Anti-depression composition and preparation method thereof

Abstract

The invention provides an anti-depression composition and a preparation method thereof. The composition is prepared from the following components in parts by mass: 1 part of saikoside D, 2 to 6 parts of saikoside A, 20 to 25 parts of paeoniflorin, 6 to 9 parts of albiflorin, 12 to 15 parts of apiose liquiritin, 2 to 4 parts of apiose isoliquiritin and 4 to 6 parts of atractylenolide I. Compared with western medicine fluoxetine and classic famous formula Xiaoyao powder which are sold on the market under the clinical equivalent dosage condition in the depression cell and depression animal experiment dimension, cell experiments and animal experiments both show that the medicine effect of the anti-depression composition is far higher than the anti-depression effect of the western medicine fluoxetine, and meanwhile, the anti-depression composition has the advantages that the anti-depression effect of the anti-depression composition is greatly improved. The medicine effect is higher than the anti-depression effect of the whole prescription of the Xiaoyao powder

Description

Technical Field

[0001] The present invention belongs to the field of antidepressants, and in particular relates to an antidepressant composition and a preparation method thereof. Background Art

[0002] Depression is one of the most common mental disorders globally, with a relatively high incidence and prevalence rate. According to statistics from the World Health Organization, approximately 3.8% of the global population suffers from depression, and its incidence is on the rise year by year. It has become an important disease that seriously affects the physical and mental health of humans in modern society. In recent years, although great research progress has been made in the treatment of depression, during the clinical treatment process, there are still characteristics such as large side effects of treatment drugs and poor patient compliance, resulting in less than satisfactory clinical medication effects.

[0003] In the early stage of depression treatment, marketed western medicines are often used. However, due to the drug side effects and drug dependence manifested clinically, the western medicines have not achieved the expected treatment goals in the treatment of depression. Summary of the Invention

[0004] In view of this, the present invention aims to overcome the defects in the prior art and proposes an antidepressant composition and a preparation method thereof.

[0005] To achieve the above object, the technical solution of the present invention is realized as follows:

[0006] The present invention provides an antidepressant composition, which comprises the following components in mass fractions:

[0007] 1 part of saikosaponin D, 2 - 6 parts of saikosaponin A, 20 - 25 parts of paeoniflorin, 6 - 9 parts of albiflorin, 12 - 15 parts of apiosylglycyrrhizin, 2 - 4 parts of apiosylisoliquiritin, 4 - 6 parts of atractylenolide I.

[0008] Preferably, the composition comprises the following components in mass fractions:

[0009] 1 part of saikosaponin D, 3 - 5 parts of saikosaponin A, 21 - 23 parts of paeoniflorin, 7 - 9 parts of albiflorin, 12 - 14 parts of apiosylglycyrrhizin, 2 - 4 parts of apiosylisoliquiritin, 4 - 6 parts of atractylenolide I.

[0010] Preferably, the composition comprises the following components in mass fractions: 1 part of saikosaponin D, 4 parts of saikosaponin A, 23 parts of paeoniflorin, 8 parts of albiflorin, 13 parts of apiosylglycyrrhizin, 2 parts of apiosylisoliquiritin, 5 parts of atractylenolide I.

[0011] Furthermore, the purity of albiflorin is 55-65%; the purity of paeoniflorin is 70-75%; the purity of apiosylglycyrrhizin is 50-55%; the purity of apiosylisoliquiritigenin is 20-30%; the purity of saikosaponin A is 35-45%; the purity of saikosaponin D is 30-35%; the purity of atractylenolide I is 10-20%.

[0012] The present invention also provides a method for preparing an antidepressant composition, which comprises the following steps: adding apiosylisoliquiritigenin to saikosaponin D, mixing evenly, then adding saikosaponin A thereto, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding apiosylglycyrrhizin thereto, mixing evenly, and then adding paeoniflorin thereto, and mixing evenly to obtain the antidepressant composition. In the above method, the components in small doses are first mixed, and then the components in slightly larger doses are sequentially added and mixed. Compared with the common mixing of multiple components or the mixing of any two components in the conventional method, the method of the present invention enables the components to be fully mixed and improves the preparation efficiency at the same time.

