Traditional Chinese medicine composition for treating anxiety and depression, preparation method and traditional Chinese medicine preparation

Abstract

The present application belongs to the technical field of medicine, and particularly relates to a traditional Chinese medicine composition for treating anxiety and depression, a preparation method and a traditional Chinese medicine preparation. The traditional Chinese medicine composition for treating anxiety and depression comprises the following raw materials and mass fractions: 925-1000 parts of Fructus Schisandrae and 25-100 parts of Wuweizi. The present application provides a traditional Chinese medicine composition for treating anxiety and depression, which has remarkable curative effect, simple prescription, easy manufacturing and almost no side effects. The present application discloses a complete large-scale production process from extraction to purification and preparation, and details the specific operation and parameters of each step. The effectiveness and safety of the traditional Chinese medicine composition are proved by pharmacodynamic and toxicological experiments. In the pharmacodynamic comparison experiment with fluoxetine hydrochloride dispersible tablets, the curative effect of the traditional Chinese medicine composition for treating anxiety and depression is better than that of fluoxetine hydrochloride dispersible tablets. Since the traditional Chinese medicine composition is a pure traditional Chinese medicine composition and has almost no side effects, the traditional Chinese medicine composition can replace chemical drugs, other Chinese patent medicines or traditional Chinese medicine decoctions, and greatly improve the life quality of patients with anxiety and depression.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a traditional Chinese medicine composition and preparation method for anxiety and depression, and a traditional Chinese medicine preparation. Background Technology

[0002] Depression is a mental disorder characterized by persistent and significant low mood, slowed thinking, cognitive impairment, and reduced willpower. It has a high relapse and suicide rate, seriously endangering human health. The World Health Organization (WHO) predicts that by 2030, depression will be the leading cause of disease burden, imposing a dual burden on patients and society, both psychologically and economically. Depression can progress to anxiety disorders, therefore, combined treatment should be considered.

[0003] Currently, depression and anxiety are mainly treated with Western medicines such as paroxetine, fluoxetine, sertraline, fluvoxamine, citalopram, and escitalopram. Although the efficacy is relatively significant, the adverse reactions are becoming increasingly prominent, seriously affecting patients' quality of life. This has made traditional Chinese medicine a hot topic in the treatment of depression and anxiety. Traditional Chinese medicine classifies depression and anxiety into categories such as "Yu Bing," "Zang Zao," and "Bai He Bing," and has thousands of years of experience in treating these conditions. Common types of depression include liver qi stagnation, heart and spleen deficiency, heart and kidney disharmony, kidney yang deficiency, and spleen and kidney yang deficiency. Currently available Chinese patent medicines for treating anxiety and depression include Jiawei Xiaoyao Wan, Chaihu Shugan Wan, Renshen Jianpi Wan, Guipi Wan, Liuwei Dihuang Wan, and Zhibai Dihuang Wan. For example, Chinese patent application CN107158099A discloses a Jiawei Xiaoyao Wan and its preparation method. The raw material components and weight ratio of Jiawei Xiaoyao Wan are as follows: Bupleurum 250-350 parts, Angelica sinensis 250-350 parts, Paeonia lactiflora 250-350 parts, Atractylodes macrocephala (stir-fried with wheat bran) 250-350 parts, Poria cocos 250-350 parts, Glycyrrhiza uralensis 200-300 parts, Paeonia suffruticosa 400-500 parts, Gardenia jasminoides (processed with ginger) 400-500 parts, and Mentha haplocalyx 50-100 parts. Chinese patent application CN106266911A discloses a ginseng spleen-strengthening pill and its preparation method. The ginseng spleen-strengthening pill is prepared from the following raw materials in parts by weight: ginseng 20-30 parts, Atractylodes macrocephala 140-160 parts, Poria cocos 40-60 parts, Dioscorea opposita 90-110 parts, Citrus reticulata peel 40-60 parts, Aucklandia lappa 10-15 parts, Amomum villosum 20-30 parts, Astragalus membranaceus 90-110 parts, Angelica sinensis 40-60 parts, Ziziphus jujuba var. spinosa 40-60 parts, and Polygala tenuifolia 20-30 parts. The above formula is complex, the manufacturing cost is high, and it is uncertain whether its efficacy is significantly better than that of Western medicine.

[0004] In conclusion, it is necessary to develop a traditional Chinese medicine composition that has significant therapeutic effects, good safety profile, and is simple to formulate and easy to manufacture. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a traditional Chinese medicine composition that has significant therapeutic effects on anxiety and depression, is simple to formulate, easy to manufacture, and has almost no side effects.

[0006] The technical solution of this invention is:

[0007] A traditional Chinese medicine composition for treating anxiety and depression, comprising the following raw materials and their mass fractions: Acanthopanax senticosus 925-1000 parts, Schisandra chinensis 25-100 parts.

[0008] Another object of the present invention is to provide a method for preparing the aforementioned traditional Chinese medicine composition for treating anxiety and depression, comprising the following steps:

[0009] (1) Mix Acanthopanax senticosus and Schisandra chinensis, crush them, and pass them through a 10-30 mesh sieve to make coarse powder;

[0010] (2) Add extractant 1 to coarse powder and stir evenly. Moisten and seal and let it stand for 1-3 hours to allow it to fully expand. Put the expanded coarse powder into a percolation tank, flatten it, add extractant 2, wait for the air between the powder particles to be expelled and ethanol liquid to flow out, close the discharge valve, cover the filter cylinder, and soak for 18-30 hours.

[0011] (3) After soaking, open the discharge valve to perform percolation. Add extractant 3 while percolating. Control the percolation speed to be 2-3 mL / min per 1000g of medicinal material. Collect the filtrate and mix it with the ethanol liquid discharged in step (2). Concentrate the mixture to obtain a concentrated liquid. Dilute the obtained concentrated liquid, let it stand, centrifuge and filter to obtain supernatant A.

[0012] (4) Pass the supernatant A obtained in step (3) through a D101 macroporous resin adsorption column with a diameter-to-height ratio of 1:2 to 1:4, add eluent 1 for elution, discard the eluent, add eluent 2 for elution, and collect eluent A; wash the adsorption column with water until neutral, add eluent 3 for elution, and collect eluent B.

[0013] (5) Adjust the pH of the eluent A obtained in step (4) to neutral, centrifuge and filter, and pass the resulting supernatant B through a polyamide resin adsorption column with a diameter-to-height ratio of 1:2 to 1:4. Wash with water until no salt is present, then elute with eluent 4, collect eluent C, concentrate and dry eluent C to obtain dry paste A; concentrate and dry eluent B obtained in step (4) to obtain dry paste B;

[0014] (6) Mix dry paste A and dry paste B, crush and sieve to obtain a traditional Chinese medicine composition for treating anxiety and depression.

[0015] Further, in step (2), the extractant 1 is an aqueous ethanol solution with a volume fraction of 70% to 90%, and the amount of extractant 1 added is 0.5 to 1 times the mass of the crude powder. The extractant 2 is added at a flow rate of 2 to 3 mL / min, and the extractant 2 is an aqueous ethanol solution with a volume fraction of 70% to 90%.

[0016] Further, in step (3), the extractant 3 is an aqueous ethanol solution with a volume fraction of 70% to 90%; the mixture is concentrated under vacuum at 40 to 80°C until the relative density at 60°C is 1.10 to 1.20 to obtain a concentrated solution; the obtained concentrated solution is diluted with 2 to 4 times the amount of water.

[0017] Further, in step (4), the eluent 1 is water, and the amount of eluent 1 added is 1 to 3 times the volume of supernatant A; the eluent 2 is a sodium hydroxide aqueous solution with a mass percentage of 0.1%, and the amount of eluent 2 added is 1 to 3 times the volume of supernatant A; the eluent 3 is an ethanol aqueous solution with a volume fraction of 70% to 90%, and the amount of eluent 3 added is 4 to 6 times the volume of D101 macroporous resin adsorption column.

[0018] Further, in step (5), the eluent 4 is an aqueous ethanol solution with a volume fraction of 70% to 90%, and the amount of eluent 4 added is 2 to 3 times the mass of the supernatant B.

[0019] Further, the specific operation of concentrating and drying the eluent C in step (5) to obtain dry paste A is as follows: the eluent C is concentrated under vacuum at 40-80°C to a concentration with a relative density of 1.20-1.30 at 60°C, and the concentration is then vacuum dried at 50-70°C until the dry water content is 3%, to obtain dry paste A; the specific operation of concentrating and drying the eluent B to obtain dry paste B is as follows: the eluent B is concentrated under vacuum at 40-80°C to a relative density of 1.20-1.30 at 60°C to obtain a concentration, and the concentration is then vacuum dried at 50-70°C until the dry water content is 3%, to obtain dry paste B.

[0020] Furthermore, it also includes mixing the obtained traditional Chinese medicine composition for treating anxiety and depression with excipients to prepare traditional Chinese medicine preparations.

