WO2010133015A1 - Pharmaceutical composition for treating depression and preparative method and use thereof - Google Patents

Abstract

A pharmaceutical composition prepared by licorice root and white peony root or extracts thereof for treating depression is provided; the said composition comprises glycyrrhizin or enoxolone, peoniflorin and albiflorin etc. The preparative method of said composition and the use of said composition for the manufacture of medicaments, health-care foods and/or nutritional agents for treating depression and complication, disorder thereof are also provided in the present invention.

Description

 Pharmaceutical composition for treating depression, preparation method and use thereof

 The invention belongs to the field of medical technology. The present invention relates to a pharmaceutical composition for treating depression and a process for the preparation thereof. The invention further relates to a method of the pharmaceutical composition for the preparation of a medicament, a health food and/or a barrier or condition. Background technique

 Depression is the main type of mood disorder and is a syndrome characterized by significant and persistent low mood. Depression is a common and frequently-occurring disease that is harmful to human health. It is a major global mental problem. 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). Information provided by the World Health Organization: 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 depression will rise in the next 20 years. It is the second most common disease in the world.

 At present, antidepressant drugs in the domestic and international markets are mainly serotonin reuptake inhibitors (SSRIs) such as Baiyoujie, Selot, Zoloft, etc., and their mechanism of action is through regulation of human monoamine neurotransmitters5. Ingredients such as serotonin alleviate depressive symptoms. These drugs have varying degrees of side effects. Studies have shown that "the complex ampoules contained in these drugs have a role in balancing the body's functions, but more often, they still can't calm the patients.", In recent years, Baiyoujie, etc. Whether the drug of depression is harmful has become a serious social problem. Among them, Selot has been found to have potential safety hazards as early as 1996. It has been recalled from the market since 2001. June 2004, New York, USA The Attorney General accused the British GlaxoSmithKline of fraudulently concealing a study linking Zelot with "increasing the risk of suicidal tendencies and behavior among adolescents" in order to make a profit. In this context, how to develop production A new generation of drugs with small side effects and significant anti-depressant effects has become a concern of the global pharmaceutical community.

In order to overcome the shortcomings of existing antidepressant drugs, the inventors combined modern medicine and modern pharmacology to study the pathogenesis and mechanism of traditional Chinese medicine in the treatment of depression, and repeated drug effects and drugs. Based on the theory of physics, the present invention has been proposed.

 To help understand the invention, some terms are defined below. The terms defined herein have the meaning commonly understood by one of ordinary skill in the art to which the invention pertains.

 The term "pharmaceutically acceptable carriers and excipients," as used herein, unless otherwise indicated, refers to those materials which are well known in the art for use as fillers or carrier materials in pills, tablets, capsules and the like. Substances are generally approved by health care professionals for this purpose and as inactive ingredients of pharmaceutical agents. Compilation of pharmaceutically acceptable carriers and excipients can be found in the Handbook of Pharmaceutical excipients, 2nd edition, by A. Wade and P. J. Weller, ed.; American Pharmaceutical Association, Washington and The

Found in the book of Pharmaceutical Press, London, 1994).

 The term "therapeutically effective amount" as used herein, unless otherwise indicated, refers to an amount of a drug that is required to produce an effective effect, which can be varied, and ultimately by the medical practitioner, depending on the route of administration, the type of formulation, the age and weight of the recipient. , and the nature and severity of the disease being treated are determined by many factors. It is an object of the present invention to provide a pharmaceutical composition for treating depression.

 Another object of the present invention is to provide a method of preparing the above pharmaceutical composition.

 It is still another object of the present invention to provide the use of the above pharmaceutical composition for the preparation of a medicament for the treatment of depression, a health food and/or a nutrient.

 It is still another object of the present invention to provide use of the above pharmaceutical composition for the preparation of a medicament, health food and/or nutrient for the simultaneous treatment of depression and a disease, disorder or condition therewith, and for providing a A method of treating depression and simultaneously treating depression, a disease, disorder or condition 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 the following active ingredients:

 1) glycyrrhizic acid or glycyrrhetinic acid;

 2) peony; and

 3) Paeoniflorin.

 Preferably, in the pharmaceutical composition, the glycyrrhizic acid or glycyrrhetic acid is 0.1-2 parts by weight, the paeoniflorin is 0.4-8 parts by weight and the paeoniflorin is 0.15-5 parts by weight;

Preferably, in the pharmaceutical composition, the glycyrrhizic acid or glycyrrhetinic acid is 0.2-0.6 parts by weight, the paeoniflorin is 1.2-3.5 parts by weight and the paeoniflorin is 0.4-2 parts by weight; Most preferably, in the pharmaceutical composition, the glycyrrhizic acid or glycyrrhetinic acid is 0.3 parts by weight, the paeoniflorin is 0.6 parts by weight and the paeoniflorin is 0.4 parts by weight.

 Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient; preferably, the pharmaceutical composition is selected from the group consisting of: an oral preparation, a parenteral preparation, a topical and inhaled preparation. And transdermal formulations;

 More preferably, the dosage form is an oral preparation selected from the group consisting of: tablets, capsules, powders, granules, pills, drops, juices and syrups;

 Further preferably, the pharmaceutically acceptable carrier or excipient is selected from the group consisting of: a disintegrant, a lubricant, a binder, a filler, a solvent, a fragrance, a sweetener, an antioxidant. In another aspect, the invention provides a method of preparing the above pharmaceutical composition, the method comprising the steps of:

 (1) crushing the licorice, extracting and watering, filtering, recovering the ethanol, concentrating and drying, and obtaining the licorice extract;

 (2) crushing the white peony, extracting with 70% ethanol solution (or water osmosis), purifying through a macroporous resin adsorption column (ME-1 or D-101, etc.), collecting the eluate, concentrating and drying, and obtaining White peony extract;

 (3) mixing the licorice extract obtained in the step (1) and the white peony extract obtained in the step (2) or pulverizing with the added auxiliary material to prepare a preparation;

 Wherein, the licorice is 5 to 100 parts by weight, and the chalk is 10 to 600 parts by weight; preferably, the licorice is 10 to 60 parts by weight, and the chalk is 40 to 200 parts by weight; most preferably The licorice is 20 parts by weight, and the chalk is 60 parts by weight. In still another aspect, the present invention provides the use of a pharmaceutical composition prepared from licorice and peony as a raw material for the preparation of a medicament for the treatment of depression, a health food and/or a nutrient. In still another aspect, the present invention provides the use of a pharmaceutical composition prepared from licorice and peony as a raw material for the preparation of a medicament, health food and/or nutrient for simultaneously treating depression and a disease, disorder or condition therewith. ;

Preferably, the disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder. In still another aspect, the present invention provides a pharmaceutical composition prepared by using a licorice extract and a white peony extract as an active ingredient, for the preparation of a medicament for treating depression, a health food and/or a nutrient; preferably, The pharmaceutical compositions are free of other active ingredients. In still another aspect, the present invention provides a pharmaceutical composition prepared by using a licorice extract and a white peony extract as active ingredients in the preparation of a medicament, a health food, and/or a medicament for simultaneously treating depression and a disease, disorder or condition complicated therewith. Use in nutraceuticals;

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

 Preferably, the pharmaceutical composition is free of other active ingredients. In still another aspect, the present invention provides the use of the pharmaceutical composition for the preparation of a medicament, health food and/or nutrient for the treatment of depression. In still another aspect, the present invention provides the use of the pharmaceutical composition for the preparation of a medicament, a health food and/or a nutrient for simultaneously treating depression and a disease, disorder or condition therewith; preferably, the disease, The 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 depression, the method comprising administering to a subject a therapeutically effective amount of a licorice extract and a white peony extract. In still another aspect, the present invention provides a method of simultaneously treating depression and a disease, disorder or condition therewith, the method comprising administering to the subject a therapeutically effective amount of a licorice extract and a white peony extract;

 Preferably, the 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 depression, the method comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition;

 The therapeutically effective daily dose of the pharmaceutical composition is preferably: glycyrrhizic acid or glycyrrhetinic acid

20mg-90mg, paeoniflorin 40mg-120mg, albiflorin 30mg-100mg; more preferably: glycyrrhizic acid or glycyrrhetinic acid 30mg-60mg, paeoniflorin 60mg-120mg, albiflorin ⁴ 0mg-80mg. In still another aspect, the present invention provides a method of simultaneously treating depression and a disease, disorder or condition therewith, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition; The therapeutically effective daily dose is preferably: glycyrrhizic acid or glycyrrhetinic acid

20mg-90mg, paeoniflorin 40mg-120mg, paeoniflorin 30mg-100mg; more preferably: Oxalic acid or glycyrrhetinic acid 30mg-60mg, paeoniflorin 60mg-120mg, paeoniflorin 40mg-80mg. Preferably, the disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder. In order to more clearly illustrate and explain the technical solutions of the present invention, the present invention will be further described below:

 The pharmaceutical composition of the present invention comprises a licorice and a white peony group, and comprises at least one selected from the following parts by weight:

 Licorice 5-100 parts by weight

 White 芍 10~600 parts by weight

 Preferably:

 Licorice 10~60 parts by weight

 White 芍 40~200 parts by weight

 More preferably:

 Licorice 20 parts by weight

 White peony 60 parts by weight

 According to the pharmaceutical composition as described above, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient;

 Preferably, the pharmaceutical composition is selected from the group consisting of: an oral preparation, a parenteral preparation, a topical and inhaled preparation, and a transdermal preparation;

 Further preferably, the dosage form is an oral preparation selected from the group consisting of tablets, capsules, powders, granules, pills, drops, juices and syrups;

 Still more preferably, the pharmaceutically acceptable carrier or excipient is selected from the group consisting of: a disintegrant, a lubricant, a binder, a filler, a solvent, a fragrance, a sweetener, an antioxidant. The pharmaceutical composition obtained by extracting the extract powder obtained by using the above-mentioned group as a raw material; or extracting the extract obtained from the licorice and the white peony by water respectively (or extracting the white peony with an aqueous solution of ethanol) The obtained extract) is obtained by mixing and pulverizing the obtained pharmaceutical composition, and preferably, the licorice extract obtained by water extraction and alcohol precipitation and the white peony extract obtained by purifying the column by water permeation.