[0013] Compared with the prior art, the present invention has the following advantages:

[0014] The antidepressant composition of the present invention is compared with the marketed western medicine antidepressant fluoxetine and the classical famous prescription Xiaoyao San under the condition of clinically equivalent doses in the dimensions of depression cell and depression animal experiments. Both the cell experiment and the animal experiment show that the antidepressant effect of the antidepressant composition is far higher than that of the western medicine fluoxetine, and at the same time, the antidepressant effect is higher than that of the whole prescription of Xiaoyao San. Description of the Drawings

[0015] Figure 1 It is the cell activity of the composition of Example 1 described in Experimental Example 1 of the present invention at different concentrations;

[0016] Figure 2 It is the pharmacodynamic activity (swimming experiment - depression behavioral test) of Example 1 described in Experimental Example 2 of the present invention on a depression animal model: among them, the positive drug is the western medicine fluoxetine, ZFL is the low dose of Example 1, ZFM is the medium dose of Example 1, ZFH is the high dose of Example 1, XYSL is the low-dose group of the whole prescription of Xiaoyao San, XYSM is the medium-dose group of the whole prescription of Xiaoyao San, and XYSH is the high-dose group of the whole prescription of Xiaoyao San;

[0017] Figure 3Pharmacodynamic activity of Example 1 described in Experimental Example 2 of the present invention on a depression animal model (tail suspension test - depression behavioral test): Among them, the positive drug is the western medicine fluoxetine, ZFL is the low dose of Example 1, ZFM is the medium dose of Example 1, ZFH is the high dose of Example 1, XYSL is the low-dose group of the whole formula of Xiaoyao San, XYSM is the medium-dose group of the whole formula of Xiaoyao San, and XYSH is the high-dose group of the whole formula of Xiaoyao San;

[0018] Figure 4 Pharmacodynamic activity of Example 1 described in Experimental Example 2 of the present invention on a depression animal model (food intake test - depression behavioral test): Among them, the positive drug is the western medicine fluoxetine, ZFL is the low dose of Example 1, ZFM is the medium dose of Example 1, ZFH is the high dose of Example 1, XYSL is the low-dose group of the whole formula of Xiaoyao San, XYSM is the medium-dose group of the whole formula of Xiaoyao San, and XYSH is the high-dose group of the whole formula of Xiaoyao San;

[0019] Figure 5 Pharmacodynamic activity of Example 1 described in Experimental Example 2 of the present invention on a depression animal model (determination of MAO-A content in the brain): Among them, the positive drug is the western medicine fluoxetine, ZFL is the low dose of Example 1, ZFM is the medium dose of Example 1, ZFH is the high dose of Example 1, XYSL is the low-dose group of the whole formula of Xiaoyao San, XYSM is the medium-dose group of the whole formula of Xiaoyao San, and XYSH is the high-dose group of the whole formula of Xiaoyao San;

[0020] Figure 6 Pharmacodynamic activity of Example 1 described in Experimental Example 2 of the present invention on a depression animal model (determination of 5-HT content in the brain): Among them, the positive drug is the western medicine fluoxetine, ZFL is the low dose of Example 1, ZFM is the medium dose of Example 1, ZFH is the high dose of Example 1, XYSL is the low-dose group of the whole formula of Xiaoyao San, XYSM is the medium-dose group of the whole formula of Xiaoyao San, and XYSH is the high-dose group of the whole formula of Xiaoyao San. Detailed implementation mode

[0021] Unless otherwise defined, the technical terms used in the following examples have the same meanings as commonly understood by those skilled in the art to which the present invention belongs. The test reagents used in the following examples are all conventional biochemical reagents unless otherwise specified; the experimental methods are all conventional methods unless otherwise specified.

[0022] The preparation methods of saikosaponin D described in the embodiments and comparative examples of the present invention include the following steps: First, put the bupleurum medicinal materials into water (the solid-liquid ratio is 1:8) and soak for 30 minutes. After heating to boiling, decoct for 30 minutes, and pour out the first filtrate for standby; then add water (the solid-liquid ratio is 1:8) to the medicinal materials and decoct. After boiling for 30 minutes, pour out the filtrate; finally, combine the two filtrates to obtain the bupleurum extract. The bupleurum extract is subjected to alcohol precipitation by the spray alcohol precipitation method. The volume ratio of the bupleurum extract to ethanol is 1:3 for alcohol precipitation, and finally the ethanol concentration in the extract is 80%. Finally, after the bupleurum alcohol precipitation solution stands for 24 hours, take the supernatant for rotary evaporation, and finally prepare the extract of bupleurum to be the sample to be measured. Next, the saikosaponin D sample components are prepared by the dynamic axial technology. The bupleurum extract is diluted with water (the solid-liquid ratio is 1:4), and rotary evaporation is carried out at 70 °C to dissolve the sample. Then, the solution sample is injected into a 1L system of Hanbang preparative liquid phase. Chromatographic column: DAC150 Kromasil C18(W), 10um. The mobile phase is A: 0.1% formic acid water, B: acetonitrile. The elution gradient reaches the range of A: 35-45%, B: 65-55%. Receive the fractions, and use HPLC (ultra-high performance liquid phase) technology to compare the collected liquid with the standard product. Confirm it as saikosaponin D, and then complete the preparation of saikosaponin D. The purity of the prepared saikosaponin D is measured by HPLC to be 30-35%.

