A nano Chinese medicine composition for improving sleep and its preparation method and application

Abstract

The invention provides a nano Chinese medicine composition for improving sleep, which is prepared from the following raw materials by weight: 8-20 parts of bupleurum, 5-15 parts of scutellaria, 5-15 parts of angelica, 10-30 parts of white peony root, 10-30 parts of Poria, 10-30 parts of stir-fried atractylodes, 10-30 parts of spiny jujube seeds, 5-20 parts of codonopsis, 10-30 parts of longan pulp, 5-15 parts of roasted liquorice, and 10-20 parts of gardenia. The above Chinese medicines are subjected to ultrafine grinding and wall breaking treatment, solvent extraction, and concentrated spray drying to form nano-scale Chinese medicine powders, which can not only increase the original efficacy, but also change the route of administration, improve the stability of the therapeutic effect, are convenient to take, have low cost, and have the effects of soothing the liver and spleen, nourishing the heart and calming the mind.

Description

Technical Field

[0001] The invention relates to a Chinese medicine composition, and in particular to a nano Chinese medicine composition capable of improving sleep, and a preparation method and application thereof. Background Art

[0002] With the acceleration of the pace of life, the incidence of insomnia is increasing. Long-term insomnia will not only reduce the quality of life and affect social functions, but also cause a series of physical and mental diseases, such as cardiovascular and cerebrovascular diseases, metabolic diseases, tumors, etc. At present, the drugs used to treat insomnia are mainly diazepines, non-benzodiazepine sedatives and hypnotics, antidepressants and antipsychotics. Long-term use of such drugs has the risk of dependence and addiction. It is urgent to actively find safe and effective treatment methods with few adverse reactions. Traditional Chinese medicine is guided by the holistic view and treats according to syndrome differentiation. It is believed that under the characteristics of modern social life, insomnia is mostly related to emotions and diet. Liver depression and spleen deficiency, and insomnia with malnutrition of the mind are the most common clinical syndromes. This type of patient clinically manifests as insomnia due to deficiency, difficulty in falling asleep, early awakening after sleeping, inability to fall asleep again, and even sleepless all night, accompanied by depression, chest and flank pain, poor appetite and loose stools, dizziness and fatigue, palpitations, etc., and is prone to fatigue and drowsiness during the day, and lack of energy, which affects work and study. This type of insomnia seriously affects people's physical and mental health. Long-term insomnia can lead to many negative effects such as fatigue and emotional disorders, which seriously affect people's health, life and work. Although there are Chinese patent medicines for the treatment of insomnia, their use is far from meeting the needs of individualized treatment, and there is no medicine available for children, teenagers, the elderly and other special groups. It is imperative to develop Chinese medicines to treat insomnia caused by liver depression, spleen deficiency and mental malnutrition.

[0003] The existing method of taking Chinese herbal medicines is mainly based on decoction, which cannot fully utilize the effective ingredients of the medicine, and has problems such as poor taste, inconvenience in taking and storage. The bagged decoction is prone to deterioration and precipitation during long-term refrigeration, which affects the taking of the decoction. Therefore, it is necessary to develop a new preparation method. Based on the above problems, the research team has developed a nano-TCM composition for the treatment of liver depression, spleen deficiency, and mental malnutrition after a long period of clinical practice experience summary, and carried out relevant animal experiments and clinical experimental research funded by the National Special Project on Inheritance and Innovation of Traditional Chinese Medicine (2022CCCX008) and the Henan Provincial Key Project on Scientific Research of Traditional Chinese Medicine (NO: 2022JDZX004), and achieved good results. Summary of the invention

[0004] In view of the above situation, in order to solve the defects of the prior art, the present invention provides a Chinese medicine composition for improving sleep and its preparation method and use, which can solve the medication problem of insomnia problems such as liver depression, spleen deficiency and mental malnutrition.

[0005] According to TCM theory, the heart governs the mind and dominates the human body's mental and emotional activities. Only when the heart is full of blood and the mind is nourished can the awakening cycle be normal. In modern life, social competition is fierce, work and study pressure is high, and what you want is not fulfilled, excessive thinking, emotional frustration, and liver depression will occur over time. Fast pace of life, irregular diet, or excessive fatigue can lead to spleen weakness. In addition, there is a restraining relationship between the liver and spleen. Wood overcomes earth. Long-term liver depression leads to spleen weakness, poor transportation and transformation, lack of qi and blood, and lack of nourishment for the heart, which will manifest as insomnia. Therefore, insomnia often manifests as liver depression and spleen deficiency, and lack of nourishment for the mind. In treatment, the liver, spleen, and heart should be coordinated, with the principle of soothing the liver and strengthening the spleen, nourishing the heart and calming the mind.

[0006] Accordingly, the technical solution of the Chinese medicine composition for improving sleep provided by the present invention is as follows:

[0007] A nano Chinese medicine composition for improving sleep, characterized in that the Chinese medicine composition is made of the following raw materials in parts by weight: 8-20 parts of bupleurum, 5-15 parts of scutellaria, 5-15 parts of angelica, 10-30 parts of white peony root, 10-30 parts of poria, 10-30 parts of stir-fried atractylodes, 10-30 parts of spiny jujube seeds, 5-20 parts of codonopsis, 10-30 parts of longan pulp, 5-15 parts of roasted liquorice, and 10-20 parts of gardenia.

