CN1416881A - Medicine composition for treating depression and its prepn - Google Patents

Abstract

The invention discloses an antidepressant pharmaceutical composition and a preparation method thereof. The pharmaceutical composition mainly comprises isoliquiritigenin, liquiritigenin, liquiritin, neohesperidin, hesperidin, isoliquiritin, 5,3'-di Hydroxy-4'-methoxydihydroflavone-7-O-β-D-glucoside, 5,7-dihydroxycoumarin, saikosaponin c, saikosaponin d, glycyrrhizic acid 11 compounds. The pharmaceutical composition firstly prepares the antidepressant active part from the water decoction of Sini powder through macroporous adsorption resin purification and separation, and then adopts various solvent separation methods and chromatographic separation techniques to separate the components of the pharmaceutical composition. The pharmaceutical composition has good antidepressant effect.

Description

A kind of antidepressant pharmaceutical composition and preparation method thereof

field of invention

The invention relates to a pharmaceutical composition and a preparation method thereof, in particular to an antidepressant pharmaceutical composition and a preparation method thereof.

Background technique

With the aggravation of various emergency factors, depression has become a common disease and a high incidence in modern society, and its incidence is rising rapidly. According to incomplete statistics, patients with depression account for 3-5% of the world's population. WHO predicts that by 2005, the incidence of depression will reach 10% of the total population, and depression will rank second among the diseases that cause the heaviest social burden in the 21st century. Currently clinically used antidepressants mainly include tricyclics, 5-HT reuptake inhibitors, 5-HT-NE reuptake inhibitors, and monoamine oxidase-A inhibitors. Many, the above-mentioned drugs that only target a single link are often difficult to achieve satisfactory curative effect. At the same time, these drugs have defects such as narrow antidepressant spectrum, large side effects, and easy relapse. In recent years, more and more attention has been paid to traditional drugs in the research and development of antidepressants at home and abroad. However, the methods and related preparations for isolating effective parts and active ingredients of drugs with antidepressant activity from pure traditional Chinese medicine preparations are rare.

Contents of the invention

The purpose of the present invention is to provide an antidepressant pharmaceutical composition; the purpose of the present invention is also to provide a preparation method of the antidepressant pharmaceutical composition.

The pharmaceutical composition of the present invention mainly includes the following compounds, and each component and proportioning ratio are as follows (by weight):

Isoliquiritigenin 8--12 parts by weight Liquiritigenin 15--25 parts by weight

Liquiritin 25--35 parts by weight Neohesperidin 3200--3400 parts by weight

Hesperidin 70-90 parts by weight Isoliquiritin 2-6 parts by weight

5,3'-dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside 1--5 parts by weight 5,7-dihydroxycoumarin 3--8 parts by weight Saikosaponin c8-16 parts by weight Saikosaponin d12--18 parts by weight Glycyrrhizic acid 20--40 parts by weight

According to the pharmaceutical method, the pharmaceutical composition of the present invention can be prepared into various clinical pharmaceutical dosage forms, including oral preparations or parenteral administration dosage forms. Said oral preparations are selected from one of tablets, capsules, pills, granules, suspensions, drop pills, and oral liquid preparations; said parenteral dosage forms are selected from injections, aerosol One of the dosage forms for bolus, suppository or subcutaneous administration.

The medicine of the present invention can also add conventional pharmaceutical excipients, such as solvents, disintegrating agents, flavoring agents, preservatives, coloring agents and the like.

The preparation method of pharmaceutical composition of the present invention:

According to the ratio of 6:6:6:9, weigh Bupleurum, Radix Glycyrrhizae, Fructus Citrus Fructus Citrus Citrifolia, Radix Paeoniae Alba, mix the 4 ingredients evenly, add water to decoct 2-3 times, combine each decoction, concentrate, pass through weak Polar macroporous adsorption resin for adsorption, after the water decoction has passed through the resin column, continue to rinse the resin column with water until the washing liquid is nearly colorless, and then use 30-80% ethanol to remove the adsorbed substances on the resin column. Elution, collect the ethanol eluent, recover the solvent under reduced pressure to dryness, and obtain the antidepressant active part of the compound Chinese medicine; pass the antidepressant active part through a polyamide column, water, 25-35% ethanol aqueous solution, 45-55% ethanol Aqueous solution and 65-75% ethanol aqueous solution were eluted sequentially, and after the solvent was recovered from each part of the eluent under reduced pressure, water eluate A, 25-35% ethanol eluate B, and 45-55% ethanol eluate were obtained respectively. C, 65-75% ethanol eluate D;

Liquiritin can be prepared by one or more of the following methods:

C is separated by silica gel column chromatography, chloroform-methanol mixed solvent gradient elution, wherein the 80～85:20～15 chloroform-methanol eluting part is separated by silica gel column chromatography, chloroform-methanol ratio is 6～8:4～ 2 mixed solvent elution to obtain liquiritin;

After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After the solvent is recovered, the chloroform extraction fraction, the ethyl acetate extraction fraction, and the n-butanol extraction fraction are obtained; the ethyl acetate fraction is separated by silica gel column chromatography, and chloroform-methanol mixed solvent gradient elution, wherein 78～85:22～15 of chloroform - The methanol eluted part is separated by polyamide column chromatography, and chloroform-methanol is eluted with a 7-9:1 mixed solvent to obtain liquiritin;

The preparation method of isoliquiritigenin:

C is separated by silica gel column chromatography, chloroform-methanol mixed solvent gradient elution, wherein the 85-95:15-5 chloroform-methanol elution part is separated by polyamide column chromatography, chloroform-methanol ratio is 7-9:1 Elution with mixed solvents to obtain isoliquiritigenin;

The preparation method of isoliquiritin:

After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After recovering the solvent, the chloroform extraction fraction, ethyl acetate extraction fraction, and n-butanol extraction fraction were obtained; the ethyl acetate fraction was separated by silica gel column chromatography, and chloroform-methanol mixed solvent gradient elution, wherein 80-85:20-15 chloroform - The methanol eluted part is separated by polyamide column chromatography, and chloroform-methanol is eluted with a 7-9:1 mixed solvent to obtain isoliquiritin;

The preparation method of glycyrrhizin:

D was separated by silica gel column chromatography, eluted with chloroform-ethyl acetate mixed solvent of 6-9:1-3, and a total of 54 parts were collected; after the 9th to 11th parts were combined, they were separated by polyamide column chromatography, and 7- 9:1 chloroform-methanol mixed solvent was eluted to obtain Liquiritigenin;

The preparation of neohesperidin can adopt one or more of the following methods:

C is separated by silica gel column chromatography, chloroform-methanol mixed solvent gradient elution, wherein the 75-85:25-15 chloroform-methanol elution part is separated by silica gel column chromatography, chloroform-methanol is 80-90:20- 10 mixed solvents were eluted, and a total of 45 parts were collected; after the 22nd to 34th parts were combined, they were separated by silica gel column chromatography, and chloroform-methanol was eluted with a 6-9:1-3 mixed solvent to obtain neohesperidin;

After B is dispersed with water, extract 4 to 6 times with chloroform, 4 to 8 times with ethyl acetate, and 6 to 10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively recovered under reduced pressure After the solvent, the chloroform extraction fraction, the ethyl acetate extraction fraction, and the n-butanol extraction fraction were obtained; the ethyl acetate fraction was separated by silica gel column chromatography, and chloroform-methanol mixed solvent gradient elution, wherein 75-80:15-20 chloroform- The part eluted with methanol was recrystallized to obtain neohesperidin;

After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After recovering the solvent, the chloroform extraction fraction, the ethyl acetate extraction fraction and the n-butanol extraction fraction were obtained; the n-butanol fraction was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed solvent gradient, wherein 70-80:30-20 The chloroform-methanol eluted part was recrystallized to obtain neohesperidin;

After A is dispersed with water, it is extracted with n-butanol for 6-10 times, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient The elution was divided into 14 parts in turn; part 4 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 7-9:1 mixed solvent to obtain neohesperidin;

The preparation method of hesperidin:

After A is dispersed with water, it is extracted with n-butanol for 6-10 times, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient Elution, divided into 14 parts in turn, part 5 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 9-12:1 mixed solvent to obtain hesperidin;

The preparation method of 5,3'-dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside:

After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After recovering the solvent, the chloroform extraction fraction, the ethyl acetate extraction fraction, and the n-butanol extraction fraction were obtained; the ethyl acetate fraction was separated by silica gel column chromatography, and chloroform-methanol mixed gradient elution, wherein 80-90:20-10 chloroform- The methanol eluted part was separated by polyamide column chromatography, and chloroform-methanol was eluted with a 7-9:1 mixed solvent to obtain 5,3'-dihydroxy-4'-methoxydihydroflavone-7-O- β-glucoside;

The preparation method of 5,7-dihydroxycoumarin:

C is separated by silica gel column chromatography, chloroform-methanol mixed gradient elution, wherein the 85-90:15-10 chloroform-methanol elution part is separated by silica gel column chromatography, chloroform-ethyl acetate is 6-9:4 -1 mixed solvent was eluted to obtain 5,7-dihydroxycoumarin;

The preparation method of saikosaponin c, saikosaponin d:

After A is dispersed with water, it is extracted with n-butanol for 6-10 times, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient Elution, fractions 9-11 were combined, separated by Sephadex LH-20 column chromatography, eluted with 90-95% ethanol to obtain saikosaponin c and saikosaponin d;

The preparation method of glycyrrhizic acid:

After A is dispersed with water, it is extracted with n-butanol for 6-10 times, the n-butanol extract is decompressed and the solvent is recovered to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient Eluted, divided into 14 parts in turn; Part 14 was separated by Sephadex LH-20 column chromatography, eluted with 90-95% ethanol to obtain glycyrrhizic acid;

The medicinal composition has good antidepressant effect, obvious effect, wide antidepressant spectrum, little side effect and not easy to relapse.