[0023] The preparation methods of saikosaponin A described in the embodiments and comparative examples of the present invention include the following steps: First, put the bupleurum medicinal materials into water (the solid-liquid ratio is 1:8) and soak for 30 minutes. After heating to boiling, decoct for 30 minutes, and pour out the first filtrate for standby; then add water (the solid-liquid ratio is 1:8) to the medicinal materials and decoct. After boiling for 30 minutes, pour out the filtrate; combine the two filtrates to obtain the bupleurum extract; the bupleurum extract is subjected to alcohol precipitation by the spray alcohol precipitation method. The volume ratio of the bupleurum extract to ethanol is 1:3 for alcohol precipitation, so that the ethanol concentration in the extract is 80%; finally, after the bupleurum alcohol precipitation solution stands for 24 hours, take the supernatant for rotary evaporation, and finally prepare the extract of bupleurum to be the sample to be measured. Next, the saikosaponin A sample components are prepared by the dynamic axial technology. The bupleurum extract is diluted with water (the solid-liquid ratio is 1:4), and rotary evaporation is carried out at 70 °C to dissolve the sample. Then, the solution sample is injected into a 1L system of Hanbang preparative liquid phase. Chromatographic column: DAC150 Kromasil C18(W), 10um. The mobile phase is A: 0.1% formic acid water, B: acetonitrile. The elution gradient reaches the range of A: 45-50%, B: 55-50%. Receive the fractions, and use HPLC (ultra-high performance liquid phase) technology to compare the collected liquid with the standard product. Confirm it as saikosaponin A, and then complete the preparation of saikosaponin A. The purity of the prepared saikosaponin A is measured by HPLC to be 35-45%.

[0024] The preparation method of paeoniflorin described in each embodiment of the present invention and the comparative example includes the following steps: first, soak the white peony medicinal material in water (solid-liquid ratio of 1:8) for 30 minutes, heat and boil and then boil for 30 minutes, pour out the first filtrate for standby use; then add water (solid-liquid ratio of 1:8) to the medicinal material for decoction, boil for 30 minutes, pour out the filtrate; combine the two filtrates to obtain a white peony extract; the white peony extract is precipitated by spray alcohol precipitation, and the volume ratio of the white peony extract to ethanol is 1:3, so that the concentration of ethanol in the extract is 80%; finally, after the white peony alcohol precipitation is left to stand for 24 hours, the supernatant is taken for rotary evaporation, and finally the white peony extract sample to be tested is prepared for the preparation of the paeoniflorin sample component. Next, the dynamic axial technology is used to prepare the paeoniflorin sample component, the white peony extract is diluted with water (solid-liquid ratio of 1:4), and rotary evaporation is performed at 70°C to dissolve the sample. Then, the solution sample was injected into the Hanbang preparative liquid phase 1L system, the chromatographic column: DAC150Kromasil C18 (W), 10um, the mobile phase was A: 0.1% formic acid water, B: acetonitrile, the elution gradient reached A: 70-75%, B was 25-30%, the fraction was received, and the collected liquid was compared with the standard by HPLC (ultra-high performance liquid phase) technology to confirm that it was paeoniflorin, and then the preparation of paeoniflorin was completed. The purity of the prepared paeoniflorin was 70-75% as determined by HPLC.