[0008] The ancient Chinese medical books and modern pharmacology studies of the above drugs are as follows:

[0009] Bupleurum is good at soothing the liver and regulating qi, and is an important medicine for regulating qi and relieving depression. "Southern Yunnan Materia Medica" says "Bupleurum can regulate the qi of the liver meridian, stop the pain of liver qi in the left rib, treat women's blood heat and burning menstruation, and regulate menstruation." Modern research shows that Bupleurum's effective ingredient, bupleurin, can be taken orally, and has a sedative effect on mice, and can prolong the sleep of pentobarbital. Bupleurin can be included in the category of central nervous system inhibitors. Scutellaria baicalensis has a bitter and cold taste and belongs to the heart and gallbladder meridians. It can relieve heat and relieve restlessness. It can assist Bupleurum, soothe the liver and relieve depression, and prevent depression from turning into fire. It can prevent diseases before they occur. As stated in "Ben Jing Shu Zheng", Bupleurum can open the qi knot, but cannot relieve the heat of the qi. Scutellaria baicalensis cooperates with Bupleurum and can clear the heat of the qi. Modern pharmacological studies have shown that Scutellaria baicalensis has the effects of harmonizing the gallbladder, relieving spasms, and sedation. Scutellaria baicalensis decoction and ethanol extract can increase the bile excretion of dogs and rabbits. Baicalin can inhibit the spontaneous activity of mice and the positive conditioned reflex of mice. This effect may be due to its strengthening of the cortical inhibition process. It can be used for hypertensive patients with increased nerve excitability and insomnia. Angelica sinensis is pungent, sweet and warm in nature. It enters the heart, liver and spleen meridians, replenishes blood and activates blood circulation, helps soften the liver and nourish the heart blood. Modern pharmacological studies have shown that Angelica sinensis has an inhibitory effect on the central nervous system. Japanese scholars have reported that Japanese Taihe Angelica sinensis volatile oil has sedative, hypnotic, analgesic, and anesthetic effects. White peony root is bitter, sour, and slightly cool. It enters the liver and spleen meridians. It is good at nourishing blood and softening the liver, relieving acute pain. The "Southern Yunnan Materia Medica" says that white peony root can "retract liver qi and pain, regulate the heart, liver, spleen and meridian blood, relax the meridians and lower qi, and stop liver qi pain." Modern pharmacological studies have shown that white peony root has a central inhibitory effect and has analgesic and sedative effects. Intraperitoneal injection of white peony root decoction can inhibit spontaneous activity in mice and enhance the hypnotic effect of sodium cyclohexylbarbital. Subcutaneous injection of peony root injection can also prolong the hypnotic time of sodium pentobarbital. Poria cocos tastes sweet, light and flat. It enters the spleen meridian and can strengthen the spleen and nourish the heart. The "Pharmacopoeia" says that Poria cocos can: "open the appetite, stop vomiting, and calm the mind." Modern studies have shown that Poria cocos has a central sedative effect. Poria cocos decoction can significantly prolong the anesthesia time compared with the control group. As the dose increases, the sedation index also increases. Atractylodes macrocephala tastes sweet, bitter and warm. It enters the spleen and stomach meridian and is good at strengthening the spleen and replenishing qi. "Compendium of Materia Medica" says that Atractylodes macrocephala is a key medicine for supporting the spleen and stomach, dispersing dampness and removing numbness, and digesting food and removing lumps. If the spleen is weak, Atractylodes macrocephala can replenish it. If the stomach is weak, Atractylodes macrocephala can help it. Modern pharmacological studies have shown that Atractylodes macrocephala can increase the contractility of gastrointestinal smooth muscle, promote weight gain in mice and enhance swimming endurance. Atractylodes macrocephala has the effect of replenishing deficiency and strengthening. Codonopsis pilosula is sweet and mild in taste and is included in the lung and spleen meridians. It can replenish the middle qi, strengthen the spleen and benefit the lungs. "Compendium of Materia Medica" says that Codonopsis pilosula can "replenish the middle qi, harmonize the spleen and stomach, and eliminate thirst." Modern research shows that Codonopsis pilosula has sedative and anti-peptic ulcer effects. Codonopsis pilosula water extract can prolong the sleep time of mice administrated with sodium pentobarbital, and has the effect of protecting the gastric mucosa against stress ulcers. Ziziphus jujuba seeds are sweet, sour, and mild in taste. They are included in the liver, gallbladder, and heart meridians. They can greatly replenish the heart and spleen, so that blood can return to the spleen and the five internal organs can be peaceful and sleep peacefully. "Compendium of Materia Medica" says: Ziziphus jujuba seeds are sweet and moist, so cooked ones are used to treat insomnia due to gallbladder deficiency, thirst, and sweating; raw ones are used to treat gallbladder heat and good sleep.Modern pharmacological research has found that Chinese jujube seeds are rich in alkaloids and have a sedative and hypnotic effect. Chinese jujube seed extract can increase the slow-wave sleep and deep sleep time of mice. Longan meat is sweet and warm, and it enters the heart and spleen meridians. It is good at nourishing the heart and spleen, replenishing qi and blood, and calming the mind. The "Southern Yunnan Materia Medica" calls it "nourishing blood and calming the mind, growing wisdom and restraining sweat, and opening the appetite and benefiting the spleen." Roasted licorice is sweet and flat in nature, and enters the heart, lungs, spleen and stomach. It can nourish the spleen and replenish qi and harmonize various medicines. Experimental studies have shown that glycyrrhetinic acid, the main component of licorice, can inhibit the excitement of the central nervous system of mice and can have a sedative and hypnotic effect. Gardenia can clear the heart and eliminate troubles. It enters the heart meridian and has the effect of clearing heart fire. It can be used to relieve symptoms such as irritability, palpitations, insomnia and dreaminess.

