; Chenpi and Atractylodes; and (2) a pharmaceutically acceptable carrier.

In an exemplary embodiment, the herbal mixture includes dried ginger, Codonopsis, and Poria. In an exemplary embodiment, the herbal mixture comprises Codonopsis, Poria, and tangerine peel. In an exemplary embodiment, the herbal mixture comprises Poria cocos, tangerine peel, and Atractylodes. In an exemplary embodiment, the herbal mixture comprises dried tangerine peel, Atractylodes Rhizoma, and dried ginger. In an exemplary embodiment, the herbal mixture includes dried ginger, dried tangerine peel, and Atractylodes. In an exemplary embodiment, the herbal mixture includes dried ginger, Codonopsis, and dried tangerine peel. In an exemplary embodiment, the herbal mixture includes dried ginger, Poria, and Atractylodes. In an exemplary embodiment, the weight ratio of the herbal mixture of Codonopsis: Chenpi: Poria is about 25-35%: 25-35%: 30-50%.

In an exemplary embodiment, the herbal mixture includes dried ginger, Codonopsis, and Atractylodes. In an exemplary embodiment, the herbal mixture includes dried ginger, Poria, and Atractylodes.

In an exemplary embodiment, the herbal mixture comprises Codonopsis, tangerine peel, and Atractylodes. In an exemplary embodiment, the herbal mixture comprises Codonopsis, Poria, and Atractylodes.

In another embodiment, the present invention discloses a pharmaceutical composition comprising (1) a herbal mixture comprising dried ginger; Codonopsis; Poria; dried tangerine peel and Atractylodes; and (2) a pharmaceutically acceptable carrier.

In an exemplary embodiment, the herbal extract comprises about 5-15% by weight dried ginger. In an exemplary embodiment, the herbal extract comprises about 15-25% by weight of Codonopsis pilosula. In an exemplary embodiment, the herbal extract comprises about 25-35 wt% Poria. In an exemplary embodiment, the herbal extract comprises about 15-25% by weight of dried tangerine peel. In an exemplary embodiment, the herbal extract contains about 10-20% by weight of Atractylodes Rhizoma. In an exemplary embodiment, the herbal extract comprises about 20-25% by weight of Codonopsis Radix; about 30-35% by weight of Poria; about 20-25% by weight of dried tangerine peel; about 15-18% by weight of Atractylodes Rhizoma and Contains about 6-12% by weight of dried ginger.

In an exemplary embodiment, the herbal mixture is a dry or lyophilized powder of at least two of the listed herbs (ie, dried ginger; Codonopsis; Poria; Chenpi and Atractylodes). In an exemplary embodiment, the herbal mixture is an aqueous or alcoholic extract of at least two of the listed herbs (ie, dried ginger; Codonopsis; Poria; Chenpi and Atractylodes).

The herbs in the herbal mixture can be raw herbs, dry powders of raw herbs, freeze-dried powders, ground powders, decoctions, crude water or alcohol extracts or extract granules. In an exemplary embodiment, the herbal mixture is prepared by aqueous extraction, in which the raw herbs (optionally ground prior to extraction for optimal extraction results) are heated in water or solvent, followed by filtration, concentration (in which liquid extraction The material is condensed by vacuum or low pressure concentration).

The National Institute on Alcohol Abuse and Alcoholism lists varying degrees of impairment as blood alcohol concentration (BAC) increases:

Mild impairment (BAC between 0.0-0.05%): mild impairment of speech, memory, concentration, coordination and balance, relaxation and somnolence.

Moderate impairment (BAC level between 0.06-0.15%): increased risk of aggression, further impairment of speech, memory, concentration, coordination, balance and driving skills.

Severe impairment (BAC level between 0.16-0.30%): significant impairment of speech, memory, attention, coordination, balance, judgment and decision-making, confusion, fainting, nausea and vomiting, loss of consciousness, mood changes, irregular breathing .