[0021] Another object of the present invention is to provide a traditional Chinese medicine preparation for treating anxiety and depression, wherein the traditional Chinese medicine preparation for treating anxiety and depression comprises the traditional Chinese medicine composition for treating anxiety and depression prepared by the above-mentioned preparation method, and pharmaceutically acceptable excipients.

[0022] Furthermore, the dosage form of the traditional Chinese medicine preparation for treating anxiety and depression is selected from granules, capsules, and tablets.

[0023] The traditional Chinese medicine composition for treating anxiety and depression provided by this invention includes Acanthopanax senticosus, the root, rhizome, and stem of the plant Acanthopanax senticosus (family Araliaceae), which has the effects of invigorating qi and strengthening the spleen, tonifying the kidneys and calming the mind. Acanthopanax senticosus possesses numerous and complex chemical components and pharmacological effects. Its chemical components mainly include active ingredients such as glycosides and flavonoids, as well as amino acids and trace elements. Its pharmacological effects include sedation, anti-stress response, anti-fatigue response, and antioxidant response. The mechanism of action of Acanthopanax senticosus in treating depression and anxiety may involve increasing the content of monoamine neurotransmitters, regulating neurotrophic factors in the brain to repair damaged neurons, and anti-oxidation and free radical scavenging.

[0024] Schisandra chinensis is the dried fruit of the Schisandra chinensis plant, belonging to the Schisandraceae family. It has a sour and sweet taste and is warm in nature. It possesses the effects of lowering enzymes and protecting the liver, astringing and consolidating, tonifying the kidneys and calming the mind, and is often used for symptoms such as neurasthenia, palpitations, and insomnia. The mechanism by which Schisandra chinensis treats depression and anxiety may involve influencing neurotransmitter secretion through the gut-brain axis, increasing the content of monoamine neurotransmitters, regulating neurotrophic factors in the brain, improving neuroinflammation, repairing damaged neurons, and possessing antioxidant and free radical scavenging properties. Furthermore, Acanthopanax senticosus and Schisandra chinensis, as traditional Chinese medicines, are widely recognized adaptogenic medicinal materials that can combat external environmental stresses such as cold, hypoxia, and physical weakness, and can exert unique neuroregulatory effects, promoting a balance between nerve excitation and depression.

[0025] In summary, the mechanisms of action of Acanthopanax senticosus and Schisandra chinensis in treating anxiety and depression through multiple targets and pathways are relatively in-depth and clear, with few side effects. Therefore, the combination of Acanthopanax senticosus and Schisandra chinensis in traditional Chinese medicine preparations has clinical value in treating depression and anxiety. The formulation is simple, easy to manufacture, and has a clear mechanism of action. Moreover, compared with Western medicine, it is very safe and has almost no side effects.

[0026] To determine the advantages of the Acanthopanax senticosus and Schisandra chinensis formulation compared to chemical drugs, the prior art of this invention conducted a comparative study on the pharmacological effects and mechanisms of the Acanthopanax senticosus and Schisandra chinensis composition extract against the Western medicine fluoxetine hydrochloride dispersible tablets. The results showed that the composition extract was more effective than fluoxetine hydrochloride dispersible tablets and had good safety.

[0027] Compared with the prior art, the present invention has the following advantages:

[0028] (1) Existing technologies for industrial-scale production of Acanthopanax senticosus and Schisandra chinensis formulations mostly involve heating and extracting Acanthopanax senticosus and Schisandra chinensis, followed by concentration, drying, and then preparation. Although this method is relatively simple to operate and the equipment is relatively inexpensive, the transfer rates of eleutheroside E and schisandrol A are low, and the decoction and preparation contain many impurities. The daily dosage is large, and the resulting preparation has poor adaptability. This invention extracts Acanthopanax senticosus and Schisandra chinensis using percolation, resulting in high transfer rates of eleutheroside E and schisandrol A. The extract is then purified using macroporous resin and polyamide resin to maximize the removal of impurities. The resulting raw material has a high content of active ingredients, and the prepared preparation has a small daily dosage and good adaptability.

[0029] (2) This invention provides a traditional Chinese medicine combination drug with significant therapeutic effects on anxiety and depression, simple formulation, easy manufacturing, and almost no side effects, as well as a complete large-scale production process. From extraction to purification to formulation, the specific operation and parameters of each step are disclosed in detail, and the efficacy and safety of the traditional Chinese medicine composition of this invention are proved through pharmacodynamic and toxicological experiments. In the pharmacodynamic comparison experiment with the Western medicine fluoxetine hydrochloride dispersible tablets, it can be seen that the efficacy of this traditional Chinese medicine composition in treating anxiety and depression is superior to that of the Western medicine fluoxetine hydrochloride dispersible tablets. Furthermore, because the traditional Chinese medicine composition of this invention is a pure traditional Chinese medicine combination with almost no side effects, it can replace chemical drugs, other traditional Chinese medicine preparations, or traditional Chinese medicine decoctions, which will greatly improve the quality of life of patients with anxiety and depression. Attached Figure Description

[0030] Figure 1 The image shows the results of HE staining of the lungs of rat No. 4106 in the high-dose group under a 100x microscope.

[0031] Figure 2 The image shows the liver of rat No. 4110 in the high-dose group, stained with HE under a microscope at 200x magnification.

[0032] Figure 3 The image shows the results of HE staining of the brain of rat No. 4010 in the high-dose group under a 12.5x microscope.

[0033] Figure 4 The image shows the results of HE staining of the spleen of rat 4105 (animal number 4105) in the high-dose group under a 40x microscope.

[0034] Figure 5 The image shows the results of HE staining of the kidneys of rats (animal number 4110) in the high-dose group under a 12.5x microscope.

[0035] Figure 6 The image shows the results of HE staining of the heart of rat No. 4106 in the high-dose group under a 12.5x microscope. Detailed Implementation

[0036] The present invention will be further described below through specific embodiments, but this is not a limitation of the present invention. Those skilled in the art can make various modifications or improvements based on the basic idea of ​​the present invention, but as long as they do not depart from the basic idea of ​​the present invention, they are all within the scope of the present invention.

[0037] Example 1: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0038] The traditional Chinese medicine composition for treating anxiety and depression includes the following raw materials and their mass proportions: 1000g of Acanthopanax senticosus and 25g of Schisandra chinensis.

[0039] The preparation method of the traditional Chinese medicine composition for treating anxiety and depression includes the following steps:

[0040] (1) Mix Acanthopanax senticosus and Schisandra chinensis, grind them, and pass them through a 10-mesh sieve to make coarse powder;

[0041] (2) Add extractant 1 to the coarse powder and stir evenly. Extractant 1 is an ethanol aqueous solution with a volume fraction of 70%. The amount of extractant 1 added is 0.5 times the mass of the coarse powder. Wet and seal and let it stand for 1 hour to allow it to fully expand. Put the expanded coarse powder into the percolation tank, flatten it, and add extractant 2 at a flow rate of 2 mL / min. The extractant 2 is an ethanol aqueous solution with a volume fraction of 70%. After the air between the powder particles is discharged and ethanol liquid flows out, close the discharge valve, cover the filter cylinder, and soak for 18 hours.

[0042] (3) After soaking, open the discharge valve to perform percolation. While percolating, add extractant 3, which is an ethanol aqueous solution with a volume fraction of 70%. Control the percolation speed to be 2 mL / min per 1000g of medicinal material. Collect the filtrate and mix it with the ethanol solution discharged in step (2). Concentrate the mixture under vacuum at 40°C until the relative density at 60°C is 1.10 to obtain a concentrated solution. Dilute the obtained concentrated solution with twice the amount of water, let it stand, centrifuge and filter to obtain supernatant A.

[0043] (4) The supernatant A obtained in step (3) is passed through a D101 macroporous resin adsorption column with a diameter-to-height ratio of 1:2. Eluent 1 is added for elution. The eluent 1 is water, and the amount of eluent 1 added is 1 times the volume of supernatant A. The eluent is discarded, and eluent 2 is added for elution. The eluent 2 is a sodium hydroxide aqueous solution with a mass percentage of 0.1%. The amount of eluent 2 added is 1 times the volume of supernatant A. Eluent A is collected. The adsorption column is washed with water until neutral. Eluent 3 is added for elution. The eluent 3 is an ethanol aqueous solution with a volume fraction of 70%. The amount of eluent 3 added is 4 times the volume of D101 macroporous resin adsorption column. Eluent B is collected.

[0044] (5) Adjust the pH of the eluent A obtained in step (4) to neutral, centrifuge and filter, pass the resulting supernatant B through a polyamide resin adsorption column with a diameter-to-height ratio of 1:2, wash with water until salt-free, and then elute with eluent 4, which is an ethanol aqueous solution with a volume fraction of 70%. The amount of eluent 4 added is twice the mass of supernatant B. Collect the eluent C, concentrate the eluent C under vacuum at 40°C, concentrate it to a concentrate with a relative density of 1.20 at 60°C, and vacuum dry the concentrate at 50°C until the dry water content is 3% to obtain dry paste A; concentrate the eluent B obtained in step (4) under vacuum at 40°C until the relative density is 1.20 at 60°C to obtain a concentrate, and vacuum dry the concentrate at 50°C until the dry water content is 3% to obtain dry paste B;

[0045] (6) Mix dry paste A and dry paste B, pulverize them and pass them through a 100-mesh sieve to obtain a traditional Chinese medicine composition for treating anxiety and depression.

