WO2010133016A1

WO2010133016A1 - Pharmaceutical composition for treating depression, preparation method and use therefor

Info

Publication number: WO2010133016A1
Application number: PCT/CN2009/001491
Authority: WO
Inventors: 张作光
Original assignee: Zhang Zuoguang
Priority date: 2009-05-21
Filing date: 2009-12-17
Publication date: 2010-11-25
Also published as: CN101890032A

Abstract

Provided in the present invention are a pharmaceutical composition for treating depression and a preparative method and use therefor. The pharmaceutical composition comprises ginsenoside Rg1, ginsenoside Rb1 and glycyrrhizic acid or glycyrrhetinic acid, and has undergone screening to arrive at the preferred ratio of components. Test-proven, the pharmaceutical composition in the present invention can prevent and treat depression and its complications.

Description

Pharmaceutical composition for treating depression, preparation method and use thereof

The present invention relates to a pharmaceutical composition for treating depression comprising ginsenoside Rgl, ginsenoside Rbl and glycyrrhizic acid (or glycyrrhetinic acid), which can be used as a pharmaceutical preparation, a nutrient and a health care product. The invention also relates to a process for the preparation of the above pharmaceutical compositions and to their use. Background technique

Depression is a common disease. According to statistics, about 25% of women in the general population have experienced depression in their lifetime, and about 10% of men have experienced depression (Zhang Chunxing: Modern Psychology) . Data from the World Health Organization (WHO): The incidence of depression in the world is about 11%. There are currently about 340 million people with depression in the world, and this number is still on the rise. The survey found that in the next 20 years, depression It will rise to the second most common disease in the world.

In the prior art, the antidepressant drugs are mainly based on Prozac, Xelote, Zoloft, etc. (SS-, SNRI, NDRI, etc. 5-HT, NE, DA reuptake inhibitors), and the mechanism of action is Regulate the content of monoamine neurotransmitters (5-HT, NE, etc.) in the human body to alleviate depressive symptoms. However, the anti-depressant drugs that have been asked have different degrees of side effects, such as: increased suicide rate, headache, dizziness, dizziness, insomnia, lethargy, tinnitus, dry mouth, anorexia, increased appetite, weight gain, blood pressure rise, gastrointestinal Discomfort, nausea, nausea, vomiting, indigestion, diarrhea, constipation, lower limb pain, skin rash, trembling, cramps, sweating, edema, loss of libido, sexual incompetence, etc. In recent years, anti-depressant drugs such as Prozac have become a serious concern in the society. The US Food and Drug Administration (FDA) in 2004 asked the pharmaceutical companies to relabel the main 32 antidepressants on the market. The side effects and warnings, and the emphasis on health care providers may increase the chances of suicide in children and adolescents. Among them, Celite was found to have security risks as early as 1996, and has been recalled from the market since 2001. In June 2004, the US Attorney General of New York accused the British GlaxoSmithKline of deceptively concealing a study related to the use of Celet and the “risk of increasing suicidal tendencies and behaviors among young people” in order to make a profit. In this context, how to develop a new generation of drugs with low side effects and significant anti-depressant effects has become a concern of the global pharmaceutical community.

In recent years, scientists in the international medical community have made new breakthroughs in the study of the pathogenesis of depression. It has been found that in addition to the treatment of depression by 5-HT, NE, and DA reuptake inhibition, it can also be regulated by Post-treatment for depression, and it represents the drug Rolipram The advent of antidepressants that make the post-receptor mechanism of action become a hot spot in the research and development of the pharmaceutical industry. Roller Pula is

Inhibitors of phosphodiesterase 4 (PDE4) have been shown to have significant antidepressant effects in clinical trials. However, due to the strong vomiting of rolita, it is forced to terminate clinical trials, however, Developed a new generation of "receptor post-action mechanism anti-depressant drugs" research and development ideas.

Thus, it is urgent to develop a drug that can effectively treat depression and is safe and has no side effects. Summary of the invention

After understanding the limitations of the prior art, the applicant has overcome the deficiencies of the existing antidepressant drugs, and the inventors have combined the principles of modern medicine and pharmacology to treat the pathogenesis and mechanism of action of traditional Chinese medicine for depression. Research and exploration, and in the spirit of perseverance, finally found a pharmaceutical composition including ginsenoside Rgl, Rbl, glycyrrhizic acid (or glycyrrhetinic acid), which can be used to treat or alleviate depression, and can be used for treatment or relief. A drug for depression and its associated diseases, disorders or conditions such as anxiety, sleep disorders and post-traumatic stress disorder.

Accordingly, it is an object of the present invention to provide a ginsenoside for the treatment of depression.

A pharmaceutical composition of Rg 1, ginsenoside Rb 1 and glycyrrhizic acid (or glycyrrhetinic acid).

Another object of the present invention is to provide a process for the preparation of the above pharmaceutical composition for treating depression.

Another object of the present invention is to provide a use of the above pharmaceutical composition, comprising preparing a medicament, a health care product or a nutrient for treating or ameliorating depression, and for treating or alleviating depression and a disease, disorder or condition complicated therewith. Use in use, and in the treatment or alleviation of depression or in the treatment or alleviation of depression and the diseases, disorders or conditions associated therewith.

In view of the above object, the present invention provides the following technical solutions:

In one aspect, the invention provides a pharmaceutical composition for treating depression, the pharmaceutical composition comprising at least two active ingredients selected from the group consisting of:

a. Ginsenoside Rgl;

b. Ginsenoside Rbl ; and

c glycyrrhizic acid or glycyrrhetinic acid;

Preferably, the pharmaceutical composition is a pharmaceutical preparation, a health care preparation or a nutritional preparation; preferably, the ginsenoside Rgl contained in the pharmaceutical composition is 20 to 100 parts by weight, and the ginsenoside Rbl is 5 to 100 weight. And the glycyrrhizic acid or glycyrrhetinic acid is 5 to 100 parts by weight; More preferably, the ginsenoside Rgl contained in the pharmaceutical composition is 60 parts by weight, the ginsenoside Rbl is 30 to 60 parts by weight, and the glycyrrhizic acid or glycyrrhetic acid is 30 to 60 parts by weight.