Life-threatening (alcohol overdose, BAC 0.31-0.45%): confusion, loss of consciousness, seizures, and hypothermia.

A hangover is a group of symptoms that occurs due to excessive drinking. Typical symptoms include fatigue, weakness, thirst, headache, muscle aches, nausea, stomach pain, dizziness, sensitivity to light and sound, anxiety, irritability, sweating, and increased blood pressure.

For example, it is desirable to administer the pharmaceutical composition shortly before, during, or shortly after social drinking of any amount of alcohol to reduce symptoms and signs of elevated BAC and to enable the drinker to sober up quickly. It is also desirable to administer the pharmaceutical compositions for the treatment and prevention of gastrointestinal and urinary system damage, such as liver, kidney or intestinal damage.

The present invention provides methods of reducing or treating the effects of alcohol consumption in an individual in need thereof by administering the pharmaceutical compositions described herein.

Non-limiting examples of drinking effects include symptoms, signs of elevated blood alcohol concentration, or an alcohol hangover.

Pharmaceutical compositions can be readily formulated or prepared using herbal mixtures and pharmaceutically acceptable carriers. A pharmaceutically acceptable carrier may contain a physiologically acceptable compound, which serves, for example, to stabilize or to increase or decrease the rate of absorption or clearance of the pharmaceutical compositions of the present invention. Physiologically acceptable compounds can include, for example, carbohydrates, such as glucose, sucrose, or polydextrose; antioxidants, such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; detergents; liposomal carriers; or other stabilizers and/or buffers. Such formulations can be prepared by a variety of techniques, including combining the herbal mixture with suitable pharmaceutically acceptable carriers such as liquid carriers, solid carriers, or both.

The pharmaceutical compositions provided herein optionally include antioxidants, buffers, bacteriostatic agents, suspending agents, thickening agents, preservatives, co-solvents, and viscosity-increasing agents or other therapeutic ingredients for the treatment of alcohol disorders, such as calcium acamprosate (acamprosate), gabapentin (gabapentin, trade name NEURONTIN) or topiramate (topiramate, trade name topamax). The carrier and other therapeutic ingredients must be acceptable in the sense of being compatible with the pharmaceutical composition and not injurious to the individual.

The pharmaceutical composition is administered in an amount effective to reduce symptoms or signs of elevated BAC in the subject. The dosage of the pharmaceutical composition administered will depend on the amount of alcohol consumed and other clinical factors, such as the weight and general condition of the individual, and the route of administration. Useful dosages of the pharmaceutical compositions provided herein are determined by comparing their in vitro activity with their in vivo activity in animal models. Methods for extrapolating effective doses in mice and other animals to humans are known in the art; see, eg, US Patent No. 4,938,949, which is incorporated herein by reference.

According to the methods provided herein, pharmaceutical compositions are delivered by any of a variety of routes, including but not limited to injection (eg, subcutaneous, intramuscular, intravenous, intraarterial, intraperitoneal, intradermal, intravitreal); dermal transdermal; transdermal; oral (eg, lozenges, powders, pills, lozenges, capsules, liquids, edible films, or any dosage form suitable for herbal medicine); implantable osmotic pumps; suppositories; aerosols Spray; topical; eye; nasal inhalation; lung inhalation or imprinted on skin. In one embodiment, the pharmaceutical composition of the present invention may be administered orally in the form of an aqueous extract.

The pharmaceutical composition can be administered in a single dose treatment or in multiple dose treatments over a period of time. The pharmaceutical compositions are conveniently administered at appropriate intervals, eg, once a day, twice a day, three times a day, four times a day, six times a day, every other day, every three days.

The methods described herein also encompass research methods and uses, including in vitro and in vivo methods of treating or reducing the effects of alcohol consumption in an individual following consumption of any amount of alcohol.

Embodiments of the invention are illustrated by the following examples, which should not be construed in any way to limit the scope of the invention. During the studies described in the examples below, unless otherwise stated, informed procedures were followed. For illustrative purposes, some procedures are described below.