Experimental Example 1: The physical and chemical constants of the above-mentioned 11 compounds were determined, and the structures were determined by spectroscopic methods.

Liquiritin white crystalline powder, mp 257～258℃, molecular formula C ₂₁ H ₂₂ O ₉ . UVλ _max nm: 276nm. ¹ HNMR (DMSO-d ₆ ) δppm: 10.58 (1H, S, 7-OH), 7.60 (1H, d, J = 8.7Hz, 5-H), 7.43 (2H, d, J = 8.7Hz, 2'6'-H), 7.05 (2H, d, J=8.7Hz, 3'5'-H), 6.50 (1H, dd, J=8.7, 2.1Hz, 6-H), 6.34 (1H, d, J =2.1Hz, 8-H), 5.52(1H, d, d, J=2.7, 12.8Hz, 2-H), 4.87(1H, d, J=6.9Hz, 1″-H), 3.47～3.08( sugarH), 2.66 (1H, dd, J=2.7, 19.8Hz, 3-Hcis). ¹³ CNMR (DMSO-d ₆ ) δppm: 78.7 (2-C), 43.2 (3-C), 190.0 (4-C ), 128.4(5-C), 110.6(6-C), 164.6(7-C), 102.6(8-C), 163.1(9-C), 113.5(10-C), 132.3(1′-C ), 128.0(2'-C), 116.1(3'-C), 157.4(4'-C), 116.1(5'-C), 128.0(6'-C), 100.3(1"-C), 73.2 (2″-C), 76.6 (3″-C), 69.7 (4″-C), 77.1 (5″-C), 60.7 (6″-C).

Isoliquiritigenin is orange-yellow granular substance, mp188～190℃(MeOH), molecular formula C ₁₅ H ₁₂ O ₄ . UVλ _max nm: 368nm. ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 13.60 (1H, S, 2′-OH), 10.67 (1H, S, 4′-OH), 10.12 (1H, S, 4-OH), 8.15 (1H , d, J = 8.7Hz, 6'-H), 7.74 (4H, m, 2, 6, α, β-H), 6.82 (2H, d, J = 9Hz, 3, 5-H), 6.39 ( 1H, dd, J = 2.1, 8.7 Hz, 5'-H), 6.26 (1H, d, J = 2.1 Hz, 3'-H). ¹³ CNMR (75MHzDMSO-d ₆ ) δppm: 191.5 (C=O), 117.4 (α-C), 144.3 (β-C), 125.7 (1-C), 131.2 (2-C), 115.8 (3-C ), 160.3(4-C), 115.8(5-C), 131.2(6-C), 113.0(1'-C), 165.8(2'-C), 102.6(3'-C), 164.9(4 '-C), 108.1 (5'-C), 132.9 (6'-C).

Isoliquiritin Yellow-green filamentous crystal, mp187～189℃ (MeOH), molecular formula C ₂₁ H ₂₂ O ₉ . UVλ _max nm: 359nm. ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 13.53 (1H, S, 2′-OH), 10.72 (1H, S, 4′-OH), 8.20 (1H, d, J=9.3Hz, 6′- C), 7.80 (4H, m, 2, 6, α, β-H), 7.09 (2H, d, J=9Hz, 3, 5-H), 6.39 (1H, dd, J=2.4, 9.3Hz, 5′-H), 6.26 (1H, d, J=2.4Hz, 3′-H), 4.97 (1H, d, 6.9Hz, 1″-H), 5.37 (1H, d, J=4.2Hz), 5.14 (1H, S), 5.07 (1H, d, J=5.7Hz), 4.61 (1H, S, OH peak on glucose), 3.34 (m) is H on sugar. ¹³ CNMR (75MHz, DMSO-d ₆ ) δppm: 191.6 (C=O), 119.1 (α-C), 143.6 (β-C), 128.4 (1-C), 130.9 (2-C), 116.5 (3-C), 159.5 (4- C), 116.5(5-C), 130.9(6-C), 113.0(1'-C), 165.2(2'-C), 102.6(3'-C), 165.9(4'-C), 108.2 (5'-C), 133.1(6'-C), 99.9(1"-C), 73.2(2"-C), 76.6(3"-C), 69.7(4"-C), 77.2(5 "-C), 60.7 (6"-C).

Liquiritigenin White needle-like clusters, mp207～208℃ (MeOH), molecular formula C ₁₅ H ₁₂ O ₄ . UVλ _max nm: 231nm, 275nm, 310nm. ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 10.56 (1H, S, 7-OH), 9.56 (1H, S, 4'-OH), 7.63 (1H, d, J=8.7Hz, 5-H) , 7.30 (2H, d, J=8.4Hz, 2′, 6′-H), 6.78 (2H, d, J=8.4Hz, 3′, 5′-H), 6.50 (1H, dd, J=8.7 , 2.4Hz, 6-H), 6.31 (1H, d, J = 2.4Hz, 8-H), 5.42 (1H, dd, J = 2.7, 12.0Hz, 2-H), 3.10 (1H, dd, J = 12.0, 17.0 Hz, 3-Htrans), 2.60 (1H, dd, J = 17.0, 2.7 Hz, 3-Hcis). ¹³ CNMR (DMSO-d ₆ ) δppm: 79.0 (2-C), 43.1 (3-C), 190.1 (4-C), 128.4 (5-C), 110.5 (6-C), 164.6 (7-C ), 102.5(8-C), 163.2(9-C), 113.5(10-C), 129.3(1'-C), 128.3(2'-C), 115.1(3'-C), 157.6(4 '-C), 115.1 (5'-C), 128.3 (6'-C).

Neohesperidin (Neohesperidin) white needle crystal, mp240～242℃ (MeOH), molecular formula C ₂₈ H ₃₄ O ₁₅ , HCl-Mg reaction is positive. UVλ _max nm: 284nm. ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 12.03 (1H, S, 5-OH), 9.13 (1H, S, 3′-OH), 6.91 (3H, m, 2′, 5′, 6′- H), 6.09 (1H, d, J=2Hz, 8-H), 6.07 (1H, d, J=2Hz, 6-H), 5.50 (1H, dd, J=2.7, 12.3Hz, 2-H) , 4.49 (1H, d, J=5.7Hz, 1-H), 3.76 (3H, S, OCH ₃ ), 3.72～3.14 (sugar H), 2.77 (1H, dd, J=17.4, 2.7Hz, 3 -Hcis), 1.14 (3H, d, J=6Hz, Rha-6- _CH3 ). ¹³ CNMR (DMSO-d ₆ ) δppm: 78.4 (2-C), 42.2 (3-C), 197.1 (4-C), 162.6 (5-C), 96.3 (6-C), 164.8 (7-C ), 95.1(8-C), 162.6(9-C), 103.3(10-C), 130.9(1'-C), 114.1(2'-C), 146.5(3'-C), 148.0(4 '-C), 112.0(5'-C), 117.8(6'-C), 100.4(1"-C), 76.9(2"-C), 76.1(3"-C), 69.6(4"- C), 77.1(5″-C), 60.4(6″-C), 97.3(1′″-C), 70.4(2-C), 70.5(3-C), 71.8(4-C ), 68.3 (5-C), 18.1 (6-C), 55.7 (OCH ₃ ).