 The licorice extract contains 0.1-2 parts by weight of glycyrrhizic acid or glycyrrhetinic acid, and the white peony extract contains 0.4 to 8 parts by weight of paeoniflorin and 0.15 to 5 parts by weight of paeoniflorin;

Preferably, the licorice extract comprising 0.2 to 0.6 parts by weight of a glycyrrhizic acid or the glycyrrhetinic acid, the albiflorin peony extract containing 1.2-3.5 parts by weight of paeoniflorin and ^0.4 to ² parts by weight ; Further preferably, the licorice extract contains 0.3 parts by weight of glycyrrhizic acid or glycyrrhetinic acid, and the white peony extract contains 0.6 parts by weight of paeoniflorin and 0.4 part by weight of paeoniflorin.

 The pharmaceutical composition of the present invention may also be directly pulverized by using glycyrrhizic acid or glycyrrhetinic acid together with paeoniflorin and paeoniflorin, and the pharmaceutical composition contains 0.1-2 parts by weight of glycyrrhizic acid or glycyrrhetinic acid, 0.4-8 by weight. a portion of paeoniflorin and 0.15 to 5 parts by weight of paeoniflorin;

 Preferably, the weight ratio of the glycyrrhizic acid or glycyrrhetinic acid to the paeoniflorin and the weight ratio of the paeoniflorin are 0.2 to 0.6: 1.2-3.5: 0.4-2;

 More preferably, the weight ratio of the glycyrrhizic acid or glycyrrhetinic acid to the paeoniflorin and the weight ratio of the paeoniflorin are 0.3:0.6:0.4.

 Another object of the present invention is to provide a method of preparing the above pharmaceutical composition.

 Method 1, the method includes:

 The pharmaceutical composition is prepared by mixing licorice and white peony (preferably ultrafine pulverization); the licorice is 5 to 100 parts by weight, and the chalk is 10 to 600 parts by weight;

 Preferably, the licorice is 10 to 60 parts by weight, and the chalk is 40 to 200 parts by weight; most preferably, the licorice is 20 parts by weight, and the chalk is 60 parts by weight.

 Method two, the method includes:

 (1) crushing the licorice, extracting and watering, filtering, recovering the ethanol, concentrating and drying, and obtaining the licorice extract;

 (2) crushing the white peony, extracting with 70% aqueous ethanol solution (or water osmosis), purifying through a macroporous resin adsorption column (ME-1 or D-101, etc.), collecting the eluate, concentrating and drying, and obtaining White peony extract;

 (3) mixing the licorice extract obtained in the step (1) and the white peony extract obtained in the step (2) or pulverizing with the added auxiliary material to prepare a preparation;

 Wherein, the licorice is 5 to 100 parts by weight, and the chalk is 10 to 600 parts by weight; preferably, the licorice is 10 to 60 parts by weight, and the chalk is 40 to 200 parts by weight; most preferably The licorice is 20 parts by weight, and the chalk is 60 parts by weight.

 Method three, the method comprises:

 The pharmaceutical composition is prepared by directly mixing and crushing glycyrrhizic acid (or glycyrrhetinic acid) with paeoniflorin and paeoniflorin;

 The pharmaceutical composition has 0.1 to 2 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid), and contains 0.4-8 parts by weight of paeoniflorin and 0.15 to 5 parts by weight of paeoniflorin;

Preferably, it contains 0.2-0.6 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid), 1.2-3.5 parts by weight of paeoniflorin and 0.4 to 2 parts by weight of paeoniflorin; Further preferably, it contains 0.3 parts by weight of glycyrrhizic acid (or glycyrrhetinic acid), 0.6 parts by weight of paeoniflorin, and 0.4 part by weight of paeoniflorin.

 The above pharmaceutical composition is a core component for achieving the object of the present invention. After the present invention is disclosed, those skilled in the art can routinely apply the above pharmaceutical composition and extract composition according to the theory of traditional Chinese medicine or related modern pharmacological theory. Addition and subtraction. Such conventional addition and subtraction is a general technical activity of those skilled in the art, as long as it is a general technical addition or subtraction based on the formulation of the pharmaceutical composition and the extract composition of the present invention, both of which are in the present invention. Within the scope of protection.

 The pharmaceutical compositions described above include a pharmaceutically acceptable carrier or excipient which can be processed into any pharmaceutically acceptable oral dosage form (tablets, capsules or powders, etc.).