[0046] Example 2: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0047] The traditional Chinese medicine composition for treating anxiety and depression includes the following raw materials and their mass proportions: Acanthopanax senticosus 975g and Schisandra chinensis 50g.

[0048] The preparation method of the traditional Chinese medicine composition for treating anxiety and depression includes the following steps:

[0049] (1) Mix Acanthopanax senticosus and Schisandra chinensis, grind them, and pass them through a 30-mesh sieve to make coarse powder;

[0050] (2) Add extractant 1 to the coarse powder and stir evenly. Extractant 1 is an ethanol aqueous solution with a volume fraction of 90%. The amount of extractant 1 added is 1 times the mass of the coarse powder. Wet and seal and let it stand for 3 hours to allow it to fully expand. Put the expanded coarse powder into the percolation tank, flatten it, and add extractant 2 at a flow rate of 3 mL / min. The extractant 2 is an ethanol aqueous solution with a volume fraction of 90%. After the air between the powder particles is discharged and ethanol liquid flows out, close the discharge valve, cover the filter cylinder, and soak for 30 hours.

[0051] (3) After soaking, open the discharge valve to perform percolation. While percolating, add extractant 3, which is an ethanol aqueous solution with a volume fraction of 90%. Control the percolation speed to 3 mL / min per 1000g of medicinal material. Collect the filtrate and mix it with the ethanol solution discharged in step (2). Concentrate the mixture under vacuum at 80°C until the relative density at 60°C is 1.20 to obtain a concentrated solution. Dilute the obtained concentrated solution with 4 times the amount of water, let it stand, centrifuge and filter to obtain supernatant A.

[0052] (4) The supernatant A obtained in step (3) is passed through a D101 macroporous resin adsorption column with a diameter-to-height ratio of 1:4. Eluent 1 is added for elution. Eluent 1 is water. The amount of eluent 1 added is 3 times the volume of supernatant A. The eluent is discarded. Eluent 2 is added for elution. Eluent 2 is a sodium hydroxide aqueous solution with a mass percentage of 0.1%. The amount of eluent 2 added is 3 times the volume of supernatant A. Eluent A is collected. The adsorption column is washed with water until neutral. Eluent 3 is added for elution. Eluent 3 is an ethanol aqueous solution with a volume fraction of 90%. The amount of eluent 3 added is 6 times the volume of D101 macroporous resin adsorption column. Eluent B is collected.

[0053] (5) Adjust the pH of the eluent A obtained in step (4) to neutral, centrifuge and filter, pass the resulting supernatant B through a polyamide resin adsorption column with a diameter-to-height ratio of 1:4, wash with water until salt-free, and then elute with eluent 4, which is an ethanol aqueous solution with a volume fraction of 90%. The amount of eluent 4 added is 3 times the mass of supernatant B. Collect the eluent C, concentrate the eluent C under vacuum at 80°C, concentrate it to a concentrate with a relative density of 1.30 at 60°C, and vacuum dry the concentrate at 70°C until the dry water content is 3% to obtain dry paste A; concentrate the eluent B obtained in step (4) under vacuum at 80°C, concentrate it to a relative density of 1.30 at 60°C to obtain a concentrate, and vacuum dry the concentrate at 70°C until the dry water content is 3% to obtain dry paste B;

[0054] (6) Mix dry paste A and dry paste B, pulverize them and pass them through a 100-mesh sieve to obtain a traditional Chinese medicine composition for treating anxiety and depression.

[0055] Example 3: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0056] The traditional Chinese medicine composition for treating anxiety and depression includes the following raw materials and their mass proportions: Acanthopanax senticosus 975g and Schisandra chinensis 50g.

[0057] The preparation method of the traditional Chinese medicine composition for treating anxiety and depression includes the following steps:

[0058] (1) Mix Acanthopanax senticosus and Schisandra chinensis, grind them, and pass them through a 20-mesh sieve to make coarse powder;

[0059] (2) Add extractant 1 to the coarse powder and stir evenly. Extractant 1 is an ethanol aqueous solution with a volume fraction of 80%. The amount of extractant 1 added is 0.8 times the mass of the coarse powder. Wet and seal and let it stand for 2 hours to allow it to fully expand. Put the expanded coarse powder into the percolation tank, flatten it, and add extractant 2 at a flow rate of 2.5 mL / min. The extractant 2 is an ethanol aqueous solution with a volume fraction of 80%. After the air between the powder particles is discharged and ethanol liquid flows out, close the discharge valve, cover the filter cylinder, and soak for 24 hours.

[0060] (3) After soaking, open the discharge valve to perform percolation. While percolating, add extractant 3, which is an 80% volume fraction ethanol aqueous solution. Control the percolation speed to be 2.5 mL / min per 1000g of medicinal material. Collect the filtrate and mix it with the ethanol solution discharged in step (2). Concentrate the mixture under vacuum at 60°C until the relative density at 60°C is 1.15 to obtain a concentrated solution. Dilute the obtained concentrated solution with 3 times the amount of water, let it stand, centrifuge and filter to obtain supernatant A.

[0061] (4) The supernatant A obtained in step (3) is passed through a D101 macroporous resin adsorption column with a diameter-to-height ratio of 1:3 and a macroporous resin weight of 495g. Eluent 1 is added for elution. Eluent 1 is water, and the amount of eluent 1 added is twice the volume of supernatant A. The eluent is discarded, and eluent 2 is added for elution. Eluent 2 is a sodium hydroxide aqueous solution with a mass percentage of 0.1%. The amount of eluent 2 added is twice the volume of supernatant A. Eluent A is collected. The adsorption column is washed with water until neutral, and eluent 3 is added for elution. Eluent 3 is an ethanol aqueous solution with a volume fraction of 80%. The amount of eluent 3 added is five times the volume of D101 macroporous resin adsorption column. Eluent B is collected.

[0062] (5) Adjust the pH of the eluent A obtained in step (4) to neutral, centrifuge and filter, pass the resulting supernatant B through a polyamide resin adsorption column with a diameter-to-height ratio of 1:3, wash with water until salt-free, and then elute with eluent 4, which is an 80% volume fraction ethanol aqueous solution. The amount of eluent 4 added is 2.5 times the mass of supernatant B. Collect the eluent C, concentrate the eluent C under vacuum at 60°C, concentrate it to a concentration with a relative density of 1.25 at 60°C, and vacuum dry the concentration at 60°C until the dry water content is 3% to obtain dry paste A; concentrate the eluent B obtained in step (4) under vacuum at 60°C until the relative density at 60°C is 1.25 to obtain a concentration, and vacuum dry the concentration at 60°C until the dry water content is 3% to obtain dry paste B;

[0063] (6) Mix dry paste A and dry paste B, pulverize them and pass them through a 100-mesh sieve to obtain a traditional Chinese medicine composition for treating anxiety and depression.

[0064] Example 4: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0065] The composition and preparation method of the traditional Chinese medicine composition for treating anxiety and depression are similar to those in Example 3.

[0066] The difference from Example 3 is that the extractant 1 in step (1) is an aqueous ethanol solution with a volume fraction of 70%.

[0067] Example 5: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0068] The composition and preparation method of the traditional Chinese medicine composition for treating anxiety and depression are similar to those in Example 3.

[0069] The difference from Example 3 is that the extractant 1 in step (1) is an aqueous ethanol solution with a volume fraction of 90%.

[0070] Example 6: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0071] The composition and preparation method of the traditional Chinese medicine composition for treating anxiety and depression are similar to those in Example 3. The difference from Example 3 is that the eluent 3 and eluent 4 are 70% ethanol aqueous solutions by volume.

[0072] Example 7: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0073] The composition and preparation method of the traditional Chinese medicine composition for treating anxiety and depression are similar to those in Example 3. The difference from Example 3 is that the eluent 3 and eluent 4 are ethanol aqueous solutions with a volume fraction of 90%.

[0074] Example 8: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0075] The traditional Chinese medicine composition for treating anxiety and depression includes the following raw materials and their mass proportions: 1000g of Acanthopanax senticosus and 25g of Schisandra chinensis.

[0076] The preparation method of the traditional Chinese medicine composition for treating anxiety and depression is similar to that in Example 3.

[0077] Example 9: A traditional Chinese medicine composition for treating anxiety and depression and its preparation method.

[0078] The traditional Chinese medicine composition for treating anxiety and depression includes the following raw materials and their mass proportions: 925g of Acanthopanax senticosus and 100g of Schisandra chinensis.

[0079] The preparation method of the traditional Chinese medicine composition for treating anxiety and depression is similar to that in Example 3.

[0080] Example 10: A traditional Chinese medicine preparation for treating anxiety and depression.