Preferably, the pharmaceutical composition as described above comprises: ginsenoside Rgl, ginsenoside Rbl and glycyrrhizic acid or glycyrrhetinic acid; more preferably, the pharmaceutical composition does not contain other active ingredients;

Further preferably, the ginsenoside Rgl in the pharmaceutical composition is 20 to 100 parts by weight, the ginsenoside Rbl is 5 to 100 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 5 to 100 parts by weight;

More preferably, the ginsenoside Rgl in the pharmaceutical composition is 60 parts by weight, the ginsenoside Rbl is 30 to 60 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 30 to 60 parts by weight. Preferably, the pharmaceutical composition as described above comprises: two active ingredients: ginsenoside Rgl and ginsenoside Rbl; preferably, the pharmaceutical composition does not comprise other active ingredients;

More preferably, the ginsenoside Rgl in the pharmaceutical composition is 20 to 100 parts by weight, and the ginsenoside Rbl is 5 to 100 parts by weight;

Further preferably, the ginsenoside Rgl in the pharmaceutical composition is 60 parts by weight, and the ginsenoside Rbl is 30 to 60 parts by weight. Preferably, the pharmaceutical composition as described above comprises: ginsenoside Rgl and glycyrrhizic acid or glycyrrhetinic acid; preferably, the pharmaceutical composition does not comprise other active ingredients;

More preferably, the ginsenoside Rgl in the pharmaceutical composition is 20 to 100 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 5 to 100 parts by weight;

Further preferably, the ginsenoside Rgl in the pharmaceutical composition is 60 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 30 to 60 parts by weight. Preferably, the pharmaceutical composition as described above comprises: ginsenoside Rbl, and glycyrrhizic acid or glycyrrhetinic acid; preferably, the pharmaceutical composition does not comprise other active ingredients;

More preferably, the ginsenoside Rbl in the pharmaceutical composition is 5 to 100 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 5 to 100 parts by weight;

Further preferably, the ginsenoside Rbl in the pharmaceutical composition is 30 to 60 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 30 to 60 parts by weight. Preferably, in the pharmaceutical composition as described above, the purity of ginsenoside Rgl is 50 to 98%, The purity of Rbl is 50 to 98% and the purity of glycyrrhizic acid or glycyrrhetinic acid is 50 to 98%; more preferably, the purity of ginsenoside Rgl in the pharmaceutical composition is 90% to 98%, and the purity of Rbl is The purity of 90%~98% and glycyrrhizic acid or glycyrrhetinic acid is 90%~98%. Preferably, the pharmaceutical composition as described above further comprises: one or more pharmaceutically acceptable carriers or additives;

Preferably, the pharmaceutical composition is selected from the group consisting of: oral preparations, parenteral preparations, topical and inhaled preparations, and transdermal preparations.

More preferably, the dosage form is an oral preparation selected from the group consisting of a tablet, a capsule, a powder, a tablet, a powder, a solution, a micro-stringing agent, a suspension, an emulsion, a granule, a pill, and a pill; Further preferably, the pharmaceutically acceptable carrier or additive is selected from the group consisting of: a disintegrant, a lubricant, a binder, a filler, a solvent, a fragrance, a sweetener, an antioxidant, a surfactant, a preservative, a correction Flavors and pigments. In another aspect, the invention provides a method of preparing a pharmaceutical composition as described above, which comprises

20-100 parts by weight of ginsenoside Rgl, 5 to 100 parts by weight of ginsenoside Rbl and 5 to 100 parts by weight of glycyrrhizic acid or glycyrrhetinic acid mixed pulverization step;

Preferably, the method further comprises mixing 20 to 100 parts by weight of ginsenoside Rgl, 5-100 parts by weight of ginsenoside Rbl, 5-100 parts by weight of glycyrrhizic acid or glycyrrhetinic acid together with a pharmaceutically acceptable carrier or additive. A step of. In still another aspect, the present invention provides the use of the pharmaceutical composition for the preparation of a pharmaceutical preparation, a health supplement or a nutraceutical for treating or ameliorating depression. In still another aspect, the present invention provides the use of the pharmaceutical composition in the preparation of a pharmaceutical preparation, a health supplement or a nutraceutical for simultaneously treating or ameliorating depression and a disease, disorder or condition complicated by depression;

Preferably, the concurrent disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder. In a further aspect, the invention provides a method of treating or ameliorating depression, the method comprising administering to a subject in need thereof a therapeutically effective amount of any of the above pharmaceutical compositions. Preferably, the therapeutically effective amount is 0.5 to 5 mg/kg/d, preferably 1 to 4 mg/kg/d, and more preferably 2 to 3 mg/kg/d. In still another aspect, the present invention provides a method of treating or ameliorating depression or for simultaneously treating or ameliorating depression and a disease, disorder or condition associated with depression, the method comprising administering to a subject in need thereof An effective amount of any of the above pharmaceutical compositions;

Preferably, the concurrent disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder.

Preferably, the therapeutically effective amount is 0.5 to 5 mg/kg/d, preferably 1 to 4 mg/kg/d, and more preferably 2 to 3 mg/kg/d. The invention can also be implemented by the following technical solutions.

The present invention provides a group of pharmaceutical compositions for the treatment of depression, which are prepared from ginsenosides Rgl, Rbl, glycyrrhizic acid (or glycyrrhetinic acid).

The pharmaceutical composition of the present invention is prepared from a raw material comprising 1 to 100 parts by weight of ginsenoside Rgl, 0.5 to 100 parts by weight of ginsenoside Rbl, and 0.5 to 100 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid).

Preferably, the pharmaceutical composition of the present invention is prepared from a raw material comprising 10 parts by weight of ginsenoside Rgl, 5-10 parts by weight of ginsenoside Rbl, and 5-10 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid).

In the pharmaceutical composition of the present invention, the purity of ginsenoside Rgl is 50 to 98%, and ginsenoside

The purity of Rbl is 50 to 98%, and the purity of glycyrrhizic acid (or glycyrrhetinic acid) is 50 to 98%.

In the pharmaceutical composition of the present invention, the preferred purity of ginsenoside Rgl is 90%, the preferred purity of ginsenoside Rbl is 90%, and the preferred purity of glycyrrhizic acid (or glycyrrhetinic acid) is 90%.

The invention also provides a method for preparing a pharmaceutical composition for treating depression:

(a) mixing and pulverizing 1-100 parts by weight of ginsenoside Rgl, ^0.5 to 100 parts by weight of ginsenoside Rbl, and ^0.5 to 100 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid), thereby obtaining the pharmaceutical composition of the present invention .

(b) 1 to 100 parts by weight of ginsenoside Rgl, 0.5-100 parts by weight of ginsenoside Rbl, 0.5-100 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid) and starch, lactose, micronized silica gel and other auxiliary materials are mixed and pulverized A pharmaceutical composition of the invention.