Hesperidin (Hesperidin) is a white crystalline powder, mp258～260°C (MeOH), molecular formula C ₂₈ H ₃₄ O ₁₅ , HCl-Mg reaction is positive. UVλ _max nm: 284nm. ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 12.02 (1H, S, 5-OH), 9.11 (1H, S, 3'-OH), 6.94 (3H, m, 2', 5, '6'- H), 6.13 (2H, m, 6, 8-H), 5.56 (1H, dd, J=3.3, 12.3Hz, 2-H), 4.98 (1H, d, J=7.5Hz, 1″-H) , 4.52 (1H, S, 1-H), 3.78 (3H, S, OCH ₃ ), 3.63～3.27 (sugar H), 2.75 (1H, dd, J=17.1, 3.3Hz, 3-Hcis), 1.08 (3H, d, J=6.2Hz, Rha-6-CH ₃ ) ^.13 CNMR (DMSO-d ₆ )δppm: 79.2(2-C), 42.9(3-C), 197.9(4-C), 163.9 (5-C), 97.2(6-C), 166.0(7-C), 96.4(8-C), 163.4(9-C), 104.2(10-C), 131.7(1'-C), 115.0 (2'-C), 147.3(3'-C), 148.7(4'-C), 112.9(5'-C), 118.8(6'-C), 101.5(1"-C), 73.8(2 ″-C), 76.3 (3″-C), 70.4 (4″-C), 77.0 (5″-C), 66.9 (6″-C), 100.3 (1-C), 71.1 (2- C), 71.5 (3-C), 72.9 (4-C), 69.2 (5-C), 18.7 (6-C), 56.5 (OCH ₃ ).

5,3′-Dihydroxy-4′-methoxyflavanone-7-O-β-D-glucose White lumpy solid, molecular formula C ₂₂ H ₂₄ O ₁₁ . ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 12.04 (1H, S, 5-OH), 9.13 (1H, S, 3′-OH), 6.91 (3H, m, 2′, 5′, 6′- H), 6.12 (2H, m, 6, 8-H), 5.49 (1H, dd, J=3.3, 12Hz, 2-H), 4.95 (1H, d, J=7.2Hz, 1″-H), 3.76 (3H, S, OCH ₃ ), 3.72～3.12 (sugar H), 2.75 (1H, dd, J=17.1, 3.3Hz, 3-Hcis). ¹³ CNMR (DMSO-d ₆ ) δppm: 78.5 (2- C), 42.2(3-C), 197.1(4-C), 162.9(5-C), 96.5(6-C), 165.3(7-C), 95.5(8-C), 162.6(9-C ), 103.3(10-C), 130.9(1'-C), 114.2(2'-C), 146.5(3'-C), 148.0(4'-C), 112.0(5'-C), 118.0 (6'-C), 99.5(1"-C), 73.0(2"-C), 76.3(3"-C), 69.5(4"-C), 77.1(5"-C), 60.5(6 "-C), 55.7 ( _OCH3 ).

5,7-Dihydroxycoumarin White needle crystal, molecular formula C ₉ H ₆ O ₄ , strong sky blue fluorescence under 365nm ultraviolet light. UVλ _max nm: 331nm. ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 10.6 (1H, S, 5-OH), 10.4 (1H, S, 7-OH), 7.93 (1H, d, J=9.6Hz, 4-H), 6.24 (1H, d, J = 2.1 Hz, 8-H), 6.16 (1H, d, J = 2.1 Hz, 6-H), 6.02 (1H, d, J = 9.6 Hz, 3-H). ¹³ CNMR (DMSO-d ₆ ) δppm: 162.0 (2-C), 108.6 (3-C), 139.6 (4-C), 156.4 (5-C), 98.2 (6-C), 160.7 (7-C ), 94.0 (8-C), 155.9 (9-C), 101.6 (10-C).

Saikosaponin c white amorphous powder, mp204～209℃, molecular formula C ₄₈ H ₇₈ O ₁₈ . FAB-MS (m/z): 965 (M ⁺ +K), 439, 421. ¹ HNMR (300MHz, C ₅ D ₅ N) δppm: 5.96 (1H, d, J=11Hz, H-12), 5.63 (1H, dd, J=11Hz, 3Hz, H-11), 4.81 (1H, d , J=9Hz, 1'"-H), 4.62(1H, d, J=9Hz, 1''-H), 4.39(1H, d, J=8Hz, 1'-H), 1.65(3H, d , 6"-H), 1.35(3H,s), 1.28(3H,s), 1.14(3H,s), 0.96(3H,s), 0.95(3H,s), 0.91(3H,s), 0.85 (3H, s). ¹³ CNMR (C ₅ D ₅ N) δppm: 38.5 (1-C), 26.5 (2-C), 89.1 (3-C), 39.7 (4-C), 55.3 (5-C), 18.5 (6- C), 31.9(7-C), 42.2(8-C), 52.9(9-C), 36.3(10-C), 132.1(11-C), 131.2(12-C), 84.0(13-C ), 45.7(14-C), 36.1(15-C), 64.1(16-C), 47.0(17-C), 52.2(18-C), 37.8(19-C), 31.7(20-C) , 34.7(21-C), 25.8(22-C), 27.9(23-C), 16.4(24-C), 17.9(25-C), 20.0(26-C), 20.9(27-C), 73.0(28-C), 33.8(29-C), 23.9(30-C), 106.7(1'-C), 75.6(2'-C), 76.8(3'-C), 79.9(4'-C C), 75.2(5'-C), 69.1(6'-C), 103.0(1"-C), 72.7(2"-C), 72.6(3"-C), 73.9(4"-C) , 70.6(5″-C), 18.2(6″-C), 105.2(1’″-C), 74.8(2’″-C), 78.5(3’″-C), 71.4(4’″- C), 78.4 (5'''-C), 62.6 (6'''-C).

Saikosaponin d white amorphous powder, mp212~216℃, molecular formula C ₄₂ H ₆₈ O ₁₃ . FAB-MS (m/z): 819 (M ⁺ +K), 455. ¹ HNMR (300MHz, C ₅ D ₅ N) δppm: 6.27 (1H, br.s, OH), 6.03 (1H, d, J=11Hz, H-12), 5.69 (1H, dd, J=11Hz, 3Hz , H-11), 5.33 (1H, d, J=8Hz, 1″-H), 4.97 (1H, d, J=8Hz, 1′-H), 1.62 (3H, s), 1.43 (3H, s ), 1.42 (3H, s), 1.35 (3H, s), 1.02 (3H, s), 0.95 (3H, s), 0.93 (3H, s). ¹³ CNMR (C ₅ D ₅ N) δppm: 38.7 ( 1-C), 26.1(2-C), 81.7(3-C), 43.8(4-C), 47.4(5-C), 18.1(6-C), 31.9(7-C), 41.9(8 -C), 53.1(9-C), 36.3(10-C), 132.0(11-C), 132.0(12-C), 85.3(13-C), 43.6(14-C), 35.5(15-C C), 77.2(16-C), 45.4(17-C), 51.4(18-C), 38.4(19-C), 31.6(20-C), 36.8(21-C), 31.3(22-C ), 64.1(23-C), 13.1(24-C), 18.9(25-C), 19.6(26-C), 18.1(27-C), 77.8(28-C), 33.8(29-C) , 24.4(30-C), 106.9(1'-C), 71.6(2'-C), 84.9(3'-C), 72.2(4'-C), 71.1(5'-C), 17.3( 6'-C), 106.8(1"-C), 75.9(2"-C), 78.8(3"-C), 71.9(4''-C), 78.5(5"-C), 62.8(6 "-C).

Glycyrrhizic acid White crystalline powder, mp170～173℃ (EeOH-H ₂ O), molecular formula C ₄₂ H ₆₂ O ₁₆ . EI-MS (m/z): 470 (M-sugar), 438, 409, 372, 303, 262, 217, 175, 136, 95. ¹ HNMR (300MHz, DMSO-d ₆ ) δppm: 5.36 (1H, S, 12-H), 4.48 (1H, 1′-H), 4.22 (1H, 1″-H), 1.13 (3H, s), 1.12(3H,s), 1.02(3H,s), 0.94(3H,s), 0.86(3H,s), 0.67(3H,s), 0.63(3H,s) ^.13 CNMR(DMSO-d ₆ ) δppm: 38.4(1-C), 26.9(2-C), 89.7(3-C), 37.3(4-C), 55.4(5-C), 17.8(6-C), 33.1(7-C) , 44.0(8-C), 62.3(9-C), 37.3(10-C), 201.8(11-C), 128.0(12-C), 171.4(13-C), 46.1(14-C), 26.7(15-C), 26.5(16-C), 32.5(17-C), 49.2(18-C), 41.6(19-C), 44.0(20-C), 31.4(21-C), 38.4 (22-C), 28.8(23-C), 16.8(24-C), 17.1(25-C), 21.0(26-C), 23.8(27-C), 28.1(28-C), 29.3( 29-C), 179.9(30-C), 104.2(1'-C), 82.8(2'-C), 75.7(3'-C), 72.3(4'-C), 76.3(5'-C ), 171.8(6′-C), 105.1(1″-C), 75.5(2″-C), 76.7(3″-C), 72.1(4″-C), 76.7(5″-C), 172.5 (6″-C).