[0081] The traditional Chinese medicine composition for treating anxiety and depression obtained in Example 3 was mixed evenly with dextrin at a mass ratio of 1:2. An 80% ethanol aqueous solution was added to the mixed mixture for granulation. The amount of ethanol aqueous solution added was 25% of the mass of the mixture. The mixture was dried and then fully mixed to obtain the final product.

[0082] Example 11: A traditional Chinese medicine preparation for treating anxiety and depression.

[0083] The traditional Chinese medicine composition for treating anxiety and depression obtained in Example 3 was mixed evenly with starch at a mass ratio of 1:3. An 80% ethanol aqueous solution was added to the mixed mixture for granulation. The amount of ethanol aqueous solution added was 30% of the mass of the mixture. The mixture was dried, mixed thoroughly, and filled into No. 1 capsules to obtain the final product.

[0084] Example 12: A traditional Chinese medicine preparation for treating anxiety and depression.

[0085] The traditional Chinese medicine composition for treating anxiety and depression obtained in Example 3 was mixed evenly with starch at a mass ratio of 1:4. An 80% ethanol aqueous solution was added to the mixed mixture for granulation. The amount of ethanol aqueous solution added was 40% of the mass of the mixture. The mixture was dried, mixed thoroughly, and compressed into tablets to obtain the final product.

[0086] Comparative Example 1: A traditional Chinese medicine composition and its preparation method

[0087] The composition and preparation method of the traditional Chinese medicine composition are similar to those in Example 3. The difference is that the extractant 1 in step (1) is an aqueous ethanol solution with a volume fraction of 60%.

[0088] Comparative Example 2: A traditional Chinese medicine composition and its preparation method

[0089] The composition and preparation method of the traditional Chinese medicine composition are similar to those in Example 3. The difference from Example 3 is that in step (4), the supernatant A obtained in step (3) is passed through an AB-8 macroporous resin adsorption column with a diameter-to-height ratio of 1:3.

[0090] Comparative Example 3: A traditional Chinese medicine composition and its preparation method

[0091] The composition and preparation method of the traditional Chinese medicine composition are similar to those in Example 3. The difference from Example 3 is that the eluent 3 and eluent 4 are ethanol aqueous solutions with a volume fraction of 60%.

[0092] Comparative Example 4: A traditional Chinese medicine composition and its preparation method

[0093] The composition and preparation method of the traditional Chinese medicine composition are similar to those in Example 3. The difference from Example 3 is that in step (4), the supernatant A obtained in step (3) is passed through a D101 macroporous resin adsorption column with a diameter-to-height ratio of 1:1.

[0094] Example 1: Comparison of dry extract yield, eleutheroside E transfer rate, and schisandrol A transfer rate

[0095] Take the supernatant A obtained in step (3) of Examples 3-5 and Comparative Example 1, and calculate the transfer rate of Acanthopanax senticosin E and the transfer rate of Schisandrin A. Acanthopanax senticosin E transfer rate % = Total amount of Acanthopanax senticosin E in supernatant A / Total amount of Acanthopanax senticosin E in medicinal material × 100%;

[0096] Schisandrin A transfer rate % = Total amount of schisandrin A in supernatant A / Total amount of schisandrin A in medicinal material × 100%.

[0097] Table 1 shows a comparison of the transfer rates of eleutheroside E and schisandrol A in supernatant A.

[0098] Table 1. Comparison of the transfer rates of eleutheroside E and schisandrol A in supernatant A

[0099] Example 3 81.20 80.52 Example 4 73.36 75.09 Example 5 80.60 79.27 Comparative Example 1 68.75 65.31

[0100] As shown in Table 1, the supernatant A obtained using Examples 3, 4, and 5 of this invention exhibits high transfer rates of eleutheroside E and schisandrol A. Among these, the supernatant A obtained in Example 3 shows the highest transfer rates of eleutheroside E and schisandrol A, making it the optimal embodiment of this invention. The transfer rates of eleutheroside E and schisandrol A in the supernatant A obtained in Example 3 are significantly better than those in Comparative Example 1. This indicates that the volume fraction of the ethanol aqueous solution in extractant 1 has a significant impact on the transfer rates of eleutheroside E and schisandrol A in supernatant A. Only by controlling the volume fraction of the ethanol aqueous solution within a suitable range can a supernatant with high transfer rates of eleutheroside E and schisandrol A be obtained.

[0101] The eluents obtained from washing with water in step (4) of Examples 3, Comparative Examples 2, and 4 were used to calculate the peak areas of eleutheroside E and schisandrol A in the water-eluted fraction. The eluents obtained from washing with alcohol in step (4) of Examples 3, Comparative Examples 2, and 4 were used to calculate the peak areas of eleutheroside E in the alcohol-eluted fraction and schisandrol A in the alcohol-eluted fraction. A comparison table of the peak areas of eleutheroside E and schisandrol A in the water-eluted fraction and the alcohol-eluted fraction is shown in Table 2.

[0102] Table 2. Comparison of peak areas of eleutheroside E and schisandrol M in water-eluted and alcohol-eluted fractions.

[0103] Area of ​​Acanthopanax senticosin E peak in water-eluting region 0.03 2.35 1.67 Area of ​​schisandrin M peak in water-eluted fraction 0.00 1.46 1.13 Area of ​​Acanthopanax senticosin E peak in alcohol elution region 268.99 150.86 174.83 Schisandrin M peak area in alcohol elution fraction 36.77 18.65 22.14

[0104] As shown in Table 2, the peak areas of eleutheroside E and schisandrol A in the water-eluted portion of the eluent obtained by washing with water in step (4) of Example 3 of the present invention are significantly lower than those in Comparative Examples 2 and 4. This indicates that the content of eleutheroside E in the eluent discarded after washing with water in step (4) of the method of the present invention is low, and the eluent discarded after washing with water does not contain schisandrol A. The peak areas of eleutheroside E and schisandrol A in the alcohol-eluted portion of the eluent obtained by washing with alcohol in step (4) of Example 3 of the present invention are significantly higher than those in Comparative Examples 2 and 4. This indicates that the content of eleutheroside E and schisandrol A in the eluent obtained by washing with alcohol in step (4) of the method of the present invention is high. Therefore, it can be seen that the type of macroporous resin and the diameter-to-height ratio of the macroporous resin adsorption column have a great influence on the separation of eleutheroside E and schisandrol A, the effective components in Acanthopanax senticosus and Schisandra chinensis. The method of this invention can increase the content of the active ingredients eleutherococcus senticosus glycoside E and schisandrol A in Acanthopanax senticosus and Schisandra chinensis.

[0105] Dry extracts A and B obtained from Examples 3, 6, 7, 8, 9, and Comparative Example 3 were used to calculate the yield of the dry extracts, the transfer rate of eleutheroside E, and the transfer rate of schisandrol A. A comparison table of the yield of the dry extracts, the transfer rate of eleutheroside E, and the transfer rate of schisandrol A is shown in Table 3.

[0106] Dry extract yield % = (weight of dry extract A + weight of dry extract B) / (weight of Acanthopanax senticosus + weight of Schisandra chinensis) × 100%;

[0107] The percentage of eleutheroside E transfer is calculated as follows: (Total amount of eleutheroside E in the herbal composition / Total amount of eleutheroside E in the herbal material) × 100%.

[0108] Schisandrol A transfer rate % = Total amount of schisandrol A in the Chinese herbal composition / Total amount of schisandrol A in the medicinal material × 100%.

[0109] Table 3 Comparison of dry extract yield, eleutheroside E transfer rate, and schisandrol A transfer rate

[0110]

[0111]

[0112] As shown in Table 3, the dry extract yield, eleutheroside E transfer rate, and schisandrol A transfer rate obtained in Examples 3, 6, 7, 8, and 9 of this invention are relatively high. Among them, the dry extract yield, eleutheroside E transfer rate, and schisandrol A transfer rate obtained in Example 3 are the highest, making it the optimal example of this invention. Compared with Comparative Example 3, the dry extract yield, eleutheroside E transfer rate, and schisandrol A transfer rate obtained in Example 3 of this invention are even higher.

[0113] Experimental Example 2: Efficacy Test

[0114] ◇Animal Grouping

[0115] After acclimatizing for one week, 180 SD rats were randomly divided into six groups: a normal control group (n=30), a model group (n=30), a group using a traditional Chinese medicine composition for treating anxiety and depression (n=30), a group using fluoxetine hydrochloride dispersible tablets (n=30), a control group 2 (n=30), and a control group 4 (n=30). The normal control group was fed normally without modeling, while the model group, the group using the traditional Chinese medicine composition for treating anxiety and depression, the group using fluoxetine hydrochloride dispersible tablets, the control group 2, and the control group 4 were used to establish a depression model.

