A compound Bupleurum Depression Relief Drops, its preparation method and its application in the preparation of drugs for the prevention and treatment of depression.

Abstract

This invention belongs to the field of traditional Chinese medicine preparation technology, specifically relating to a preparation method of Compound Bupleurum Depression-Relieving Droplets and its application in antidepressant drugs. This droplet preparation simplifies and optimizes the drug composition based on Xiaoyao San, while employing solid dispersion technology to improve the dissolution rate of poorly soluble components. The use of a composite matrix and nano-flow aids optimizes the droplet's shape and drug loading. Sustained-release coating technology meets the sustained-release requirements of circadian rhythm depression, while also improving the dissolution rate and stability of the active ingredients and possessing sustained-release characteristics, making it suitable for the prevention and treatment of depression.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine preparation technology, specifically relating to a preparation method of Compound Bupleurum Depression-Relieving Droplets and its application in antidepressant drugs. Through optimized extraction processes, formulation techniques, and quality control methods, these drops significantly improve the dissolution and stability of the active ingredients and possess sustained-release properties, making them suitable for the prevention and treatment of depression. Background Technology

[0002] Depression is a mental illness characterized by low mood, loss of interest, and cognitive impairment. Its global prevalence exceeds 3.5% and is increasing annually. While first-line antidepressants (such as SSRIs and SNRIs) have some efficacy, they suffer from slow onset of action (2-4 weeks), side effects (insomnia, sexual dysfunction, gastrointestinal reactions), and drug tolerance, leading to poor patient adherence.

[0003] Traditional Chinese medicine (TCM) compound formulas, due to their synergistic mechanism of action involving multiple components and targets, demonstrate unique advantages in improving depressive symptoms. Classic formulas such as Xiaoyao San (containing Bupleurum, Angelica sinensis, Paeonia lactiflora, Atractylodes macrocephala, Poria cocos, and Glycyrrhiza uralensis) regulate emotions through "soothing the liver and relieving depression," and clinical studies have confirmed that they can significantly reduce Hamilton Depression Rating Scale (HAMD) scores. However, traditional dosage forms (decoctions, powders, and ordinary pills) have certain limitations, such as low component dissolution rates: effective components such as saikosaponins and paeoniflorin are encapsulated by plant cell walls, resulting in insufficient extraction efficiency through decoction; poor stability: volatile components, such as Bupleurum volatile oil, are easily oxidized and lost, with a significant decrease in content during storage; and inconvenient administration: decoctions need to be prepared and used immediately, and pills are difficult to swallow, making them unsuitable for the needs of modern patients.

[0004] As a novel traditional Chinese medicine preparation, drop pills have advantages such as high drug loading capacity, rapid onset of action, and easy portability. However, their application in antidepressant compound preparations still faces the following technical challenges: (1) Outdated extraction process: Traditional water extraction relies on long-term decoction, which consumes a lot of energy and destroys heat-sensitive components; single extraction technology does not completely release intracellular components wrapped in cellulose, resulting in a low extraction rate of effective components. (2) Insufficient formulation performance: Single matrix selection: Most drop pills use polyethylene glycol (PEG) as a single matrix, which leads to uneven drug distribution and low dissolution of effective components; lack of functional design: Existing drop pills are mainly for immediate release, which cannot meet the needs of patients with diurnal symptom fluctuations (such as severe symptoms in the morning and mild symptoms in the evening) for sustained-release administration.

[0005] To address the above problems, this invention proposes a systematic solution:

[0006] First, this invention improves upon the classic Chinese medicine formula Xiaoyao San, selecting Bupleurum chinense, known for its liver-soothing and mood-lifting effects, as the principal herb to dispel liver heat and stagnation. It is supplemented with Paeonia lactiflora to nourish blood and soften the liver; the two work synergistically to dynamically regulate the liver's functions of dispersing and storing blood, improving the pathogenesis of liver stagnation and blood deficiency. Simultaneously, Poria cocos is added, working in conjunction with Glycyrrhiza uralensis to strengthen the spleen and eliminate dampness, breaking the pathological chain of "liver stagnation → spleen deficiency → phlegm-dampness." This invention simplifies and optimizes the formula based on Xiaoyao San, concentrating the medicinal power on the core pathogenesis, focusing on core drug pairs and effective components, simplifying quality control indicators, and promoting standardized quality control. This better meets international regulatory requirements for herbal compound formulas—"clearly defined components, clear mechanisms, and controllable quality"—and contributes to the internationalization of traditional Chinese medicine. Experimental data from this invention show that the reduced formula significantly improves the treatment of depression.