The preferred ratio of the three raw materials in the above preparation method is composed of 10 parts by weight of ginsenosides.

Rgl is prepared from 5 to 10 parts by weight of ginsenoside Rbl and 5 to 10 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid). Preferably, the pharmaceutical composition of the present invention can be processed into a dosage form selected from the group consisting of a tablet, a capsule, a powder, a tablet, a powder, a solution, a microclay, a suspension, an emulsion, a granule, and a drop. One of the oral pharmaceutical dosage forms of pills, pills and pharmaceutics.

Preferably, the pharmaceutical compositions of the invention may comprise a pharmaceutically acceptable carrier or additive. Preferably, the pharmaceutical compositions of the invention are also useful in the manufacture of nutraceuticals and nutrients.

The solution and the prescription of the present invention are the results of intensive research and exploration by the inventors, according to the pathology and pharmacology theory of modern medical treatment of depression, especially the anti-depression drug target research combined with the post-receptor mechanism, after a large number of Animal experiments prove that: Ginsenoside Rgl, Ginsenoside Rbl, Glycyrrhizin (or glycyrrhetinic acid) are cAMP phosphodiesterase inhibitors, which are compatible with each other and synergistically act through the blood-brain barrier to rapidly activate cAMP in the hippocampus. The PKA pathway enhances the expression of CREB and BDNF, so it has significant antidepressant function. In addition, although ginsenoside Rgl, ginsenoside Rbl, glycyrrhizin (or glycyrrhetinic acid) each have a certain degree of antidepressant effect, the results of orthogonal experiment of mouse tail suspension show that the preferred ratio of the three The antidepressant effect of the composition (i.e., the preferred pharmaceutical composition of the invention) is significantly stronger than any of the monomers. Accordingly, the present invention includes a pharmaceutical composition of ginsenoside Rgl, ginsenoside Rbl, and glycyrrhizic acid (or glycyrrhetinic acid) having a significant improvement in antidepressant effect relative to ginsenoside Rgl, ginsenoside Rbl, glycyrrhizic acid (or glycyrrhetinic acid) Effect.

As we all know, ginseng and licorice are common medicinal materials and foods used in traditional Chinese medicine supplements. It has been proved that the compatibility and ration of ginseng and licorice are compatible with ginseng and licorice for thousands of years of consumption and clinical use. Ginsenoside Rgl, ginsenoside Rbl, glycyrrhizic acid (or glycyrrhetinic acid), a combination of drugs or health supplements, with the same side effects, can be taken for a long time, for the treatment of conditioning.

It can be seen that the post-receptor antidepressant pharmaceutical composition provided by the present invention is characterized in that the three raw materials of the prescription, ginsenoside Rgl, ginsenoside Rbl and glycyrrhizic acid (or glycyrrhetinic acid) are high. The monomer extract of purity (50~98%) has clear functional ingredients, clear mechanism of action, and can be quantified. Therefore, the quality of the drug prepared by the drug is controllable, stable, effective, safe, easy to take, and It works quickly.

The pharmaceutical composition provided by the present invention is a core formulation for achieving the object of the present invention. After the disclosure of the present invention, those skilled in the art can routinely add or subtract the above pharmaceutical composition according to the theory of traditional Chinese medicine or related modern pharmacological theory. Reduction. Such conventional addition and subtraction is a general technical activity of those skilled in the art, and any general technical addition or subtraction performed on the basis of the formulation of the pharmaceutical composition of the present invention is within the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, in which:

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a flow chart showing the preparation method of the pharmaceutical composition in Example 1 and Example 3 of the present invention. Figure 2 is a flow chart showing the preparation method of the pharmaceutical composition in Example 1 of the present invention.

Fig. 3 is a graph showing the results of a mouse forced swimming test performed in Example 6 of the present invention.

Fig. 4 is a graph showing the results of cAMP content in rat hippocampal tissue measured in Example 7 of the present invention. 5A is a gel electrophoresis diagram of an experiment for cAMP-dependent PKA activity in rat hippocampus tissue measured in Example 7 of the present invention, wherein the lanes are from left to right 1-4 saline group (group A); 5-8 paroxetine Group (Group B); 9-11 The pharmaceutical composition group of the present invention (Group C); 12 positive control samples; 13 negative control samples).

Fig. 5B is a graph showing the results of differences in cAMP-dependent PKA activity in rat hippocampus tissues measured in Example 7 of the present invention.

Fig. 6 is a graph showing the results of changes in p-CREB content in rat hippocampus tissues measured in Example 7 of the present invention.

Fig. 7 is a graph showing the results of PDE activity in rat hippocampus tissues measured in Example 7 of the present invention. The best way to implement the invention

The invention will be further illustrated by the following figures and specific examples. However, the examples are only intended to illustrate the invention and are not intended to limit the scope of the invention. The experimental methods for the specific experimental conditions are not indicated in the following examples, usually in accordance with conventional conditions, or in accordance with the conditions recommended by the manufacturer. Example 1: Preparation of the pharmaceutical composition of the present invention

This embodiment provides a preferred preparation method of the pharmaceutical composition of the present invention, and the specific scheme is shown in FIG. As shown in the method flow chart of Figure 1, 100 g of ginsenoside Rgl, which has been prepared to a purity of 95%, is directly prepared.

(purchased from Jilin Agricultural University Huadi Bio-Biotechnology Co., Ltd.), 100g ginsenoside Rbl (purchased from Jilin Agricultural University Huadi Bio-Biotechnology Co., Ltd.) with purity of 96.97% and 100g glycyrrhizic acid with purity of 95.2% (purchased from Xi'an Fujie Pharmaceutical Co., Ltd.) mixed pulverization, that is, 300 g of the pharmaceutical composition provided by the present invention. Example 2: Preparation of the pharmaceutical composition of the present invention

This embodiment provides another preferred preparation method of the pharmaceutical composition of the present invention. figure 2.

As shown in the method flow chart of Figure 2, directly prepared 100g of ginsenoside Rgl (purchased from Shanghai Eternal Biotechnology Co., Ltd.) with a purity of 90%, 100g of ginsenoside Rbl with a purity of 90% (purchased from Shanghai Eternal Biotechnology Co., Ltd.) Company), 100g of glycyrrhizic acid (purchased from Xi'an Fujie Pharmaceutical Co., Ltd.) with a purity of 90%, mixed with 20g starch, 10g of lactose, 10g of micro-silica gel, to make 1000 capsules, which is provided by the present invention. Pharmaceutical composition. Example 3: Preparation of the pharmaceutical composition of the present invention

This embodiment provides a preferred preparation method of the pharmaceutical composition of the present invention, and the specific scheme is the same as that of the embodiment 1. See FIG.