 The scope of application of the pharmaceutical composition of the present invention: it can be formulated into a botanical preparation, a health food and/or a nutrient for the treatment of depression. The pharmaceutical composition of the present invention is useful for treating depression and diseases, disorders or conditions associated therewith. Preferably, the disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder. Compared with the prior art, the present invention has the following distinct advantages:

 The main active ingredients of anti-depressant of Radix Paeoniae Alba are paeoniflorin and paeoniflorin, in which paeoniflorin inhibits ATPase and increases AC activity to enhance cAMP content in cytoplasm; the main active constituents of licorice anti-depression are glycyrrhizic acid and licorice Coumarin and the like are cAMP phosphodiesterase inhibitors. The white peony and licorice group can activate the cAMP-PKA pathway in the brain region by multiple targets, and promote the phosphorylation of CREB, which enhances the anti-suppressive effect.

 Although both the white peony root extract and the licorice extract have a certain degree of anti-depressant function, the results of the orthogonal experiment of the mouse tail suspension prove that the pharmaceutical composition of the present invention is superior to the single white peony in the antidepressant effect. Extract, licorice extract.

 In addition, the traditional Chinese medicine prescription Zhigan Tang is also only used in the prescription of white peony and licorice, but its indications do not include depression, and the prescription ratio of 芍甘汤 and its preparation method are also related to the medicine of the present invention. Different from the composition, the pharmaceutical composition of the present invention has obvious advantages over the glutinous soup in terms of composition, preparation and indications.

BRIEF DESCRIPTION OF THE DRAWINGS

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

 1 is a process flow diagram of a method for preparing a pharmaceutical composition of the present invention using licorice and chalk as raw materials;

2 is a preparation of the medicament of the present invention directly using glycyrrhizic acid, paeoniflorin and paeoniflorin as raw materials Process flow diagram of the method of the composition;

 Fig. 3 is a process flow diagram showing a method of directly mixing licorice and white peony to prepare a pharmaceutical composition of the present invention. The best way to implement the invention

 The invention is further illustrated below in conjunction with specific embodiments. However, the examples are intended to be illustrative only and not to limit the scope of the invention. The experimental methods for the specific 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 method for preparing a pharmaceutical composition of the present invention using licorice and chalk as a raw material, the operation comprising the following steps:

 Referring to Figure 1, 15 kg of licorice (in accordance with the 2005 edition of the Pharmacopoeia of the People's Republic of China) was broken, and water was extracted 3 times, each time being 2, 2, 1.5 hours, and the amount of water added was 12, 10, 10 times. Liquid, recovered extract, to medicinal material: extract is 1: 1.5, add ethanol to 75%, cold, filter, recover ethanol, to relative density of 1.20-1.30 (70 °C), concentrate at 70 °C under reduced pressure , got 3 kg of licorice extract (containing 10% glycyrrhizic acid);

 Referring to Figure 1, 150 kg of white peony (according to the 2005 edition of the Pharmacopoeia of the People's Republic of China) was crushed and heated under reflux with a 70% aqueous solution of ethanol for 3 times. The solvent weight of the three times was 5 times that of the white peony, 4 Double, 3 times, concentrated ethanol recovery, dilute the paste to 4 times the volume, filtered to obtain a ME-1 type macroporous resin adsorption column on the clear liquid (provided by the Key Laboratory of Bioactive Materials Education, Nankai University), with 1 bed volume/hour Flow rate adsorption, first rinsed with 4 times bed volume of water, then rinsed with 2 times bed volume of 10% ethanol, then rinsed with 30% ethanol, collected 2, 3, 4, 5 bed volume of eluent, concentrated Dry, get 4.5 kg of white peony extract (the ash yield is 3%, containing 48.13% paeoniflorin, 31.16% paeoniflorin);

 The two extracts obtained in the above steps are mixed and pulverized to obtain the pharmaceutical composition of the present invention. Example 2 Preparation of the pharmaceutical composition of the present invention

 This embodiment provides a preferred method for preparing a pharmaceutical composition of the present invention directly from a licorice extract and a white peony extract, the operation comprising the steps of:

Referring to Figure 2, 3 g of glycyrrhizic acid (or glycyrrhetinic acid) with a purity of 90% is directly mixed with 12 g of paeoniflorin with a purity of 90% and 6 g of paeoniflorin with a purity of 90%. A pharmaceutical composition of the invention. Example 3 Preparation of the pharmaceutical composition of the present invention

 This example provides a preferred method of preparing a pharmaceutical composition of the present invention directly by mixing licorice and white peony, the operation comprising the steps of:

 Referring to Figure 3, the pharmaceutical composition of the present invention was obtained by directly mixing 1.5 kg of licorice with 14 kg of white peony and then super-pulverizing. Experimental Example 4 Effects of extracts of Angelica sinensis (B) and Glycyrrhiza uralensis (G) on the time of mouse tail suspension

 This embodiment is a preferred combination of the pharmaceutical composition of the present invention having an antidepressant effect, according to the effect of the white peony extract (B) and the licorice extract (G) of the pharmaceutical composition of the present invention on the time of suspension of the mouse. .