[0007] Secondly, this invention innovates the traditional extraction process of Chinese medicine by using solid dispersion technology to improve the solubility of poorly soluble components (such as saikosaponins). At the same time, the use of composite matrix and nano-flow aids optimizes the pellet formation and drug loading. The sustained-release coating technology meets the sustained-release requirements of circadian rhythm depression. These improvements significantly enhance the technical effect of traditional pellet technology by integrating modern extraction technology, formulation process and intelligent control methods. Summary of the Invention

[0008] The purpose of this invention is to provide a compound Bupleurum Depression Relief Drops, its preparation method, and its application in the preparation of drugs for the prevention and treatment of depression.

[0009] To achieve the above objectives, the present invention adopts the following technical solution:

[0010] In a first aspect, the present invention provides a compound Bupleurum Depression-Relieving Dripping Pill, which is made from the following raw materials in parts by weight: 10-30 parts Bupleurum, 5-20 parts White Peony Root, 5-20 parts Poria, 1-10 parts Licorice Root, and a pharmaceutically acceptable dripping pill matrix.

[0011] More preferably, the weight ratio of the raw materials is: Bupleurum chinense 15-25 parts, Paeonia lactiflora 8-15 parts, Poria cocos 8-15 parts, and Glycyrrhiza uralensis 3-6 parts.

[0012] More preferably, the weight ratio of the raw materials is: 20 parts Bupleurum, 15 parts White Peony Root, 10 parts Poria, and 5 parts Licorice.

[0013] Secondly, the present invention provides a method for preparing Compound Bupleurum Depression-Relieving Dripping Pills, characterized by comprising the following steps:

[0014] (1) Weigh out Bupleurum, White Peony Root, Poria, and Licorice according to the ratio, and extract using microwave-assisted water extraction-compound enzymatic hydrolysis technology:

[0015] a. Grind the medicinal materials into 40-80 mesh, add 8-12 times the total weight of the medicinal materials in water, extract with microwave power of 300-600W for 10-20 minutes, and filter to obtain the first extract;

[0016] b. Add a compound enzyme solution of cellulase and pectinase to the residue, with a weight ratio of cellulase to pectinase of 1:1-3, and enzymatically hydrolyze for 30-60 minutes at pH 4.5-5.5 and 45-55℃.

[0017] c. After enzymatic hydrolysis, the residue is boiled again with water 1-2 times, all filtrates are combined, and concentrated under reduced pressure to an extract with a relative density of 1.15-1.30;

[0018] (2) Mix the extract and the pellet matrix at a weight ratio of 1:1-4, add 50%-150% of the weight of the extract solid dispersion carrier, melt at 60-80℃ and homogenize by high-speed shearing at a speed of 8000-12000 rpm for 3-5 minutes. The solid dispersion carrier is selected from at least one of poloxamer 188 and copovidone S630.

[0019] (3) The molten material is dripped into the cooling liquid through a pelletizing machine. The dripping speed is controlled at 20-40 drops / minute, the dripping head temperature is 70-85℃, and the cooling liquid is a mixture of liquid paraffin and polydimethylsiloxane at 0-5℃ with a volume ratio of 2-4:1. After pelleting, the pellets are dried in a gradient. The first stage is drying at 40-50℃ for 1-2 hours, and the second stage is vacuum drying at 25-30℃ until the water content is ≤3%.

[0020] Further, the amount of the compound enzyme solution added in step (1) is 0.5-1.5% of the weight of the residue. After enzymatic hydrolysis, it is separated by ultrafiltration membrane to retain macromolecular impurities with a molecular weight cutoff of 5000-10000 Da.

[0021] More preferably, the pellet matrix in step (2) is a composite matrix of polyethylene glycol 6000 and glyceryl behenate in a weight ratio of 3-5:1, and 0.1%-0.5% of nano silica by weight of the extract is added as a flow aid.

[0022] Furthermore, in step (3), an online infrared monitoring system is used to adjust the temperature of the dripper in real time during the dripping process, and the viscosity of the melt is controlled within the range of 200-500 mPa·s.

[0023] Furthermore, it also includes step (4) of microcapsule coating of the pellets: using an ethanol solution of ethyl cellulose and hydroxypropyl methylcellulose (weight ratio 1:2-4) for fluidized bed coating, controlling the weight gain to 2%-5%, to form a sustained-release layer.

[0024] Thirdly, this invention provides the application of Compound Bupleurum Depression-Relieving Dripping Pills in the preparation of drugs for the prevention and treatment of depression.