The following experimental examples adopt the drug screening method of the mouse forced swimming model, the mouse tail suspension model, the rat forced swimming model, and the rat chronic stress depression model. active. Experimental Example 2: Drug Screening Experiment of Mice Forced Swimming and Tail Suspension Model

1. Animals and grouping: 45 male Kunming tertiary mice, weighing 20-25 g, were raised in groups of 15. Divided into control group, Sini powder extract size dose group, a total of 6 groups. Administered from 8:00 to 10:00am every day for a total of 3 weeks. Administration was given 24h, 5h, and 1h before forced swimming and tail suspension tests.

2. Behavioral test method: forced swimming test: 10cm of water at 21-23°C was placed in the graduated cylinder, and an opaque plate was placed between the graduated cylinders to prevent the mice from seeing each other. One hour after the last dosing, the mice were put into the measuring cylinder and stayed for 6 minutes, and the immobility time of the last 4 minutes was calculated, and the four mice were measured at the same time. Criteria for judging immobility: stop struggling, stand vertically, float without moving, and only do some necessary activities to keep its head out of the water.

Tail-suspension test: 1 hour after the last drug injection, place a rod-shaped object horizontally, and fix it with adhesive tape at a distance of 30 mm from the tail tip of the mouse, and separate it with a plate in the middle to prevent each other from seeing each other. Mice were approximately 100 mm from the nearest object. Hang for 6 minutes, record the immobility time after 4 minutes, and measure 5 animals each time.

3. Results: The effective parts of large doses of Sini Powder significantly reduced the immobility time of mice forced to swim.

Table 1 Effects of drugs on the immobility time of the forced swimming model in mice

  Groups   Number of cases   Changes in immobility time compared with the blank group

(n) (s) (%)

Control 15 158.35±32.84 - Large dose of effective part of Sini powder 15 79.58±38.94 ^** -50 Small dose of effective part of Sini powder 15 132.78±46.85 -16

^* p<0.05 vs. control group, ^** p<0.01 vs. control group

      Table 2 Effects of Drugs on the Immobility Time of the Mouse Tail Suspension Model

  Groups   Number of Cases   Changes in immobility time compared with the blank group

(s) (%)

Blank 15 113.08±24.15 -High dose of effective part of Sini Powder 15 75.80±37.85 -32 Small dose of effective part of Sini Powder 15 91.71 ±30.07 - 9

^** p<0.01, compared with the control group

4 Conclusion:

The effective part of Sini powder has antidepressant effect. Experimental Example 3: The effect of the effective part of Sini Powder on the forced swimming model of rats

1. Animals and grouping: 30 male SD rats, weighing 180 g. They were randomly divided into 2 groups: 15 rats in the control group and 15 rats in the effective part of Sini powder (antidepressant active part) group. Animals in each group were housed in 3 cages, with 5 animals in each cage. Oral gavage was carried out at 8-9 hours every morning to make the animals adapt to the stimulation of scratching during administration.

2. Experimental method: intragastric administration for 12 days, and on the 13th day of the experiment, the rats were put into a container with a height of 40 cm and a diameter of 18 cm, which contained 20 cm of water at 25°C, forced to swim for 15 minutes, and dried after being taken out. On the 14th day, 23.5h, 5h, and 1h before the second forced swimming were administered respectively. On the 14th day of the experiment, the rats were forced to swim for the second time, and the immobility time of the rats was observed. After 60 minutes of forced swimming, 5 animals in the same cage were given intraperitoneal injection of 1% sodium pentobarbital (80 mg/kg) rapidly within 5 minutes, blood was collected from the abdominal aorta after anesthesia, centrifuged to obtain serum, and cortisol was measured . The thoracotomy was fixed by cardiac perfusion, and the blood was flushed with 100ml of normal saline, and then perfused with pre-cooled (4°C) 4% paraformaldehyde phosphate buffer (PB, 0.1mol/L, pH7.4). Irrigation, followed by slow irrigation. The brain was taken out, post-fixed with 4% paraformaldehyde for 2 hours, and then the brain tissue block was put into 20% sucrose phosphorus buffer (PB, 0.1mol/L, pH7.4) at 4°C overnight. After the tissue block completely sank to the bottom, 40 μm serial coronal sections were made with a frozen microtome, and the expression of C-FOS positive neurons was detected by immunohistochemistry.

3. Results:

3.1 Compared with the control group, the effective part of Sini powder can significantly shorten the immobility time of rats in forced swimming.

Table 3 The effect of the effective part of Sini powder on the forced swimming rat model

Group Number of cases Immobility time (s) Control group 14 138.77±32.90 Sini San effective part group 14 109.42±32.60 ^*

^* p<0.005, compared with control group

3.2 The effect on serum cortisol, FST has a significant effect on the serum cortisol content of rats, cortisol in the control group remains at a high level after FST 1 hour, and the prevention of the effective part of Sini powder group can significantly reduce its content.

Table 4 Effects of effective parts of Sini powder on serum cortisol in rat forced swimming model

Groups Number of Cases Serum Cortisol Concentration

(NG/ML) The control group 8 7.185 ± 2.607 Four inverse scattered effective parts 8 4.287 ± 0.987

3.3 Effects on the expression of C-FOS protein in brain neurons

Forced swimming can increase the expression of C-FOS positive neurons in the brain of rats, while the effective part of Sini Powder can reduce the expression of C-FOS positive neurons in the prefrontal cortex and hippocampal CA3 area of rats.

4 Conclusion:

It is confirmed that the effective part of Sini powder has antidepressant effect on the forced swimming model. Experimental Example 4: Effects of effective fractions of Sini Powder on chronic stress depression model in rats

1. Animals and methods: 18 SD rats, weighing 180-200 g. Grouping: Adaptive feeding for one week, during which sucrose water consumption and open field experiments were carried out, according to the results, they were divided into 3 groups semi-randomly. The control group had 6 rats per cage, and the model group was raised alone. On the 21st day, the animals were decapitated, and the brains of the rats were taken, and the hippocampus was isolated in an ice dish and weighed. Determination of changes in monoamine neurotransmitters and their metabolites by high-performance liquid-phase electrochemical detection.

2. Results: The content ratio of dopamine transmitters and their metabolites in the hippocampus of rats in the model group decreased, and the effective fraction of Sini Powder had an effect on dopamine metabolism, and could significantly increase the horizontal and vertical movements of the depression model rats.

Table 5 Ratio of hippocampal dopamine transmitter content

DA/DOPAC control group 6 6.65 ± 1.73 model group 6 2.49 ± 4.62 Four reverse scattered valid parts group 6 26.62 ± 5.67

Table 6 The behavioral effects of Sini powder and its effective parts on the depression model of rats

Grouping Number of Cases Horizontal Movement Vertical Movement

Model group 10 11.47±10.33 3.65±3.99 Sini powder effective part group 10 39.11±33.10 6.28±5.03

The following embodiments can all achieve the effects of the above experimental examples. Example 1

Isoliquiritigenin 10mg Liquiritinin 20mg Liquiritin 30mg

Neohesperidin 3305mg Hesperidin 80mg Isoliquiritin 10mg

Glycyrrhizic acid 30mg Saikosaponin d 13mg

5,3'-Dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside 2mg

5,7-dihydroxycoumarin 5mg saikosaponin c12mg made into 360 capsules, each containing 9.4mg (0.33g of the effective part) 2 times a day, 3 capsules each time. Example 2

Isoliquiritigenin 10mg Liquiritinin 20mg Liquiritin 30mg

Neohesperidin 3305mg Hesperidin 80mg Isoliquiritin 10mg

Glycyrrhizic acid 30mg Saikosaponin d 13mg

5,3'-Dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside 2mg

5,7-dihydroxycoumarin 5mg saikosaponin c12mg made into 360 tablets, each

Tablets contain 9.4mg (0.33g of effective parts) and take 2 times a day, 3 tablets each time. Example 3

Isoliquiritigenin 10mg Liquiritinin 20mg Liquiritin 30mg

Neohesperidin 3305mg Hesperidin 80mg Isoliquiritin 10mg

Glycyrrhizic acid 30mg Saikosaponin d 13mg

5,3'-dihydroxy-4'-methoxydihydroflavone-7-O-β-D-glucoside 2mg 5,7-dihydroxycoumarin 5mg saikosaponin c12mg made into 360 drop pills, each Capsules contain 9.4mg (0.33g of effective parts) and take 2 times a day, 3 capsules each time. Example 4

Isoliquiritigenin 10mg Liquiritinin 20mg Liquiritin 30mg

Neohesperidin 3305mg Hesperidin 80mg Isoliquiritin 10mg

Glycyrrhizic acid 30mg Saikosaponin d 13mg

5,3'-dihydroxy-4'-methoxydihydroflavone-7-O-β-D-glucoside 2mg 5,7-dihydroxycoumarin 5mg saikosaponin c12mg made into suppositories 360 capsules, each Capsules contain 9.4mg (0.33g of effective parts) used twice a day, 3 capsules each time Example 5

