Anti-depression composition containing compound licorice tablets and application of anti-depression composition

Abstract

The invention belongs to the field of antidepressant drugs, and discloses an antidepressant composition containing compound liquorice tablets and application of the antidepressant composition. The composition obtained by compounding the compound licorice tablets and various traditional Chinese medicine extracts has the effects of soothing liver-qi stagnation, calming heart and tranquilizing mind, nourishing blood and regulating liver, and treating qi and blood at the same time, can significantly improve depressive symptoms caused by LPS stimulation, and has a good anti-depression effect. On the basis, a fingerprint control mode based on the dual-wavelength high performance liquid chromatography standard characteristics of the compound liquorice tablets is established. According to the method, macro qualitative similarity and macro quantitative similarity are used for checking the change range of the overall component content and the quality consistency control condition of the compound liquorice tablets. The quality consistency of the compound liquorice tablets is finally not less than 0.95 in terms of macro qualitative similarity of 220nm and 260nm high performance liquid chromatography fingerprints, the macro quantitative similarity is between 85% and 115%, and the compound liquorice tablets have the effect of improving the depression behavior of mice caused by LPS (Lipopolysaccharide).

Description

Technical Field

[0001] The invention belongs to the field of antidepressant drugs, and more specifically relates to an antidepressant composition containing compound licorice tablets and application thereof. Background Art

[0002] Depression is a common affective mental disorder, a mental illness with significant and lasting changes in mood (elevation or depression) as the basic clinical manifestation, often accompanied by corresponding changes in thinking and behavior. Clinically, it can be divided into many types, such as endogenous depression, psychogenic depression, secondary depression, etc.; with low mood as the core symptom, manifested as pessimism and disappointment, loss of interest in daily life, mental depression, and loss of appetite, causing great distress to the patient's life and family.

[0003] At present, there are three main types of Western medicine for the treatment of depression: monoamine oxidase inhibitors, tricyclic antidepressants and heterocyclic antidepressants. Monoamine oxidase inhibitors were the first antidepressants discovered in the 1950s. Because of their serious toxic and side effects, they were quickly replaced by tricyclic antidepressants. Tricyclic antidepressants are the most widely used antidepressants, including amitriptyline, doxepin and imipramine, etc., which are suitable for various depressions, but they are prone to withdrawal syndrome due to sudden discontinuation of the drug. Heterocyclic antidepressants include mianserin, fluoxetine, paroxetine and sertraline. Depression is a chronic disease. There are many problems with long-term use of Western antidepressant drugs, such as slow onset, high price and obvious side effects, such as cardiac toxicity, liver toxicity and other serious allergic reactions. Traditional Chinese medicine believes that depression belongs to the category of "depression", "epilepsy" and "organ dryness", and the onset of depression is mostly caused by emotional injury. Emotional factors are the main factors of depression. Emotional factors cause disease, and the liver is the first to be affected. The development of the disease affects the spleen and stomach in the middle jiao. Since traditional Chinese medicine has the characteristics of multiple components, multiple targets and complex compatibility, the therapeutic effect of existing traditional Chinese medicine needs to be improved.

[0004] Fufang Gancao Pian is made by drying licorice extract, grinding it, adding sodium benzoate and opium powder and mixing them evenly to make granules, adding camphor and star anise oil dissolved in a small amount of ethanol, mixing and pressing them into tablets, and then using them to relieve cough and expectorate. At present, there is no research on Fufang Gancao Pian in other disease fields, and the existing technology does not control the dual-wavelength fingerprint spectrum in Fufang Gancao Pian, so its therapeutic effect cannot be reflected.

[0005] Therefore, how to provide an antidepressant composition containing compound licorice tablets and its application is a problem that needs to be solved urgently by those skilled in the art. Summary of the invention

[0006] In order to overcome the shortcomings and deficiencies in the prior art, the present invention provides an antidepressant composition containing compound licorice tablets and application thereof.

[0007] In order to achieve the above object, the present invention adopts the following technical solution: An antidepressant composition containing a compound licorice tablet, the antidepressant composition comprising the compound licorice tablet and a traditional Chinese medicine extract; the mass ratio of the compound licorice tablet to the traditional Chinese medicine extract is 8-12:2-5; The Chinese medicine extract comprises the following raw materials in parts by weight: 30-40 parts of bupleurum, 15-25 parts of saffron, 15-25 parts of millettia reticulata, and 8-12 parts of spiny jujube seeds.

[0008] Preferably, the compound licorice tablets include the following raw materials: licorice extract powder, opium powder or poppy fruit extract, camphor, star anise oil, sodium benzoate, 95% ethanol, magnesium oxide, talc, microcrystalline cellulose, silicon dioxide, and magnesium stearate.