[0010] In the present invention, Bupleurum is bitter in taste, slightly cold and enters the liver and gallbladder meridians, and is good at regulating liver qi and relieving liver depression, so it is the main medicine; Angelica sinensis and White Peony Root nourish liver blood, assist in soothing the liver, and also nourish the heart blood and assist in calming the mind, so it is the minister medicine; Codonopsis pilosula, Atractylodes macrocephala, Poria cocos invigorate the spleen and replenish qi, longan pulp and Ziziphus jujuba seed nourish the heart and calm the mind, and Scutellaria baicalensis clears away depression and heat, so they are all adjuvant medicines; Licorice is mild in nature and harmonizes the various medicines, so it is the guiding medicine.

[0011] The traditional method of making water decoctions usually involves soaking the Chinese medicine in water, decocting it quickly with high heat, then decocting it slowly with low heat after boiling, and then separating the drug residue from the liquid medicine before letting the patient take the liquid medicine. This method does not utilize the drug residue, so it is difficult to utilize the active ingredients in the medicine. In addition, the bagged water decoctions made by this method have a short shelf life when stored in a refrigerator. Therefore, it is necessary to improve the production process of traditional water decoctions.

[0012] To this end, the present invention proposes a method for preparing a nano Chinese medicine composition for improving sleep, which is characterized by:

[0013] 1) Weigh and wash: take 8-20 parts of bupleurum, 5-15 parts of scutellaria, 5-15 parts of angelica, 10-30 parts of white peony root, 10-30 parts of poria, 10-30 parts of stir-fried atractylodes, 10-30 parts of spiny jujube seeds, 5-20 parts of codonopsis, 10-30 parts of longan pulp, 5-15 parts of roasted liquorice root, 10-20 parts of gardenia, and wash with clean water;

[0014] 2) Cleaning and pulverizing: putting the cleaned medicine in step 1) into a dryer for drying, putting the dried medicine into an ultrafine pulverizer for pulverizing, and pulverizing the medicine into a Chinese medicine powder of ≤1.5 μm;

[0015] 3) Solvent extraction: Add the Chinese medicine powder obtained in step 2) into a solvent at a solid-liquid ratio of 1:8, and boil for 1.5 hours. The solvent is 70% ethanol solution, and reflux is used;

[0016] 4) vacuum distillation: adding the extract obtained in step 3) to an external circulation vacuum concentration tank, selecting a pressure of 0.06 kPa, and performing vacuum distillation to concentrate;

[0017] 5) Spray drying: The slurry obtained by concentrating in step 4) is fed into a high-speed centrifugal spray dryer, and the operating temperature is selected to be 160-180° C.;

[0018] 6) Granulation: The spray-dried particles in step 5) are fed into a FL-60 fluidized bed granulator and granulated at 75°C.

[0019] In the above preparation method, an ultrafine pulverizer is first used to pulverize the drug before solvent extraction, which increases the contact area between the drug and the solvent and can better promote the dissolution of the active ingredients of the drug. Secondly, since the powder after pulverization by the ultrafine pulverizer is very fine, there is no problem of separation of drug residues and liquid. Moreover, since the medicinal materials have been fully pulverized by the ultrafine pulverizer and there is fine powder, there is no need to add other granulating agents in the subsequent spray drying granulation process, which can reduce the negative impact of additives on the human body. Compared with the traditional Chinese medicine decoction method, the drug residue has finally entered the product in disguise, which can achieve the purpose of making full use of the effective ingredients in the Chinese medicinal materials and reducing resource waste.

[0020] After searching the literature for the active ingredients and extraction methods of the above-mentioned traditional Chinese medicines, it is believed that the use of 70% ethanol solution can better take into account the extraction of ketones, glycosides, volatile oils and other components in the above-mentioned medicinal materials, and the use of ethanol solution for extraction is also easy to concentrate and dry by heating in subsequent operations, and will not cause extractant residues and traditional Chinese medicine odor to the final granular product.