Weigh 2.4kg of Bupleurum, 2.4kg of Radix Glycyrrhiza, 2.4kg of Citrus aurantium, and 3.6kg of Paeoniae Alba in proportion, mix them evenly, add water to decoct twice, and add 100L of water for the first time, decoct for 2 hours; add 80L of water for the second time , decocted for 1.5 hours; combined 2 decoctions, concentrated to 60L; through the weak polar AB-8 macroporous adsorption resin, the resin volume is 15L, for adsorption, the adsorption flow rate is 2L/h; After passing through the resin column, continue to rinse the resin column with 8 times the volume of the resin, about 120L of water until the washing liquid is nearly colorless, and then use 8 times the volume of the resin, about 120L of 50% ethanol to remove the adsorbed substances on the resin column. Elution, the elution flow rate is 2L/h; 50% ethanol eluate is collected, the solvent is recovered under reduced pressure to dryness, and 707.4 g of the effective antidepressant fraction of the present invention is obtained. Example 6

Weigh 2.4kg of Bupleurum bupleuri, 2.4kg of Zhigancao, 2.4kg of Citrus aurantii, 3.6kg of Radix Paeoniae Alba, 10.8kg of the 4 herbs in proportion, mix them evenly, add water to decoct 2 to 3 times, combine each decoction, and concentrate , through the weak polarity AB-8 type macroporous adsorption resin for adsorption, after the water decoction has passed through the resin column, continue to wash the resin column with water until the washing liquid is almost colorless, and then use 40-70% ethanol to The substance adsorbed on the resin column is eluted, the ethanol eluent is collected, and the solvent is recovered under reduced pressure to dryness, and the antidepressant active part of the compound Chinese medicine is obtained. Take 120g of the antidepressant active part of the present invention, pass through a polyamide column, elute with water, 30% ethanol aqueous solution, 50% ethanol aqueous solution and 70% ethanol aqueous solution successively, after each part of the eluent decompresses and reclaims the solvent, respectively obtains water-washed Eluate E55g, 30% ethanol eluate F40g, 50% ethanol eluate H10g, 70% ethanol eluate K5g.

Preparation of Liquiritin:

H was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed gradient of 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, and 70:30, respectively, 100 mL each, and a total of Collect 100 fractions, of which: the 55th to 63rd fractions eluted with 83:17 chloroform-methanol were separated by silica gel column chromatography, eluted with 7:3 mixed solvent of chloroform-methanol to obtain 5 mg of liquiritin;

After F was dispersed by adding 500mL of water, it was extracted 5 times with chloroform successively, each time with 200mL of chloroform, a total of 1000mL; ethyl acetate was extracted 6 times, each time with 250mL of ethyl acetate, a total of 1500mL; n-butanol was extracted 8 times, each time with 250 mL of n-butanol, a total of 2000 mL; chloroform extract, ethyl acetate extract, and n-butanol extract were recovered under reduced pressure to obtain 0.5 g of chloroform extract, 3.65 g of ethyl acetate extract, and 7.0 g of n-butanol extract g; the aqueous solution after n-butanol extraction was dried under reduced pressure to obtain 0.55 g of the water part; 3.65 g of the ethyl acetate part was separated by silica gel column chromatography, and the ratio of chloroform-methanol was 100:0, 98:2, 95:5, 90: 10, 87:13, 85:15, 82:18, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed gradient elution, each 100mL, A total of 240 fractions were collected, among which: the 98th to 126th fraction eluted with 82:18 chloroform-methanol was separated by polyamide column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain 15 mg of glucoside;

After F was dispersed by adding 500mL of water, it was extracted 5 times with chloroform successively, each time with 200mL of chloroform, a total of 1000mL; ethyl acetate was extracted 6 times, each time with 250mL of ethyl acetate, a total of 1500mL; n-butanol was extracted 8 times, each time with 250 mL of n-butanol, a total of 2000 mL; chloroform extract, ethyl acetate extract, and n-butanol extract were recovered under reduced pressure to obtain 0.5 g of chloroform extract, 3.65 g of ethyl acetate extract, and 7.0 g of n-butanol extract g; the aqueous solution after n-butanol extraction was dried under reduced pressure to obtain 0.55 g of the water part; 7.0 g of the n-butanol part was separated by silica gel column chromatography, and the ratio of chloroform-methanol was 90:10, 87:13, 85:15, 83: 17, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed solvent gradient elution, each 100mL, a total of 160 parts were collected, of which: 87:13 The 62nd to 82nd parts of the chloroform-methanol elution fraction were separated by polyamide column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain 10 mg of liquiritin;

The preparation method of isoliquiritigenin:

H was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed gradient of 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, and 70:30, respectively, 100 mL each, and a total of Collect 100 copies, of which:

The 90:10 chloroform-methanol eluted fractions 9-14 were separated by polyamide column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain 10 mg of isoliquiritigenin;

Preparation of isoliquiritin:

After F was dispersed by adding 500mL of water, it was extracted 5 times with chloroform successively, each time with 200mL of chloroform, a total of 1000mL; ethyl acetate was extracted 6 times, each time with 250mL of ethyl acetate, a total of 1500mL; n-butanol was extracted 8 times, each time with 250 mL of n-butanol, a total of 2000 mL; chloroform extract, ethyl acetate extract, and n-butanol extract were decompressed and recovered solvent respectively to obtain 0.5 g of chloroform extract, 3.65 g of ethyl acetate extract, and 7.0 g of n-butanol extract g; the aqueous solution after n-butanol extraction was dried under reduced pressure to obtain 0.55 g of the water part; 3.65 g of the ethyl acetate part was separated by silica gel column chromatography, and the ratio of chloroform-methanol was 100:0, 98:2, 95:5, 90: 10, 87:13, 85:15, 82:18, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed gradient elution, 100 mL each, A total of 240 fractions were collected, among which: the 98th to 126th fractions eluted with 82:18 chloroform-methanol were separated by polyamide column chromatography and eluted with 9:1 mixed solvent of chloroform-methanol to obtain 3 mg of isoliquiritin;

Licorice preparation:

K was separated by silica gel column chromatography, and 8:2 chloroform-ethyl acetate mixed solvent was eluted, each part was 40 mL, and a total of 54 parts were collected; after the 9th to 11th parts were combined, they were separated by polyamide column chromatography, 9: 1 in chloroform-methanol mixed solvent to obtain 15 mg of liquiritigenin;

Preparation of neohesperidin:

H was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed gradient of 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, and 70:30, respectively, 100 mL each, and a total of Collect 100 fractions, of which: the 75th to 85th fractions eluted with 80:20 chloroform-methanol were separated by silica gel column chromatography, eluted with 85:15 mixed solvent of chloroform-methanol, each fraction was 10 mL, and a total of 45 fractions were collected; After the 22nd to 34th fractions were combined, they were separated by silica gel column chromatography and eluted with a 8:2 mixed solvent of chloroform-methanol to obtain 5 mg of neohesperidin;

After F was dispersed by adding 500mL of water, it was extracted 5 times with chloroform successively, each time with 200mL of chloroform, a total of 1000mL; ethyl acetate was extracted 6 times, each time with 250mL of ethyl acetate, a total of 1500mL; n-butanol was extracted 8 times, each time with 250 mL of n-butanol, a total of 2000 mL; chloroform extract, ethyl acetate extract, and n-butanol extract were recovered under reduced pressure to obtain 0.5 g of chloroform extract, 3.65 g of ethyl acetate extract, and 7.0 g of n-butanol extract g; the aqueous solution after n-butanol extraction was dried under reduced pressure to obtain 0.55 g of the water part; 3.65 g of the ethyl acetate part was separated by silica gel column chromatography, and the ratio of chloroform-methanol was 100:0, 98:2, 95:5, 90: 10, 87:13, 85:15, 82:18, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed gradient elution, each 100mL, A total of 240 fractions were collected, among which: the 183rd to 205th fractions eluted with 78:22 chloroform-methanol were recrystallized to obtain 400 mg of neohesperidin;

After F was dispersed by adding 500mL of water, it was extracted 5 times with chloroform successively, each time with 200mL of chloroform, a total of 1000mL; ethyl acetate was extracted 6 times, each time with 250mL of ethyl acetate, a total of 1500mL; n-butanol was extracted 8 times, each time with 250 mL of n-butanol, a total of 2000 mL; chloroform extract, ethyl acetate extract, and n-butanol extract were recovered under reduced pressure to obtain 0.5 g of chloroform extract, 3.65 g of ethyl acetate extract, and 7.0 g of n-butanol extract g; the aqueous solution after n-butanol extraction was dried under reduced pressure to obtain 0.55 g of the water part; 7.0 g of the n-butanol part was separated by silica gel column chromatography, and chloroform-methanol (90:10, 87:13, 85:15, 83: 17, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50) mixed solvent gradient elution, each 100mL, a total of 160 parts were collected. Among them: chloroform-methanol (75:25) eluted part (115th to 135th part) was recrystallized to obtain 900 mg of neohesperidin;