[0025] The beneficial effects of this invention are as follows: (1) This invention improves the classic Chinese medicine formula Xiaoyao San, and simplifies and optimizes the drugs based on the classic formula, focusing the medicinal power on the core pathogenesis, focusing on the core drug pair and effective components, simplifying quality control indicators, promoting the standardization of quality control, and helping the internationalization of Chinese medicine. The reduced formula has a significant effect on the prevention and treatment of depression. (2) This invention uses drop pills to replace traditional Chinese medicine preparations, which have higher bioavailability, better stability, and are more convenient to take, improving the poor taste of traditional Chinese medicine preparations. (3) Solid dispersion technology is used to improve the dissolution of poorly soluble components (such as saikosaponins). At the same time, the use of composite matrix and nano-flow aids optimizes the drop pill's formability and drug loading. The sustained-release coating technology meets the sustained-release requirements of diurnal rhythm depression. The above improvements significantly enhance the technical effect of traditional drop pill technology by integrating modern extraction technology, preparation process and intelligent control methods. Attached Figure Description

[0026] Figure 1 Dissolution curve Detailed Implementation

[0027] The present invention will now be clearly and completely described with reference to specific embodiments, but the present invention is not limited thereto. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0028] The active pharmaceutical ingredients or excipients used in this invention, as well as the reagents or instruments used without specifying the manufacturer, are all conventional products that can be obtained commercially.

[0029] In this invention, terms such as "further," "particularly," and "preferred" are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0030] In this invention, percentage content refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures, unless otherwise specified.

[0031] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains, and the terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to limit the invention.

[0032] Example 1: Preparation of Compound Bupleurum Droplets

[0033] Weigh out 20g of Bupleurum chinense, 15g of Paeonia lactiflora, 10g of Poria cocos, and 5g of Glycyrrhiza uralensis. Grind the herbs to 60 mesh. Add 12 times the total weight of water and extract using a microwave at 600W for 20 minutes. Filter to obtain the first extract. Add a compound enzyme solution of cellulase and pectinase to the residue. The amount of the compound enzyme solution added is 1.5% of the weight of the residue, and the weight ratio of cellulase to pectinase is 1:3. [The process is carried out at pH...] 5.5 Enzymatic hydrolysis was performed at 45℃ for 60 minutes. After hydrolysis, the mixture was passed through an ultrafiltration membrane to separate macromolecular impurities with a molecular weight cutoff of 5000 Da. The residue after hydrolysis was boiled twice with water, and all filtrates were combined and concentrated under reduced pressure to an extract with a relative density of 1.15. The extract was mixed with a composite matrix of polyethylene glycol 6000 and glyceryl behenate at a weight ratio of 1:2. Poloxamer 188 (50% by weight of the extract) and nano-silica (0.5% by weight of the extract) were added as flow aids. The mixture was melted at 80℃ and homogenized by high-speed shearing at 12000 rpm for 5 minutes. The weight ratios of polyethylene glycol 6000 and glyceryl behenate were measured. The ratio of the molten material to the coolant is 3:1. The molten material is dripped into the coolant through a pelletizing machine at a dripping rate of 40 drops / minute and a dripper temperature of 85℃. The coolant is a mixture of liquid paraffin and polydimethylsiloxane at 0℃ with a volume ratio of 2:1. After pellet formation, the pellets are subjected to gradient drying. The first stage is drying at 50℃ for 2 hours, and the second stage is vacuum drying at 30℃ until the water content is ≤3%. During the pelleting process, an online infrared monitoring system is used to adjust the dripper temperature in real time and control the viscosity of the molten material within the range of 200 mPa·s. The pellets are then coated with microcapsules: an ethanol solution of ethyl cellulose and hydroxypropyl methylcellulose (weight ratio 1:2) is used for fluidized bed coating, increasing the weight by 5% and forming a slow-release layer.

[0034] Example 2: Preparation of Compound Bupleurum Droplets

[0035] Weigh out 20g of Bupleurum chinense, 15g of Paeonia lactiflora, 10g of Poria cocos, and 5g of Glycyrrhiza uralensis. Grind the herbs to 60 mesh. Add 12 times the total weight of water and extract using a microwave at 600W for 20 minutes. Filter to obtain the first extract. Add a compound enzyme solution of cellulase and pectinase to the residue. The amount of the compound enzyme solution added is 1.5% of the weight of the residue, and the weight ratio of cellulase to pectinase is 1:3. [The process is carried out at pH...] 5.5 Enzymatic hydrolysis was performed at 45℃ for 60 minutes. After hydrolysis, the mixture was passed through an ultrafiltration membrane to separate macromolecular impurities with a molecular weight cutoff of 5000 Da. The residue after hydrolysis was boiled twice with water, and all filtrates were combined and concentrated under reduced pressure to an extract with a relative density of 1.15. The extract was mixed with a composite matrix of polyethylene glycol 6000 and glyceryl behenate at a weight ratio of 1:2. Poloxamer 188 (50% by weight of the extract) and nano-silica (0.5% by weight of the extract) were added as a flow aid. The mixture was melted at 80℃ and homogenized by high-speed shearing at a rotation speed of 8000 r. The process involved dripping the molten material into a cooling liquid using a pelletizing machine. The dripping speed was controlled at 40 drops / minute, the dripper temperature was 85℃, and the cooling liquid was a mixture of liquid paraffin and polydimethylsiloxane at 0℃ with a volume ratio of 2:1. After pellet formation, the pellets underwent gradient drying. The first stage involved drying at 50℃ for 2 hours, and the second stage involved vacuum drying at 30℃ until the moisture content was ≤3%. During the pelleting process, an online infrared monitoring system was used to adjust the dripper temperature in real time and control the viscosity of the molten material within the range of 200 mPa·s.