As shown in the method flow chart of Figure 1, directly prepared 100g of ginsenoside Rgl (purchased from Shanghai Eternal Biotechnology Co., Ltd.) with purity of 95%, and 50g of ginsenoside Rbl (purchased from Shanghai Eternal Biotechnology Co., Ltd.) And 50 g of glycyrrhetic acid (purchased from Xi'an Fujie Pharmaceutical Co., Ltd.) having a purity of 95% was mixed and pulverized to obtain 200 g of the pharmaceutical composition provided by the present invention. Example 4: Effect of the pharmaceutical composition of the present invention on mouse tail suspension experiment

This example provides the effect of the pharmaceutical composition of the present invention and the components thereof on the tail suspension test of mice, that is, the ginsenoside Rgl (90% purity), ginsenoside Rbl (90% purity), glycyrrhizic acid were carried out in this example. (90% purity) Screening studies on antidepressant effects of various combinations to screen out the optimal combination of anti-depression of ginsenoside Rgl, ginsenoside Rbl and glycyrrhizic acid. The specific experiment is detailed below.

1. Experimental materials

Animals: ICR mice, male, weighing 20 ± lg, purchased from the Vitallihua Animal Experimental Center. Test drug: ginsenoside Rgl (90% purity, purchased from Shanghai Eternal Biotechnology Co., Ltd.), ginsenoside Rbl (90% pure, purchased from Shanghai Eternal Biotechnology Co., Ltd.), glycyrrhizic acid (90% purity, purchased from Xi'an) Fujie Pharmaceutical Co., Ltd.);

Positive drugs: Fluoxetine hydrochloride capsule (Baiyoujie), Lilly Suzhou Pharmaceutical Co., Ltd. product, batch number: A333341-070608, Chinese medicine Zhunji J20030017, daily dosage, 20mg/曰.

2, laboratory equipment

JZ type 300g tension transducer (Gaobeidian Xinhang Accumulation Equipment Co., Ltd.), Medlab biological signal acquisition and processing system (Nanjing Meiyi Technology Co., Ltd.)

3. Experimental methods 1) Orthogonal design scheme of ginsenoside Rgl (A drug), ginsenoside Rbl (B drug), glycyrrhizic acid (C drug) combination

The dose and combination of ginsenoside Rgl (A drug), ginsenoside Rbl (B drug) and glycyrrhizic acid (C drug) were selected by orthogonal test using the time of mouse tail suspension as an indicator.

The factor level table is shown in Table 1, and the orthogonal test table is shown in Table 2.

Table 1 Factor Level Table

L9(3 ⁴ ) orthogonal test table

The dosing schedules according to Factor Level Table 1 and Orthogonal Test Table 2 are shown in Table 3.

Table 3 Table of dosage schedules (mg/kg/d)

Dosing group number A drug B drug C drug fluoxetine hydrochloride group 0 0 0 0

Group 1 10 10 10

Group 2 10 5 5

Group 3 10 0 0

Group 4 5 10 5

Group 5 5 5 0

Group 6 5 0 10

Group 7 0 10 0

Group 8 0 5 10 Group 9 0 0 0

Group 10 3.5

2) group administration

In each group of experiments, normal mice were randomly divided into 11 groups according to their body weight. Each group was 20 blank control group (administration group 0), 1-9 group, positive drug fluoxetine hydrochloride capsule (to ί 10 group) . Each group was administered at 0.2 ml/10 g body weight for 2 consecutive days, once a day.

3) Test method

Each of the above groups was administered continuously for 2 days, and the experiment was carried out 1 hour after the administration on the second day. The tail end of the mouse (2 cm from the tip of the tail) was fixed with a tape on the line of the 100 g tension transducer, and the head was placed in an upside down state. The head was about 15 cm away from the test bench, and two animals were tested at the same time. Separated by cardboard. The transducer is connected to the Medlab biosignal acquisition and processing system. After 2 minutes of adaptation, the results within 4 minutes are recorded and the immobile state is converted to time (seconds, s).

4, the experimental results

The results of the mouse tail suspension experiment are shown in Table 4.

Table 4 Results of mouse tail suspension experiment

Dosage (mg/kg/d)

20 113.4士 42.4

A drug 10mg + B lOmg + C % lOmg 20 63.0 ± 30.8** A; 10mg + B 5mg + C: ) 5mg 20 69.0 ± 41.2** A; 10mg + B % Omg + C: 5 Omg 20 78.0 ± 31.8** A 3⁄4 5mg+ B芗lOmg + C: ) 5mg 20 74.1士30.1** A 3⁄4 5mg + B i 5mg + C ί / Omg 20 84.2士 45.3* A; 5mg + B ί Omg+ C lOmg 20 78.1士40.4** A ί Omg + B lOmg + C 3⁄4 Omg 20 86.5 ± 39.2* A; 3⁄4 Omg + B ί 5mg + C lOmg 20 81.4 ± 38.4* A 3⁄4 Omg + B Omg + C ί / 5mg 20 87.9 ± 44.1*

Flufluoxetine 3.5mg 20 63.9 ± 42.9** In Table 4, compared with the blank group, *Ρ<0·05 * *Ρ<0·01

4 2 1 2 3 74.0

5 2 2 3 1 84.2

6 2 3 1 2 78.1

7 3 1 3 2 86.5

8 3 2 1 3 81.4

9 3 3 2 1 87.9 Not moving Kl 70.0 74.5 74.1 78.3

K2 78.8 78.2 77.0 77.9

K3 85.2 81.3 82.9 77.8

R 15.2 6.8 8.8 0.5 It can be seen from the experimental treatment results in Table 5: 1 The best combination is A1B1C1 and A1B2C2, namely ginsenoside Rgl 10mg + ginsenoside Rbl 10mg + glycyrrhizic acid 10mg, and ginsenoside Rgl 10mg + ginsenoside Rbl 5mg + glycyrrhizic acid 5mg; Rgl has the greatest contribution; 3 alone ginsenoside Rgl 10mg, ginsenoside Rbl 10mg or glycyrrhizic acid 5mg have anti-experimental depression effect, but the screening experiment data shows that the effect is decreasing. 4 Any combination of two and two (ginsenoside Rgl 5mg + ginsenoside Rbl 5mg, ginsenoside Rgl 5mg + glycyrrhizic acid 10mg and ginsenoside Rbl 5mg + glycyrrhizic acid 10mg) showed that there is also a certain anti-experimental depression effect.