After F was dispersed by adding 500mL of water, it was extracted 5 times with chloroform successively, each time with 200mL of chloroform, a total of 1000mL; ethyl acetate was extracted 6 times, each time with 250mL of ethyl acetate, a total of 1500mL; n-butanol was extracted 8 times, each time with 250 mL of n-butanol, a total of 2000 mL; chloroform extract, ethyl acetate extract, and n-butanol extract were recovered under reduced pressure to obtain 0.5 g of chloroform extract, 3.65 g of ethyl acetate extract, and 7.0 g of n-butanol extract g; the aqueous solution after n-butanol extraction was dried under reduced pressure to obtain 0.55 g of the water part; 7.0 g of the n-butanol part was separated by silica gel column chromatography, and the ratio of chloroform-methanol was 90:10, 87:13, 85:15, 83: 17, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed solvent gradient elution, each 100mL, a total of 160 parts were collected, of which: E was dispersed with 600mL of water Finally, extract with n-butanol 8 times with 300 mL of n-butanol each time, a total of 2400 mL. After the n-butanol extract is decompressed and recovers the solvent, 7.55 g of the n-butanol extract is obtained. After the n-butanol extracted aqueous solution is dried under reduced pressure 6.45 g of the water part; 7.55 g of the n-butanol part were separated by silica gel column chromatography, and the ratio of chloroform-methanol-water was 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5:0.1, 80 :5:0.1, 75:5:0.1, 70:5:0.1, 65:5:0.1, 60:5:0.1, 55:5:0.1, 50:5:0.1, 40:5:0.1, 35:5 : 0.1, 30: 5: 0.1 mixed gradient elution, divided into 14 parts in turn; Part 4 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 9: 1 mixed solvent to obtain 2 g of neohesperidin;

Preparation of hesperidin:

After adding 600mL of water to disperse, extract with n-butanol 8 times with 300mL of n-butanol each time, totaling 2400mL, and recover the solvent from the n-butanol extract under reduced pressure to obtain 7.55g of n-butanol extract, the aqueous solution after n-butanol extraction After drying under reduced pressure, 6.45 g of the water part was obtained; 7.55 g of the n-butanol part were separated by silica gel column chromatography, and the ratio of chloroform-methanol-water was 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5 :0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5:0.1, 60:5:0.1, 55:5:0.1, 50:5:0.1, 40:5:0.1 , 35:5:0.1, 30:5:0.1 mixed gradient elution, divided into 14 parts in turn; Part 4 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain neohesperidin 2 g; Part 5 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 11:1 mixed solvent to obtain 80 mg of hesperidin;

Preparation of 5,3'-dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside:

After F was dispersed by adding 500mL of water, it was extracted 5 times with chloroform successively, each time with 200mL of chloroform, a total of 1000mL; ethyl acetate was extracted 6 times, each time with 250mL of ethyl acetate, a total of 1500mL; n-butanol was extracted 8 times, each time with 250 mL of n-butanol, a total of 2000 mL; chloroform extract, ethyl acetate extract, and n-butanol extract were recovered under reduced pressure to obtain 0.5 g of chloroform extract, 3.65 g of ethyl acetate extract, and 7.0 g of n-butanol extract g; the aqueous solution after n-butanol extraction was dried under reduced pressure to obtain 0.55 g of the water part; 3.65 g of the ethyl acetate part was separated by silica gel column chromatography, and the ratio of chloroform-methanol was 100:0, 98:2, 95:5, 90: 10, 87:13, 85:15, 82:18, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed gradient elution, each 100mL, A total of 240 fractions were collected, among which: the 92nd to 97th fractions eluted with 85:15 chloroform-methanol were separated by polyamide column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain 5,3'-di Hydroxy-4'-methoxyflavanone-7-O-β-glucoside 2mg;

Preparation of 5,7-dihydroxycoumarin:

H was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed gradient of 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, and 70:30, respectively, 100 mL each, and a total of Collect 100 parts, of which: 88:12 chloroform-methanol elution fractions 19-22 were separated by silica gel column chromatography, chloroform-ethyl acetate was eluted with 8:2 mixed solvent to obtain 5,7-dihydroxy Coumarin 5mg;

Preparation of saikosaponin c:

After adding 600mL of water to disperse, extract with n-butanol 8 times with 300mL of n-butanol each time, totaling 2400mL, and recover the solvent from the n-butanol extract under reduced pressure to obtain 7.55g of n-butanol extract, the aqueous solution after n-butanol extraction After drying under reduced pressure, 6.45 g of the water part was obtained; 7.55 g of the n-butanol part were separated by silica gel column chromatography, and the ratio of chloroform-methanol-water was 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5 :0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5:0.1, 60:5:0.1, 55:5:0.1, 50:5:0.1, 40:5:0.1 , 35:5:0.1, 30:5:0.1 mixed gradient elution, parts 9 to 11 were combined, separated by Sephadex LH-20 column chromatography, eluted with 95% ethanol to obtain saikosaponin c12mg;

Preparation of saikosaponin d:

After adding 600mL of water to disperse, extract with n-butanol 8 times with 300mL of n-butanol each time, totaling 2400mL, and recover the solvent from the n-butanol extract under reduced pressure to obtain 7.55g of n-butanol extract, the aqueous solution after n-butanol extraction After drying under reduced pressure, 6.45 g of the water part was obtained; 7.55 g of the n-butanol part were separated by silica gel column chromatography, and the ratio of chloroform-methanol-water was 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5 :0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5:0.1, 60:5:0.1, 55:5:0.1, 50:5:0.1, 40:5:0.1 , 35:5:0.1, 30:5:0.1 mixed gradient elution, parts 9 to 11 were combined, separated by Sephadex LH-20 column chromatography, and eluted with 95% ethanol to obtain saikosaponin d 15 mg;

Preparation of Glycyrrhizic Acid:

After adding 600mL of water to disperse, extract with n-butanol 8 times with 300mL of n-butanol each time, totaling 2400mL, and recover the solvent from the n-butanol extract under reduced pressure to obtain 7.55g of n-butanol extract, the aqueous solution after n-butanol extraction After drying under reduced pressure, 6.45 g of the water part was obtained; 7.55 g of the n-butanol part were separated by silica gel column chromatography, and the ratio of chloroform-methanol-water was 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5 :0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5:0.1, 60:5:0.1, 55:5:0.1, 50:5:0.1, 40:5:0.1 , 35:5:0.1, 30:5:0.1 mixed gradient elution, divided into 14 parts in turn; Part 14 was separated by Sephadex LH-20 column chromatography, eluted with 95% ethanol, and 30 mg of glycyrrhizic acid was obtained;

Claims (11)