Example

Example 1: Case Study on the Effects of Pharmaceutical Compositions on Mice

The purpose of this study is to study the protective effect of pharmaceutical compositions S1-S11 on improving alcoholic liver injury in a mouse model of alcoholic liver disease. .

Table 1

Numbering	pH	Codonopsis	Atractylodes	tangerine peel	Poria	dried ginger
S1	4.26	√	√	√	√	√
S2	4.27	√	√	√	-	-
S3	4.85	√	√	-	√	-
S4	5.15	√	√	-	-	√
S5	4.02	√	-	√	√	-
S6	4.32	√	-	√	-	√
S7	4.83	√	-	-	√	√
S8	3.78	-	√	√	√	-
S9	4.23	-	√	√	-	√
S10	4.84	-	√	-	√	√
S11	3.77	-	-	√	√	√

1. Alcoholic liver disease in mice

Fifty-two eight-week-old C57BL/6 male mice were acclimated to a pair-fed liquid diet (control diet without alcohol) for one week, and then the mice were divided into alcohol-fed and pair-fed groups (normal control group); the mice in the alcohol-fed group received a Lieber-DeCarli liquid diet (a high-fat alcoholic liquid diet) with an alcohol concentration of 1-5% (1-5% daily increase), and the mice were on a liquid diet 5% alcohol was fed ad libitum for 5 weeks; pair-fed mice served as a normal control group and received a calorie-matched liquid diet (liquid vehicle was redistilled water (ddH2O)) for ₅ weeks.

2. Treatment

At the beginning of the disease induction period, the vehicle (ddH2O ₎ or the pharmaceutical composition was administered orally (PO) once a day (QD) to the mice, and the mice were treated for 35 days or 5 weeks. The dose volume was 12.3 mL/kg (body weight) and day 1 of treatment was indicated as day 0. The experimental design is shown in Table 2 below.

Table 2

group	quantity	diet	treat	dose	path		Dosing frequency
1	4	Liquid diet control group	Carrier (ddH2O)	12.3mL/kg	PO	QD x 35
2	4	Lieber-DeCarli Liquid Diet	Carrier (ddH2O)	12.3mL/kg	PO	QD x 35
3	4	Lieber-DeCarli Liquid Diet	S1	12.3mL/kg	PO	QD x 35
4	4	Lieber-DeCarli Liquid Diet	S2	12.3mL/kg	PO	QD x 35
5	4	Lieber-DeCarli Liquid Diet	S3	12.3mL/kg	PO	QD x 35
6	4	Lieber-DeCarli Liquid Diet	S4	12.3mL/kg	PO	QD x 35
7	4	Lieber-DeCarli Liquid Diet	S5	12.3mL/kg	PO	QD x 35
8	4	Lieber-DeCarli Liquid Diet	S6	12.3mL/kg	PO	QD x 35
9	4	Lieber-DeCarli Liquid Diet	S7	12.3mL/kg	PO	QD x 35
10	4	Lieber-DeCarli Liquid Diet	S8	12.3mL/kg	PO	QD x 35
11	4	Lieber-DeCarli Liquid Diet	S9	12.3mL/kg	PO	QD x 35
12	4	Lieber-DeCarli Liquid Diet	S10	12.3mL/kg	PO	QD x 35
13	4	Lieber-DeCarli Liquid Diet	S11	12.3mL/kg	PO	QD x 35

11 groups of pharmaceutical compositions, including S1 to S11, were tested on mice with alcoholic liver disease to evaluate the protective effect on alcoholic liver injury. Animals in group 1 received a pair-fed liquid diet and vehicle (ddH ₂ O) as a normal control group; animals in group 2 received a Lieber-DeCarli liquid diet with vehicle (ddH ₂ O) as a disease control group; animals in group 3 to Group 13 animals received Lieber-DeCarli liquid diet and pharmaceutical compositions S1-S11, respectively.