[0116] ◇Establishment of a depression model

[0117] A depression model was established by administering CUMS stimulation to the model group, the traditional Chinese medicine composition group for treating anxiety and depression, the comparative group 2, the comparative group 4, and the fluoxetine hydrochloride dispersible tablet group. The modeling process lasted for 42 days. The CUMS stimulation method is shown in Table 4 below:

[0118] Table 4 CUMS Stimulation Methods

[0119]

[0120]

[0121] rat administration

[0122] Normal control group (30 animals): No stimulation was given during the experiment. During the administration of medication, 0.9% physiological saline was administered at a dose of 1 mL / 100g once a day. This was done continuously by gavage for 14 days.

[0123] Model group (30 rats): Rats were subjected to group housing and chronic, unpredictable, moderate-intensity stimulation, followed by administration of 0.9% saline at 1 mL / 100g once daily for 14 consecutive days.

[0124] The traditional Chinese medicine composition for treating anxiety and depression (30 rats): Rats were first housed in groups and subjected to chronic, unpredictable, moderate-intensity stimulation. The traditional Chinese medicine composition for treating anxiety and depression obtained in Example 3 was diluted with water to a solution of 0.4 g / mL, and then administered by gavage at a dose of 1 mL / 100 g once daily for 14 consecutive days.

[0125] The administration methods for Comparative Examples 2 and 4 were similar to those for the traditional Chinese medicine composition groups used to treat anxiety and depression. The difference was that the traditional Chinese medicine compositions obtained in Comparative Examples 2 and 4 were diluted with water to a solution of 0.4 g / mL, and then administered by gavage at a dose of 1 mL / 100 g once a day for 14 consecutive days.

[0126] The group of rats receiving fluoxetine hydrochloride dispersible tablets (n=30) was first housed in groups and subjected to chronic, unpredictable, moderate-intensity stimulation. Fluoxetine hydrochloride dispersible tablets were ground into powder and prepared into a 4 g / mL solution, which was then administered by gavage at a dose of 1 mL / 100 g once a day for 14 consecutive days.

[0127] ◇Behavioral testing

[0128] All behavioral experiments were conducted during the rats' light exposure period. Before each experiment, the rats were placed in the behavioral room for 2 hours to acclimatize. During each experiment, after each rat was tested, it was wiped with 70% alcohol to prevent the odor from affecting the next rat. Each rat was held by hand for a few minutes daily before the behavioral experiments to allow it to acclimatize and prevent fear during the experiments from affecting the results; this acclimatization period lasted at least 4 days.

[0129] B1 Sugar Water Preference Experiment

[0130] Rats were acclimatized to two water bottles in their cages for two days. On the first day, both bottles contained ordinary drinking water. On the second day, both bottles contained 1% sucrose solution (to eliminate the influence of fear of novelty). During this process, the animals had free access to food and water. After the acclimatization period, a sucrose preference test was conducted. At 12:00 on the day of the test, both bottles were removed. At 19:00, the two bottles were replaced with the new drinking water and 1% sucrose solution, respectively, until 22:00, and the amount of sucrose solution consumed was recorded. During the recording process, at 20:30, the positions of the sucrose solution and the ordinary water were changed to avoid positional preference. Both bottles were weighed before and after the test, and the sucrose preference index was calculated as follows: Sucrose preference index (%) = (Sucrose solution consumption / (Sucrose solution + Ordinary water consumption)) × 100%.

[0131] B2 Open Field Test

[0132] An open field test was conducted, recording the movement behavior of rats in an open field (60×60×45cm) for 10 minutes. The experiment was carried out under dim lighting, with no shadows in the open field. The time the rats remained stationary in the open field was recorded to analyze their motor function and anxiety-like mood disorders.

[0133] B3 Elevated Cross Maze Experiment

[0134] An elevated cross maze experiment was conducted, recording the time and frequency of rats entering the open (50×10cm) and closed (50×10cm) arms of the maze. The recording time was 10 minutes. During the experiment, the rats were placed in the central area (10×10cm), with their heads facing the open arms upon release. The experiment was conducted under dim lighting, ensuring no shadows in the closed arms. If a rat fell from the elevated maze during the experiment, that rat was removed from the experiment. The percentage of times rats entered the open arms was calculated as: (Number of times rats entered the open arms / (Number of times rats entered the open arms + Number of times rats entered the closed arms)) × 100%.

[0135] The results of the efficacy test are shown in Table 5.

[0136] B4 ELISA method for detecting TNF-α, IDO, 5-HT and DA levels in hippocampal tissue

[0137] ◇After the behavioral studies, tissue samples were collected, and the levels of TNF-α (tumor necrosis factor-α), IDO (indoleamine 2,3-dioxygenase), 5-HT (serotonin), and DA (dopamine) in rat hippocampal tissue were detected using ELISA. Fresh hippocampal tissue from each group of rats was collected, homogenized on ice, and centrifuged at 4500 rpm for 10 min. The supernatant was collected, and the levels of TNF-α, IDO, 5-HT, and DA in the hippocampal tissue of each group of rats were detected using enzyme-linked immunosorbent assay (ELISA), strictly following the instructions of the kit. The comparison of TNF-α, IDO, 5-HT, and DA levels in hippocampal tissue is shown in Table 6.

[0138] Table 5. Results of the efficacy test

[0139]

[0140]

[0141] Table 5 shows that in the saccharide preference experiment, the saccharide preference index of the model group rats was significantly lower than that of the normal control group, indicating that the depression model was successfully established. The saccharide preference index of the fluoxetine hydrochloride dispersible tablet group and the traditional Chinese medicine combination group for treating anxiety and depression was close to that of the normal control group. The saccharide preference index of the traditional Chinese medicine combination group for treating anxiety and depression was significantly higher than that of comparative groups 2 and 4. In the open field test, the immobility time of the model group rats in the open field was significantly lower than that of the normal control group, indicating that the depression model was successfully established. The immobility time of the fluoxetine hydrochloride dispersible tablet group in the open field was close to that of the model group. The immobility time of the traditional Chinese medicine combination group for treating anxiety and depression was close to that of the normal control group. The immobility time of the traditional Chinese medicine combination group for treating anxiety and depression in the open field was significantly higher than that of comparative groups 2 and 4. In the elevated cross maze experiment, the percentage of rats in the model group entering the open arm was significantly lower than that in the normal control group, indicating that the depression model was successfully established. The percentage of rats entering the open arm in the fluoxetine hydrochloride dispersible tablet group was similar to that in the model group. The percentage of rats entering the open arm in the traditional Chinese medicine combination group for treating anxiety and depression was similar to that in the normal control group. However, the percentage of rats entering the open arm in the traditional Chinese medicine combination group for treating anxiety and depression was significantly higher than that in comparative group 2 and comparative group 4.

[0142] In summary, the traditional Chinese medicine composition provided by this invention is more effective than the Western medicine fluoxetine hydrochloride dispersible tablets in treating anxiety and depression. Furthermore, since the traditional Chinese medicine composition of this invention is a pure traditional Chinese medicine combination with almost no side effects, it can replace chemical drugs, other traditional Chinese medicine preparations, or traditional Chinese medicine decoctions, which will greatly improve the quality of life of patients with anxiety and depression.

[0143] Table 6 Comparison of TNF-α, IDO, 5-HT, and DA levels in hippocampal tissue

[0144]

[0145] As shown in Table 6, the TNF-α and IDO levels in the traditional Chinese medicine composition group, the fluoxetine hydrochloride dispersible tablet group, and the normal control group were all lower than those in the model group. The TNF-α and IDO levels in the traditional Chinese medicine composition group were lower than those in the fluoxetine hydrochloride dispersible tablet group but higher than those in the normal control group. The TNF-α and IDO levels in the traditional Chinese medicine composition group were lower than those in comparative groups 2 and 4. The 5-HT and DA levels in the traditional Chinese medicine composition group, the fluoxetine hydrochloride dispersible tablet group, and the normal control group were all higher than those in the model group. The 5-HT and DA levels in the traditional Chinese medicine composition group were higher than those in the fluoxetine hydrochloride dispersible tablet group but lower than those in the normal control group. The 5-HT and DA levels in the traditional Chinese medicine composition group were higher than those in comparative groups 2 and 4. Therefore, the efficacy of the traditional Chinese medicine composition of this invention in treating anxiety and depression is superior to that of the fluoxetine hydrochloride dispersible tablet group.

[0146] Experimental Example 3: Toxicological Experiment

[0147] ◇Group: SD rats, 4 groups (normal control group and low, medium and high dose groups), a total of 120 rats, 30 rats in each group, half male and half female.

[0148] ◇Administration method / frequency:

[0149] Formulation of the composition

[0150] Preparation method: Calculate the total dosage and required solution volume based on the weight of the rats in the treatment group. Take an appropriate amount of the traditional Chinese medicine composition obtained in Example 3 and dilute it with purified water to prepare the required concentration.

[0151] Preparation frequency: Prepare once a day.

[0152] Precautions for drug delivery preparations: Shake well before use.

[0153] Table 7. Administration method / frequency for each group

[0154]

[0155] The normal control group was given drinking water, while the low, medium, and high dose groups were given the traditional Chinese medicine composition.