1. an antidepressant pharmaceutical composition, characterized in that the pharmaceutical composition is mainly composed of the following compounds: Isoliquiritigenin 8--12 parts by weight Liquiritigenin 15--25 parts by weight Liquiritin 25--35 parts by weight Neohesperidin 3200--3400 parts by weight Hesperidin 70-90 parts by weight Isoliquiritin 2-6 parts by weight 5,3'-dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside 1--5 parts by weight 5,7-Dihydroxycoumarin 3--8 parts by weight Saikosaponin c 8--16 parts by weight Saikosaponin d 12--18 parts by weight Glycyrrhizic acid 20--40 parts by weight 2. The pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition mainly consists of the following compounds: 10 parts by weight of isoliquiritigenin 20 parts by weight of liquiritigenin 30 parts by weight of liquiritin Neohesperidin 3305 parts by weight Hesperidin 80 parts by weight Isoliquiritin 10 parts by weight Glycyrrhizic acid 30 parts by weight Saikosaponin d 13 parts by weight 2 parts by weight of 5,3'-dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside 5,7-dihydroxycoumarin 5 parts by weight Saikosaponin c 12 parts by weight 3. The preparation method of each compound in the pharmaceutical composition of claim 1 or 2, characterized in that the method is: According to the ratio of 6:6:6:9, weigh Bupleurum, Radix Glycyrrhizae, Fructus Citrus Fructus Citrus Citrifolia, Radix Paeoniae Alba, mix the 4 ingredients evenly, add water to decoct 2-3 times, combine each decoction, concentrate, pass through weak Polar macroporous adsorption resin for adsorption, after the water decoction has passed through the resin column, continue to rinse the resin column with water until the washing liquid is nearly colorless, and then use 30-80% ethanol to remove the adsorbed substances on the resin column. Elution, collect the ethanol eluent, recover the solvent under reduced pressure to dryness, and obtain the antidepressant active part of the compound Chinese medicine; pass the antidepressant active part through a polyamide column, water, 25-35% ethanol aqueous solution, 45-55% ethanol Aqueous solution and 65-75% ethanol aqueous solution were eluted sequentially, and after the solvent was recovered from each part of the eluent under reduced pressure, water eluate A, 25-35% ethanol eluate B, and 45-55% ethanol eluate were obtained respectively. C, 65-75% ethanol eluate D, and then purified from eluates A, B, C, and D to obtain the pure products of each compound. 4. the preparation method of each compound in the pharmaceutical composition as claimed in claim 3 is characterized in that the method is: According to the ratio of 6:6:6:9, weigh Bupleurum, Radix Glycyrrhizae, Fructus Citrus Fructus Citrus Citrifolia, Radix Paeoniae Alba, mix the 4 ingredients evenly, add water to decoct 2-3 times, combine each decoction, concentrate, pass through weak Polar macroporous adsorption resin for adsorption, after the water decoction has passed through the resin column, continue to rinse the resin column with water until the washing liquid is nearly colorless, and then use 40-70% ethanol to remove the adsorbed substances on the resin column. Elution, collect the ethanol eluate, recover the solvent under reduced pressure to dryness, and obtain the antidepressant active site of the compound traditional Chinese medicine; pass the antidepressant active site through a polyamide column, water, 30% ethanol aqueous solution, 50% ethanol aqueous solution and 70% ethanol aqueous solution The aqueous ethanol solution was eluted sequentially, and after the solvent was recovered from each part of the eluent under reduced pressure, the water eluate A, the 30% ethanol eluate B, the 50% ethanol eluate C, and the 70% ethanol eluate D were respectively obtained. Then, the pure products of each compound were obtained through purification from the eluates A, B, C, and D respectively. 5. The preparation method according to claim 3 or 4, characterized in that the purification process of each compound comprises the following methods: Liquiritin can be used in one or more of the following ways: C is separated by silica gel column chromatography, chloroform-methanol mixed solvent gradient elution, wherein the 80～85:20～15 chloroform-methanol eluting part is separated by silica gel column chromatography, chloroform-methanol ratio is 6～8:4～ 2 mixed solvent elution to obtain liquiritin; After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After the solvent is recovered, the chloroform extraction fraction, the ethyl acetate extraction fraction, and the n-butanol extraction fraction are obtained; the ethyl acetate fraction is separated by silica gel column chromatography, and chloroform-methanol mixed solvent gradient elution, wherein 78～85:22～15 of chloroform - The methanol eluted part is separated by polyamide column chromatography, and chloroform-methanol is eluted with a 7-9:1 mixed solvent to obtain liquiritin; The preparation method of isoliquiritigenin: C is separated by silica gel column chromatography, chloroform-methanol mixed solvent gradient elution, wherein the 85-95:15-5 chloroform-methanol elution part is separated by polyamide column chromatography, chloroform-methanol ratio is 7-9:1 Elution with mixed solvents to obtain isoliquiritigenin; The preparation method of isoliquiritin: After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After recovering the solvent, the chloroform extraction fraction, ethyl acetate extraction fraction, and n-butanol extraction fraction were obtained; the ethyl acetate fraction was separated by silica gel column chromatography, and chloroform-methanol mixed solvent gradient elution, wherein 80-85:20-15 chloroform - The methanol eluted part is separated by polyamide column chromatography, and chloroform-methanol is eluted with a 7-9:1 mixed solvent to obtain isoliquiritin; The preparation method of glycyrrhizin: D was separated by silica gel column chromatography, eluted with chloroform-ethyl acetate mixed solvent of 6-9:1-3, and a total of 54 parts were collected; after the 9th to 11th parts were combined, they were separated by polyamide column chromatography, and 7- 9:1 chloroform-methanol mixed solvent was eluted to obtain Liquiritigenin; The preparation of neohesperidin can adopt one or more of the following methods: C is separated by silica gel column chromatography, chloroform-methanol mixed solvent gradient elution, wherein the 75-85:25-15 chloroform-methanol elution part is separated by silica gel column chromatography, chloroform-methanol is 80-90:20- 10 mixed solvents were eluted, and a total of 45 parts were collected; after the 22nd to 34th parts were combined, they were separated by silica gel column chromatography, and chloroform-methanol was eluted with a 6-9:1-3 mixed solvent to obtain neohesperidin; After B is dispersed with water, extract 4 to 6 times with chloroform, 4 to 8 times with ethyl acetate, and 6 to 10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively recovered under reduced pressure After the solvent, the chloroform extraction fraction, the ethyl acetate extraction fraction, and the n-butanol extraction fraction were obtained; the ethyl acetate fraction was separated by silica gel column chromatography, and chloroform-methanol mixed solvent gradient elution, wherein 75-80:15-20 chloroform- The part eluted with methanol was recrystallized to obtain neohesperidin; After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After recovering the solvent, the chloroform extraction fraction, the ethyl acetate extraction fraction and the n-butanol extraction fraction were obtained; the n-butanol fraction was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed solvent gradient, wherein 70-80:30-20 The chloroform-methanol eluted part was recrystallized to obtain neohesperidin; After A is dispersed with water, it is extracted with n-butanol for 6-10 times, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient The elution was divided into 14 parts in turn; part 4 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 7-9:1 mixed solvent to obtain neohesperidin; The preparation method of hesperidin: After A is dispersed with water, it is extracted with n-butanol for 6-10 times, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient Elution, divided into 14 parts in turn, part 5 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 9-12:1 mixed solvent to obtain hesperidin; The preparation method of 5,3-dihydroxy-4'-methoxyflavanone-7-O-β-D-glucoside: After B is dispersed with water, extract 4-6 times with chloroform, 4-8 times with ethyl acetate, and 6-10 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively decompressed After recovering the solvent, the chloroform extraction fraction, the ethyl acetate extraction fraction, and the n-butanol extraction fraction were obtained; the ethyl acetate fraction was separated by silica gel column chromatography, and chloroform-methanol mixed gradient elution, wherein 80-90:20-10 chloroform- The fraction eluted with methanol was separated by polyamide column chromatography, and chloroform-methanol was eluted with a 7-9:1 mixed solvent to obtain 5,3'-dihydroxy-4'-methoxyflavanone-7-0- β-glucoside; The preparation method of 5,7-dihydroxycoumarin: C is separated by silica gel column chromatography, chloroform-methanol mixed gradient elution, wherein the 85-90:15-10 chloroform-methanol elution part is separated by silica gel column chromatography, chloroform-ethyl acetate is 6-9:4 -1 mixed solvent was eluted to obtain 5,7-dihydroxycoumarin; The preparation method of saikosaponin c, saikosaponin d: After A is dispersed with water, it is extracted with n-butanol for 6-10 times, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient Elution, fractions 9-11 were combined, separated by Sephadex LH-20 column chromatography, eluted with 90-95% ethanol to obtain saikosaponin c and saikosaponin d; The preparation method of glycyrrhizic acid: After A is dispersed with water, it is extracted with n-butanol for 6-10 times, the n-butanol extract is decompressed and the solvent is recovered to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and the chloroform-methanol-water mixed gradient Eluted, divided into 14 parts in turn; Part 14 was separated by Sephadex LH-20 column chromatography, eluted with 90-95% ethanol to obtain glycyrrhizic acid; 6. The preparation method according to claim 5, characterized in that the purification process of each compound comprises the following methods: Liquiritin can be used in one or more of the following ways: C was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed gradient of 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, and 70:30, and a total of 100 parts were collected. Among them: 83:17 chloroform-methanol