[0009] Preferably, the preparation method of the Chinese medicine extract comprises the following steps: 1) Weigh the raw materials according to the above weight parts and set aside; 2) drying and crushing the radix bupleuri, saffron, millettia reticulata, and spiny jujube seeds, passing through a 100-150 mesh sieve to obtain a powder, mixing the powder with a 70-85% ethanol aqueous solution, extracting, filtering, and obtaining a filtrate; 3) The filtrate is evaporated to remove ethanol, and concentrated under reduced pressure to obtain an extract with a relative density of 1.2-1.3 at 25°C, which is freeze-dried to obtain a Chinese medicine extract.

[0010] Preferably, in step 2), the mass ratio of the powder to the ethanol aqueous solution is 1:8-15, the extraction temperature is 70-85°C, and the extraction time is 80-120 min; The freeze-drying temperature in step 3) is -80~-40°C, the vacuum degree is 100~150Pa, and the time is 10~16h.

[0011] The present invention also provides the use of the antidepressant composition in the preparation of antidepressant drugs. The compound licorice tablets are subjected to quality control by collecting dual-wavelength fingerprints using high performance liquid chromatography, which comprises the following steps: (1) Preparation of test solution: Take compound licorice tablets, accurately weigh, grind, accurately weigh 4 tablets, place in a 50 mL volumetric flask fixed to volume with extraction solvent, accurately weigh, ultrasonicate, let stand to room temperature, accurately weigh again, make up the lost weight with extraction solvent, shake well, filter through 0.45 μm filter membrane, and take the filtrate to obtain; (2) Preparation of reference solution: Take appropriate amounts of sodium benzoate reference substance and ammonium glycyrrhizinate reference substance, weigh them accurately, add methanol to make a mixed reference solution containing 160 μg sodium benzoate and 800 μg ammonium glycyrrhizinate per 1 mL, shake well, and obtain the solution. (3) Determination of HPLC chromatographic conditions: octadecyl silica gel column, 0.2% phosphoric acid-water solution containing 0.005 mol / L sodium heptane sulfonate as mobile phase A, 9:1 acetonitrile-methanol mixture as mobile phase B, gradient elution, detection wavelengths at 220 nm and 260 nm, column temperature at 35°C, flow rate at 1.0 mL per minute, injection volume at 10 μL; (4) HPLC fingerprint evaluation method: The test solution and the reference solution are measured according to the chromatographic conditions of step (3), and analyzed and compared to obtain the HPLC fingerprint of the compound licorice tablets composed of the common characteristic peaks of the samples, and the macro-qualitative similarity and macro-quantitative similarity between the characteristic spectrum of the test solution and the characteristic spectrum of the reference solution are evaluated.

[0012] Preferably, the extraction solvent in step (1) is an 80% V / V methanol aqueous solution containing 0.5% V / V phosphoric acid; The ultrasonic treatment was performed at a power of 240 W, a frequency of 40 kHz, a temperature of 45° C., and a time of 10 min.

[0013] Preferably, the chromatographic column in step (3) is of COSMOSIL 5C18-MS-II, with a column length of 25 cm, an inner diameter of 4.6 mm, and a particle size of 5 µm.

[0014] Preferably, the conditions for the gradient elution in step (3) are: 0~10min, the volume percentage of mobile phase A is 96%→79%, and the volume percentage of mobile phase B is 4%→21%; 10~20min, the volume percentage of mobile phase A is 79%→65%, and the volume percentage of mobile phase B is 21%→35%; 20~32min, the volume percentage of mobile phase A is 65%→47%, and the volume percentage of mobile phase B is 35%→53%; 32~45min, the volume percentage of mobile phase A is 47%→18%, and the volume percentage of mobile phase B is 53%→82%; 45~50min, the volume percentage of mobile phase A is 18%→15%, and the volume percentage of mobile phase B is 82%→85%; 50~55min, the volume percentage of mobile phase A is 15%→96%, and the volume percentage of mobile phase B is 85%→4%.

[0015] Preferably, at a detection wavelength of 220 nm, the fingerprint of the compound licorice tablets includes 31 common peaks, of which peak No. 10 is morphine, peak No. 14 is liquiritigenin, peak No. 15 is codeine phosphate, peak No. 16 is sodium benzoate, peak No. 21 is glycyrrhizin, and peak No. 28 is glycyrrhizic acid.

[0016] Preferably, at a detection wavelength of 260 nm, the fingerprint of the compound licorice tablets includes 27 common peaks, of which peak No. 10 is liquiritigenin, peak No. 11 is sodium benzoate, peak No. 16 is glycyrrhizin, and peak No. 23 is glycyrrhizic acid.