[0036] Example 3: Preparation of Compound Bupleurum Droplets

[0037] Weigh out 30g of Bupleurum chinense, 5g of Paeonia lactiflora, 20g of Poria cocos, and 10g of Glycyrrhiza uralensis. Grind the herbs to 60 mesh. Add water equal to 8 times the total weight of the herbs. Extract using a microwave at 300W for 20 minutes. Filter to obtain the first extract. Add a compound enzyme solution of cellulase and pectinase to the residue. The amount of the compound enzyme solution added is 0.5% of the weight of the residue. The weight ratio of cellulase to pectinase is 1:3. [The process is carried out at pH...] 4.5. Enzymatic hydrolysis was performed at 55℃ for 30 minutes. After enzymatic hydrolysis, the mixture was passed through an ultrafiltration membrane to separate macromolecular impurities with a molecular weight cutoff of 10000 Da. The residue after enzymatic hydrolysis was boiled again with water, and all filtrates were combined and concentrated under reduced pressure to an extract with a relative density of 1.30. The extract was mixed with a composite matrix of polyethylene glycol 6000 and glyceryl behenate at a weight ratio of 1:2. Poloxamer 188 (50% by weight of the extract) and nano-silica (0.5% by weight of the extract) were added as flow aids. The mixture was melted at 80℃ and homogenized by high-speed shearing at 12000 rpm for 3 minutes. The weight ratios of polyethylene glycol 6000 and glyceryl behenate were... The ratio of 5:1 was used to drop the molten material into the cooling liquid through a pelletizing machine, controlling the dropping rate at 40 drops / minute and the dropper temperature at 85℃. The condensate was a mixture of liquid paraffin and polydimethylsiloxane at 0℃ with a volume ratio of 2:1. After pellet formation, the pellets underwent gradient drying. The first stage was drying at 50℃ for 2 hours, and the second stage was vacuum drying at 30℃ until the moisture content was ≤3%. During the dropping process, an online infrared monitoring system was used to adjust the dropper temperature in real time and control the viscosity of the molten material within the range of 200 mPa·s. The pellets were then coated with microcapsules: a fluidized bed coating was performed using an ethanol solution of ethyl cellulose and hydroxypropyl methylcellulose (weight ratio 1:4) to increase the weight by 5% and form a slow-release layer.

[0038] Example 4: Preparation of Compound Bupleurum Droplets

[0039] Weigh out 10g of Bupleurum chinense, 8g of Paeonia lactiflora, 15g of Poria cocos, and 6g of Glycyrrhiza uralensis. Grind the herbs to 60 mesh. Add water equal to 10 times the total weight of the herbs. Extract using a microwave at 600W for 10 minutes. Filter to obtain the first extract. Add a compound enzyme solution of cellulase and pectinase to the residue. The amount of the compound enzyme solution added is 1.5% of the weight of the residue, and the weight ratio of cellulase to pectinase is 1:2. Set the pH to [missing value]. 5.5 Enzymatic hydrolysis was performed at 45℃ for 60 minutes. After hydrolysis, the mixture was passed through an ultrafiltration membrane to separate macromolecular impurities with a molecular weight cutoff of 5000 Da. The residue after hydrolysis was boiled twice with water, and all filtrates were combined and concentrated under reduced pressure to an extract with a relative density of 1.15. The extract was mixed with a composite matrix of polyethylene glycol 6000 and glyceryl behenate at a weight ratio of 1:2. Poloxamer 188 (50% by weight of the extract) and nano-silica (0.5% by weight of the extract) were added as flow aids. The mixture was melted at 80℃ and homogenized by high-speed shearing at 8000 rpm for 5 minutes. The weight ratio of polyethylene glycol 6000 to glyceryl behenate was [not specified in the original text]. 4:1; The molten material is dripped into the cooling liquid through a pelletizing machine at a dripping rate of 40 drops / minute, with a dripper temperature of 85℃. The condensate is a mixture of liquid paraffin and polydimethylsiloxane at 0℃, with a volume ratio of 2:1. After pellet formation, the pellets undergo gradient drying: the first stage is drying at 50℃ for 2 hours, and the second stage is vacuum drying at 30℃ until the moisture content is ≤3%. During the pelleting process, an online infrared monitoring system is used to adjust the dripper temperature in real time and control the viscosity of the molten material within the range of 200 mPa·s. The pellets are then coated with microcapsules: an ethanol solution of ethyl cellulose and hydroxypropyl methylcellulose (weight ratio 1:3) is used for fluidized bed coating, increasing the weight by 5% and forming a slow-release layer.