[0025] The preparation method of paeoniflorin described in each embodiment of the present invention and the comparative example includes the following steps: first, soak the white peony medicinal material in water (solid-liquid ratio of 1:8) for 30 minutes, heat and boil for 30 minutes, and pour out the first filtrate for standby use; then add water (solid-liquid ratio of 1:8) to the medicinal material for decoction, and pour out the filtrate after boiling for 30 minutes; combine the two filtrates to obtain a white peony extract; the white peony extract is precipitated by spray alcohol precipitation, and the volume ratio of the white peony extract to ethanol is 1:3, so that the concentration of ethanol in the extract is 80%; finally, after the white peony alcohol precipitation is left to stand for 24 hours, the supernatant is taken for rotary evaporation, and finally the white peony extract sample to be tested is prepared for the preparation of paeoniflorin sample components. Next, the dynamic axial technology is used to prepare the paeoniflorin sample components, the white peony extract is diluted with water (solid-liquid ratio of 1:4), and rotary evaporation is performed at 70°C to dissolve the sample. Then, the solution sample was injected into the Hanbang preparative liquid phase 1L system, the chromatographic column: DAC150 Kromasil C18 (W), 10um, the mobile phase was A: 0.1% formic acid water, B: acetonitrile, the elution gradient reached A: 65-70%, B was 30-35%, the fraction was received, and the collected liquid was compared with the standard by HPLC (ultra-high performance liquid phase) technology to confirm that it was paeoniflorin, and then the preparation of paeoniflorin was completed. The purity of the prepared paeoniflorin was 55-65% as determined by HPLC.

[0026] The extraction methods of liquiritin described in the embodiments and comparative examples of the present invention include the following steps: First, put the licorice medicinal materials into water (the solid-liquid ratio is 1:8) and soak for 30 minutes, then boil and decoct for 30 minutes, and pour out the first filtrate for standby; then add water (the solid-liquid ratio is 1:8) to the medicinal materials and decoct, after boiling for 30 minutes, pour out the filtrate; combine the two filtrates to obtain the licorice extract; the licorice extract is subjected to alcohol precipitation by spray alcohol precipitation, and the volume ratio of the licorice extract to ethanol is 1:3 for alcohol precipitation, finally making the ethanol concentration in the extract 80%; finally, after the licorice alcohol precipitation solution stands for 24 hours, take the supernatant for rotary evaporation, and finally prepare the extract of licorice to be the sample to be tested. Next, the liquiritin sample components are prepared by the dynamic axial technique. The licorice extract is diluted with water (the solid-liquid ratio is 1:4) and rotary evaporated at 70°C to dissolve the sample. Then, the solution sample is injected into a 1L system of Hanbang preparative liquid phase, chromatographic column: DAC150 Kromasil C18(W), 10um, mobile phase A: 0.1% formic acid water, B: acetonitrile, the elution gradient reaches the range of A: 45-50%, B: 55-50%, collect the fractions, and compare the collected liquid with the standard product by HPLC (ultra-high performance liquid chromatography) technology to confirm it as liquiritin, and then complete the preparation of liquiritin. The purity of the prepared liquiritin is determined by HPLC to be 50-55%.

[0027] The preparation methods of isoliquiritin apioside in the embodiments and comparative examples of the present invention include the following steps: First, put the licorice medicinal materials into water (the solid-liquid ratio is 1:8) and soak for 30 minutes, boil by heating and then decoct for 30 minutes, pour out the first filtrate for standby; then add water (the solid-liquid ratio is 1:8) to the medicinal materials for decoction, after boiling for 30 minutes, pour out the filtrate; combine the two filtrates to obtain the licorice extract; the licorice extract is subjected to alcohol precipitation by the spray alcohol precipitation method, and the volume ratio of the licorice extract to ethanol is 1:3 for alcohol precipitation, finally making the ethanol concentration in the extract 80%; finally, after the licorice alcohol precipitation solution stands for 24 hours, take the supernatant for rotary evaporation, and finally prepare the extract of licorice as the sample to be measured. Next, use the dynamic axial technology to prepare the isoliquiritin apioside sample components, dilute the licorice extract with water (the solid-liquid ratio is 1:4), and perform rotary evaporation at 70°C to dissolve the sample. Then, inject the solution sample into the Hanbang preparative liquid phase 1L system, chromatographic column: DAC150 Kromasil C18(W), 10um, mobile phase A: 0.1% formic acid water, B: acetonitrile, the elution gradient reaches the range of A: 40 - 45%, B: 60 - 55%, collect the fractions, and use HPLC (ultra-high performance liquid phase) technology to compare the collected liquid with the standard product, confirm it as isoliquiritin apioside, and thus complete the preparation of isoliquiritin apioside. The purity of the prepared isoliquiritin apioside is measured by HPLC to be 20 - 30%.