[0156] The medication was administered orally via gavage for 28 days, once daily and 7 times per week, with a recovery period of 28 days.

[0157] ◇Clinical symptom observation:

[0158] General clinical symptoms: During the adaptation period, administer once daily; during the 28-day administration period, administer once before and once after administration; during the 28-day recovery period, weigh yourself once daily; once before administration, and once or twice a week after administration (including the recovery period), and before dissection.

[0159] Food intake: once a week after administration (including the recovery period).

[0160] Ophthalmological examination: at the end of the medication period and during the recovery period.

[0161] Clinical examination: At the end of the drug administration period and before necropsy during the recovery period, routine hematological, coagulation, blood biochemistry, and urinalysis tests were performed. Hematological test indicators are shown in Table 8, coagulation test indicators are shown in Table 9, blood biochemistry test indicators are shown in Table 10, and urinalysis test indicators are shown in Table 11. The experimental results of each group of rats after drug administration are shown in Tables 12-31.

[0162] (1) Hematological testing

[0163] Take approximately 1 mL of blood, anticoagulate with EDTA-K2, and test the specified indicators.

[0164] Table 8 Hematological Detection Indicators

[0165] Red blood cell count (RBC) <![CDATA[10 6 / μL]]> Hemoglobin (HGB) g / dL Hematocrit (HCT) ％ Mean corpuscular volume (MCV) fL Mean corpuscular hemoglobin (MCH) pg Mean corpuscular hemoglobin concentration (MCHC) g / dl Reticulocyte percentage (%Retic) ％ White blood cell count (WBC) <![CDATA[10 3 / μL]]> Neutrophil percentage (%Neut) ％ Lymphocyte percentage (%Lymph) ％ Mononuclear cell percentage (%Mono) ％ Percentage of eosinophils (%Eos) ％ Basophil percentage (%Baso) ％ Platelet count (PLT) <![CDATA[10 3 / μL]]>

[0166] (2) Coagulation time index detection

[0167] Take about 2 mL of blood, anticoagulate with sodium citrate, centrifuge (3000 rpm, 10 min), and then take plasma to measure the specified indicators.

[0168] Table 9 Coagulation Test Indicators

[0169] Prothrombin time (PT) s Activated partial thromboplastin time (APTT) s

[0170] (3) Blood biochemistry tests

[0171] Take about 3 mL of blood, put it in a test tube, centrifuge (3000 rpm, 10 min), and then collect the serum for testing the specified indicators.

[0172] Table 10 Blood Biochemical Detection Indicators

[0173] Aspartate aminotransferase (AST) IU / L IFCC method Alanine aminotransferase (ALT) IU / L IFCC method Alkaline phosphatase (ALP) IU / L IFCC method Gamma-glutamyl transferase (GGT) IU / L IFCC method (BUN) mmol / L Urease, glutamate dehydrogenase method Creatinine (CREA) μmol / L Creatinase, HMMPS method Total protein (TP) g / L Biuret method Albumin (ALB) g / L Bromocresol Green Method Glucose (GLU) mmol / L Hexokinase, glucose-6-phosphate Total bilirubin (TBIL) μmol / L Vanadic acid oxidation method Total cholesterol (TC) mmol / L cholesterol oxidase, HMMPS Triglycerides (TG) mmol / L GPO, HMMPS method, de-free glycerol method Creatine kinase (CK) IU / L IFCC method <![CDATA[Na + ]]> mmol / L Ion-selective electrode method <![CDATA[K + ]]> mmol / L Ion-selective electrode method <![CDATA[Cl - ]]> mmol / L Ion-selective electrode method

[0174] (4) Urine analysis

[0175] Urine collection: At the end of the drug administration period and the end of the recovery period, the animals to be dissected were placed in metabolic cages, fasted but allowed water overnight, and urine was collected for about 13 hours.

[0176] Urine analysis: After collecting urine, specific indicators are tested.

[0177] Table 11 Urine Analysis Detection Indicators

[0178] color Light source refraction method Specific gravity (SG) Light source refraction method pH value pH indicator method Proteinuria (PRO) Protein error method for pH indicators Glucose in urine (GLU) Enzymatic methods (GOD, POD methods) Urinary ketone bodies (KET) Alkaline sodium nitroprusside method White blood cells (LEU) Leukocyte esterase activity assay BLD (Blank Blood in the Eye) Peroxidase-like activity of hemoglobin (Hb) Urobilinogen (URO) Diazo coupling method Urinary bilirubin (BIL) Diazo coupling method Nitrite (NIT) Oil method

[0179] Table 12 Effects of repeated-dose toxicity studies of the oral-oral-gavage combination on hematological parameters in rats (end of administration; male; n = 10; x ± s)

[0180]

[0181]

[0182] Table 13 Effects of repeated oral-oral gavage toxicity tests on hematology in rats (end of administration; female; n = 10; x ± s)

[0183] WBC (×10 / μL) 6.07±2.11 6.34±1.75 5.98±1.51 6.43±1.40 RBC (×10 / μL) 7.40±0.45 7.30±0.26 7.03±0.47 7.24±0.22 HGB(g / dL) 15.7±0.7 15.9±0.6 15.5±0.8 15.6±0.4 HCT (%) 43.4±2.0 43.8±1.7 42.8±2.1 43.0±1.0 MCV(fL) 58.7±2.0 60.1±1.4 60.9±2.0* 59.4±0.9 MCH(pg) 21.2±0.7 21.8±0.5 22.1±0.9* 21.6±0.3 MCHC (g / dl) 36.1±0.3 36.2±0.4 36.3±0.6 36.3±0.5 PLT (×10 / μL) 1,107±149 1,127±93 1,120±439 1,139±135 Retic (%) 2.43±0.89 2.90±0.36 2.66±0.59 2.78±0.48 Neut(%) 11.8±5.5 11.3±3.4 9.8±2.9 11.7±3.3 Lymph (%) 85.4±5.5 85.3±3.8 87.3±3.2 85.4±3.2 Mono (%) 1.3±0.2 1.5±0.7 1.3±0.4 1.3±0.2 Eos(%) 0.8±0.3 1.0±0.6 0.7±0.3 0.9±0.3 Baso (%) 0.2±0.1 0.2±0.1 0.2±0.1 0.2±0.1

[0184] Note: *P<0.05 compared with the normal control group.

[0185] Table 14 Effects of repeated oral-oral gavage toxicity tests on hematology in rats (recovery period; males; n=5; x±s)

[0186]

[0187]

[0188] Table 15 Effects of repeated oral-oral gavage toxicity tests on hematology in rats (recovery period; female; n=5; x±s)

[0189] WBC (×10 / μL) 5.33±2.28 5.97±1.42 5.14±0.53 5.77±0.79 RBC (×10 / μL) 7.04±0.30 7.06±0.24 6.98±0.33 7.08±0.11 HGB(g / dL) 16.0±0.6 15.4±0.3 15.6±0.5 15.8±0.4 HCT (%) 40.4±1.2 39.8±0.8 40.7±1.7 40.7±0.6 MCV(fL) 57.4±1.1 56.5±0.8 58.3±1.4 57.5±1.0 MCH(pg) 22.6±0.4 21.8±0.5 22.4±0.6 22.4±0.6 MCHC (g / dl) 39.5±0.3 38.6±0.5* 38.4±0.4** 38.9±0.5 PLT (×10 / μL) 1,001±51 982±109 1,035±123 949±35 Retic (%) 3.39±0.56 3.41±0.34 3.62±0.66 3.16±0.46 Neut(%) 14.9±3.9 14.2±6.5 12.3±5.7 14.6±8.1 Lymph (%) 80.6±3.8 82.4±6.2 84.8±6.0 82.3±8.5 Mono (%) 2.2±0.3 1.6±0.5 1.4±0.4* 1.5±0.6 Eos(%) 1.4±0.5 0.9±0.3 0.9±0.4 0.8±0.3 Baso (%) 0.2±0.1 0.2±0.1 0.2±0.0 0.2±0.1

[0190] Note: Compared with the normal control group, *P<0.05, **P<0.01.

[0191] Table 16 Effects of the oral gavage composition on male coagulation parameters in a 1-month repeated-dose toxicity test in rats (end of administration; n = 10; x ± s)

[0192] PT(s) 9.3±0.5 9.4±0.5 9.2±0.4 9.2±0.4 INR 0.81±0.4 0.81±0.4 0.79±0.03 0.79±0.03 APTT(s) 1.57±1.2 16.4±0.9 16.2±1.2 16.1±0.8

[0193] Table 17 Effects of the oral gavage composition on female coagulation parameters in a 1-month repeated-dose toxicity test in rats (end of administration; n = 10; x ± s)

[0194] PT(s) 7.4±0.3 7.3±0.2 7.4±0.2 7.2±0.2 INR 0.65±0.02 0.64±0.01 0.65±0.02 0.64±0.02 APTT(s) 14.0±0.9 14.5±0.6 14.0±0.9 13.9±0.6

[0195] Table 18 Effects of oral gavage composition on male coagulation parameters in rats after 1 month of repeated administration (recovery period; n = 5; x ± s)

[0196] PT(s) 8.5±0.4 8.7±0.4 8.9±0.5 8.7±0.6 INR 0.74±0.03 0.76±0.03 0.77±0.04 0.75±0.05 APTT(s) 16.4±1.4 16.4±0.7 16.2±1.1 16.2±1.3

[0197] Table 19 Effects of oral gavage composition on female coagulation parameters in a 1-month repeated-dose toxicity test in rats (recovery period; n=5; x±s)

[0198] PT(s) 7.4±0.4 7.6±0.1 7.8±0.4 7.5±0.2 INR 0.65±0.03 0.67±0.01 0.68±0.03 0.66±0.02 APTT(s) 14.1±0.6 15.4±0.8* 15.1±0.7 15.6±0.5**

[0199] Note: Compared with the normal control group, *P<0.05, **P<0.01.