[0021] Compared with the prior art, the above-mentioned Chinese medicine composition and the corresponding extraction method have the following beneficial effects:

[0022] 1. The formula is scientific and reasonable, easy to take, low cost, has the effects of soothing the liver and strengthening the spleen, nourishing the heart and calming the mind, and has a significant effect on the treatment of insomnia caused by liver depression and spleen deficiency, and mental malnutrition. It has a definite effect and is safe to use. It is an innovation in the treatment of insomnia caused by liver depression and spleen deficiency, mental malnutrition, and the formula harmonizes the three organs of the heart, liver, and spleen, and can hit the pathogenesis of liver depression and spleen deficiency, mental malnutrition, and has a good clinical effect. It belongs to the innovation of pathogenesis theory and therapeutic drug innovation. This prescription is safe and effective, and can be applied to adolescents, the elderly and other patients with insomnia caused by liver depression and spleen deficiency, mental malnutrition, and fills the gap of no medicine available for special populations.

[0023] 2. The proposed preparation method of the composition greatly overcomes the shortcomings of the preparation and use of water decoctions, is conducive to the preservation of medicines, improves the efficacy of medicines, is safer to use, and is more convenient for patients to use. The alcohol reflux decoction method used can greatly improve the extraction efficiency of the effective active ingredients of the Chinese medicine composition. No other solvents except ethanol and water are introduced in the extraction. The solvent can be removed by drying in the end, ensuring that the final medicine is not contaminated by the extraction solvent. The ultrafine grinding process in the preparation method avoids the production of drug residues and realizes the full utilization of medicinal materials. Compared with the traditional preparation method, it is an innovation in medication.

[0024] 3. After rigorous animal experiments and human clinical trials, the nano Chinese medicine composition for improving sleep prepared by the present invention has generally better improvement effects on related sleep memory, mood and other indicators of experimental mice and insomniacs than existing Western medicines for treating sleep amnesia, and has fewer side effects and is safer to use, which has incomparable advantages. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Comparison of Nissl staining results of mice in each experimental group (Note: A normal group B model group C western medicine group D control 1 E control 2 F control 3 G Example 3) DETAILED DESCRIPTION

[0026] The preparation examples and control examples of the Chinese medicine of the present invention are shown in Table 1 below

[0027] Table 1 Preparation Example and Control Example Settings

[0028]

[0029] The difference between Example 2-3 and Example 1 is that the material configuration is different, the extraction method used is the same, and the new preparation method proposed by the present invention is adopted. In Control 1, two drugs, Bupleurum and Scutellaria, are missing. The formula of Control 2 is not within the scope of the medicinal material ratio required for protection. The difference between Control 3 and Example 3 is that it adopts the preparation method of traditional medicinal decoction, and the specific operation method is as follows: Bupleurum, Scutellaria, Angelica, White Peony, Poria, Stir-fried Atractylodes, Ziziphus Ziziphi Spinosae, Codonopsis, Longan Meat, and Roasted Licorice are mixed together, put into a medicine pot, add water to immerse, soak for 30-60min, boil over high heat until boiling, then use slow fire to decoct for 20-30min, filter, obtain filtrate, add water to the filter residue to immerse, boil over high heat until boiling, then use slow fire to decoct for 20-30min, filter, obtain filtrate, merge the two filtrates, and mix well.

[0030] Experimental evidence of effectiveness:

[0031] (1) Animal experiments

[0032] 70 8-week-old SPF female Balb / c mice were selected, and the individual weight difference was controlled within the range of 23±2g. Modeling grouping: randomly divided into 7 groups, namely ① Example 3; ② Control 1; ③ Control 2; ④ Control 3; ⑤ Western medicine group (R-zopiclone group); ⑥ Model group; ⑦ Normal group.

[0033] Except for the 10 mice in the normal group, the other groups were first intraperitoneally injected with PCPA (300 mg / kg / d, once a day) for 3 consecutive days to obtain insomnia mice. Administration: Groups ①-④ were gavaged with 16.8 g / kg / d of the Chinese medicine prepared by the formula configuration of Example 3, Control 1, Control 2, and Control 3, respectively, where Control 3 needed to be prepared by concentration and drying after the water decoction was prepared to make solid particles equal to that of Example 3 for mice to take, and the Western medicine group ⑤ was given 0.2 mg / kg / d of right zopiclone, which was configured in physiological saline and gavaged every day at a dose of 16.8 g / kg / d. The normal group and the model group were gavaged with an equal volume of physiological saline every day. The above-mentioned drugs, except for physiological saline, were configured into liquid solutions (configured with physiological saline) at a volume of 0.1 ml / 10 g (mouse body weight) and then gavaged, and the drugs were administered once a day for 14 consecutive days. Morris water maze test was performed on the 9th day after administration to observe the effects of each group of drugs on learning and memory of insomniac mice. Pentobarbital sodium induced sleep test was performed on the 13th day after administration to observe sleep-wake status. Sucrose preference experiment was performed on the 14th day to observe the depression of mice.

[0034] The Morris water maze consists of a circular pool, a movable platform hidden under the water, and an automatic image acquisition and processing system. The animal (mice) is placed in the water with its head facing the pool wall, and the placement position is randomly selected from the four starting positions of east, west, south, and north. The time from the animal being placed in the water to finding the underwater platform is recorded. The escape latency is the time required for the animal to successfully find the platform for the first time after entering the water each time. Its length represents the quality of the animal's spatial learning and memory ability. A short latency indicates that the animal has good learning and memory ability. The time in the quadrant where the platform is located is an indicator for evaluating the animal's learning ability. The longer the animal is active in this quadrant, the better the animal's spatial memory ability is.