 Practical purpose

 A preferred combination of antidepressant effects of B (white peony extract in Example 1) and G ( licorice extract G in Example 1) was screened by orthogonal experiments designed.

 2. Experimental materials

 2.1 Experimental animals: ICR mice, male, weighing 18-22 g, provided by the Ministry of Animals and Animals of Capital Medical University, certificate number scxk (Beijing) 2005-0006.

 2.2 Practical drugs:

 Test drugs: B (white peony extract in Example 1), G ( licorice extract in Example 1); positive drug: fluoxetine hydrochloride 百 (baiyou solution), purchased from Lilly Suzhou Pharmaceutical Co., Ltd. Co., Ltd., batch number: A333341-070608, Chinese medicine Zhunzi J20030017, daily dosage, 20mg/day.

 2.3 Experimental equipment:

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

 3. Experimental methods

 3.1 Orthogonal design of B and G drug combinations

 正交 The orthogonal test method is used, and the dose and combination of the B drug and the G drug are optimized based on the time when the mouse is suspended. The factor level table and the orthogonal test table are shown in Table 1 and Table 2.

Table 1 Factor Level Table

 2 40 20

3 0 0

 1 \乡\乡乡乡乡乡乡 & ^合合^合会4.. 1 1 1 1

 About about about ^ about ^

 2 1 2 2 2

3 1 3 3 3

4 2 1 2 3

5 2 2 3 1

6 2 3 1 2

7 3 1 3 2

8 3 2 1 3

9 3 3 2 1 According to the factor level table and the orthogonal test table, the dosing schedule is shown in Table 3: Table 3 Dosing schedule

 Medicine ^! B medicine /kg/d G medicine / kg/d

 Group 1 B medicine 80mg G medicine 40mg

 2 groups B medicine 80mg G medicine 20mg

 3 groups B medicine 80mg G medicine Omg

 4 groups B medicine 40mg G medicine 40mg

 5 groups B medicine 40mg G medicine 20mg

 6 groups B medicine 40mg G medicine Omg

 Group 7 B medicine Omg G medicine 40mg

 Group 8 B medicine Omg G medicine 20mg

 9 groups B medicine Omg G medicine Omg

3.2 group administration

150 normal mice, randomly divided into 10 groups according to body weight, each group of ⁵ , that is, 1-9 groups, positive drug fluoxetine hydrochloride capsules (3.5 mg / kg / d), blank control group (administered 9 groups are equivalent to blank groups). Each group was administered at 0.2 ml/10 g body weight for 2 consecutive days.

 3.3 Test methods

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 on the line of the lOOg tension transducer with a tape to make it fall. In the suspended state, the head is about 15 cm away from the test bench, and two animals are tested at the same time, separated from each other by cardboard. The transducer is connected to the Medlab biosignal acquisition and processing system. After 2 min, the small suspension time within 4 min is recorded, and the immobile state is converted into time (s).

 And the second, first and second rats

 4. Experiment ^W245678 (31 results

 s: group group group group group group

 See Table 4 and Table 5 for the experimental results.

 Table 4 Results of mouse tail suspension experiment

 M dose (mg/kg) number of samples immobility time (x ± s )

 - 14 112.21 ± 38.95

3.5mg 15 62.57 ± 19.79**

B medicine 80mg + G medicine 40mg 15 70.03 + 24.12**

B medicine 80mg + G medicine 20mg 14 79.50 ± 33.02**

B medicine 80mg + G medicine Omg 13 85.46 + 34.98*

B medicine 40mg + G medicine 40mg 14 78.75 + 31.92**

B medicine 40mg + G medicine 20mg 14 90.21 ± 29.97

B medicine 40mg + G medicine Omg 15 88.87 ± 28.17

 B medicine Omg + G medicine 40mg 14 83.82 ± 33.60*

B medicine Omg + G medicine 20mg 15 98.42 soil 40.72 Note: Compared with the blank group, * P <0.05, **P<0.01

 2 1 2 2 2 79.50

 3 1 3 3 3 85.46

 4 2 1 2 3 78.75

 5 2 2 3 1 90.21

 6 2 3 1 2 88.87

 7 3 1 3 2 83.82

 8 3 2 1 3 98.42

 9 3 3 2 1 112.21

 234.99 232.6 257.32 272.45

 257.83 268.13 270.46 252.19

 294.45 286.54 259.49 262.63

59.46 53.94 13.14 20.26 According to the experimental results, the best condition for the range analysis is B1G1, which is a combination of 80 mg of B medicine and 40 mg of G medicine. In the orthogonal table, the first group is the best condition, and in each administration group, the administration is the first. The group had the shortest time of suspension and had a significant difference compared with the blank control group (PO.01).

The combination of B1G2 (B medicine 80mg+G medicine 20mg) and B2G1 (B medicine 40mg+G medicine 40mg) can also shorten the time of mouse tail suspension, which is significantly different from the blank control group ( PO.01).