Referring to Figure 1, significant weight loss was observed in alcohol-fed mice; while no significant body weight changes were observed in the treated group compared to the vehicle control group (Group 2).

Referring to Figures 2 and 3, at the end of the study (Day 34), the mean liver/body weight ratio was 5.78%, while the normal control group (Group 1) was 3.55% (p<0.05); and the kidney/body weight ratio The mean was 1.35% compared to 1.01% in the normal control group (Group 1) (p<0.05). It can be seen that the liver/body weight ratio and the kidney/body weight ratio of the mice fed with alcohol were significantly increased.

In addition, referring to Figure 2, with treatment group S1 (Group 3), S2 (Group 4), S3 (Group 5), S5 (Group 7), S6 (Group 8) or S10 (Group 12) After treatment, the liver/body weight ratio was significantly decreased (p<0.05) compared to the vehicle control group (Group 2).

3. Serum Chemistry Measurements

Mice were fasted overnight before blood collection on days 0 and 34, and blood samples were collected for serum chemistry analysis 4 hours after the last dose. Serum chemical parameters include AST (Aspartate aminotransferase, Aspartate aminotransferase), ALT (Alanine aminotransferase, Alanine aminotransferase), total cholesterol (T-CHO), triglyceride (TG) and blood glucose (GLU) , where AST, ALT, CHO and TG were measured using a Hitachi 7180 analyzer, while GLU was measured using a blood glucose meter.

Referring to Figure 4 and Figure 5, the blood chemistry analysis on the 34th day showed that compared with the normal control group (group 1), the serum AST and ALT levels of the mice fed with alcohol were significantly increased, and the AST value was 122.5 ± 29.6U /L, while the normal control group (group 1) was 80.8 ± 3.5 U/L; ALT value was 49.7 ± 6.3 U/L, while the normal control group (group 1) was 33.9 ± 3.0 U/L (p < 0.05 ). However, no significant differences in serum AST and ALT were observed in the treated groups when compared to the vehicle control group (Group 2).

Referring to Figure 6, at the start of treatment (day 0), treatment groups S1 (group 3), S3 (group 5), S5 (group 7), S7 (group 9) or S9 (group 11) Serum total cholesterol (T-CHO) levels were significantly increased (p<0.05) compared to the vehicle control group (group 2); and at the end of the study (day 34), compared with the normal control group (group 1) Compared with alcohol-fed mice, the serum T-CHO level was significantly increased (p<0.05). However, no significant increase in serum T-CHO was found in the treated group compared to the vehicle control group (Group 2).

Referring to Figure 7, compared to the vehicle control group (Group 2), the treatment groups S2 (Group 4), S3 (Group 5), S5 (Group 7), S6 (Group 8) and S9 (Group 11) Group) serum triglyceride (TG) levels were significantly lower (p < 0.05); while at the end of the study (day 34), alcohol-fed mice had serum TG levels compared to normal controls (group 1) Significantly lower (p<0.05); however, no significant changes in serum TG were found in the treated group compared to the vehicle control group (Group 2).

Referring to Figure 8, at the start of treatment (day 0), serum glucose levels in treatment groups S1 (group 3), S2 (group 4), or S5 (group 7) were comparable to the vehicle control group (group 2). ratio was significantly improved (p<0.05); while at the end of the study (day 34), the alcohol-fed mice in the treatment group S8 (group 10) had significantly higher serum glucose levels compared to the vehicle control group (group 2) decreased (p<0.05).

4. Measurement of liver total cholesterol (T-CHO) and triglyceride levels

At the end of the study on day 34, liver tissue was collected and extracted with PBS; the tissue was analyzed for total cholesterol (T-CHO) and triglycerides (TG) by using a Hitachi 7180 analyzer.

Referring to Figures 9 and 10, the liver total cholesterol (T-CHO) and triglyceride (TG) levels were increased in alcohol-fed mice. Compared with the vehicle control group (group 2), liver T-CHO levels were significantly reduced in the treatment groups S7 (group 9), S8 (group 10), S9 (group 11) and S11 (group 13) ( p<0.05); while the liver TG levels were significantly decreased (p<0.05) in the treatment group S8 (group 10) compared to the vehicle control group (group 2).