[0035] Sodium pentobarbital induced sleep test: Sodium pentobarbital is a commonly used sleep inducing agent that causes animals to fall asleep by inhibiting the activity of the central nervous system. The sleep latency refers to the time it takes for mice to fall asleep after being injected with sodium pentobarbital. The shorter the sleep latency, the better the sedative effect. Sleep duration refers to the duration of sleep in mice. The longer the sleep duration, the better the sedative effect. If the drug is used on the basis of sodium pentobarbital injection, the sleep latency of the pentobarbital group + drug group is shorter and the sleep time is longer than that of the pentobarbital group alone, indicating that the drug has a better effect in improving sleep.

[0036] Sucrose preference test: It is a commonly used animal behavior experiment, which evaluates the animal's sucrose preference by providing the experimental animals with two choices: sucrose-containing liquid and ordinary water. Under normal circumstances, animals prefer to drink sucrose water, but when the animals are depressed, their interest in the favorite things will decrease, and the amount of sucrose water they drink will decrease. The percentage of decrease is used to observe the depression of mice. The greater the decrease, the more severe the depression.

[0037] After the above behavioral experiments, 3 mice from each group were perfused with 10% paraformaldehyde through the heart, and the blood was quickly washed away. The tissue was pre-fixed before the mice died to avoid autolysis of brain tissue, which was used for subsequent Nissl staining to observe pathological changes in hippocampal tissue. For the remaining mice, fresh hypothalamic tissue homogenate was taken on ice, and the 5-hydroxytryptamine (5-TH) content of the hypothalamic tissue of each group of mice was detected by competitive enzyme-linked immunosorbent assay (ELISA). 5-hydroxytryptamine (5-TH) is a neurotransmitter related to emotions, and its level can be used as a reference index for evaluating the body's pleasure. Generally, a high concentration indicates a strong sense of pleasure.

[0038] 1. Water maze experiment

[0039] The results are shown in Table 1. Compared with the normal group, the model group had a significant increase in escape latency and a significant decrease in target quadrant residence time; compared with the model group, the Chinese medicine group ①-④ and the Western medicine group ⑤ had a significant decrease in escape latency, an increase in the number of platform crossings, and an increase in target quadrant residence time, indicating that compared with the model group, both the Chinese medicine and Western medicine intervention groups can effectively improve the learning and memory and spatial memory abilities of insomnia mice. However, compared with the Western medicine group, the escape latency and target quadrant residence time data of the Chinese medicine group were closer to those of the normal group, indicating that the improvement of the Chinese medicine group was better than that of the Western medicine right-zopiclone group. In the Chinese medicine group ①-④, the escape latency, platform crossing times, and platform quadrant residence time data of Example 3 were closer to those of the normal group, and had the best therapeutic effect on insomnia mice.

[0040] Table 1 Escape latency, number of platform crossings, and platform quadrant residence time of mice in each group

[0041] Experimental Group Medication Escape latency(s) Number of platform crossings Platform quadrant dwell time (s) ① Example 3 19.41±15.63 1.33±1.16 20.78±9.25 ② Comparison 1 28.31±14.84 1.32±1.16 13.42±2.63 ③ Comparison 2 23.48±17.02 1.33±0.58 19.81±7.59 ④ Comparison 3 25.85±20.70 1.20±1.30 15.06±24.30 ⑤ Western medicine group 32.28±22.98 1.10±1.30 11.56±5.08 ⑥ Model Group 43.72±22.40 0.25±0.50 9.92±4.58 ⑦ Normal group 15.85±20.60 1.00±1.23 22.78±8.26

[0042] 2. Pentobarbital sodium synergistic sleep test

[0043] Each mouse was given 35 mg kg -1Sodium pentobarbital was used to induce sleep in mice, and the sleep latency and sleep duration of each group of mice (6 mice in each group) were observed. The results are shown in Table 2. Compared with the normal group of mice, the sleep latency of the model group mice without treatment drug intervention was longer, and the sleep time was significantly reduced. Compared with the model group, the sleep latency of the Chinese medicine group ①-④ and the Western medicine group ⑤ were improved. The improvement effect of the sleep latency of the Chinese medicine group ①-④ was better than that of the Western medicine group, and only the experimental group ① and the control 2 were better than the Western medicine group in terms of sleep time. Compared with the normal group, the sleep latency and sleep time of the Chinese medicine group ① Example 3 were closer to those of the normal group, indicating that its intervention effect on insomnia mice was the most ideal.