[0017] Preferably, the macro qualitative similarity of the compound licorice tablets at detection wavelengths of 220nm and 260nm is not less than 0.95, and the macro quantitative similarity is between 85-115%.

[0018] It can be seen from the above technical solution that, compared with the prior art, the present invention provides an antidepressant composition containing compound licorice tablets and its application, which has the following beneficial effects: The present invention compounds compound licorice tablets with a variety of Chinese medicine extracts, and the resulting composition has the effects of soothing the liver and relieving depression, calming the mind and calming the nerves, nourishing blood and regulating the liver, and treating qi and blood at the same time, which can improve the patient's depressive state, relieve anxiety, regulate mental state, and have significant therapeutic effects. According to animal experiments, the composition can significantly improve the depressive symptoms caused by LPS stimulation and has a good antidepressant effect. On this basis, an antidepressant control method model and an evaluation method for the overall quality control of compound licorice tablets are obtained, and a standard characteristic fingerprint control mode based on dual-wavelength high-performance liquid chromatography of compound licorice tablets is established. The present invention is based on the study of 35 batches of quantitative HPLC fingerprints of compound licorice tablets and forms its dual-wavelength high-performance liquid chromatography characteristic fingerprint. Different from the fingerprint inspection method of the Chinese Pharmacopoeia, it is an antidepressant control method based on the overall quality control of compound licorice tablets. The present invention uses macro qualitative similarity and macro quantitative similarity to check the variation range and quality consistency control of the overall component content of compound licorice tablets. The quality consistency of compound licorice tablets is ultimately based on the macro-qualitative similarity of the 220nm and 260nm HPLC fingerprints, which should be no less than 0.95, and the macro-quantitative similarity should be between 85% and 115%. Compound licorice tablets that meet these conditions have an improving effect on LPS-induced depressive behavior in mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0020] Figure 1 Figures AG in the figure are the results of the antidepressant test of compound licorice tablets on normal mice; A: mouse movement trajectory, B: total movement distance, C: center area distance, D: center area time, E: average speed, F: TST immobility time, G: FST immobility time; Figure 1 HI in the figure is the test result of compound licorice tablets on LPS-induced depression model in mice; where H: TST immobility time, I: FST immobility time.

[0021] Figure 2 This is the 220nm characteristic fingerprint of compound licorice tablets.

[0022] Figure 3 This is the 260nm characteristic fingerprint of compound licorice tablets.

[0023] Figure 4 This is an overlay of HPLC fingerprints of 35 batches of compound licorice tablets at 220nm.

[0024] Figure 5 This is an overlay of HPLC fingerprints of 35 batches of compound licorice tablets at 260nm. DETAILED DESCRIPTION

[0025] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Example 1 An antidepressant composition, comprising a compound licorice tablet and a traditional Chinese medicine extract; the mass ratio of the compound licorice tablet to the traditional Chinese medicine extract is 10:3; The compound licorice tablets are produced by Guangzhou Yuehua Pharmaceutical Co., Ltd., with the approval number of National Medicine Standard H44021404 and the drug batch number of 230903. The raw material formula of the compound licorice tablets is shown in Table 1.

[0027] Table 1: Ratio of raw materials for compound licorice tablets

[0028] The preparation method of the compound licorice tablet is as follows: sodium benzoate is taken and crushed into 65 meshes. Licorice extract powder, sodium benzoate, opium powder, and microcrystalline cellulose are accurately weighed and placed in a wet mixing granulator. After dry mixing for 5 minutes, 95% ethanol is added for granulation under the conditions of low stirring speed and low chopping speed for 2 minutes until the granulation is completed. The wet granules are dried to 3.0% to 6.0% moisture at a temperature of 55 to 65°C according to the "CT-C Hot Air Circulation Oven Standard Operating Procedure", and the dry granules are sieved with a 10-mesh sieve. Accurately weigh the formula amount of camphor and star anise oil, place them in a clean container and mix them evenly, and add an appropriate amount of ethanol to dissolve the camphor according to the dissolution of the camphor. Magnesium oxide, silicon dioxide, and talc are placed in a wet mixing granulator, dry mixed for 5 minutes, and then a mixture of ethanol, camphor, and star anise oil is added under the conditions of low stirring speed, low chopping speed, and 2 minutes, and the mixture is continued for 5 minutes until uniform. The whole granules, the prescribed amount of magnesium stearate, the mixed camphor and star anise oil, and the mixed materials of magnesium oxide, silicon dioxide, and talcum powder are placed in a three-eccentric mixer and mixed for 30 minutes to obtain a granular intermediate product, which is tableted after passing the inspection, and then packaged and packaged to obtain the compound licorice tablets.

[0029] The Chinese medicine extract comprises the following raw materials in parts by weight: 35 parts of bupleurum, 20 parts of saffron, 20 parts of millettia reticulata, and 10 parts of spiny jujube seeds.