[0028] The preparation methods of atractylenolide I in the embodiments and comparative examples of the present invention include the following steps: First, put the atractylodes rhizome medicinal materials into water (the solid-liquid ratio is 1:8) and soak for 30 minutes, boil by heating and then decoct for 30 minutes, pour out the first filtrate for standby; then add water (the solid-liquid ratio is 1:8) to the medicinal materials for decoction, after boiling for 30 minutes, pour out the filtrate; combine the two filtrates to obtain the atractylodes rhizome extract; the atractylodes rhizome extract is subjected to alcohol precipitation by the spray alcohol precipitation method, and the volume ratio of the atractylodes rhizome extract to ethanol is 1:3 for alcohol precipitation, finally making the ethanol concentration in the extract 80%; finally, after the atractylodes rhizome alcohol precipitation solution stands for 24 hours, take the supernatant for rotary evaporation, and finally prepare the extract of atractylodes rhizome as the sample to be measured. Next, use the dynamic axial technology to prepare the atractylenolide I sample components, dilute the atractylodes rhizome extract with water (the solid-liquid ratio is 1:4), and perform rotary evaporation at 70°C to dissolve the sample. Then, inject the solution sample into the Hanbang preparative liquid phase 1L system, chromatographic column: DAC150 Kromasil C18(W), 10um, mobile phase A: 0.1% formic acid water, B: acetonitrile, the elution gradient reaches the range of A: 35 - 40%, B: 65 - 60%, collect the fractions, and use HPLC (ultra-high performance liquid phase) technology to compare the collected liquid with the standard product, confirm it as atractylenolide I, and thus complete the preparation of atractylenolide I. The purity of the prepared atractylenolide I is measured by HPLC to be 10 - 20%.

[0029] The present invention will be described in detail below in conjunction with embodiments.

[0030] Example 1

[0031] An antidepressant composition, the composition comprising the following components by mass fraction: 1 part of saikosaponin D, 4 parts of saikosaponin A, 23 parts of paeoniflorin, 8 parts of albiflorin, 13 parts of apiosylglycyrrhizin, 2 parts of apiosylisoliquiritin, and 5 parts of atractylenolide I.

[0032] The preparation method of the antidepressant composition comprises the following steps: adding apiosylisoliquiritin to saikosaponin D, mixing evenly, then adding saikosaponin A thereto, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding apiosylglycyrrhizin thereto, mixing evenly, and then adding paeoniflorin thereto, and mixing evenly to obtain the antidepressant composition.

[0033] Example 2

[0034] An antidepressant composition, the composition comprising the following components by mass fraction: 1 part of saikosaponin D, 5 parts of saikosaponin A, 23 parts of paeoniflorin, 8 parts of albiflorin, 13 parts of apiosylglycyrrhizin, 2 parts of apiosylisoliquiritin, and 5 parts of atractylenolide I.

[0035] The preparation method of the antidepressant composition comprises the following steps: adding apiosylisoliquiritin to saikosaponin D, mixing evenly, then adding saikosaponin A thereto, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding apiosylglycyrrhizin thereto, mixing evenly, and then adding paeoniflorin thereto, and mixing evenly to obtain the antidepressant composition.

[0036] Example 3

[0037] An antidepressant composition, the composition comprising the following components by mass fraction: 1 part of saikosaponin D, 4 parts of saikosaponin A, 23 parts of paeoniflorin, 5 parts of albiflorin, 13 parts of apiosylglycyrrhizin, 2 parts of apiosylisoliquiritin, and 5 parts of atractylenolide I.

[0038] The preparation method of the antidepressant composition comprises the following steps: adding apiosylisoliquiritin to saikosaponin D, mixing evenly, then adding saikosaponin A thereto, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding apiosylglycyrrhizin thereto, mixing evenly, and then adding paeoniflorin thereto, and mixing evenly to obtain the antidepressant composition.

[0039] Comparative Example 1 (The ratio of saikosaponin A is outside the protection scope)

[0040] An antidepressant composition, which comprises the following components in mass fractions: Saikosaponin D 1 part, Saikosaponin A 10 parts, Paeoniflorin 23 parts, Albiflorin 8 parts, Glucosylglycyrrhizin 13 parts, Isoglucosylglycyrrhizin 2 parts, Atractylenolide I 5 parts.