[0040] Comparative Example 1

[0041] Weigh out 20g of Bupleurum chinense, 15g of Paeonia lactiflora, 10g of Poria cocos, and 5g of Glycyrrhiza uralensis. Grind the herbs into 60 mesh and add 12 times the total weight of the herbs in water. Decoction is carried out 3 times, 1 hour each time. The decoction is concentrated under reduced pressure. The resulting extract is used as control group 1.

[0042] Comparative Example 2

[0043] Weigh out 20g of Bupleurum chinense, 15g of Paeonia lactiflora, 10g of Poria cocos, and 5g of Glycyrrhiza uralensis. Grind the herbs into 60 mesh. Add water equal to 12 times the total weight of the herbs and extract using a microwave at 600W for 20 minutes. Filter to obtain the first extract. Add water to the residue and continue extraction twice more using microwave assistance. Filter the extracts. Combine the extracts and concentrate under reduced pressure. The resulting extract is used as control group 2.

[0044] To verify the beneficial effects of this invention, a series of experimental examples and comparative experiments are presented below. The experiments include evaluation of the extraction rate of the active ingredient, formulation quality, stability, pharmacodynamics, and sustained-release characteristics.

[0045] Experiment Example 1: Comparison Experiment of Active Ingredient Extraction Rate

[0046] Experimental Methods: The product of Example 1 of this invention was used as experimental group 1. The extracts obtained from comparisons 1 and 2 were used as control groups 1 and 2, respectively. Commercially available Xiaoyao Pills (National Drug Approval Number Z43020469, specification: 360 pills / bottle) were used as control group 3. The contents of bupleuropeptide a (the main active ingredient of Bupleurum chinense) and paeoniflorin (the main active ingredient of Paeonia lactiflora) in the extract were determined by high performance liquid chromatography (HPLC). Chromatographic conditions: C18 column, acetonitrile-water gradient elution mobile phase, detection wavelength 210 nm. The extraction rate was calculated as follows: Extraction rate (%) = (weight of active ingredients in the extract / total weight of active ingredients in the medicinal material) × 100%. The results are shown in Table 1 below.

[0047] Table 1 Extraction rate of active ingredients

[0048]

[0049] *p<0.01

[0050] The results showed that the extraction rates of saikosaponin a and paeoniflorin using the method of this invention were significantly higher than those of the control group, indicating that the microwave-assisted water extraction-compound enzymatic hydrolysis combined technology can effectively disrupt plant cell walls, release intracellular components, and improve extraction efficiency. Traditional water extraction had the lowest extraction rate; while microwave-assisted water extraction showed some improvement, compound enzymatic hydrolysis further enhanced the extraction effect.

[0051] Experimental Example 2: Quality Evaluation of the Droplet Formulation in Example 1

[0052] Experimental Methods: Test Samples: The droplets prepared in Example 1 of this invention were used as Experimental Group 1, the droplets prepared in Example 2 of this invention were used as Experimental Group 2, and commercially available Xiaoyao Pills (National Drug Approval Number Z43020469, Specification: 360 pills / bottle) were used as Control Group 1. The appearance of the droplets (roundness, size uniformity) was observed, and the yield rate (number of qualified droplets / total number of droplets × 100%) was calculated; the content of saikosaponin a in each droplet (mg / pill) and the drug loading were determined; at the same time, the hardness and friability of the pills were determined using a hardness tester and a friability tester.

[0053] The dissolution test method according to the 2020 edition of the Chinese Pharmacopoeia was adopted, using water as the dissolution medium (900 mL), at a temperature of 37 ± 0.5℃ and a rotation speed of 50 rpm. Samples of 5 mL were taken at 10, 30, 60, and 120 minutes, with simultaneous addition of isothermal and equal volumes of medium. The solution was filtered through a 0.45 μm microporous membrane, and the content of saikosaponin a in the filtrate was determined by high-performance liquid chromatography (HPLC). The cumulative dissolution rate (%) was calculated. Chromatographic conditions: C18 column (4.6 mm × 250 mm, 5 μm), mobile phase: acetonitrile-water (gradient elution), flow rate: 1.0 mL / min, detection wavelength: 210 nm. Results are as follows: Figure 1 As shown in Table 2-3 below:

[0054] Table 2. Results of Formulation Quality Evaluation*

[0055]

[0056] *p<0.05

[0057] Table 3 Cumulative Dissolution Test Results*

[0058]

[0059] *p<0.01

[0060] The results showed that the experimental group had significantly higher yield, drug loading, and dissolution rate than the control group (p < 0.01), indicating that the composite matrix (polyethylene glycol 6000 and glycerol behenate), nano-silica flow aid, and solid dispersion technology used in this invention effectively optimized the pellet formation and drug release. Experimental group 2 had a faster dissolution rate, suitable for immediate release requirements; experimental group 1 had stable dissolution, consistent with sustained-release design. The control group showed poor pellet formation and low dissolution rate, demonstrating the limitations of a single matrix and traditional processes.