[0030] The preparation method of the Chinese medicine extract comprises the following steps: 1) Weigh the raw materials according to the above weight parts and set aside; 2) drying and crushing the radix bupleuri, saffron, millettia reticulata, and spiny jujube seeds, passing through a 120-mesh sieve to obtain a powder, mixing the powder with a 75% ethanol aqueous solution, extracting, and filtering to obtain a filtrate; 3) The filtrate is evaporated to remove ethanol, and concentrated under reduced pressure to obtain an extract with a relative density of 1.2 at 25°C, which is freeze-dried to obtain a Chinese medicine extract.

[0031] In step 2), the mass ratio of the powder to the ethanol aqueous solution is 1:12, the extraction temperature is 80°C, and the extraction time is 100 min; The freeze-drying temperature in step 3) is -50°C, the vacuum degree is 120Pa, and the time is 12h.

[0032] Example 2 An antidepressant composition, comprising a compound licorice tablet and a traditional Chinese medicine extract; the mass ratio of the compound licorice tablet to the traditional Chinese medicine extract is 8:2; The Chinese medicine extract comprises the following raw materials in parts by weight: 30 parts of bupleurum, 15 parts of saffron, 15 parts of millettia reticulata, and 8 parts of spiny jujube seeds. The preparation method of the Chinese medicine extract is the same as that of Example 1.

[0033] The compound liquorice tablet is the same as in Example 1.

[0034] Example 3 An antidepressant composition, comprising a compound licorice tablet and a traditional Chinese medicine extract; the mass ratio of the compound licorice tablet to the traditional Chinese medicine extract is 12:5; The Chinese medicine extract comprises the following raw materials in parts by weight: 40 parts of bupleurum, 25 parts of saffron, 25 parts of millettia reticulata, and 12 parts of spiny jujube seeds. The preparation method of the Chinese medicine extract is the same as that of Example 1.

[0035] The compound liquorice tablet is the same as in Example 1.

[0036] Comparative Example 1 A compound liquorice tablet, same as that of Example 1.

[0037] Comparative Example 2 A Chinese medicine extract comprises the following raw materials in parts by weight: 35 parts of bupleurum, 20 parts of saffron, 20 parts of millettia reticulata, and 10 parts of spiny jujube seeds. The preparation method of the Chinese medicine extract is the same as that of Example 1.

[0038] Comparative Example 3 An antidepressant composition, which is different from Example 1 in that Bupleurum is removed.

[0039] Comparative Example 4 An antidepressant composition, which is different from Example 1 in that saffron is removed.

[0040] 1. Animal Experiment 1: P1. Purpose of the test The improving effects of the products of various examples and comparative examples on the depressive behavior of mice induced by lipopolysaccharide were investigated.

[0041] P2. Experimental Animals Kunming mice, 5 mice per group, weighing 18-22g, half male and half female.

[0042] P3. Test Principle Depression is a common mental illness, mainly manifested by low mood, anhedonia, and abnormal sleep. Lipopolysaccharide (LPS) is a cell wall component of Gram-negative bacteria, which can not only induce peripheral inflammatory processes, but also cause inflammatory responses in the central nervous system. Intraperitoneal injection of LPS causes inflammatory responses in the central nervous system, thereby causing depressive-like behaviors and cognitive changes in mice. Therefore, it can be used to establish an animal model of immune activation-induced depressive-like behaviors. The LPS-induced mouse depression model was used to detect the antidepressant effect of the product, and the therapeutic effect was measured by the changes in the immobility time in the forced swimming and tail suspension tests.

[0043] P4. Experimental Grouping Kunming mice were randomly divided into a blank group, a positive group, groups 1 to 3 of Examples, and groups 1 to 4 of Comparative Examples. The corresponding products were gavaged to each group of Examples and Comparative Examples at a dose of 200 mg / kg, and the blank group and the positive group were gavaged with an equal amount of normal saline once a day for 30 days. After 30 days, 3 mg / kg LPS solution was intraperitoneally injected into each group of mice, but the blank group was injected with an equal amount of normal saline. After 24 hours, the test was carried out.

[0044] P5. Test methods (1) Tail suspension test (TST) The mice were suspended from a metal rod at a height of 50 cm above the table. The mice were considered immobile only if their limbs did not move. At the beginning of the adaptation test, the mice were suspended for 2 minutes. The immobility time of the mice was measured from the 3rd to the 6th minute of the test.

[0045] (2) Forced swimming test (FST) Mice were placed individually in 4 plastic cylindrical water tanks (30 cm high and 11 cm in diameter) filled with water (24 ± 1 °C) at a depth of 25 cm for 6 min. Each mouse was judged as immobile when it stopped struggling and remained motionless in the water. Immobility time was measured in the last 4 min of the test.