[0041] The preparation method of the antidepressant composition comprises the following steps: adding Isoglucosylglycyrrhizin to Saikosaponin D, uniformly mixing, then adding Saikosaponin A thereto, uniformly mixing, then adding Atractylenolide I thereto, uniformly mixing, then adding Albiflorin thereto, uniformly mixing, then adding Glucosylglycyrrhizin thereto, uniformly mixing, and then adding Paeoniflorin thereto, and uniformly mixing to obtain the antidepressant composition.

[0042] Comparative Example 2 (the ratio of Paeoniflorin is outside the protection range)

[0043] An antidepressant composition, which comprises the following components in mass fractions: Saikosaponin D 1 part, Saikosaponin A 4 parts, Paeoniflorin 25 parts, Albiflorin 8 parts, Glucosylglycyrrhizin 13 parts, Isoglucosylglycyrrhizin 2 parts, Atractylenolide I 5 parts.

[0044] The preparation method of the antidepressant composition comprises the following steps: adding Isoglucosylglycyrrhizin to Saikosaponin D, uniformly mixing, then adding Saikosaponin A thereto, uniformly mixing, then adding Atractylenolide I thereto, uniformly mixing, then adding Albiflorin thereto, uniformly mixing, then adding Glucosylglycyrrhizin thereto, uniformly mixing, and then adding Paeoniflorin thereto, and uniformly mixing to obtain the antidepressant composition.

[0045] Comparative Example 3 (without Saikosaponin A)

[0046] An antidepressant composition, which comprises the following components in mass fractions: Saikosaponin D 1 part, Paeoniflorin 23 parts, Albiflorin 8 parts, Glucosylglycyrrhizin 13 parts, Isoglucosylglycyrrhizin 2 parts, Atractylenolide I 5 parts.

[0047] The preparation method of the antidepressant composition comprises the following steps: adding Isoglucosylglycyrrhizin to Saikosaponin D, uniformly mixing, then adding Atractylenolide I thereto, uniformly mixing, then adding Albiflorin thereto, uniformly mixing, then adding Glucosylglycyrrhizin thereto, uniformly mixing, and then adding Paeoniflorin thereto, and uniformly mixing to obtain the antidepressant composition.

[0048] Comparative Example 4 (without Paeoniflorin)

[0049] An antidepressant composition, which comprises the following components in mass fractions: Saikosaponin D 1 part, Saikosaponin A 4 parts, Albiflorin 8 parts, Glucosylglycyrrhizin 13 parts, Isoglucosylglycyrrhizin 2 parts, Atractylenolide I 5 parts.

[0050] The preparation method of the antidepressant composition comprises the following steps: adding apiosylglycyrrhizin to saikosaponin D, mixing evenly, then adding saikosaponin A thereto, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding apiosylglycyrrhizin thereto, and mixing evenly to obtain the antidepressant composition.

[0051] Comparative Example 5 (the ratios of apiosylglycyrrhizin and apiosylglycyrrhizin are both outside the protection scope)

[0052] An antidepressant composition, which composition comprises the following components in mass fractions: 1 part of saikosaponin D, 4 parts of saikosaponin A, 23 parts of paeoniflorin, 8 parts of albiflorin, 15 parts of apiosylglycyrrhizin, 5 parts of apiosylglycyrrhizin, and 5 parts of atractylenolide I.

[0053] The preparation method of the antidepressant composition comprises the following steps: adding apiosylglycyrrhizin to saikosaponin D, mixing evenly, then adding saikosaponin A thereto, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding apiosylglycyrrhizin thereto, and mixing evenly, then adding paeoniflorin thereto, and mixing evenly to obtain the antidepressant composition.

[0054] Comparative Example 6 (without apiosylglycyrrhizin and apiosylglycyrrhizin)

[0055] An antidepressant composition, which composition comprises the following components in mass fractions: 1 part of saikosaponin D, 4 parts of saikosaponin A, 23 parts of paeoniflorin, 8 parts of albiflorin, and 5 parts of atractylenolide I.

[0056] The preparation method of the antidepressant composition comprises the following steps: adding saikosaponin A to saikosaponin D, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding paeoniflorin thereto, and mixing evenly to obtain the antidepressant composition.

[0057] Comparative Example 7 (the ratios of saikosaponin D and atractylenolide I are both outside the protection scope)

[0058] An antidepressant composition, which composition comprises the following components in mass fractions: 1 part of saikosaponin D, 4 parts of saikosaponin A, 23 parts of paeoniflorin, 8 parts of albiflorin, 13 parts of apiosylglycyrrhizin, 2 parts of apiosylglycyrrhizin, and 1 part of atractylenolide I.