5. Measurement of liver enzymes

The level of glutathione peroxidase (GPx) in the liver was measured by ELISA, and the results are shown in Table 3 below.

table 3

6. Microscopic Evaluation

At the time of dissection of mice, tissue samples were collected and preserved in 10% neutral buffered formalin (NBF); livers, kidneys and intestines were trimmed, paraffin embedded, sectioned and treated with H (hematoxylin) and E (eosin) stained and examined microscopically (Leica DM2700M) by a veterinary pathologist blinded to the nature of the treatment.

6.1 Semi-quantitative scoring of hepatic steatosis lesions

The severity grading system of hepatic steatosis lesions under the microscope is shown in Table 4 below, and the scores of each group after treatment are shown in Table 5 and Figure 11 below.

Table 4

Fraction		grade
0	Normal (0%)
1	Minimum (<10%)
2	Slight (10 to 33%)
3	Moderate (33 to 66%)
4	Severe (66～100%)

table 5

^# p<0.05 Group 2 vs. Group 1

^* p<0.05 Group 7 (the weight ratio of Codonopsis, Poria and tangerine peel is 14:20:14) vs. Group 2

Referring to Figure 11, compared with the vehicle control group (group 2), the Lieber-DeCarli liquid diet-fed mice treated with treatment group S5 (group 7) had significantly lower hepatic steatosis lesion scores (p<0.05; one-way ANOVA); therefore, the results indicated that treatment group S5 (group 7) protected mice from alcohol-induced increases in hepatic steatosis and improved liver pathology in a mouse model of alcoholic liver disease.

6.2 Semiquantitative scoring of histopathological observations

According to the reference materials published in Toxicologic Pathology (Shackelfold et al., 2002), the severity grading system standard of other microscopic lesions (liver, kidney and intestine) is shown in Table 6 below, and the scores of each group after treatment are shown in the table below 7 is shown.

Table 6

Fraction		grade
0	normal
1	Minimum (<10%)
2	Slight (10 to 39%)
3	Moderate (40～79%)
4	Severe (80～100%)

Table 7

^# p<0.05 Group 2 vs. Group 1

^* p<0.05 Group 7 vs. Group 2

As can be seen from Table 7, in the liver, local minimal mononuclear cell infiltration was observed (2/4 males in group 1, 3/4 males in group 2, 1/4 males in group 4). , 3/4 male animals in group 5, 1/4 male animals in group 6, 1/4 male animals in group 7, 3/4 male animals in group 8, 1/4 male animals in group 9 , 1/4 male animals in group 10, 2/4 male animals in group 11, 2/4 male animals in group 12 and 2/4 male animals in group 13). There was no statistical difference in the severity of microscopic changes between the treated and vehicle control groups (Group 2) (p>0.05; one-way ANOVA).

As can be seen from Table 7, in the kidneys, cortical focal mild chronic progressive nephropathy (1/4 male animals in group 4), minimal hyperplasia of eosinophils in cortical tubules (1 in group 3) were observed. /4 males), minimal hyperplasia of eosinophils in cortical tubules (1/4 males in group 5), minimal to mild local tubular casts (1/4 males in group 5 and 7 in group 7) 1/4 males), focal minimal tubular dilatation (1/4 males in Group 5 and 1/4 males in Group 9), and focal minimal mineralization of papillary tubules (1 in Group 7) /4 male animals). There was no statistical difference in the severity of microscopic changes between the treated and vehicle control groups (Group 2) (p>0.05; one-way ANOVA).

As can be seen in Table 7, in the gut, focal minimal to slight degeneration/necrosis was observed (1/4 males in group 5 and 1/4 males in group 10). There was no statistical difference in the severity of microscopic changes between the treated and vehicle control groups (Group 2) (p>0.05; one-way ANOVA).