[0046] P6. Test results The results are shown in Table 2.

[0047] Table 2: Test results

[0048] It can be seen that LPS stimulation in the positive group significantly increased the immobility time of mice. The antidepressant composition of the present invention can significantly shorten the immobility time of the tail suspension test and the forced swimming test of mice, among which the improvement effect of the composition of Example 3 is the best. The present invention uses the compound licorice tablets and the Chinese medicine extract in combination, and the therapeutic effect is better than that of using them alone, and has a good antidepressant effect.

[0049] 2. Chromatographic experiment: The compound licorice tablet of the present invention adopts high performance liquid chromatography to collect dual wavelength fingerprint spectrum for quality control, comprising the following steps: (1) Preparation of test solution: Take compound licorice tablets, accurately weigh, grind, accurately weigh 4 tablets, place in a 50 mL volumetric flask fixed to volume with extraction solvent, accurately weigh, ultrasonicate, let stand to room temperature, accurately weigh again, make up the lost weight with extraction solvent, shake well, filter through 0.45 μm filter membrane, and take the filtrate to obtain; (2) Preparation of reference solution: Take appropriate amounts of sodium benzoate reference substance and ammonium glycyrrhizinate reference substance, weigh them accurately, add methanol to make a mixed reference solution containing 160 μg sodium benzoate and 800 μg ammonium glycyrrhizinate per 1 mL, shake well, and obtain the solution. (3) Determination of HPLC chromatographic conditions: octadecyl silica gel column, gradient elution, detection wavelengths of 220 nm and 260 nm, column temperature of 35 °C, flow rate of 1.0 mL per minute, injection volume of 10 μL; (4) HPLC fingerprint evaluation method: The test solution and the reference solution were measured according to the chromatographic conditions of step (3), and the analysis and comparison were performed to obtain the HPLC fingerprint of the compound licorice tablets composed of the common characteristic peaks of the samples. The macro-qualitative similarity and macro-quantitative similarity between the characteristic spectrum of the test solution and the characteristic spectrum of the reference solution were evaluated using the traditional Chinese medicine quantitative fingerprint software.

[0050] The extraction solvent in step (1) is an 80% V / V methanol aqueous solution containing 0.5% V / V phosphoric acid; The ultrasonic treatment was performed at a power of 240 W, a frequency of 40 kHz, a temperature of 45° C., and a time of 10 min.

[0051] The model of the chromatographic column in step (3) is COSMOSIL 5C18-MS-II, with a column length of 25 cm, an inner diameter of 4.6 mm, and a particle size of 5 µm.

[0052] In step (3), a 0.2% phosphoric acid-water solution containing 0.005 mol / L sodium heptane sulfonate is used as mobile phase A, and an acetonitrile-methanol mixture with a volume ratio of 9:1 is used as mobile phase B.

[0053] The conditions for the gradient elution in step (3) are shown in Table 3.

[0054] Table 3: Gradient elution conditions

[0055] At 220nm detection wavelength: The fingerprint of compound licorice tablets includes 31 common peaks, with glycyrrhizic acid as the reference peak, of which peak 10 is morphine, peak 14 is liquiritin, peak 15 is codeine phosphate, peak 16 is sodium benzoate, peak 21 is glycyrrhizin, and peak 28 is glycyrrhizic acid, see Figure 2 .

[0056] The relative retention time values ​​of each characteristic peak are: 0.08 (peak 1), 0.09 (peak 2), 0.19 (peak 3), 0.24 (peak 4), 0.26 (peak 5), 0.29 (peak 6), 0.36 (peak 7), 0.37 (peak 8), 0.43 (peak 9), 0.45 (peak 10), 0.46 (peak 11), 0.48 (peak 12), 0.49 (peak 13), 0.50 (peak 14), 0.52 (peak 15), 0.61 (peak 16), 0.63 (peak 17), 0.65 (peak 18), 0.66 (peak 19), 0.67 (peak 20), 0.71 (peak 21), 0.77 (peak 22), 0.87 (peak 23), 0.90 (peak 24), 0.93 (peak 25), 0.94 (peak 26), 0.96 (peak 27), 1.00 (peak 28, S), 1.06 (peak 29), 1.27 (peak 30), 1.30 (peak 31).