[0200] Table 20 Blood biochemical tests in the 1-month repeated-dose toxicity test of the oral-oral gavage composition in rats (end of administration; male; n=10; x±s)

[0201]

[0202]

[0203] Note: *P<0.05 compared with the normal control group.

[0204] Table 21 Blood biochemical tests in the 1-month repeated-dose toxicity test of the oral-oral gavage composition in rats (end of administration; female; n=10; x±s)

[0205]

[0206]

[0207] Note: Compared with the normal control group, *P<0.05, **P<0.01.

[0208] Table 22 Blood biochemical tests in the 1-month repeated-dose toxicity test of the oral-oral gavage composition in rats (recovery period; male; n=5; x±s)

[0209]

[0210]

[0211] Note: *P<0.05 compared with the normal control group.

[0212] Table 23 Blood biochemical tests in the repeated-dose toxicity test of the oral-oral-gavage composition in rats after 1 month (recovery period; female; n=5; x±s)

[0213] ALT (IU / L) 27.0±4.8 30.4±6.5 25.6±2.7 26.0±2.5 AST (IU / L) 123.2±17.8 116.7±11.2 106.9±9.7 100.9±21.6 TP (g / L) 64.8±3.4 65.8±1.7 63.7±1.2 66.5±5.4 ALB (g / L) 28.9±2.1 28.1±0.9 28.6±0.7 29.3±3.1 TBIL (μmol / L) 0.9±0.2 1.0±0.3 1.1±0.2 0.9±0.2 ALP (IU / L) 102.7±13.6 95.9±11.2 101.3±28.0 85.7±23.5 GGT (IU / L) 0.8±0.2 0.9±0.3 1.1±0.4 0.9±0.2 GLU (mmol / L) 7.26±1.10 6.99±0.79 7.30±0.58 8.23±1.44 BUN (mmol / L) 6.70±1.40 7.76±0.97 8.17±1.61 7.69±0.94 CREA (μmol / L) 29±6 28±1 30±6 32±6 TC (mmol / L) 1.69±0.58 1.64±0.17 1.66±0.25 1.42±0.48 TG (mmol / L) 0.17±0.08 0.18±0.06 0.14±0.05 0.12±0.04 CK(IU / L) 850.7±181.7 895.0±333.4 639.2±66.9 583.5±199.2 K+ (mmol / L) 4.18±0.27 4.30±0.37 4.22±0.20 4.19±0.15 Na+ (mmol / L) 143.2±1.0 143.6±1.4 143.6±0.3 143.7±0.7 Cl- (mmol / L) 107.0±1.2 105.7±1.6 107.1±0.8 107.8±1.5

[0214] Table 24. Urine test results of the oral gavage composition in rats after one month of repeated administration (end of administration period, males).

[0215]

[0216]

[0217]

[0218] Note: A urinalysis result of (-) indicates a negative result, a result of (+) indicates a positive result, a result of (+-) indicates a weak positive result, and a result of (1+) indicates that the concentration of the substance is slightly higher than the normal value.

[0219] Table 25. Urine test results of the oral gavage composition in rats after one month of repeated administration (end of administration period, males).

[0220]

[0221]

[0222] Table 26. Urine toxicity test results of the oral gavage composition in rats after 1 month of repeated administration (end of administration period, females).

[0223]

[0224]

[0225] Note: A urinalysis result of (-) indicates a negative result, a result of (+) indicates a positive result, and a result of (+-) indicates a weak positive result.

[0226] Table 27. Urine test results of the oral gavage composition in rats after one month of repeated administration (end of administration period, females).

[0227] normal control group 1.018±0.006 6.7±0.3 low-dose group 1.015±0.009 6.8±0.3 medium dose group 1.015±0.008 6.8±0.4 High-dose group 1.018±0.005 6.9±0.2

[0228] Table 28. Urine toxicity test results of the oral gavage composition in rats after 1 month of repeated administration (recovery period, males).

[0229]

[0230]

[0231] Note: A urinalysis result of (-) indicates a negative result, a result of (+) indicates a positive result, a result of (+-) indicates a weak positive result, and a result of (1+) indicates that the concentration of the substance is slightly higher than the normal value.

[0232] Table 29. Urine toxicity test results of the oral gavage composition in rats after 1 month of repeated administration (recovery period, males).

[0233] normal control group 1.019±0.009 7.0±0.0 low-dose group 1.020±0.005 7.1±0.2 medium dose group 1.018±0.005 6.9±0.2 High-dose group 1.022±0.003 7.2±0.3

[0234] Table 30. Urine test results of repeated-dose toxicity test of the oral-oral-gavage composition in rats after 1 month (recovery period, female).

[0235]

[0236]

[0237] Note: A urinalysis result of (-) indicates a negative result, a result of (+) indicates a positive result, a result of (+-) indicates a weak positive result, and a result of (1+) indicates that the concentration of the substance is slightly higher than the normal value.

[0238] Table 31. Urine toxicity test results of rats after one month of repeated oral gavage administration of the composition (recovery period, female).

[0239] normal control group 1.016±0.008 6.9±0.2 low-dose group 1.021±0.007 6.8±0.4 medium dose group 1.021±0.012 6.9±0.2 High-dose group 1.019±0.007 6.7±0.4

[0240] The following conclusions can be drawn from the data in Table 12-31 above:

[0241] As shown in Tables 12-15, in terms of hematology, at the end of the administration period, the MCV and MCH of female rats in the medium-dose group were higher, and during the recovery period, the MCHC of female rats in the low- and medium-dose groups and the Mono% of the medium-dose group were lower, but there was no obvious dose-response relationship or time-response relationship, and it was not considered to be related to the administration.

[0242] As can be seen from Table 16-19, regarding coagulation indicators, during the recovery period, the APTT of female rats in the low- and high-dose groups was higher, but there was no obvious dose-response relationship or time-response relationship. Furthermore, the indicators of each dose group during the administration period were not significantly different from those of the normal control group. Therefore, the above abnormalities are not considered to be related to the administration.

[0243] As shown in Tables 20-23, in terms of blood biochemistry, at the end of the treatment period, TG was lower in the medium-dose group of male rats; Na+ was higher in the high-dose group of female rats; GLU was lower in the low- and medium-dose groups; and BUN was lower in the low-, medium-, and high-dose groups. During the recovery period, Cl- was lower in the medium-dose group of male rats. No significant differences were observed in any of the other dose groups compared to the normal control group of the same sex at the same time point. These abnormal changes were mostly sporadic and isolated, with no clear dose-response relationship, and mostly fluctuated within the normal range. Therefore, they are considered to have only statistical significance and no biological significance.

[0244] As can be seen from Tables 24-31, in terms of urine testing, there were no significant differences in the various test indicators of rats in each dosage group compared with the normal control group of the same sex at the end of the administration period and the recovery period.

[0245] ◇Dissection: Surviving animals were dissected twice (after administration and after the recovery period).

[0246] Routine gross anatomical examination and femur length measurement were performed, along with organ weight: Major organs were weighed after animal sacrifice, and organ coefficients were calculated. Experimental results are shown in Tables 32-35, and... Figures 1-6 As shown.

[0247] Table 32 Statistical results of organ weight and organ index (end of drug administration / male, n=10, x±s)

[0248]

[0249]

[0250] Body weight, organ weight: g, organ-to-body ratio, organ-to-brain ratio: %.

[0251] Note: Compared with the normal control group, *P<0.05, **P<0.01.

[0252] Table 33 Statistical results of organ weight and organ index (end of drug administration / female, n=10, x±s)

[0253]

[0254]

[0255] Note: Body weight and organ weight: g; organ-to-body ratio and organ-to-brain ratio: %.

[0256] Table 34 Statistical results of organ weight and organ index (end of recovery period / male, n=5, x±s)

[0257]

[0258]

[0259] Note: Body weight and organ weight: g; organ-to-body ratio and organ-to-brain ratio: %.

[0260] Note: *P<0.05 compared with the normal control group.

[0261] Table 35 Statistical results of organ weight and organ index (end of recovery period / female, n=5, x±s)

[0262]

[0263]

[0264] Body weight, organ weight (g), organ-to-body ratio, organ-to-brain ratio (%).