Therefore, the comparison of the above orthogonal experiments suggests that the combination of ginsenoside Rgl 10mg, ginsenoside Rbl 10mg or 5mg, and glycyrrhizic acid 10mg or 5mg has a remarkable and stable antidepressant effect. In addition, ginsenoside Rgl, ginsenoside Rbl or glycyrrhizic acid alone may be antidepressant, but not as effective or stable as the combination of the three or any combination of the two (the combination of the drugs in the first embodiment and the second embodiment of the present invention) The materials are prepared according to the above optimized ratios). Example 5: Further screening of different ratios of antidepressant effects of the pharmaceutical combinations of the invention

Based on the results obtained in Example 4, the anti-depressant effects of various combinations of ginsenoside Rgl, ginsenoside Rbl and glycyrrhizic acid in the pharmaceutical composition were further selected by the mouse tail suspension experiment. And determine, the specific experiment is detailed below.

1. Experimental materials

Positive drugs: Fluoxetine hydrochloride capsule (Baiyoujie), Lilly Suzhou Pharmaceutical Co., Ltd. product, batch number: A333341-070608, Chinese medicine Zhunzi J20030017, daily dosage, 20mg/day. 2, laboratory equipment

3. Experimental methods

1) group administration

Normal mice were randomly divided into 3 groups according to body weight, 20 rats in each group, namely, blank control group and 1-6 groups (for ginsenoside Rgl (A drug), ginsenoside Rbl (B drug), glycyrrhizic acid (C) The different combinations of drugs, as shown in Table 6), the positive drug fluoxetine hydrochloride (3.5mg/kg/d). Each group was administered at 0.2 ml/10 g body weight for 2 consecutive days.

2) Test method

4, the experimental results

The results of the mouse tail suspension experiment are shown in Table 6.

Table 6 Results of mouse tail suspension experiment

Compared with the blank group, *P<0.05 * *P<0.0 1 The test results showed that the combination of ginsenoside Rgl 5mg, ginsenoside Rbl O.lmg and glycyrrhizic acid O. lmg, ginsenoside Rgl 5mg and ginsenoside Rbl 0.5 The combination of mg, ginsenoside Rgl 4mg and glycyrrhizic acid 0.18mg, ginsenoside Rbl 5mg and glycyrrhizic acid 0.15mg combined anti-depression was not obvious, compared with the blank group, there was no significant difference (P > 0.05). The combination of ginsenoside Rgl 10mg and glycyrrhizic acid 5mg, ginsenoside Rbl 5mg and glycyrrhizic acid 5mg had significant antidepressant effects, which was significantly different from the blank group (P < 0.01 or 0.05). Example 6: Effect of the pharmaceutical composition of the present invention on forced swimming test in mice

This example provides the effect of the pharmaceutical composition of the present invention on the immobility time of chronic stress-suppressed mice in a forced swimming test in mice. The specific experimental process is detailed below.

1. Experimental materials

Experimental animals: 12 KM mice, male (20±2g), purchased from Institute of Experimental Animals, Chinese Academy of Medical Sciences (certificate No. SCXK (Beijing) 2004-0001), animals were kept in light and dark (12 h: 12 h) In the clean animal room, eat freely. Animals were acclimated to the environment in the animal room and experiments were started 3 days later.

Test drug: The pharmaceutical composition prepared in Example 3 of the present invention is divided into a low dose group

(30mg/kg) and high dose group (60mg/kg).

2, the experimental method

The drug was administered once a day for 2 days, once a day, and 1 hour after the morning of the second day, the mice were placed in a glass jar with a water depth of 10 cm and a diameter of 14 cm at a water temperature of 25 ° C for 5 minutes. The cumulative time of the rat in the water. Each group of animals was tested in parallel.

3, the experimental results

The experimental results are shown in Figure 3. It can be seen from Fig. 3 that the pharmaceutical composition provided by the invention can shorten the immobility time of the mice in the forced swimming test, compared with the control group, *Ρ<0.05, **Ρ<0.01, the average value of SD, n =12. The low dose group showed a better trend, but there was no significant difference between the two dose groups. Example 7: The pharmaceutical composition of the present invention is applied to the hippocampus of normal rats cAMP-PKA-CREB

(p-CREB) pathway effects

This example provides the effect of the pharmaceutical composition of the present invention on the cAMP-PKA-CREB (p-CREB) pathway in the hippocampus of normal rats, through cAMP, cAMP-dependent protein kinase (PKA), cAMP response element Detection of changes in the binding protein (CREB or p-CREB) and phosphodiesterase (PDE) levels. The specific experimental process is detailed below.

1. Experimental materials

Experimental animals: Healthy male SD rats weighing 180-200 g, 70, purchased from Beijing Vittoni China Animal Testing Center.

Real horse reagent: Parameter CAMP Assay Kit, KGE002 (American R&D Systems, Inc.); DuoSet IC Human/Mouse/Rat Phospho-CREB (S 133) ELISA Kit,

DYC2510-2 (American R&D Systems, Inc.); PepTag Assay for Non-Radioactive Detection of cAMP-Dependent Protein Kinase Kit, V5340 (Promega Corporation, USA); PDE-Glo Phosphodiesterase Assay Kit, V1361 (Promega Corporation, USA); Pierce BCA Protein Assay Kit, 23227, (Thermo, USA); Paroxetine Hydrochloride (Batch No.: 08030078, purchased from Sino-US Tianjin Shike Pharmaceutical Co., Ltd.); Controls such as adenosine monophosphate and adenosine were purchased from China Pharmaceutical and Biological Products Co., Ltd. NaH ₂ P0 ₄ , Na ₂ HP0 ₄ , KH ₂ P0 ₄ , KC1, NaCl, MgCl ₂ , Tris Base, Tris-HCl and other reagents are all cell culture grade biochemical reagents, purchased from Sigma, USA; E-64, APROTININ, LEUPEPTIN, Pepstatin A, PMSF, NaF, EDTA, EGTA, DTT, NaV04, Sodium pyrophosphate, agarose, glycerol and other reagents are all pure grade, purchased from BioBasic, Canada; acetonitrile, sterol (chromatographically pure, German MERCK) Company); Ultrapure water (MilliQ pure water, homemade).