Taken together, the overall results suggest that the pharmaceutical composition S5 protects mice from alcohol-induced increases in hepatic steatosis and ameliorates liver pathology in the aforementioned mouse models.

7. Statistical analysis

All values are presented as mean ± standard deviation (S.D.) or standard error of the mean (SEM); unpaired Student's t-test was used in the normal group (group 1) and vehicle group (group 2) groups), and one-way ANOVA and Dunnett's test were used to compare the vehicle group (group 2) and the other treatment groups (groups 3 to 13) for Statistical Analysis. Differences were considered statistically significant when the p-value was less than 0.05 (p<0.05).

Example 2: Case Study on the Effects of Pharmaceutical Compositions on Humans

A study involving 25 volunteer individuals was conducted to evaluate the effect of the pharmaceutical compositions of the present invention on elevated BAC. Twenty-one men and four women, aged 31 to 50, were enrolled in the study. Table 8 shows the basic demographic details of the selected individuals.

Table 8. Demographic details of selected individuals

A pharmaceutical composition (hereinafter referred to as "CGW3 formulation") comprises an extract of a herbal mixture comprising about 5-15% by weight of dried ginger; about 15-25% by weight of Codonopsis; about 25-35% by weight of Poria cocos ; about 15-25% by weight of dried tangerine peel; and about 10-20% by weight of Atractylodes.

Each individual was examined under each of the following 3 conditions:

1) Alcohol consumption without CGW3 preparation (control setting)

2) Consume 120ml of CGW3 preparation 30 minutes before and 30 minutes after drinking.

3) Once a day for 15 days before drinking, and 30 minutes before and 30 minutes after drinking, ingest 120ml of CGW3 preparation.

Each male individual drank 147ml of alcohol and each female drank 73.5ml of alcohol. After drinking alcohol, the following examinations and assessments are performed by a neurologist:

(a) On-site sobriety tests, including finger-nose test, counting test (counting with eyes closed) and Romberg balance test.

(b) Self-administered questionnaires 1 hour, 2 hours and 24 hours after drinking. The 1-hour post-drinking questionnaire addresses alcohol-induced hangover symptoms such as dizziness, fever, sweating, headache, abdominal pain, diarrhea, itching and rash, and abdominal pain. The 2-hour post-drinking questionnaire addressed alcohol-induced hangover symptoms, including incoherent speech, altered level of consciousness, confusion, unsteady gait, blurred vision, and hand tremors. The 24-hour drinking questionnaire addresses the following alcohol-induced hangover symptoms: decreased work performance, concentration or memory, nausea, vomiting, abdominal discomfort, back pain, head and neck pain, dry eyes, insomnia, and lethargy.

(c) Serological tests: glucose-6-phosphate dehydrogenase (G6PD), alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamate aminotransferase (γ-GT) ), creatinine and aldosterone.

As shown in Figure 12 and Table 9, individuals who took the CGW3 formulation 15 times before drinking performed significantly better on the finger-nose test, counting test and maintained better balance compared to the control setting. Additionally, individuals who took the CGW3 formulation once before drinking alcohol also had significantly more correct counting tests and maintained better balance compared to the control setting. The results showed that at least one intake of CGW3 preparations before drinking alcohol significantly improved performance on non-standardized field sobriety tests (finger-nose, counting, and balance tests).

Table 9. Effects of CGW3 formulations on field sobriety test performance

Table 10 and Figures 13 to 15 show that one or 15 ingestion of CGW3 formulations prior to drinking reduced all alcohol-induced hangover symptoms. Specifically, taking the CGW3 formulation 15 times before drinking significantly reduced dizziness, confusion and improved concentration or memory the next day compared to the control setting. Compared to the control setting, taking the CGW3 formulation once before drinking significantly reduced incoherence, altered levels of consciousness, and improved work performance the next day.