[0044] Table 2 Sleep latency and sleep duration of mice in each group

[0045] Experimental Group Medication Sleep latency (min) Sleep time (min) ① Example 3 4.88±0.17 72.64±4.98 ② Comparison 1 5.64±0.33 51.20±1.59 ③ Comparison 2 5.72±0.2 68.51±1.16 ④ Comparison 3 5.9±0.17 55.35±1.32 ⑤ Western medicine group 6.32±0.17 66.38±2.05 ⑥ Model Group 9.64±0.86 29.14±3.68 ⑦ Normal group 4.41±0.73 78.64±4.98

[0046] 3. Results of sucrose preference experiment

[0047] Referring to Table 3, the results of the sucrose preference experiment show that compared with the normal group, the sugar intake rate of the model group was significantly reduced, compared with the model group, the sugar intake rates of the Chinese medicine groups ①-④ and the western medicine group ⑤ increased, and compared with the western medicine group, the sugar intake rates of the Chinese medicine groups ①-④ were significantly improved, especially the sugar intake rate of Example 3 was closer to that of the normal group, indicating that Example 3 has a great effect on improving the diet of mice, and the depression is not obvious.

[0048] Table 3 Sugar intake rate of mice in each group after gavage

[0049] Experimental Group Medication Sugar intake rate (%) ① Example 3 91±0.03 ② Comparison 1 85±0.01 ③ Comparison 2 87±0.02 ④ Comparison 3 81±0.01 ⑤ Western medicine group 78±0.05 ⑥ Model Group 75±0.01 ⑦ Normal group 93±0.04

[0050] 4. Nissl staining results

[0051] Nissl staining results Figure 1 Show: The hippocampal pyramidal neurons in the normal group A are neatly arranged and compact, with regular morphology. Compared with the normal group, the hippocampal pyramidal neurons in the model group B are disordered and sparsely arranged, the Nissl staining is deepened, and there is neuronal loss. Compared with the model group, the number of hippocampal pyramidal neurons in the western medicine group C and the Chinese medicine group DG increased, and the arrangement was more neat. Among them, the color depth of the group G under the microscope was closer to the normal group, but there were still differences in morphology and compactness from the normal group.

[0052] 5. Results of 5-TH in the hypothalamus of mice in each group

[0053] The hypothalamus of mice in each group was taken, and the ELSH kit was used to detect the 5-TH content in the hypothalamus. The results showed that compared with the normal group, the 5-TH content in the model group was significantly reduced, and compared with the model group, the 5-TH content in the Chinese medicine group ①-④ and the Western medicine group was significantly increased. Compared with the Western medicine group, the 5-TH content in the Chinese medicine group ①-④ was higher, among which the 5-TH value of Example 3 was higher, which was closer to that of the normal group mice, indicating that the mental state of the mice under the intervention of Example 3 was closer to that of the normal group, and the mood was more pleasant.

[0054] Table 4 5-TH content in the hypothalamus of mice in each group

[0055] Experimental Group Medication 5-TH value (OD value) ① Example 3 65.56±1.23 ② Comparison 1 58.21±2.25 ③ Comparison 2 63.37±5.44 ④ Comparison 3 60.31±1.57 ⑤ Western medicine group 60.19±0.81 ⑥ Model Group 55.67±0.79 ⑦ Normal group 67.97±1.99

[0056] The above series of mouse experiments prove that the nano Chinese medicine composition ratio proposed in the present invention can improve the learning and cognitive functions of insomnia mice, effectively increase the sleep time of insomnia mice, improve the sleep latency and sleep duration, as well as their appetite and depression, and improve their mood. Compared with the Chinese medicine controls 1-3, the improvement advantage of the nano Chinese medicine composition ratio proposed in the present invention is more obvious. Most indicators of Example 3 are better than those of the Chinese medicine control group and the Western medicine group.

[0057] (2) Clinical trials

[0058] 1. Case selection criteria

[0059] Insomnia diagnostic criteria: Refer to the diagnostic criteria for insomnia in the third edition of the Chinese Classification and Diagnostic Criteria for Mental Disorders (CCMD-3). ① Insomnia as the only symptom. ② Dissatisfaction with sleep quality and time. ③ Occurring at least 3 times a week for at least 1 month. ④ Exclude secondary insomnia caused by other serious physical illnesses or mental problems.

[0060] TCM symptom standard: Insomnia adopts the diagnostic standard of insomnia-liver depression, spleen deficiency and mental malnutrition type. It is formulated with reference to the diagnostic standard of "Insomnia" liver depression, spleen deficiency and mental malnutrition type in the "TCM Disease Diagnosis and Efficacy Standard (1994)" issued by the State Administration of Traditional Chinese Medicine. Main symptoms: insomnia, depression / worry / anxiety, loose stools, and loose stools and diarrhea will worsen when the mood is depressed or the diet is not careful; Secondary symptom one: chest tightness, sighing, flank pain; Secondary symptom two: fatigue, sallow complexion, poor appetite, abdominal distension and bowel sounds, sticky and uncomfortable stools or dry and loose stools, tongue: pale tongue with white fur, pulse: stringy or fine stringy pulse. Patients with one of the above main symptoms and one or more secondary symptoms, and with corresponding tongue and pulse, can be diagnosed as insomnia of liver depression, spleen deficiency and mental malnutrition type.

[0061] Anyone with the above symptoms will be used as the criteria for case selection.