Test drug: The pharmaceutical composition prepared in Example 1 of the present invention.

Positive drugs: paroxetine hydrochloride tablets, Lilly Suzhou Pharmaceutical Co., Ltd. products, daily dose, 20mg / day.

2, experimental equipment

FlexStation 3 Multi-Purpose Plate Analyzer (Molecular Devices, USA)

Corporation. ); Waters600E High Performance Liquid Chromatograph (Quaternary Pump, Online Degasser, Autosampler, Column Thermostat, UV Detector, Waters, USA); Refrigerated Centrifuge (BECKMAN, USA); Slurry (U TRASOU D TECHNOLOGY, USA); Electrophoresis (Beijing Liuyi Instrument Factory); Gel Imager (SYN GENE); ULTRA LOW Ultra-low Temperature Refrigerator (SANYO, Japan); mLine single-channel pipette, 8 channels Pipette (Biohit, Finland).

3. Experimental methods

1) Administration

Three days after adaptive feeding, the rats were randomly divided into 3 groups, which were labeled as: group A (blank control saline group), group B (positive drug paroxetine group) and group C (prepared by test drug example 1). Pharmaceutical composition group).

The paroxetine hydrochloride tablets were crushed and mixed with ultrapure water to a certain concentration. The rats were intragastrically administered at a dose of 5 mg/kg. The pharmaceutical composition prepared in Example 1 was formulated with water to a certain concentration. The solution was administered to rats at a dose of 30 mg/kg; the blank control saline group was given an equal volume of 0.9% saline; all rats were weighed and labeled with picric acid before administration, all at 37 °C. The drug was administered after 30 minutes of preheating.

2) Sampling

Eight hours after administration of the three groups of A, B and C, the rats were anesthetized with ether, the femoral artery was exsanguinated, and the brain was decapitated on ice. The hippocampus tissue was divided and finely cut into three parts, which were placed in the pre-numbered 1.5. The mL color cap screw is stored in the frozen tube, accurately weighed and quickly put into liquid nitrogen for quick freezing for 15 minutes, and then stored in a -80 °C refrigerator for later use.

4. Detection of experimental samples

1) Determination of cAMP content in rat hippocampus

Thaw the hippocampal tissue samples, rinse with a small amount of physiological saline, and add the cell lysate provided by the kit at a ratio of 1:20 (g: mL) (diluted with 5 times concentrated solution), homogenized by ultrasonic homogenizer After 30 s, centrifugation at 1000 °C for 5 minutes at 4 ° C, the supernatant was placed in a pre-numbered 1.5 mL color Eppendorf centrifuge tube and placed in an ice box for testing. Samples were taken using the US R&D Systems Parameter CAMP Assay ELISA kit. The sample was returned to room temperature and the cAMP content of the sample was determined by competitive ELISA according to the kit instructions. The OD value was measured at 450 nm using a multiwell plate analyzer, and the cAMP content in the sample was calculated from the standard curve.

2) Determination of cAMP-dependent PKA activity in rat hippocampus

The hippocampal tissue samples were thawed, rinsed with a small amount of saline, and then added to the PKA extraction buffer in a ratio of 1:10 (g: mL) (according to the formula in the kit;), homogenized by an ultrasonic homogenizer for 30 seconds, 4° Centrifuge at 100 rpm for 5 minutes at C. The supernatant was placed in a pre-coded 1.5 mL color Eppendorf centrifuge tube and placed in an ice box for testing. Detection and analysis were performed using Prop PepTag Assay for Non-Radioactive Detection of cAMP-Dependent Protein Kinase kit. Premixed reagents were added to the pre-coded 200μ PCR occlusion tube according to the kit instructions. 9 L of each sample was added to each tube, vortexed, centrifuged, reacted at room temperature for 30 minutes, and placed in a PCR machine. The enzyme was inactivated at 98 ° C for 5 minutes. In the experiment, positive control and negative control tubes were set according to the requirements of the manual, and the experiment was carried out. A 0.8% agarose gel was prepared, and 10 μL of each sample after the enzyme reaction was added to the comb hole of the gel, and electrophoresis was carried out for 30 minutes at 100 V, 130 mA, and the electrophoresis solution was 50 mM Tris-HCl (pH 8.0) buffer. After electrophoresis, the agarose gel was taken out, photographed by a gel imager, and then placed on an ultraviolet analyzer. The phosphorylated PipTag A1 peptide spots were cut out and placed in a pre-numbered 1.5 mL cap screw. In the tube. Heat to agarose gel, dilute to 250μ with ultrapure water, quickly remove 125 L and add pre-numbered 1.5mL In a color Eppendorf centrifuge tube, add the sol solution provided by the 75μΙ^ kit and 50μΙ glacial acetic acid, vortex and mix, take 200μ into the 96-well microtiter plate, and use the negative control tube agarose as the blank control. Fluorescence analysis was performed on a multiwell plate analyzer. The excitation wavelength was set at 568 nm and the emission wavelength was 592 nm. PKA activity is expressed in terms of sample fluorescence intensity.

3) Determination of total protein in rat hippocampus

In order to more accurately calibrate the amount of p-CREB protein per mg of protein in the sample, the total protein content of the sample needs to be determined. The supernatant after centrifugation of the tissue homogenate in the p-CREB assay was diluted 25 times with PBS, and then used as a test sample, and the OD value was measured at 562 nm using a perforated plate analyzer according to the Pierce BCA Protein Assay Kit reagent specification. Using bovine serum albumin (BSA) as a standard, the total protein content in the sample was calculated from the standard curve.

4) Determination of p-CREB in rat hippocampus

Samples were tested using the American R&D Systems DuoSet IC Human/Mouse/Rat Phospho-CREB (S 133) ELISA kit. The hippocampal tissue samples were thawed, rinsed with a small amount of physiological saline, and added to the tissue extract slurry at a ratio of 1:20 (g: mL) (according to the IC DELUENT 6# formula in the kit;), electronic ultrasonic homogenizer homogenate 30 The cells were centrifuged at 1000 °C for 5 minutes at 4 ° C. The supernatant was placed in a pre-coded 1.5 mL color Eppendorf centrifuge tube and placed in an ice box for testing. The sample was returned to room temperature during the assay, and the p-CREB content in the sample was determined by double-anti-sandwich ELISA according to the kit instructions. The OD value was measured at 450 nm using a multiwell plate analyzer, and the p-CREB content in the sample was calculated from the standard curve.