Table 10. Effects of CGW3 formulations on alcohol-induced hangover symptoms

Table 11 and Figures 16-18 show the effect of CGW3 formulations on liver function tests, kidney function tests, and G6PD. Compared with the control setting, taking CGW3 preparations once before drinking significantly reduced serum levels of G6PD and γ-GT, and taking CGW3 preparations 15 times before drinking significantly reduced serum levels of ALT and AST, and noted a downward trend in serum creatinine levels .

Table 11 Effects of CGW3 preparations on liver and kidney function tests

Example 3: In vivo studies on the effect of pharmaceutical compositions

The effects of CGW3 preparations on the intestinal capsule and glomeruli of mice were evaluated.

[Corrected 24.04.2022 according to Rule 26]
8-10 week old male C57BL/6 mice with a body weight of about 20 grams (purchased from Taiwan Animal Center and housed at Chang Gung University Animal Center, IACUC number CGU108-009) were used in the study. The study included the following 5 study groups of 10 mice each.

1. Regular control diet: Mice were fed Lieber-DeCarli diet (221.78 g of Lieber-DeCarli standard diet powder stirred in 1 liter of distilled water) by oral gavage for 4 weeks. One week before and throughout the regular control diet, 12.3 ml/kg body weight/day of distilled water was administered by oral gavage.

[Corrected 24.04.2022 according to Rule 26]
2. Liquid alcohol diet: According to the "Evaluation Method of Liver Health Care Efficacy of Healthy Food" published by Taiwan Food and Drug Administration in 2016, the mice were fed by oral gavage. A Lieber-DeCarli alcoholic diet (132.18 g Lieber-DeCarli standard diet powder and 67 ml 95% alcohol in 933 ml distilled water) for 4 weeks was used to induce intestinal and kidney damage. One week before and throughout the liquid alcohol diet, 12.3 ml/kg body weight/day of distilled water was administered by oral gavage.

3. Liquid alcohol diet + 4.1 ml/kg body weight/day of CGW3 formulation: Mice were fed a Lieber-DeCarli alcohol diet by oral gavage for 4 weeks. 4.1 ml/kg body weight/day of the CGW3 formulation (equivalent to 0.33 mg/kg body weight/day in humans) was administered by oral gavage once a day for a total of 5 weeks one week before and throughout the liquid alcohol diet.

4. Liquid alcohol diet + 12.3 ml/kg body weight/day of CGW3 formulation: Mice were fed a Lieber-DeCarli alcohol diet by oral gavage for 4 weeks. One week before and throughout the liquid alcohol diet, 12.3 ml/kg body weight/day of the CGW3 formulation (equivalent to 1 mg/kg body weight/day in humans) was administered by oral gavage once a day for 5 weeks.

5. Liquid alcohol diet + 36.9 ml/kg body weight/day of CGW3 formulation: Mice were fed a Lieber-DeCarli alcohol diet by oral gavage for 4 weeks. One week before and throughout the liquid alcohol diet, 36.9 ml/kg body weight/day of the CGW3 formulation (equivalent to 3 mg/kg body weight/day in humans) was administered by oral gavage once a day for 5 weeks.

The protective effect of the CGW3 formulation on the intestinal envelope and glomeruli was assessed after 5 weeks of administration of the CGW3 formulation.

Pathological examination

After mice were sacrificed, jejunum and kidney tissues were fixed in 10% formalin and stained according to the hematoxylin and eosin protocol. Staining results: the nucleus is blue, and the cytoplasm and interstitial tissue are red.

result:

Figure 19, Panel B shows that jejunal villi were damaged in mice fed a liquid alcohol diet compared to mice on a regular control diet (Figure 19, Panel A). Jejunal villus damage was reversed or attenuated by regular feeding of the CGW3 formulation (Figure 19, panels C to E).

Figure 20, Panel B shows glomerular damage in mice fed a liquid alcohol diet compared to mice on a regular control diet (Figure 20, Panel A). The glomerulus is a type of kidney tissue that filters the raw urine that is encapsulated in Bowman's capsule. The main function of the glomerulus is to filter plasma to remove waste products and to form urine. Glomerular damage was reversed or attenuated by regular feeding of the CGW3 formulation (Figure 20, panels C to E).