[0057] The relative peak area specifications of each characteristic peak are: 0.45 (peak 1), 0.35 (peak 2), 0.15 (peak 3), 0.04 (peak 4), 0.07 (peak 5), 0.18 (peak 6), 0.84 (peak 7), 0.09 (peak 8), 0.16 (peak 9), 0.67 (peak 10), 0.65 (peak 11), 0.70 (peak 12), 1.68 (peak 13), 1.13 (peak 14), 0.25 (peak 15), 2.81 (peak 16), 0.36 (peak 17), 0.22 (peak 18), 0.50 (peak 19), 0.42 (peak 20), 0.67 (peak 21), 0.18 (peak 22), 0.07 (peak 23), 0.13 (peak 24), 0.25 (peak 25), 0.06 (peak 26), 0.13 (peak 27), 1.00 (peak 28, S), 0.21 (peak 29), 0.11 (peak 30), 0.82 (peak 31).

[0058] At 260nm detection wavelength: The fingerprint of compound licorice tablets includes 27 common peaks, among which peak 10 is liquiritin, peak 11 is sodium benzoate, peak 16 is glycyrrhizin, and peak 23 is glycyrrhizic acid. Figure 3 .

[0059] The relative retention time values ​​of each characteristic peak are: 0.09 (peak 1), 0.19 (peak 2), 0.24 (peak 3), 0.28 (peak 4), 0.36 (peak 5), 0.43 (peak 6), 0.46 (peak 7), 0.48 (peak 8), 0.49 (peak 9), 0.51 (peak 10), 0.61 (peak 11), 0.63 (peak 12), 0.65 (peak 13), 0.66 (peak 14), 0.68 (peak 15), 0.71 (peak 16), 0.77 (peak 17), 0.87 (peak 18), 0.90 (peak 19), 0.94 (peak 20), 0.94 (peak 21), 0.96 (peak 22), 1.00 (peak 23, S), 1.06 (peak 24), 1.27 (peak 25), 1.30 (peak 26), 1.49 (peak 27).

[0060] The relative peak area specifications of each characteristic peak are: 0.09 (peak 1), 0.03 (peak 2), 0.02 (peak 3), 0.02 (peak 4), 0.06 (peak 5), 0.02 (peak 6), 0.01 (peak 7), 0.09 (peak 8), 0.13 (peak 9), 0.08 (peak 10), 0.08 (peak 11), 0.08 (peak 12), 0.04 (peak 13), 0.15 (peak 14), 0.04 (peak 15), 0.06 (peak 16), 0.03 (peak 17), 0.07 (peak 18), 0.02 (peak 19), 0.05 (peak 20), 0.06 (peak 21), 0.04 (peak 22), 1.00 (peak 23, S), 0.20 (peak 24), 0.01 (peak 25), 0.63 (peak 26), 0.02 (peak 27).

[0061] The present invention uses 35 batches of compound licorice tablet samples to obtain quantitative fingerprints, and then forms characteristic fingerprints. The specific information of each sample is shown in Table 4. Figure 4 and Figure 5 shown.

[0062] Table 4: Sample details No. Drug number Drug batch number Manufacturer Approval No. S1 YH-S1 230101 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S2 YH-S2 230102 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S3 YH-S3 230103 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S4 YH-S4 230104 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S5 YH-S5 230105 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S6 YH-S6 230106 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S7 YH-S7 230107 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S8 YH-S8 230201 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S9 YH-S9 230202 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S10 YH-S10 230203 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S11 YH-S11 230204 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S12 YH-S12 230205 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S13 YH-S13 230903 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S14 YH-S14 230904 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S15 YH-S15 230905 Guangzhou Yuehua Pharmaceutical Co., Ltd. National Medicine Approval No. H44021404 S16 SW-S1 23051701 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S17 SW-S2 23051901 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S18 SW-S3 23052001 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S19 SW-S4 23061501 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S20 SW-S5 23062001 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S21 SW-S6 23062601 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S22 SW-S7 23062801 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S23 SW-S8 23061901 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S24 SW-S9 23070101 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S25 SW-S10 23070401 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S26 SW-S11 23070501 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S27 SW-S12 23070701 Shenwei Pharmaceutical (Zhangjiakou) Co., Ltd. National Medicine Approval No. H13022025 S28 TH-S1 20211017 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 S29 TH-S2 20211018 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 S30 TH-S3 20211019 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 S31 TH-S4 20211020 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 S32 TH-S5 20211101 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 S33 TH-S6 20211102 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 S34 TH-S7 20211103 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 S35 TH-S8 20211104 Harbin Taihua Pharmaceutical Co., Ltd. National Medicine Approval No. H23020708 The similarity evaluation results are shown in Table 5.

[0063] Table 5: Similarity evaluation results of dual-wavelength characteristic fingerprints of 35 batches of compound licorice tablets

[0064] The quality consistency of compound licorice tablets should be ultimately based on the macro qualitative similarity of the 220nm and 260nm fingerprints, which should not be less than 0.95, and the macro quantitative similarity should be between 85% and 115%.