[0061] Experimental Example 3: Formulation Stability Experiment

[0062] Experimental Methods: The droplets prepared in Example 1 of this invention were used as Experimental Group 1. Accelerated stability testing was conducted according to the guidelines for stability testing in the 2020 edition of the Chinese Pharmacopoeia (temperature 40℃±2℃, relative humidity 75%±5%), with samples taken at 0, 1, 2, and 3 months. Appearance, moisture content, saiko saponin a content, and paeoniflorin content were measured. The results are shown in Table 4 below:

[0063] Table 4 Stability Experiment*

[0064]

[0065] *p<0.05

[0066] Experimental results show that within 3 months of accelerated testing, the content of the active ingredient in the pellets of this invention remained stable (decline rate <5%), and the moisture content met the requirements, indicating that this invention significantly improved the stability of the formulation through gradient drying and sustained-release coating technology.

[0067] Experimental Example 4: Evaluation of the antidepressant effect of the droplet formulation of the present invention

[0068] Animal model: SPF-grade male Kunming mice, weighing 18-22g, were used in the experiment after one week of acclimatization under standard conditions (temperature 22±2℃, humidity 50±10%, 12 / 12-hour light / dark cycle). Main reagents and instruments: sucrose, ELISA kits (5-HT, CORT, BDNF, TNF-α), forced swimming test apparatus, tail suspension test apparatus, and open field test apparatus. Test samples: sustained-release pills prepared in Example 1 were used as experimental group 1; commercially available Xiaoyao pills (National Drug Approval Number Z43020469, specification: 360 pills / bottle) were used as control group 3; positive control drug: fluoxetine hydrochloride capsules (trade name: Prozac).

[0069] Establishment of a mouse model of depression caused by chronic unpredictable mild stress (CUMS): For 5 weeks, 1-2 different mild stressors were randomly applied daily, such as 24h of water / food deprivation, 24h of cage tilt at 45°, 24h of moist bedding, 10min of horizontal shaking, day-night reversal, and 1min of tail clamping. All stressors were not replicated during the experimental period.

[0070] Experimental Methods: 120 mice were randomly divided into 6 groups of 20 mice each. Blank Control Group: Normally fed without stress, and administered an equal volume of physiological saline daily by gavage. Model Control Group: Underwent CUMS modeling and administered an equal volume of physiological saline daily by gavage. Positive Control Group: Underwent CUMS modeling and administered fluoxetine hydrochloride (10 mg / kg) daily by gavage. Low-Dose Group: Underwent CUMS modeling and administered the present invention's pills (equivalent to 1 g / kg of raw drug) daily by gavage. High-Dose Group: Underwent CUMS modeling and administered the present invention's pills (equivalent to 2 g / kg of raw drug) daily by gavage. Traditional Formulation Group: Underwent CUMS modeling and administered commercially available Xiaoyao pills (equivalent to 2 g / kg of raw drug) daily by gavage.

[0071] Dosing regimen: Starting from week 3 of modeling, each group was administered the above-mentioned medication once daily for 4 consecutive weeks. The control group and the model group underwent simultaneous grasping and gavage procedures.

[0072] Behavioral Testing Methods: Sugar Water Preference Test: Mice were trained with sugar water before the experiment. During the formal test, each mouse was housed individually and given two pre-weighed bottles of liquid (1% sucrose solution and pure water). After 24 hours, the bottles were swapped to avoid positional preference. After another 24 hours, the sugar water preference rate was calculated: Sugar water preference rate (%) = Sugar water consumption / (Sugar water consumption + Pure water consumption) × 100%. Forced Swimming Test: Mice were placed in a transparent glass cylinder (25cm high, 10cm in diameter), with a water depth of 10cm and a water temperature of 23±1℃. The cumulative immobility time (i.e., the time spent cessating struggle, merely floating, or making slight movements to keep the head above water) was recorded for the last 4 minutes within a 6-minute period. Tail Suspension Test: The mouse's tail was fixed 2cm from the end to a horizontal lever with tape, causing the mouse to hang upside down 15cm above the table. The experimental environment was closed, and the cumulative immobility time (i.e., the state of complete stillness, cessation of struggle) for the last 4 minutes within a 6-minute period was recorded.