[0059] The preparation method of the antidepressant composition comprises the following steps: adding apiosylglycyrrhizin to saikosaponin D, mixing evenly, then adding saikosaponin A thereto, mixing evenly, then adding atractylenolide I thereto, mixing evenly, then adding albiflorin thereto, mixing evenly, then adding apiosylglycoside thereto, mixing evenly, and then adding paeoniflorin thereto, and mixing evenly to obtain the antidepressant composition.

[0060] Experimental Example 1: Depression cell model

[0061] PC-12 cells are preserved in DMEM basal medium supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. The PC-12 cells are cultured in a humidified incubator at 37 °C and 5% CO 2 . Corticosterone injury can be used as an effective method for establishing in vitro and in vivo depression models. Digest the PC-12 cells in the logarithmic growth phase and blow them into a single-cell suspension, adjust the cell density to 4×10 5 / mL, and place them in a 96-well plate. When the PC-12 cells grow to 80-90% confluence, conduct the experiment. Add 100 μL of corticosterone solution (384 μmol / L) to the wells, and place it in a conventional cell incubator for another 24 hours to construct a corticosterone injury model. Then add the tested drug to the corticosterone injury model. By adding the dye CCK8 and detecting the absorbance at 450 nm, the cell viability of the depression cells can be calculated. The pharmacodynamic activities of the compositions obtained in Examples 1-3 and Comparative Examples 1-7 on the depression cell model are shown in Table 1. By comparing the cell viabilities of the drug administration group and the model group, it can be seen which drugs have a strong ability to improve depression cells.

[0062] Table 1: Pharmacodynamic activities

[0063]

[0064] As shown in Table 1, the composition of Example 1 has the strongest ability to improve cell viability on the depression cell model. Based on the depression model activity of 47.69%, adding the composition of Example 1 increases the cell viability of the depression-active cells to 75.28%; in Example 2, the proportion of saikosaponin A is changed, and the cell viability of the depression-active cells is 56.23%. Compared with Example 1, the ability to improve the effect of depression cells is weaker, indicating the importance of saikosaponin A in the system, which seriously affects the antidepressant efficacy of the overall formula; in Example 3, the addition amount of albiflorin is adjusted, and the cell viability of the depression-active cells is 65.88%. Compared with Example 1, the ability to improve the effect of depression cells is relatively weak; the above results show that the change in the proportion of albiflorin has a relatively small impact on the pharmacodynamic effect of the overall formula, while the change in the proportion of the saikosaponin A component has a relatively large impact on the pharmacodynamic effect of the overall formula.

[0065] After adding the compositions of Comparative Example 1 and Comparative Examples 3-4, the ability to improve depressive cells will decline sharply, indicating that as important components, the change in the ratio of saikosaponin A and paeoniflorin will greatly reduce the ability of depressive active cells; in Comparative Examples 5-7, by changing the ratios of liquiritin apioside, isoliquiritin apioside, saikosaponin D and atractylenolide I, it was found that the antidepressant activity decreased, but not significantly, indicating that the above components have relatively little impact on the efficacy of the overall formula.

[0066] The cell activities of the composition obtained in Example 1 at different concentrations are as Figure 1 shown. Figure 1 The results show that the composition and the antidepressant efficacy show a concentration-dependent type. As the concentration of the composition increases, its ability to improve depression becomes stronger.

[0067] Experimental Example 2 Depressed animal model

[0068] 90 male C57BL / 6 mice were randomly assigned to 9 groups, including a control group, a model group, a positive drug (fluoxetine) group, a low-dose composition group (ZFL), a medium-dose composition group (ZFM), a high-dose composition group (ZFH), a low-dose Xiaoyaosan group (XYSL), a medium-dose Xiaoyaosan group (XYSM), and a high-dose Xiaoyaosan group (XYSH), with 10 mice in each group. From the first day, lipopolysaccharide (LPS) was intraperitoneally injected once every three days. The control group was gavaged with an appropriate amount of normal saline. The gavage dosage for 14 consecutive days was 0.1 mL / 10 g, and thus the establishment of the animal depression model was completed. On the 15th and 16th days, animal depression behavior tests (swimming test, tail suspension test, and feeding test) were conducted, and the body weights of the mice were weighed every day before drug administration during the experiment. On the 17th day, the body weights of the mice were weighed before sampling, and the mice were anesthetized with an appropriate amount of isoflurane, and after the mice were sacrificed, the brain tissues were taken. After the brain tissues were ground, the supernatant was extracted, and an Elisa kit was used to measure the content changes of monoamine oxidase A (MAO-A) and 5-hydroxytryptamine (5-HT) in the brain.