[0065] 3. Animal Experiment 2: P1. Purpose of the test To investigate the improving effect of compound licorice tablets on lipopolysaccharide-induced depressive behavior in mice.

[0066] P2. Experimental Animals Kunming mice, 5 mice per group, weighing 18-22g, half male and half female.

[0067] P3. Test Principle Normal mice and LPS-induced mouse depression models were used to detect the antidepressant effect of compound licorice tablets, and the efficacy was measured by open field test, forced swimming test and tail suspension test.

[0068] P4. Experimental Grouping The compound licorice tablets used in this experiment were produced by Guangzhou Yuehua Pharmaceutical Co., Ltd., with the approval number of National Medicine Standard H44021404 and the drug batch number of 230903. This experiment first conducted an antidepressant test after administration to normal mice (1) as a preliminary screening of the antidepressant efficacy. On the basis of experiment (1), an LPS-induced mouse depression model was established and an antidepressant test was conducted (2). The specific steps are as follows: (1) Kunming mice were randomly divided into a control group (Control), a fluoxetine group (18 mg / kg), and three compound licorice tablet groups (121.64 mg / kg, 243.27 mg / kg, and 486.54 mg / kg). Each group was gavaged with the corresponding drug, and the control group was gavaged with an equal amount of normal saline once a day for 34 days. After 34 days, the open field test, tail suspension test, and forced swimming test were evaluated every 3 hours in sequence.

[0069] (2) Kunming mice were randomly divided into a control group (Control), an LPS group, a fluoxetine group (18 mg / kg), and three compound licorice tablet groups (121.64 mg / kg, 243.27 mg / kg, and 486.54 mg / kg). Each group was gavaged with the corresponding drug, and the LPS group and the control group were gavaged with an equal amount of normal saline once a day for 34 days. After 34 days, each group of mice was intraperitoneally injected with 3 mg / kg LPS solution, but the control group was injected with an equal amount of normal saline. After 24 hours, the tail suspension test and forced swimming test were evaluated every 3 hours in sequence.

[0070] P5. Test methods 5.1 Open field test (OFT) The open area was square (50×50 cm) and divided into a peripheral area and a central area (21×21 cm). The mouse was placed in a corner of the peripheral area. The spontaneous activity of the mouse was recorded for 5 minutes. After each test, the open area was washed with water to remove olfactory cues.

[0071] 5.2 Tail suspension test (TST) Same as animal experiment 1.

[0072] 5.3 Forced swimming test (FST) Same as animal experiment 1.

[0073] P6. Test results Using normal mice and LPS-induced depression mouse models, the antidepressant effects of compound licorice tablets were examined by evaluating the behavioral changes of mice in OFT, TST, and FST.

[0074] The results showed that in OFT, LPS stimulation could reduce the total distance, center area distance, center area time and average speed of mice. The total distance of mice in the open field increased significantly in each dose group of compound licorice tablets ( Figure 1 B in the figure). It can also significantly increase the center zone time, center zone distance and average speed of LPS-stimulated mice ( Figure 1 In addition, LPS stimulation can significantly prolong the immobility time of mice in FST and TST ( Figure 1 Compound licorice tablets significantly improved the immobility symptoms of mice induced by LPS in FST and TST ( Figure 1 FI in ).

[0075] P7. Conclusion In summary, qualified compound licorice tablets can improve the depressive symptoms caused by LPS stimulation, indicating that compound licorice tablets have a certain antidepressant effect.

[0076] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. An antidepressant composition containing compound licorice tablets, characterized in that: The antidepressant composition comprises a compound licorice tablet and a traditional Chinese medicine extract; the mass ratio of the compound licorice tablet to the traditional Chinese medicine extract is 8-12:2-5; The Chinese medicine extract comprises the following raw materials in parts by weight: 30-40 parts of bupleurum, 15-25 parts of saffron, 15-25 parts of millettia reticulata, and 8-12 parts of spiny jujube seeds.

2. The antidepressant composition containing compound licorice tablets according to claim 1, characterized in that: The compound licorice tablet comprises the following raw materials: licorice extract powder, opium powder or poppy fruit extract, camphor, star anise oil, sodium benzoate, 95% ethanol, magnesium oxide, talc, microcrystalline cellulose, silicon dioxide and magnesium stearate.

3. The antidepressant composition containing compound licorice tablets according to claim 1, characterized in that: The preparation method of the Chinese medicine extract comprises the following steps: 1) Weigh the raw materials according to the above weight parts and set aside; 2) drying and crushing the radix bupleuri, saffron, millettia reticulata, and spiny jujube seeds, passing through a 100-150 mesh sieve to obtain a powder, mixing the powder with a 70-85% ethanol aqueous solution, extracting, filtering, and obtaining a filtrate; 3) The filtrate is evaporated to remove ethanol, and concentrated under reduced pressure to obtain an extract with a relative density of 1.2-1.3 at 25°C, which is freeze-dried to obtain a Chinese medicine extract.