[0265] As shown in Tables 32-35, gross anatomical examination revealed that all animals survived to the planned dissection, and no obvious abnormalities were found in any group. At the end of the administration period, the organ weights of the kidneys and livers in the medium-dose group and the thymus in the low-dose group were lower, while the testicular weights in the medium-dose group were higher, but no dose-response relationship was observed. No corresponding histopathological changes were observed in the weight changes of any organ. Due to the lack of dose compliance or associated histopathological changes, the changes in organ weight and coefficients at the end of the administration and recovery periods were considered unrelated to the toxicity of the test substance. Histopathological examination revealed no histopathological changes related to the test substance in any of the administered groups. The pathological changes observed in the test animals in each group, including the control group, included mild focal inflammatory cell infiltration in the liver and other lesions in some animals. Because these lesions were single, occasional, sporadic, or lacked dose compliance, they were all spontaneous lesions of the corresponding age group in SD rats. The analysis concluded that they belonged to the common spontaneous background lesions of this species and were not significantly related to the test substance.

[0266] The image shows the 100x magnification photograph of the lungs of rat No. 4106 in the high-dose group after HE staining. Figure 1 As shown; the liver of rat (animal number 4110) in the high-dose group, stained with hematoxylin and eosin (HE) and taken under a microscope at 200x magnification. Figure 2 As shown; the brain of rat No. 4010 in the high-dose group, stained with HE and photographed under a microscope at 12.5x magnification. Figure 3 As shown; the spleen of rat No. 4105 in the high-dose group was stained with HE and photographed under a microscope at 40x magnification. Figure 4 As shown; the kidney of rat (animal number 4110) in the high-dose group, stained with hematoxylin and eosin (HE) and taken under a microscope at 12.5x magnification. Figure 5 As shown; the heart of rat No. 4106 in the high-dose group, stained with HE and photographed under a microscope at 12.5x magnification. Figure 6 As shown. By Figure 1It can be seen that no obvious abnormalities were observed in the lungs of rat 4106 in the high-dose group. Figure 2 It can be seen that no obvious abnormalities were observed in the livers of rats (animal number 4110) in the high-dose group. Figure 3 It can be seen that no obvious abnormalities were observed in the brains of rats (animal number 4010) in the high-dose group. Figure 4 It can be seen that no obvious abnormalities were observed in the spleen of rat 4105 in the high-dose group. Figure 5 It can be seen that no obvious abnormalities were observed in the kidneys of rats (animal number 4110) in the high-dose group. Figure 6 It can be seen that no obvious abnormalities were observed in the hearts of rats in the high-dose group, animal number 4106.

[0267] ◇Histopathological examination

[0268] Routine toxicological examinations were performed on organs, all animals in the normal control group and high-dose group, all animals that were near death or had already died during the experiment, target organs in the low-dose group, medium-dose group, and all animals in the recovery period, and lesion sites in all animals. The experimental results are shown in Tables 36-38.

[0269] Table 36. Histopathological examination results (end of drug administration, male rats)

[0270]

[0271]

[0272]

[0273] Table 37. Histopathological examination results (at the end of the drug administration period, female rats)

[0274]

[0275]

[0276]

[0277]

[0278]

[0279] Table 38. Histopathological examination results (end of recovery period, rats)

[0280]

[0281]

[0282]

[0283] Note: In Tables 36-38, "N" indicates no abnormality; "1" indicates a slight abnormality; "-" indicates no content; and "√" indicates an abnormality.

[0284] The toxicological results (Tables 36-38) show that rats were administered the traditional Chinese medicine composition for treating anxiety and depression obtained in Example 3 via gavage for one month at experimental doses of 1.06, 2.12, and 4.24 g / kg, approximately 20, 40, and 80 times the human clinical dose, respectively. No target organ toxicity or toxic reactions related to the test substance were observed. Under the experimental conditions, the no obvious adverse event observed (NOAEL) was 4.24 g / kg, approximately 80 times the human clinical dose. This indicates that the traditional Chinese medicine composition provided by this invention has high safety.

Claims

1. A method for preparing a traditional Chinese medicine composition for treating anxiety and depression, characterized in that, Includes the following steps: (1) Mix Acanthopanax senticosus and Schisandra chinensis, crush them, and pass them through a 10-30 mesh sieve to make coarse powder; (2) Add extractant 1 to coarse powder and stir evenly. Moisten and seal and let it stand for 1-3 hours to allow it to fully expand. Put the expanded coarse powder into a percolation tank, flatten it, add extractant 2, wait until the air between the powder particles is discharged and ethanol flows out, close the discharge valve, cover the filter cylinder, and soak for 18-30 hours. (3) After soaking, open the discharge valve to percolate, add extractant 3 while percolating, control the percolation speed to be 2~3 mL / min per 1000g of medicinal material, collect the filtrate and mix it with the ethanol liquid discharged in step (2), concentrate the mixture to obtain the concentrate, dilute the obtained concentrate, let it stand, centrifuge and filter to obtain the supernatant A. (4) Pass the supernatant A obtained in step (3) through a D101 macroporous resin adsorption column with a diameter-to-height ratio of 1:2 to 1:4, add eluent 1 for elution, discard the eluent, add eluent 2 for elution, and collect eluent A; wash the adsorption column with water until neutral, add eluent 3 for elution, and collect eluent B. (5) Adjust the pH of the eluent A obtained in step (4) to neutral, centrifuge and filter, pass the supernatant B through a polyamide resin adsorption column with a diameter-to-height ratio of 1:2 to 1:4, wash with water until no salt is present, then elute with eluent 4, collect eluent C, concentrate and dry eluent C to obtain dry paste A; concentrate and dry eluent B obtained in step (4) to obtain dry paste B; (6) Mix dry paste A and dry paste B, pulverize and sieve to obtain a traditional Chinese medicine composition for treating anxiety and depression; The traditional Chinese medicine composition for treating anxiety and depression is made from the following raw materials in parts by weight: Acanthopanax senticosus 925-1000 parts, Schisandra chinensis 25-100 parts; In step (2), the extractant 1 is an aqueous ethanol solution with a volume fraction of 70% to 90%. The amount of extractant 1 added is 0.5 to 1 times the mass of the crude powder. Extractant 2 is added at a flow rate of 2 to 3 mL / min. The extractant 2 is an aqueous ethanol solution with a volume fraction of 70% to 90%. The extractant 3 in step (3) is an aqueous ethanol solution with a volume fraction of 70%~90%; In step (4), the eluent 1 is water, and the amount of eluent 1 added is 1 to 3 times the volume of supernatant A; the eluent 2 is a sodium hydroxide aqueous solution with a mass percentage of 0.1%, and the amount of eluent 2 added is 1 to 3 times the volume of supernatant A; the eluent 3 is an ethanol aqueous solution with a volume fraction of 70% to 90%, and the amount of eluent 3 added is 4 to 6 times the volume of the D101 macroporous resin adsorption column. In step (5), the eluent 4 is an aqueous ethanol solution with a volume fraction of 70% to 90%, and the amount of eluent 4 added is 2 to 3 times the mass of the supernatant B.

2. The method for preparing the traditional Chinese medicine composition for treating anxiety and depression according to claim 1, characterized in that, In step (3), the mixture is concentrated under vacuum at 40~80℃ until the relative density at 60℃ is 1.10~1.20 to obtain a concentrated solution; the obtained concentrated solution is diluted with 2~4 times the amount of water.

3. The method for preparing the traditional Chinese medicine composition for treating anxiety and depression according to claim 1, characterized in that, The specific steps for concentrating and drying the eluent C in step (5) to obtain dry paste A are as follows: Eluent C is concentrated under vacuum at 40-80°C to a concentration with a relative density of 1.20-1.30 at 60°C. The concentrate is then vacuum dried at 50-70°C until the dry moisture content is 3%, yielding dry paste A. Eluent B is concentrated and dried to obtain dry paste B. The specific steps are as follows: Eluent B is concentrated under vacuum at 40-80°C to a relative density of 1.20-1.30 at 60°C, yielding a concentrate. The concentrate is then vacuum dried at 50-70°C until the dry moisture content is 3%, yielding dry paste B.

4. The method for preparing the traditional Chinese medicine composition for treating anxiety and depression according to any one of claims 1-3, characterized in that, It also includes mixing the obtained traditional Chinese medicine composition for treating anxiety and depression with excipients to prepare traditional Chinese medicine preparations.

5. A traditional Chinese medicine preparation for treating anxiety and depression, characterized in that, The traditional Chinese medicine preparation for treating anxiety and depression includes the traditional Chinese medicine composition for treating anxiety and depression prepared by the preparation method according to any one of claims 1-3, and pharmaceutically acceptable excipients.

6. The traditional Chinese medicine preparation for treating anxiety and depression according to claim 5, characterized in that, The dosage form of the traditional Chinese medicine preparation for treating anxiety and depression is selected from granules, capsules, and tablets.

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