[0062] 2. Inclusion and Exclusion Criteria

[0063] 1) Inclusion criteria: ① Meet the diagnostic criteria for primary insomnia in the Chinese Classification and Diagnostic Criteria of Mental Disorders (CCMD-3); ② Meet the TCM diagnostic criteria for insomnia due to liver depression and fire transformation; ③ PSQI score ≥ 7 points, HAMD score > 8 points; ④ Voluntary participation and signing of informed consent.

[0064] 2) Exclusion criteria: ① Those with physical illness or (and) other mental disorders; ② Women who are pregnant or breastfeeding; ③ Those with insomnia caused by work and rest habits; ④ Those with insomnia caused by systemic diseases (such as pain, fever, cough, surgery, etc.), external environmental interference and other mental illnesses; ⑤ Those caused by drugs.

[0065] 3) Criteria for case exclusion and experiment termination: ① Subjects who did not take the designated medication as prescribed or took additional medications that affected sleep or had to take other medications that affected sleep for personal reasons; ② Subjects with incomplete clinical data records; ③ Subjects who had serious adverse reactions during the observation and could not continue the study.

[0066] 3. Treatment options

[0067] Control group: Estazolam tablets (Shanghai Xinyi Pharmaceutical Factory, specification: 1 mg, national medicine standard H31021534) + basic treatment (education + cognitive therapy)

[0068] Treatment group 1: using the Chinese medicine composition in Example 3 + basic treatment (education + cognitive therapy)

[0069] Main outcome measure: Pittsburgh Sleep Quality Index (PSQI). Each component is scored on a 0-3 scale, and the total score of each component is the total PSQI score, which ranges from 0 to 21. The higher the score, the worse the sleep quality. It can be evaluated (sleep quality, sleep onset time, sleep time, sleep efficiency, sleep disorders, hypnotic drugs, daytime dysfunction, etc.).

[0070] Secondary outcome indicators: ① Quantitative scoring of TCM symptom grading; refer to the TCM symptom grading quantitative table for insomnia with liver depression, spleen deficiency and malnutrition of the mind in the "Standards for Diagnosis and Efficacy of TCM Diseases and Syndromes" formulated by the State Administration of Traditional Chinese Medicine in 1994; the more points, the better the effect.

[0071] ②Hamilton Depression Rating Scale (HAMD).Hamilton Depression Rating Scale (HAMD) is an important tool for doctors to assess the severity of patients' depressive symptoms. The higher the score, the more severe the depression.

[0072] 4. Efficacy evaluation criteria

[0073] 1) Pittsburgh Sleep Quality Inventory score: The evaluation criteria are formulated by calculating the score reduction rate according to the PSQI scale scoring criteria. Score reduction rate = (score before treatment - score after treatment) / score before treatment * 100%. ① Cured: Sleep time returns to normal or the sleep time is > 6 hours, the sleep is deeper, and the energy is full after waking up, and the score reduction rate is > 75%. ② Significantly effective: Sleep is significantly improved, sleep is deeper, and sleep time is extended to > 3 hours, and the score reduction rate is 51%-75%. ③ Effective: Symptoms have improved to a certain extent, and sleep time is extended compared to before, but < 3 hours, and the score reduction rate is 25%-50%. ④ Ineffective: Insomnia symptoms have not improved or have worsened, and the score reduction rate is < 25%.

[0074] 2) Assessment of TCM syndrome scale: The scoring standard was developed with reference to the "Guidelines for Clinical Research of New Chinese Medicines". Each major symptom was scored as 0, 2, 4, or 6 points according to the severity, and each secondary symptom was scored as 0, 1, 2, or 3 points according to the severity. The TCM symptom scores before and after treatment were compared to determine the changes.

[0075] 3) Hamilton Depression Rating Scale score change

[0076] 4) Safety evaluation: Both groups of patients underwent complete blood routine, urine routine, stool routine, liver and kidney function and electrocardiogram examinations before and after treatment.

[0077] 5. Statistical Analysis

[0078] SPSS 18.0 software was used for analysis, and all data were entered twice. P < 0.05 was considered statistically significant. Measurement data were described by mean ± standard deviation (x ± s), and the comparison of normal distribution was performed by t test, the comparison of non-normal distribution was performed by rank sum test, and the comparison of counts was performed by χ2 test.

[0079] 6. Results

[0080] A total of 58 subjects were included in this study, of which 1 patient dropped out of the treatment group because the patient refused to take the medicine, and 1 patient dropped out of the control group because of residual symptoms during the day after taking the medicine. Finally, 28 patients were included in the experimental group and 28 patients were included in the control group. The basic information of the two groups of patients was not statistically different after testing, as shown in Table 5.

[0081] Table 5 Basic data statistics

[0082] Experimental group (n=28) Control group (n=28) <![CDATA[X 2 / t value]]> P Age (standard deviation) 38.75(13.67) 39.25(13.26) \ 0.866 Gender (male / female) 13 / 15 14 / 14 0.072 0.788

[0083] 1) Comparison of PSQI scores between the treatment group and the control group before and after treatment

[0084] As shown in Table 6, the PSQI score indicates that both Western medicine and Chinese medicine can improve sleep. After 4 weeks of treatment, the efficacy of Chinese medicine is better than that of the Western medicine group, and the difference is statistically significant.