Example 4: Case study on the administration of pharmaceutical compositions after drinking red wine

Two adult volunteers were found to drink 250mL of red wine as the control group, and took the S5 pharmaceutical composition containing three ingredients of Codonopsis, Poria and tangerine peel (the weight ratio of Codonopsis, Poria and tangerine peel is 25-35%: 30-50% : 25-35%) and 250mL of red wine as the experimental group 1, and taking the pharmaceutical composition containing the three components of Codonopsis, Poria and Green Skin (the weight ratio of Codonopsis, Poria and Green Skin is 25-35%: 30-50%: 25 -35%) and 250mL of red wine as experimental group 2 to test to confirm the difference in alcohol concentration reduction between the addition of tangerine peel or green peel to the pharmaceutical composition. The tester (model HOHOGA-AT576-AlcoREAL) was used for measurement; the average alcohol concentration of the two volunteers is shown in Table 12 below.

Table 12

As can be seen from Table 12, the alcohol concentration measured by the experimental group 1 after 30 minutes is lower than that of the control group and the experimental group 2; and the alcohol concentration of the experimental group 1 is 0 after 90 minutes, that is, the alcohol tester cannot measure it. It can be seen from the above results that the anti-alcoholic effect of containing tangerine peel in the pharmaceutical composition of the present invention is better than that of the composition containing green peel, and the alcohol concentration can be reduced to 0 in a shorter time.

Claims (13)

1. Use of a pharmaceutical composition in the preparation of a medicament for treating or reducing the effects of alcohol consumption in an individual, the pharmaceutical composition comprising:

   (1) A herbal mixture comprising:

   Party ginseng;

   Poria; and

   tangerine peel, and

   (2) A pharmaceutically acceptable carrier.
2. The use according to claim 1, wherein in the herbal mixture, the weight ratio of Codonopsis: tangerine peel: Poria is 25-35%: 25-35%: 30-50%.
3. The use of claim 1, wherein the herbal mixture further comprises dried ginger and Atractylodes Rhizoma.
4. The use according to claim 3, wherein the herbal mixture comprises 5-15% by weight of dried ginger, 15-25% by weight of Codonopsis, 25-35% by weight of Poria cocos, 15-25% by weight of dried tangerine peel and 10-20 wt% Atractylodes.
5. The use of claim 1 to claim 4, wherein the pharmaceutical composition is administered prior to drinking alcohol.
6. 4. The use of claim 1 to claim 4, wherein the drinking effect is selected from the group consisting of symptoms of elevated blood alcohol concentration and alcohol hangover.
7. A pharmaceutical composition comprising:

   (1) A herbal mixture comprising:

   Party ginseng;

   Poria; and

   tangerine peel, and

   (2) A pharmaceutically acceptable carrier.
8. The pharmaceutical composition according to claim 7, wherein in the herbal mixture, the weight ratio of Codonopsis: tangerine peel: Poria is 25-35%: 25-35%: 30-50%.
9. The pharmaceutical composition of claim 7, wherein the herbal mixture further comprises dried ginger and Atractylodes Rhizoma.
10. The pharmaceutical composition of claim 9, wherein the herbal mixture comprises 5-15% by weight of dried ginger, 15-25% by weight of Codonopsis, 25-35% by weight of Poria, 15-25% by weight of Chenpi and Atractylodes 10-20 wt%.
11. A use of the pharmaceutical composition according to claim 7 to claim 10 for preparing a medicament for treating or reducing hepatic cell inflammation and fat accumulation.
12. A use of the pharmaceutical composition according to claim 7 to claim 10 for preparing a medicament for treating or reducing damage to small intestinal mucosa or intestinal function.
13. A use of the pharmaceutical composition according to claim 7 to claim 10 for preparing a medicament for treating or reducing glomerular damage or renal function damage.

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