4. The antidepressant composition containing compound licorice tablets according to claim 3, characterized in that: In step 2), the mass ratio of the powder to the ethanol aqueous solution is 1:8-15, the extraction temperature is 70-85°C, and the extraction time is 80-120 min; The freeze-drying temperature in step 3) is -80~-40°C, the vacuum degree is 100~150Pa, and the time is 10~16h.

5. Use of the antidepressant composition according to any one of claims 1 to 4 in the preparation of an antidepressant drug, characterized in that: The quality control of compound licorice tablets was carried out by collecting dual-wavelength fingerprints using high performance liquid chromatography, which included the following steps: (1) Preparation of test solution: Take compound licorice tablets, accurately weigh, grind, accurately weigh 4 tablets, place in a 50 mL volumetric flask fixed to volume with extraction solvent, accurately weigh, ultrasonicate, let stand to room temperature, accurately weigh again, make up the lost weight with extraction solvent, shake well, filter through 0.45 μm filter membrane, and take the filtrate to obtain; (2) Preparation of reference solution: Take appropriate amounts of sodium benzoate reference substance and ammonium glycyrrhizinate reference substance, weigh them accurately, add methanol to make a mixed reference solution containing 160 μg sodium benzoate and 800 μg ammonium glycyrrhizinate per 1 mL, shake well, and obtain the solution. (3) Determination of HPLC chromatographic conditions: octadecyl silica gel column, 0.2% phosphoric acid-water solution containing 0.005 mol / L sodium heptane sulfonate as mobile phase A, 9:1 acetonitrile-methanol mixture as mobile phase B, gradient elution, detection wavelengths at 220 nm and 260 nm, column temperature at 35°C, flow rate at 1.0 mL per minute, injection volume at 10 μL; (4) HPLC fingerprint evaluation method: The test solution and the reference solution are measured according to the chromatographic conditions of step (3), and analyzed and compared to obtain the HPLC fingerprint of the compound licorice tablets composed of the common characteristic peaks of the samples, and the macro-qualitative similarity and macro-quantitative similarity between the characteristic spectrum of the test solution and the characteristic spectrum of the reference solution are evaluated.

6. Use of the antidepressant composition according to claim 5 in the preparation of antidepressant drugs, characterized in that: The extraction solvent in step (1) is an 80% V / V methanol aqueous solution containing 0.5% V / V phosphoric acid; The ultrasonic treatment was performed at a power of 240 W, a frequency of 40 kHz, a temperature of 45° C., and a time of 10 min.

7. Use of the antidepressant composition according to claim 5 in the preparation of antidepressant drugs, characterized in that: The conditions for the gradient elution in step (3) are: 0~10min, the volume percentage of mobile phase A is 96%→79%, and the volume percentage of mobile phase B is 4%→21%; 10~20min, the volume percentage of mobile phase A is 79%→65%, and the volume percentage of mobile phase B is 21%→35%; 20~32min, the volume percentage of mobile phase A is 65%→47%, and the volume percentage of mobile phase B is 35%→53%; 32~45min, the volume percentage of mobile phase A is 47%→18%, and the volume percentage of mobile phase B is 53%→82%; 45~50min, the volume percentage of mobile phase A is 18%→15%, and the volume percentage of mobile phase B is 82%→85%; 50~55min, the volume percentage of mobile phase A is 15%→96%, and the volume percentage of mobile phase B is 85%→4%.

8. Use of the antidepressant composition according to claim 5 in the preparation of antidepressant drugs, characterized in that: At a detection wavelength of 220nm, the fingerprint of compound licorice tablets includes 31 common peaks, among which peak No. 10 is morphine, peak No. 14 is liquiritigenin, peak No. 15 is codeine phosphate, peak No. 16 is sodium benzoate, peak No. 21 is glycyrrhizin, and peak No. 28 is glycyrrhizic acid.

9. Use of the antidepressant composition according to claim 5 in the preparation of antidepressant drugs, characterized in that: At a detection wavelength of 260nm, the fingerprint of compound licorice tablets includes 27 common peaks, among which peak No. 10 is liquiritigenin, peak No. 11 is sodium benzoate, peak No. 16 is glycyrrhizin, and peak No. 23 is glycyrrhizic acid.

10. Use of the antidepressant composition according to claim 5 in the preparation of antidepressant drugs, characterized in that: At detection wavelengths of 220nm and 260nm, the macro-qualitative similarity of compound licorice tablets was no less than 0.95, and the macro-quantitative similarity was between 85-115%.

Images