eluted fractions 55-63 were separated by silica gel column chromatography, chloroform-methanol was eluted with 7:3 mixed solvent to obtain liquiritin; After B was dispersed with water, it was extracted 5 times with chloroform, 6 times with ethyl acetate, and 8 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract were respectively decompressed to recover the solvent to obtain chloroform extraction Part, ethyl acetate extraction part, n-butanol extraction part; ethyl acetate part was separated by silica gel column chromatography, chloroform-methanol ratio 100:0, 98:2, 95:5, 90:10, 87:13, 85 : 15, 82: 18, 80: 20, 78: 22, 75: 25, 70: 30, 65: 35, 60: 40, 50: 50 mixed gradient elution, a total of 240 parts were collected, of which: 82: 18 The 98th to 126th part of the chloroform-methanol elution fraction was separated by polyamide column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain liquiritin; After B was dispersed with water, it was extracted 5 times with chloroform, 6 times with ethyl acetate, and 8 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract were respectively decompressed to recover the solvent to obtain chloroform extraction Part, ethyl acetate extraction part, n-butanol extraction part; n-butanol part was separated by silica gel column chromatography, chloroform-methanol ratio 90:10, 87:13, 85:15, 83:17, 80:20, 78 : 22, 75: 25, 70: 30, 65: 35, 60: 40, 50: 50 mixed solvent gradient elution, a total of 160 parts were collected, of which: 87: 13 chloroform-methanol elution fraction 62nd to 82nd , separated by polyamide column chromatography and eluted with a 9:1 mixed solvent of chloroform-methanol to obtain liquiritin. Isoliquiritigenin can be used in the following ways: C was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed gradient of 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, and 70:30, and a total of 100 parts were collected. Among them: the 90:10 chloroform-methanol eluted fractions 9-14 were separated by polyamide column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain isoliquiritigenin. The following methods can be used for isoliquiritin: After B is dispersed with water, it is extracted 5 times with chloroform, 6 times with ethyl acetate, and 8 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract are respectively reduced After recovering the solvent, the chloroform extraction fraction, the ethyl acetate extraction fraction, and the n-butanol extraction fraction were obtained; the ethyl acetate fraction was separated by silica gel column chromatography, and the chloroform-methanol ratio was 100:0, 98:2, 95:5, 90: 10, 87:13, 85:15, 82:18, 80:20, 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed gradient elution, a total of 240 parts were collected , wherein: 82:18 chloroform-methanol eluted fractions 98-126 were separated by polyamide column chromatography, and chloroform-methanol was eluted with a 9:1 mixed solvent to obtain isoliquiritin. Liquiritigenin can be used in the following method: D is separated by silica gel column chromatography, and 8:2 chloroform-ethyl acetate mixed solvent is eluted, and a total of 54 parts are collected; the 9th to 11th parts are combined and separated by polyamide column chromatography. The 9:1 chloroform-methanol mixed solvent was eluted to obtain liquiritigenin. Neohesperidin can be used in one or more of the following ways: C was separated by silica gel column chromatography, and chloroform-methanol was eluted with a mixed gradient of 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, and 70:30, and a total of 100 parts were collected. Among them: the 80:20 chloroform-methanol elution fraction, the 75th to 85th fraction, was separated by silica gel column chromatography, and the chloroform-methanol was eluted with a 85:15 mixed solvent, and a total of 45 fractions were collected; after the 22nd to 34th fractions were combined, Separation by silica gel column chromatography, elution with chloroform-methanol mixed solvent of 8:2, to obtain neohesperidin; After B was dispersed with water, it was extracted 5 times with chloroform, 6 times with ethyl acetate, and 8 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract were respectively decompressed to recover the solvent to obtain chloroform extraction Part, ethyl acetate extraction part, n-butanol extraction part; ethyl acetate part was separated by silica gel column chromatography, chloroform-methanol ratio 100:0, 98:2, 95:5, 90:10, 87:13, 85 : 15, 82: 18, 80: 20, 78: 22, 75: 25, 70: 30, 65: 35, 60: 40, 50: 50 mixed gradient elution, a total of 240 parts were collected, of which: 78: 22 The 183rd to 205th fractions eluted with chloroform-methanol were recrystallized to obtain neohesperidin; After B was dispersed with water, it was extracted 5 times with chloroform, 6 times with ethyl acetate, and 8 times with n-butanol; the chloroform extract, ethyl acetate extract, and n-butanol extract were respectively decompressed to recover the solvent to obtain chloroform extraction Part, ethyl acetate extraction part, n-butanol extraction part; n-butanol part 7.0g was separated by silica gel column chromatography, chloroform-methanol ratio 90:10, 87:13, 85:15, 83:17, 80:20 , 78:22, 75:25, 70:30, 65:35, 60:40, 50:50 mixed solvent gradient elution, a total of 160 parts were collected, of which: the 75:25 chloroform-methanol elution fraction No. 115～ 135 parts were recrystallized to obtain neohesperidin; After A is dispersed with water, it is extracted with n-butanol 8 times each time, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, and chloroform-methanol-water :2:0.1, 95:5:0.1, 90:5:0.1, 85:5:0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5:0.1, 60:5 : 0.1, 55: 5: 0.1, 50: 5: 0.1, 40: 5: 0.1, 35: 5: 0.1, 30: 5: 0.1 mixed gradient elution, which were divided into 14 parts in turn; part 4 was used for silica gel column Chromatographic separation, chloroform-methanol was eluted with a 9:1 mixed solvent to obtain neohesperidin. The following method can be used for hesperidin: After A is dispersed with water, it is extracted with n-butanol 8 times each time, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography. Chloroform-methanol-water at 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5:0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65: 5:0.1, 60:5:0.1, 55:5:0.1, 50:5:0.1, 40:5:0.1, 35:5:0.1, 30:5:0.1 mixed gradient elution, divided into 14 parts in turn , Part 5 was separated by silica gel column chromatography, and chloroform-methanol was eluted with a 11:1 mixed solvent to obtain hesperidin. 5,3'-dihydroxy-4'-methoxydihydroflavone-7-O-β-D-glucoside can be used in the following method: After B is dispersed with water, it is extracted 5 times with chloroform and 6 times with ethyl acetate , n-butanol extraction 8 times; chloroform extract, ethyl acetate extract, and n-butanol extract were decompressed and recovered solvent respectively to obtain chloroform extract, ethyl acetate extract, n-butanol extract; ethyl acetate part Separation by silica gel column chromatography, chloroform-methanol ratio 100:0, 98:2, 95:5, 90:10, 87:13, 85:15, 82:18, 80:20, 78:22, 75:25 , 70:30, 65:35, 60:40, 50:50 mixed gradient elution, a total of 240 parts were collected, of which: the 92nd to 97th part of the 85:15 chloroform-methanol elution fraction was subjected to polyamide column chromatography Separation, chloroform-methanol was eluted with a 9:1 mixed solvent to obtain 5,3'-dihydroxy-4'-methoxyflavanone-7-O-β-glucoside. 5,7-dihydroxycoumarin can be separated by the following method: C is separated by silica gel column chromatography, and chloroform-methanol is respectively 90:10, 88:12, 85:15, 83:17, 80:20, 75:25, 70:30 mixed gradient elution, a total of 100 parts were collected, among which: the 19th to 22nd parts eluted with 88:12 chloroform-methanol, separated by silica gel column chromatography, and washed with 8:2 mixed solvent of chloroform-ethyl acetate Off to get 5,7-dihydroxycoumarin. Saikosaponin c can be obtained by the following method: After A is dispersed with water, it is extracted 8 times with n-butanol, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, chloroform -methanol-water in 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5:0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5 : 0.1, 60: 5: 0.1, 55: 5: 0.1, 50: 5: 0.1, 40: 5: 0.1, 35: 5: 0.1, 30: 5: 0.1 mixed gradient elution, fractions 9 to 11 were combined, and the Sephadex LH-20 column chromatography separation, 95% ethanol elution, to obtain saikosaponin c. The following methods can be used for saikosaponin d: After A is dispersed with water, it is extracted 8 times with n-butanol, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, chloroform -methanol-water in 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5:0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5 : 0.1, 60: 5: 0.1, 55: 5: 0.1, 50: 5: 0.1, 40: 5: 0.1, 35: 5: 0.1, 30: 5: 0.1 mixed gradient elution, fractions 9 to 11 were combined, and the Sephadex LH-20 column chromatography and elution with 95% ethanol to obtain saikosaponin d. Glycyrrhizic acid can be obtained by the following method: After A is dispersed with water, it is extracted 8 times with n-butanol, and the n-butanol extract is decompressed to recover the solvent to obtain the n-butanol extraction part, and the n-butanol part is separated by silica gel column chromatography, chloroform-methanol - water at 98:2:0.1, 95:5:0.1, 90:5:0.1, 85:5:0.1, 80:5:0.1, 75:5:0.1, 70:5:0.1, 65:5:0.1 , 60:5:0.1, 55:5:0.1, 50:5:0.1, 40:5:0.1, 35:5:0.1, 30:5:0.1 mixed gradient elution, divided into 14 parts in turn; Separation by Sephadex LH-20 column chromatography and elution with 95% ethanol to obtain glycyrrhizic acid. 7. The use of the pharmaceutical composition according to claim 1 or 2 in the preparation of antidepressant drugs. 8. The application according to claim 6, characterized in that anti-depression refers to increasing the content ratio of dopamine transmitters and their metabolites or reducing the expression of C-FOS positive neurons in brain neurons. 9. The pharmaceutical composition according to claim 1 or 2, characterized in that pharmaceutically acceptable dressings or excipients can also be added. 10. The pharmaceutical composition according to claim 8, characterized in that the dressing or excipient refers to solvent, disintegrant, flavoring agent, preservative, coloring agent and the like. 11. The pharmaceutical composition according to claim 1 or 2, characterized in that it can be made into a clinically acceptable dosage form. 12. The pharmaceutical composition according to claim 11, characterized in that the dosage form is tablet, capsule, pill, granule, suspension, drop pill, oral liquid preparation injection, aerosol, suppository.