5) Determination of phosphodiesterase (PDE) activity

Use Promega Bioluminescent (POT-Glo)

The Phosphodiesterase Assay kit measures the effect of the test drug on PDE activity in rat hippocampus.

(1) Preparation of liquid medicine

The test drug (ie, the pharmaceutical composition prepared in Example 1, Group C) was prepared into a content.

Group C low dose (0.02mg/mL), group C medium dose (0.05mg/mL) and group C high dose (1.0mg/mL) three dose groups.

(2) sample preparation

Take a certain amount of hippocampus tissue, rinse with a small amount of normal saline, add PDE-Glo Reaction Buffer ( Tris-HCl 40mM, MgC12 lOmM, BSA O. lmg/ml, in a ratio of 1: 10 (g: mL). PMSF lmM, leupetin 2μΜ/ηΛ, aprotinin 2μΜ/ηΛ, E-64 2μΜ/πύ, ), homogenized by electron ultrasonicizer, centrifuged at 14000 rpm for 30 minutes at 4°C, and the supernatant was taken as an enzyme solution and used. (3) Determination by bioluminescence

According to the method provided in the kit instructions, the enzyme reaction solution is partially added with a solution of 1.5 μM enzyme solution for 2.5 μM; a substrate solution containing 2 μηιο1 cAMP is added at 2.5 μM, mixed, and reacted at 37 ° C for 30 minutes. Then, add 2.5 μΙ of the reaction termination solution containing the strong inhibitor of PDE, IBMX, and mix; add 2.5 L of the test solution, mix and react at room temperature for 20 minutes; finally add the luminescent reagent lO L, react at room temperature for 10 minutes and then in the multifunctional microporous Test on the board analyzer.

5, the experimental results

1) cAMP content in rat hippocampus

The cAMP concentration in the rat hippocampal tissue homogenate determined by ELISA was divided by the weight of the weighed tissue sample to obtain the cAMP content in the hippocampus, expressed as "pmol/g tissue", and the results are shown in Fig. 4.

As can be seen from Fig. 4, the content of cAMP in the hippocampus of rats was changed after 8 hours of administration, the group C (the pharmaceutical composition group prepared by the test drug example 1) and the eight groups (the blank control saline group), 8 groups. Compared with the positive drug paroxetine group, the content of CAMP in rat hippocampus was significantly increased, *P<0.05, n=10.

2) cAMP-dependent PKA activity in rat hippocampus

After the enzyme reaction, the sample was subjected to agarose gel electrophoresis and photographed by a gel imager. A typical gel electrophoresis pattern in the cAMP-dependent PKA activity experiment is shown in Figure 5A, for which a rough analysis is performed. The phosphorylated PepTag-Al peptide has a negative charge and moves toward the positive electrode; the unphosphorylated PepTag-Al peptide has a positive charge and moves toward the negative electrode to separate the two. The higher the brightness of the phosphorylated A1 peptide spot moving toward the positive electrode, indicating that the higher the phosphorylation level, the higher the cAMP-dependent PKA activity in the sample. As can be seen from Fig. 5A, the brightness of the positive direction direction of the group C (test drug) sample was higher than that of the group A (saline) and the group B (paroxetine).

The agarose gel spots were cut, and the volume was adjusted after melting, and the cAMP-dependent PKA activity in the hippocampus of rats was measured by fluorescence. The experimental results expressed by fluorescence intensity are shown in Fig. 5B. As can be seen from the figure, there was a significant difference in cAMP-dependent PKA activity in hippocampus of different groups of rats after administration for 8 hours, group C (the pharmaceutical composition group prepared by the test drug example 1) and eight groups (blank control) Compared with group B (positive drug paroxetine group), cAMP-dependent PKA activity in rat hippocampus was significantly increased, *P<0.05, n=10.

3) Total protein content in rat hippocampus

The concentration of total protein in the rat hippocampal tissue homogenate was determined by BCA method to calibrate the amount of P-CREB per total protein. The results are shown in Table 6. Table 6 Total protein concentration of rat hippocampal tissue homogenate (g/mL)

Group A (blank control) Group B (paroxetine) Group C (test drug)

1 308.627 310.458 289.965

2 303.134 318.697 268.697

3 312.218 333.697 295.246

4 311.937 341.162 302.218

5 306.162 324.613 283.204

6 306.796 318.275 307.007

7 314.542 304.754 277.430

8 307.077 310.458 287.359

9 297.570 301.514 295.528

10 276.796 293.627 315.880 Average ±SD 304.486±10.877 315.725±14.646 292.254±14.066

4) p-CREB content in rat hippocampus

Determination of the double-antibody sandwich ELISA rat hippocampal tissue samples hook slurry concentration p-CREB, and to p-CREB (pg) / total protein (μ _β) represents p-CREB in the hippocampus of rats tissue, results are shown 6.

As can be seen from Fig. 6, the content of p-CREB in the hippocampus of rats was changed 8 hours after administration, (group (the pharmaceutical composition group prepared by the test drug example 1) and eight groups (the blank control saline group) Compared with the sputum group (positive drug paroxetine group), the content of p-CREB in rat hippocampus was increased, but there was no statistically significant difference, η=10.

5) Effect of drugs on PDE activity in rat hippocampus

The activity of PDE in hippocampus and the inhibition of PDE activity in vitro were determined by bioluminescence. The PDE activity was expressed by the luminescence intensity. Compared with the sputum group (the blank control saline group), the luminescence intensity was higher, indicating that the PDE activity was higher; the luminescence intensity was lower, indicating that the PDE activity was inhibited as shown in Fig. 7. As can be seen from Fig. 7, compared with the group A blank control, the medium dose group (0.05 mg/mL) provided by the present invention significantly inhibited PDE activity, *P<0.05, n=4.

From the above results, it can be seen that the middle dose group of the pharmaceutical composition of the present invention can significantly inhibit the activity of phosphodiesterase in rat hippocampus, since the phosphodiesterase is an inactivated enzyme of cAMP, which can inhibit the rat hippocampus after being inhibited. The cAMP content in the tissue is elevated. After 8 hours of intragastric administration in rats, the pharmaceutical composition prepared in Example 1 of the present invention was compared with the saline group and the paroxetine group, and the hippocampus tissue of the brain was compared. The cAMP content increased significantly; cAMP-dependent PKA activity was significantly enhanced; p-CREB content was also mentioned! 3⁄4.