[0073] Sample collection and biochemical index detection: After the behavioral test, blood was collected from 10 mice in each group, and the serum was separated by centrifugation (3500 rpm, 15 min) after standing. The serum levels of 5-hydroxytryptamine (5-HT), corticosterone (CORT), brain-derived neurotrophic factor (BDNF), and tumor necrosis factor-α (TNF-α) were measured by ELISA strictly following the kit instructions.

[0074] Experimental results:

[0075] Table 5 Behavioral test results

[0076]

[0077] Note: Compared with the control group, **p<0.01; compared with the model group, *p<0.05, #p<0.01

[0078] Behavioral test results showed that, compared with the control group, the model group mice had a significantly lower sucrose preference rate and significantly prolonged forced swimming and tail-suspended immobility time, indicating successful replication of the CUMS depression model. The high-dose group of this invention significantly reversed the behavioral deficits induced by CUMS in all behavioral indicators, with effects comparable to the positive control drug fluoxetine, and even showing a superior trend in sucrose preference rate. The antidepressant effect of the high-dose group of this invention was significantly better than that of the low-dose group, indicating that its efficacy is dose-dependent. Furthermore, at the same dosage of raw herbs, the antidepressant effect of the high-dose group of this invention was comprehensively superior to that of the Xiaoyao pill group, demonstrating the significant advantages of the dripping pill dosage form of this invention in improving bioavailability and efficacy.

[0079] Table 6. Results of serum biochemical marker detection

[0080]

[0081] Note: Compared with the control group, **p<0.01; compared with the model group, *p<0.05, #p<0.01

[0082] Experimental results showed that the model group exhibited a regulatory effect on monoamine neurotransmitters: 5-HT levels were significantly reduced, while the high-dose group of this invention restored them to near-normal levels, with an effect comparable to fluoxetine and superior to Xiaoyao Wan. This reflects its classic antidepressant mechanism of "soothing the liver and relieving depression." Simultaneously, the model group showed abnormally elevated CORT levels, indicating overactivation of the hypothalamus-pituitary-adrenal (HPA) axis. The high-dose group of this invention effectively reduced CORT levels significantly, suggesting that it exerts its antidepressant effect by alleviating stress response. Furthermore, the model group showed decreased BDNF levels and increased TNF-α levels. The high-dose group of this invention significantly increased BDNF and decreased TNF-α, indicating that its antidepressant effect is also related to multi-target mechanisms such as promoting nerve repair and inhibiting neuroinflammation. This effect is superior to fluoxetine (which has less impact on BDNF and TNF-α), highlighting the synergistic advantages of multi-component and multi-target traditional Chinese medicine compound.

[0083] Based on the above experimental data, the Compound Bupleurum Depression-Relieving Dripping Pills of this invention exhibited significant antidepressant activity in the CUMS mouse depression model, with efficacy comparable to the first-line clinical drug fluoxetine, and showed potential advantages in improving anhedonia (saccharide preference). Its mechanism of action involves multiple pathways, including regulating monoamine neurotransmitters, inhibiting excessive activation of the hypothalamic-pituitary-adrenal axis, promoting neurotrophic effects, and anti-inflammatory effects, reflecting the multi-target synergistic therapeutic characteristics of traditional Chinese medicine compound formulas. Furthermore, at the same dosage of raw herbs, the antidepressant effect of the dripping pills of this invention is superior to that of traditional Xiaoyao Pills, which strongly demonstrates that this invention significantly improves the bioavailability and efficacy of the drug through advanced formulation technology, achieving the technical goal of modernizing traditional Chinese medicine.

[0084] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit it. After reading this specification, those skilled in the art may make modifications to these embodiments without departing from the core design of the present invention. Such modifications are all within the scope of protection of the present invention and should fall within the scope of protection defined by the claims of the present invention.