 B1G3 (80 mg alone) and B3G1 (40 mg G alone) also shortened the time of mouse tail suspension, which was significantly different from the blank control group (PO.05).

 5 Conclusion

 5.1 Administration by the intragastric administration for 2 days, the combination of the pharmaceutical composition B1G1 (white peony extract 80 mg + licorice extract 40 mg), the pharmaceutical composition B1G2 (white peony extract 80 mg + licorice extract 20 mg) Or the combination of the pharmaceutical composition B2G1 (white peony root extract 40mg + licorice extract 40m) can significantly shorten the time of mouse tail suspension (P<0.01, compared with the blank group), and has significant antidepressant effect. .

 5.2 Administration by intragastric administration for 2 days, although the extracts of Radix Paeoniae Alba and Radix Glycyrrhizae were also effective (P<0.05, compared with the blank group), the pharmaceutical composition of the present invention (optimization of extracts of Radix Paeoniae Alba and licorice extract) The antidepressant effect of the combination combination B1G1, B1G2, B2G1) was more obvious and more stable (PO.01, compared with the blank group). Experimental Example 5 Effect of Gadolin chinensis on Forced Swimming Experiment in Mice

 1 Experimental materials

 1.1 Experimental animals

 ICR mice, male, weighing 22.0 ± 2g, secondary, provided by the Department of Laboratory Animal Science, Capital Medical University, Beijing.

 1.2 Real 3 whole medicine

 Test drug: The pharmaceutical composition of the present invention prepared in accordance with Example 1 is a capsule, and the positive drug: paroxetine (Selite), a product of Sino-US Tianjin Shike Pharmaceutical Co., Ltd.

 1.3 Experimental instruments

 GM222 type electronic thermometer, stopwatch.

 2. Experimental methods and results

 2.1 Dose design

 The large dose of the test drug is 120mg kg/d, the medium dose is 60mg kg/d, and the small dose is

 2.2 group administration

The mice were randomized into groups of 10: (1) High dose group of test drugs (UOmg*!^- ¹ , Oral administration, administration 2d); (2) medium dose group (60mg *kg-l, intragastric administration, administration for 2d); (3) small dose group (SOmg *]^- ^{1 of the} test drug Oral administration, administration 2d); (4) positive drug paroxetine group Gmg kg- ¹ , gavage, administration 2d).

 2.3 Experimental methods

 The mice were placed in a glass jar with a water depth of 10 cm and a diameter of 14 cm, and the water temperature was 25 ° C. The cumulative immobility time of the mice in the water was recorded for 5 minutes.

 2.4 Statistical processing

 The experimental data were expressed as ^ ±, and the experimental results were analyzed by SPSS 11.5 statistical software for analysis of variance. 2.5 Experimental results

 See Table 6 for the experimental results.

 Table 6 芍 嚢 嚢 嚢 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠 小鼠17. 27* 芍Gold 嚢 high dose group 10 79. 17 ± 39. 33* 芍 胶囊 capsule medium dose group 10 64. 58 ± 24, 00* 芍 胶囊 capsule small dose group 10 128. 00 ± 19. 34 and Model group comparison, * P<0.05

 3 Conclusion

 According to the above experiment, it can be seen that the pharmaceutical composition of the present invention, the ruthenium glutamate, the middle dose group and the paroxetine group can significantly shorten the cumulative time of forced swimming in mice, the middle dose group and the high dose group and the saline group ( There is a significant difference in the model group), so that it can be inferred that the pharmaceutical composition of the present invention, guanidine gum, has an anti-experimental depression function. Experimental Example 6 Effect of Gadolin chinensis on Monoamine Neurotransmitter in Rats with Chronic Stress

 1 Experimental materials

 1.1 Experimental drugs

 Test drug: The pharmaceutical composition of the present invention is guanidine gum; positive drug: fluoxetine hydrochloride (Prozac), provided by Lilly Suzhou Pharmaceutical Co., Ltd. (batch number: Chinese medicine standard J20030017).

 1.2 real 3 佥 animals

 Wistar male rats, weighing 220g ~ 240g, were provided by Beijing Vital Lihua Company. All animals were purchased one week in advance and routinely reared.

1.3 Instruments and reagents Norepinephrine NE (Serva), dopamine DA (Fluka), adrenaline (E), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), high vanillic acid (HVA) , 3, 4-dihydroxyphenylacetic acid (DOPAC), 3, 4-dihydroxybenzylamine (DHBA) (both Sigma), di-n-butylamine (Shanghai Chemical Reagent Purchasing Station imported package), B-8 ion For reagents (Tianjin Chemical Reagent 2), sterol (excellent grade, Beijing Chemical Plant), other reagents are domestically analyzed.