[0085] Table 6 Comparison of PSQI scores between the treatment group and the control group before and after treatment

[0086] Number of cases Pre-treatment score After treatment score Treatment Group 28 16.72±1.82 <![CDATA[7.23±3.95 *# ]]> Control group 28 16.52±1.75 <![CDATA[9.65±3.64 * ]]>

[0087] Note: * Compared with the group before treatment, P < 0.05; # Compared with the control group after treatment, P<0.05

[0088] 2) Comparison of clinical efficacy between the treatment group and the control group after treatment

[0089] As shown in Table 7, statistical data analysis showed that the total clinical effective rate of the Chinese medicine composition in Example 3 was 89.29%, and the total clinical effective rate of the control group was 67.86%. The results of the rank sum test analysis were statistically significant (P<0.05), indicating that the clinical efficacy of the treatment group was higher than that of the control group.

[0090] Table 7 Comparison of clinical efficacy between the treatment group and the control group

[0091] Number of cases get well Significant effect efficient invalid Total Effectiveness Treatment Group 28 8 10 7 3 89.26％ Control group 28 4 6 9 9 67.84％

[0092] Note: The clinical efficacy of the two groups of data was compared by rank sum test, Z=-2.272, P=0.023<0.05, which was statistically significant.

[0093] 3) Comparison of the difference in the scores of the TCM symptom grading quantitative table between the two groups

[0094] As shown in Table 8, before treatment, the TCM syndrome scores of insomnia between the two groups were compared, P=0.174>0.05, and there was no statistically significant difference between the two groups; after treatment, the TCM syndrome scores of insomnia between the two groups were compared, P=0.01<0.05, and the difference was statistically significant.

[0095] Table 8 Comparison of TCM syndrome scores of insomnia between the treatment group and the control group before and after treatment

[0096] Number of cases Pre-treatment score After treatment score Treatment Group 28 13.35±1.42 5.86±2.59 Control group 28 13.26±1.41 8.43±2.10

[0097] 4) Comparison of HAMD scores between the two groups after treatment

[0098] As shown in Table 9, before treatment, the HAMD scores of the two groups were compared, P = 0.317> 0.05, and there was no statistically significant difference between the two groups; after treatment, the HAMD scores of the two groups were compared, P = 0.035< 0.05, and the difference was statistically significant.

[0099] Table 9 Comparison of HAMD scores between the treatment group and the control group before and after treatment

[0100] Number of cases Pre-treatment score After treatment score Treatment Group 28 15.18±1.11 <![CDATA[7.91±2.61 *# ]]> Control group 28 15.22±1.30 9.37±2.95

[0101] 5) Safety evaluation

[0102] During the treatment process, one case of dizziness occurred in the control group, which was relieved on its own without any treatment. No adverse reactions occurred in all participants during the medication period.

[0103] The results of clinical experiments showed that the PSQI score, overall efficacy, TCM symptom score and HAMD score of the treatment group were better than those of the control group, and the difference was statistically significant (P < 0.05), indicating that the clinical efficacy of the present invention is significant, it can improve the symptoms of liver depression and spleen deficiency, and mental malnutrition in insomnia, and it has good safety.

[0104] It should be understood that although the present specification is described in terms of implementation methods, this narrative style of the specification is only for the sake of clarity, and those skilled in the art should take the specification as a whole to form other implementation methods that can be understood by those skilled in the art, which should also fall within the scope of protection of the present invention.

Claims

1. A method for preparing a Chinese medicine composition for improving sleep, Features: 1) Weigh and wash: Take 10 parts of Bupleurum, 10 parts of Scutellaria, 10 parts of Angelica, 10 parts of White Peony, 20 parts of Poria, 20 parts of Stir-fried Atractylodes, 20 parts of Ziziphus jujuba seeds, 10 parts of Codonopsis, 20 parts of Longan pulp, 10 parts of Roasted Licorice, 15 parts of Gardenia, and wash with clean water; 2) Crushing: putting the washed medicine in step 1) into a dryer for drying, and putting the dried medicine into an ultrafine grinder for crushing, and crushing the medicine into a Chinese medicine powder of ≤1.5 μm; 3) Solvent extraction: the Chinese medicine powder obtained in step 2) is added with 70% ethanol solution at a solid-liquid ratio of 1:8, and refluxed for 1.5 hours to obtain an extract; 4) Vacuum distillation: adding the extract obtained in step 3) to an external circulation vacuum concentration tank, performing vacuum distillation and concentration; 5) Spray drying: The slurry obtained by concentration in step 4) is fed into a high-speed centrifugal spray dryer, and the operating temperature is selected to be 160-180°C; 6) Granulation: The spray-dried material in step 5) is fed into a boiling granulator and granulated at 75°C.

2. A Chinese medicine composition granule obtained according to the preparation method according to claim 1.

3. Use of the Chinese medicine composition particles obtained by the preparation method according to claim 1 in preparing medicines, Features: The Chinese medicine composition granules can be used to prepare a preparation for improving sleep function.

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