Claims

1. A compound Chaihu Jieyu drop pill, characterized in that, The compound Chaihu Jieyu drop pills are prepared from the following raw medicinal materials by weight: Chaihu 10-30 parts, Baishao 5-20 parts, Fuling 5-20 parts, and Gancao 1-10 parts, and a pharmaceutically acceptable drop pill matrix, and the preparation steps include the following: (1) The Chaihu, Baishao, Fuling, and Gancao are weighed according to the ratio, and a microwave-assisted water extraction-complex enzymatic hydrolysis combined technology is used for extraction: a. The medicinal materials are ground to 40-80 mesh, 8-12 times the weight of water of the total medicinal materials is added, and extraction is performed under a microwave power of 300-600 W for 10-20 minutes, and a first extraction liquid is obtained by filtration; b. The medicinal residue is added with a complex enzyme solution of cellulase and pectinase, the weight ratio of cellulase to pectinase is 1:1-3, and enzymatic hydrolysis is performed at pH 4.5-5.5 and 45-55°C for 30-60 minutes; c. The medicinal residue after enzymatic hydrolysis is boiled with water for 1-2 times, all the filtrates are combined, and the filtrate is concentrated under reduced pressure to a relative density of 1.15-1.30 of the extract; (2) The extract and the drop pill matrix are mixed according to a weight ratio of 1:1-4, 50%-150% of the weight of the extract is added as a solid dispersion carrier, and the mixture is melted and homogenized at a high speed at 60-80°C, the rotation speed is 8000-12000 rpm, and the time is 3-5 minutes, and the solid dispersion carrier is at least one selected from poloxamer 188 and copovidone S630; (3) The melt is dropped into a cooling liquid through a drop pill machine, the dropping speed is controlled to be 20-40 drops per minute, the temperature of the drop head is 70-85°C, the condensate is a mixture of liquid paraffin and polydimethylsiloxane at a volume ratio of 2-4:1, and after the drop pills are formed, gradient drying is performed, the first stage is drying at 40-50°C for 1-2 hours, and the second stage is vacuum drying at 25-30°C until the water content is ≤3%.

2. The compound Chaihu drop pill of claim 1, characterized in that, The weight ratio of the raw medicinal materials is as follows: Chaihu 15-25 parts, Baishao 8-15 parts, Fuling 8-15 parts, and Gancao 3-6 parts.

3. The compound Chaihu drop pill of claim 1, characterized in that, The weight ratio of the raw medicinal materials is as follows: Chaihu 20 parts, Baishao 15 parts, Fuling 10 parts, and Gancao 5 parts.

4. The preparation method of the compound Chaihu Jieyu Dropping Pills of any one of claims 1-3, characterized in that The preparation steps include the following: (1) The Chaihu, Baishao, Fuling, and Gancao are weighed according to the ratio, and a microwave-assisted water extraction-complex enzymatic hydrolysis combined technology is used for extraction: a. The medicinal materials are ground to 40-80 mesh, 8-12 times the weight of water of the total medicinal materials is added, and extraction is performed under a microwave power of 300-600 W for 10-20 minutes, and a first extraction liquid is obtained by filtration; b. The medicinal residue is added with a complex enzyme solution of cellulase and pectinase, the weight ratio of cellulase to pectinase is 1:1-3, and enzymatic hydrolysis is performed at pH 4.5-5.5 and 45-55°C for 30-60 minutes; c. The medicinal residue after enzymatic hydrolysis is boiled with water for 1-2 times, all the filtrates are combined, and the filtrate is concentrated under reduced pressure to a relative density of 1.15-1.30 of the extract; (2) The extract and the drop pill matrix are mixed according to a weight ratio of 1:1-4, 50%-150% of the weight of the extract is added as a solid dispersion carrier, and the mixture is melted and homogenized at a high speed at 60-80°C, the rotation speed is 8000-12000 rpm, and the time is 3-5 minutes, and the solid dispersion carrier is at least one selected from poloxamer 188 and copovidone S630; (3) The molten material is dropped into the cooling liquid through the drop pill machine, the drop speed is controlled at 20-40 drops per minute, the drop head temperature is 70-85℃, the condensate is a mixed liquid of liquid paraffin and polydimethylsiloxane with a volume ratio of 2-4:1, and the drop pill is formed and then dried by gradient drying, the first stage is dried at 40-50℃ for 1-2 hours, and the second stage is vacuum dried at 25-30℃ until the water content is ≤3%.

5. The preparation method of claim 4, wherein: In step (1), the addition amount of the complex enzyme solution is 0.5-1.5% of the weight of the residue, and after enzymolysis, the macromolecular impurities are separated by ultrafiltration membrane with a molecular weight cutoff of 5000-10000 Da.

6. The preparation method of claim 4, wherein: In step (2), the drop pill matrix is a complex matrix of polyethylene glycol 6000 and glycerol behenate with a weight ratio of 3-5:1, and 0.1%-0.5% of nano-silicon dioxide by weight of the extract is added as a flow aid.

7. The preparation method of claim 4, wherein: In step (3), an online infrared monitoring system is used to real-time control the drop head temperature during the drop process, and the viscosity of the molten material is controlled within the range of 200-500 mPa·s.

8. The preparation method of claim 4, further comprising step (4) of coating the drop pills: using an ethanol solution of ethyl cellulose and hydroxypropyl methyl cellulose for fluidized bed coating, controlling the weight gain of 2%-5% to form a sustained release layer, and the weight ratio of ethyl cellulose to hydroxypropyl methyl cellulose is 1:2-4.

9. The use of the compound Chaihu Jieyu drop pills of any one of claims 1-8 in the preparation of a drug for preventing and treating depression. ​

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