 2 Experimental methods

 2.1 Grouping and administration

 72 normal rats, after 24 hours of water inadequacy, were given 1% sucrose water and measured the consumption within 1 hour. According to the consumption of sucrose water, it was randomly divided into 6 groups, 12 in each group, namely normal control group, model group, positive drug solution group (3.5mg kg/d), and glutinous gum high (100mg/kg/d). , medium (50mg kg/d), low (25mg/kg/d) dose group. Simultaneous administration of the model was performed once a day for 21 days. Each group was administered at a dose of 1.0 ml/100 g body weight, and the body weight was weighed once a week.

 2.2 Modeling

 The rats in the control group were raised, and the rats in the medication group and the model group were raised in each cage. Reference literature method (Xu Jing, Li Xiaoqiu, Establishment and Evaluation of Chronic Stress Depression Model, Chinese Journal of Behavioral Medicine, 2003, 12 (1): 14-16), medication group (administered simultaneously) and model group rats accepted 21d various random stresses, including 4°C ice water swimming (5min), 45°C hot drying (5min), tail (lmin), wet feeding (wet litter), day and night reversal (24h), ban Food (24h), water-free (24h) and other stimuli.

 2.3 Sample preparation and detection methods

 The animals were killed by decapitation, and the brain was quickly dissected on the ice. The forehead cortex was taken. The tissue was weighed and frozen in a frozen tube with liquid nitrogen, and then stored in a -70 °C refrigerator until the measurement.

 According to the weight of the brain tissue, pre-cooled 0.1mol / L perchloric acid (containing 0.3mM EDTA disodium and 0.5mM sodium sulfite), 2 μg / m LDHBA, ultrasonic homogenization, l lOOOrpm centrifugation lOmin, supernatant use night For neurotransmitter determination.

 The high performance liquid chromatography-electrochemical detector system (HPLC-ECD type, purchased from ESA Biosciences, Inc., USA) was used to determine the color: the color i column was 4 χ 150 mm.

Nova-pakC18, 5 μ m (filled by National Color Language Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences); mobile phase is 50 mM citric acid-sodium acetate buffer pH 3.5 (containing 1.0 mM B-8 ion pair) Reagent, 1.8 mM di-n-butylamine, 0.3 mM EDTA disodium, 4% methanol), flow rate 1.0 mL/min, glassy carbon working electrode, detection cell voltage +0.75 V; 3,4-dihydroxybenzylamine DHBA For the standard, the main components in the sample were quantified by the internal standard method. 2.4 Statistical analysis Experimental data were expressed as mean soil standard deviation ( ^ ± s ). SPSS13.0 statistical software was used to compare each group with one-way ANOVA. The significance level was 0.05 and 0.01. As standard.

 3 Experimental results

 The results showed that after 21 days of continuous administration, compared with the blank group, the monoamine neurotransmitters in the model group had lower levels of norepinephrine and serotonin (p<0.05), compared with the model group. The monoamine neurotransmitters in the middle and high dose groups of the capsules and the positive drug fluoxetine hydrochloride increased the levels of norepinephrine and serotonin (PO.05). See Table 7 and Table 8. Table 7 Effect of guanidine gum on norepinephrine in the brain (士 s )

 Group dose (; mg/kg) number of samples NE

 Blank group - 9 398.47 ± 111.11* model group - 11 159.20 ± 49.31 positive group 3.5mg/kg/d 8 440.88 ± 152.58* 芍 嚢 嚢 low dose group 25mg/kg/d 11 174.59 + 70.35 芍甘 capsule medium dose group 50mg/kg/d 11 345.19 ± 133.43* 芍Gold 嚢 high dose group 100mg/kg/d 10 271.48 ± 102.50 Compared with the model group, *P<0.05, **P<0.01

Table 8 ^p芍 嚢 嚢 嚢 脑 脑 脑 脑 x x x x x x x x x x x x x x x x x x x x x x x x x x

 Blank group - 8 340.11 ± 75.88* model group - 11 158.27 + 90.34 positive group 3.5mg/kg/d 10 272.19 ± 93.42* 芍 嚢 嚢 low dose group 25mg/kg/d 11 188.43 ± 167.91 芍 嚢 嚢Group 50mg/kg/d 8 270.77 soil 96.43* 芍Gold 嚢 high dose group lOOmg/kg/d 9 258.86 + 87.55* Compared with the model group, *P<0.05, **P<0.01

4 Conclusion

 The results of the experimental study showed that the content of 5-HT and NE in the brain of rats with chronic stress was significantly decreased. The medium and high dose group of the pharmaceutical composition of the present invention can significantly increase the content of 5-HT and NE in the brain, suggesting the drug of the present invention. The composition may act as an antidepressant by elevating the expression of monoamine neurotransmitters in the brain.