By the present example, it was confirmed that the pharmaceutical composition provided by the present invention can exert pharmacological action through the second messenger cAMP cell signal transduction pathway, and can rapidly activate cAMP-PKA-CREB (p-CREB) after administration for 8 hours. Pathway (significantly different from the blank control saline group and the positive drug paroxetine group, *P < 0.05), while the same control drug, paroxetine, failed to activate cAMP-PKA-CREB (p-CREB) )path.

Claims

Rights request

A pharmaceutical composition for treating depression, characterized in that the pharmaceutical composition comprises at least two active ingredients selected from the group consisting of:

a. Ginsenoside Rgl;

b. Ginsenoside Rbl ; and

c glycyrrhizic acid or glycyrrhetinic acid;

The ginsenoside Rgl contained in the pharmaceutical composition is 20 to 100 parts by weight, the ginsenoside Rbl is 5 to 100 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 5 to 100 parts by weight;

Preferably, the pharmaceutical composition comprises 60 parts by weight of ginsenoside Rgl, 30 to 60 parts by weight of ginsenoside Rbl, and 30 to 60 parts by weight of glycyrrhizic acid or glycyrrhetinic acid.

The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises three active ingredients of ginsenoside Rgl, ginsenoside Rbl and glycyrrhizic acid or glycyrrhetinic acid; preferably, the pharmaceutical composition Does not contain other active ingredients;

The ginsenoside Rgl in the pharmaceutical composition is 20 to 100 parts by weight, the ginsenoside Rbl is 5 to 100 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 5 to 100 parts by weight;

Preferably, the ginsenoside Rgl in the pharmaceutical composition is 60 parts by weight, the ginsenoside Rbl is 30 to 60 parts by weight, and the glycyrrhizic acid or glycyrrhetic acid is 30 to 60 parts by weight.

The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises two active ingredients of ginsenoside Rgl and ginsenoside Rbl; preferably, the pharmaceutical composition does not comprise other active ingredients;

The ginsenoside Rgl in the pharmaceutical composition is 20 to 100 parts by weight, and the ginsenoside Rbl is 5 to 100 parts by weight;

More preferably, the ginsenoside Rgl in the pharmaceutical composition is 60 parts by weight, and the ginsenoside Rbl is 30 to 60 parts by weight.

The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises two active ingredients of ginsenoside Rgl and glycyrrhizic acid or glycyrrhetinic acid; preferably, the pharmaceutical composition does not contain other active ingredients Ingredient

The ginsenoside Rgl in the pharmaceutical composition is 20 to 100 parts by weight, and glycyrrhizic acid or licorice The secondary acid is 5 to 100 parts by weight;

Preferably, the ginsenoside Rgl in the pharmaceutical composition is 60 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 30 to 60 parts by weight.

The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises two active ingredients: ginsenoside Rbl, and glycyrrhizic acid or glycyrrhetinic acid; preferably, the pharmaceutical composition does not contain other ingredients Active ingredient

The ginsenoside Rbl in the pharmaceutical composition is 5 to 100 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 5 to 100 parts by weight;

Preferably, the ginsenoside Rbl in the pharmaceutical composition is 30 to 60 parts by weight, and the glycyrrhizic acid or glycyrrhetinic acid is 30 to 60 parts by weight.

The pharmaceutical composition according to any one of claims 1 to 5, wherein the ginsenoside Rgl in the pharmaceutical composition has a purity of 50 to 98%, a purity of Rbl of 50 to 98%, and The purity of glycyrrhizic acid or glycyrrhetinic acid is 50-98%;

Preferably, the ginsenoside Rgl in the pharmaceutical composition has a purity of 90% to 98%, a purity of Rbl of 90% to 98%, and a purity of glycyrrhizic acid or glycyrrhetinic acid of 90% to 98%.

The pharmaceutical composition according to any one of claims 1 to 6, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers or additives;

More preferably, the dosage form is an oral preparation selected from the group consisting of a tablet, a capsule, a powder, a tablet, a powder, a solution, a micro-stringing agent, a suspension, an emulsion, a granule, a pill, and a pill; Further preferably, the pharmaceutically acceptable carrier or additive is selected from the group consisting of: a disintegrant, a lubricant, a binder, a filler, a solvent, a fragrance, a sweetener, an antioxidant, and the like.

A method of preparing the pharmaceutical composition according to any one of claims 1 to 7, which comprises 20 to 100 parts by weight of ginsenoside Rgl, 5 to 100 parts by weight of ginsenoside Rbl and 5 to 100 parts by weight of glycyrrhizic acid or licorice a step of mixing and pulverizing the hypoacid;

Preferably, the method further comprises 20 to 100 parts by weight of ginsenoside Rgl, 5-100 parts by weight of ginsenoside Rbl, 5-100 parts by weight of glycyrrhizic acid or glycyrrhetinic acid, and a pharmaceutically acceptable carrier or The additive is mixed and pulverized.

9. Use of a pharmaceutical composition according to any one of claims 1 to 7 for the preparation of a pharmaceutical preparation, a health supplement or a nutraceutical for the treatment or alleviation of depression.

10. Use of a pharmaceutical composition according to any one of claims 1 to 7 for the preparation of a pharmaceutical preparation, a health supplement or a nutraceutical for the simultaneous treatment or alleviation of depression and a disease, disorder or condition complicated by depression;

The use according to claim 9 or 10, characterized in that the depression is a depression associated with inflammation, preferably: stroke depression, rheumatoid arthritis associated depression and/or geriatric depression .

12. A method of treating or ameliorating depression, the method comprising administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical composition of any one of claims 1 to 7;

Preferably, the therapeutically effective amount is 0.5 to 5 mg/kg/d, preferably 1 to 4 mg/kg/d, and more preferably 2 to 3 mg/kg/d.

13. A method for simultaneously treating or ameliorating depression and a disease, disorder or condition associated with depression, the method comprising administering to a subject in need thereof a therapeutically effective amount of any one of claims 1 to 7. Said pharmaceutical composition;

Preferably, the concurrent disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder;

More preferably, the therapeutically effective amount is 0.5 to 5 mg/kg/d, preferably 1 to 4 mg/kg/d, and further preferably 2 to 3 mg/kg/d.

The method according to claim 12 or 13, wherein the depression is depression associated with inflammation, preferably: stroke depression, rheumatoid arthritis associated depression and/or geriatric depression .