[0069] The composition in Example 1 was measured on the depressed animal model. First, the depression behavior was measured. It can be Figure 2 seen that the immobile time in the swimming test of both the composition group and the whole extract group of Xiaoyaosan was less than that of the positive drug fluoxetine (marketed western medicine). The less the immobile time in the swimming test indicates the stronger the ability to improve antidepressant activity. In addition, the immobile time in the swimming test of Example 1 was less than that of the whole extract group of Xiaoyaosan. It can be Figure 3 seen that in the tail suspension test, the less the immobile time in the tail suspension test, the stronger the ability to improve depression. The results show that the immobile time in the tail suspension test of the composition in Example 1 was less than that of the marketed western medicine fluoxetine group and the whole extract group of Xiaoyaosan. It can be Figure 4It can be seen that in the feeding experiment, the shorter the feeding time, the stronger the ability to improve depression. The results show that the feeding time of the composition in Example 1 is less than that of the marketed western medicine fluoxetine group and the total extract of Xiaoyaosan group. Modern medical research shows that when depression is improved, the content of MAO-A (monoamine oxidase A) in the brain will decrease, and the content of 5-HT (5-hydroxytryptamine) will increase. From Figure 5 It can be seen that the content of MAO-A in the brain of the composition in Example 1 is less than that of the total extract of Xiaoyaosan group and is equivalent to that of the fluoxetine group. From Figure 6 It can be seen that the content of 5-HT in the brain of the composition in Example 1 is more than that of the total extract of Xiaoyaosan group and the fluoxetine group.

[0070] In summary, the composition of Example 1 has the best antidepressant effect both in the cell model and in the antidepressant animal model. Compared with the marketed western medicine fluoxetine, it has a better ability to improve depression, and compared with the total formula of Xiaoyaosan, it has a stronger ability to improve depression. This composition can reduce the dosage of the medicine taken by patients, with a small amount per dose and few dosing times, while improving the antidepressant effect and having few side effects.

[0071] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An antidepressant composition, characterized in that: The composition includes the following components by mass fraction: 1 part of saikosaponin D, 2-6 parts of saikosaponin A, 20-25 parts of paeoniflorin, 6-9 parts of paeoniflorin, 12-15 parts of apiosyl liquiritigenin, 2-4 parts of apiosyl isoliquiritigenin, 4-6 parts of atractylodes lactone I.

2. The antidepressant composition according to claim 1, characterized in that: The composition comprises the following components by mass fraction: 1 part of saikosaponin D, 3-5 parts of saikosaponin A, 21-23 parts of paeoniflorin, 7-9 parts of paeoniflorin, 12-14 parts of apiosyl liquiritigenin, 2-4 parts of apiosyl isoliquiritigenin, 4-6 parts of atractylodes lactone I.

3. The antidepressant composition according to claim 2, characterized in that: The composition comprises the following components in mass fractions: 1 part of saikosaponin D, 4 parts of saikosaponin A, 23 parts of paeoniflorin, 8 parts of paeoniflorin, 13 parts of apiolactone, 2 parts of apiolactone isoliquiritigenin and 5 parts of atractylodes lactone I.

4. The antidepressant composition according to claim 1, characterized in that: The purity of the described paeoniflorin is 55-65%; the purity of the described paeoniflorin is 70-75%; the purity of the described apiosyl liquiritin is 50-55%; the purity of the described apiosyl isoliquiritin is 20-30%; the purity of the described saikosaponin A is 35-45%; the purity of the described saikosaponin D is 30-35%; the purity of the described atractylodes lactone I is 10-20%.

5. The method for preparing the antidepressant composition according to any one of claims 1 to 4, characterized in that: The method comprises the following steps: adding apiosyl isoliquiritigenin to saikosaponin D, adding saikosaponin A after mixing evenly, adding atractylodes lactone I after mixing evenly, adding peoniflorin after mixing evenly, adding apiosyl isoliquiritigenin after mixing evenly, adding peoniflorin after mixing evenly, and mixing evenly to obtain the antidepressant composition.

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