Claims

Rights request A pharmaceutical composition for treating depression, characterized in that the pharmaceutical composition contains the following active ingredients:  4) glycyrrhizic acid or glycyrrhetinic acid;  5) peony; and  6) Paeoniflorin. The pharmaceutical composition according to claim 1, wherein in the pharmaceutical composition, the glycyrrhizic acid or glycyrrhetinic acid is 0.1 to 2 parts by weight, and the paeoniflorin is 0.4 to 8 parts by weight. And the paeoniflorin is 0.15-5 parts by weight;  Preferably, in the pharmaceutical composition, the glycyrrhizic acid or glycyrrhetinic acid is 0.2-0.6 parts by weight, and the paeoniflorin is 1. 2-3.5 parts by weight and the paeoniflorin is 0.4-2 parts by weight;  Most preferably, in the pharmaceutical composition, the glycyrrhizic acid or glycyrrhetinic acid is 0.3 parts by weight, the paeoniflorin is 0.6 parts by weight and the paeoniflorin is 0.4 parts by weight. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient;  Preferably, the pharmaceutical composition is selected from the group consisting of: an oral preparation, a parenteral preparation, a topical and inhaled preparation, and a transdermal preparation;  More preferably, the dosage form is an oral preparation selected from the group consisting of: tablets, capsules, powders, granules, pills, drops, juices and syrups;  Further preferably, the pharmaceutically acceptable carrier or excipient is selected from the group consisting of: a disintegrant, a lubricant, a binder, a filler, a solvent, a fragrance, a sweetener, an antioxidant. A method of producing the pharmaceutical composition according to any one of claims 1 to 3, characterized in that the method comprises the steps of:  (1) crushing the licorice, extracting and watering, filtering, recovering the ethanol, concentrating and drying, and obtaining the licorice extract;  (2) crushing the white peony, extracting with 70% ethanol solution (or water osmosis), purifying through a macroporous resin adsorption column (ME-1 or D-101, etc.), collecting the eluate, concentrating and drying, and obtaining White peony extract; (3) mixing the licorice extract obtained in the step (1) and the white peony extract obtained in the step (2) or pulverizing with the added auxiliary material to prepare a preparation; Wherein, the licorice is 5 to 100 parts by weight, and the chalk is 10 to 600 parts by weight; preferably, the licorice is 10 to 60 parts by weight, and the chalk is 40 to 200 parts by weight; most preferably The licorice is 20 parts by weight, and the chalk is 60 parts by weight. 5. Use of a pharmaceutical composition prepared from licorice and peony as a raw material for the preparation of a medicament for the treatment of depression, a health food and/or a nutrient. 6. Use of a pharmaceutical composition prepared from licorice and peony as a raw material for the preparation of a medicament, a health food and/or a nutrient for simultaneously treating depression and a disease, disorder or condition therewith; preferably, The disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder. 7. Use of a pharmaceutical composition prepared by using a licorice extract and a white peony extract as an active ingredient for the preparation of a medicament for treating depression, a health food and/or a nutrient;  Preferably, the pharmaceutical composition is free of other active ingredients. 8. Use of a pharmaceutical composition prepared from a licorice extract and a white peony extract as an active ingredient for the preparation of a medicament, health food and/or nutrient for simultaneously treating depression and a disease, disorder or condition therewith ;  Preferably, the disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder; and/or  Preferably, the pharmaceutical composition is free of other active ingredients. Use of the pharmaceutical composition according to any one of claims 1 to 3 for the preparation of a medicament for the treatment of depression, a health food and/or a nutrient. 10. Use of a pharmaceutical composition according to any one of claims 1 to 3 for the preparation of a medicament, health food and/or nutrient for the simultaneous treatment of depression and a disease, disorder or condition therewith; preferably, The disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder. 11. A method of treating depression, the method comprising administering to a subject a therapeutically effective amount of a licorice extract and a white peony extract. 12. A method of simultaneously treating depression and a disease, disorder or condition associated therewith, Characterized in that the method comprises administering to the subject a therapeutically effective amount of a licorice extract and a white peony extract;  Preferably, the disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder. 13. A method of treating depression, the method comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition of any of claims 1-3;  The therapeutically effective daily dose of the pharmaceutical composition is preferably: glycyrrhizic acid or glycyrrhetinic acid 20 mg-90 mg, paeoniflorin 40 mg-120 mg, paeoniflorin 30 mg-100 mg; more preferably: glycyrrhizic acid or glycyrrhetinic acid 30 mg-60 mg, paeoniflorin 60 mg-120 mg, paeoniflorin 40 mg-80 mg. A method for simultaneously treating depression and a disease, disorder or condition therewith, characterized in that the method comprises administering to the subject a therapeutically effective amount of the pharmaceutical composition according to any one of claims 1-3. Object  The therapeutically effective daily dose of the pharmaceutical composition is preferably: glycyrrhizic acid or glycyrrhetinic acid  20 mg-90 mg, paeoniflorin 40 mg-120 mg, paeoniflorin 30 mg-100 mg; more preferably: glycyrrhizic acid or glycyrrhetinic acid 30 mg-60 mg, paeoniflorin 60 mg-120 mg, paeoniflorin 40 mg-80 mg.  Preferably, the disease, disorder or condition is selected from the group consisting of anxiety, sleep disorders and post-traumatic stress disorder.