Pharmaceutical composition for treating insomnia (heart-spleen deficiency type) and hypomnesis and preparation method thereof

Abstract

The invention discloses a pharmaceutical composition for insomnia (heart-spleen deficiency type) and hypomnesis and a preparation method of the pharmaceutical composition, and relates to the technical field of medicines.The composition is prepared from acanthopanax, astragalus membranaceus, lucid ganoderma, ginseng leaves, schisandra chinensis and gastrodia elata, the prescription mainly aims at the pathogenesis of insomnia caused by heart-spleen deficiency, the synergistic effect is achieved from multiple aspects, and the curative effect is good. The traditional Chinese medicine composition has the effect of treating insomnia caused by heart and spleen, and is a comprehensive and effective treatment scheme. Meanwhile, the effect of improving memory is also achieved. The prescription provided by the invention can be prepared into granules, oral preparations and the like. The pharmaceutical composition disclosed by the invention has a remarkable insomnia treatment effect which is superior to that of Western medicines such as eszopiclone tablets and estazolam tablets, the sleep inducing time is shortened, the sleep time of a patient with difficulty in falling asleep can be effectively shortened, meanwhile, the sleep time is optimally prolonged, the insomnia treatment effect is remarkable, and the situations of somnolence and non-awakening are avoided; the memory can also be improved. Meanwhile, no dependence is caused, and the insomnia is fundamentally treated.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss and a preparation method thereof. Background Art

[0002] Insomnia is a type of disease characterized by the inability to get normal sleep due to emotional and dietary injuries, or after illness and old age, insufficient endowment, guilty conscience and other causes, which cause the mind to be unnourished or uneasy, resulting in frequent inability to get normal sleep. High work pressure during the day, excessive nerve tension, and the inability to relax when sleeping are all causes of insomnia. Insomnia manifests as difficulty falling asleep, not being able to fall asleep, waking up early, being unable to fall asleep again after waking up, frequently waking up from nightmares, feeling that you have nightmares all night, not being able to eliminate fatigue, and recovering physical strength and energy after sleeping. In severe cases, you can't sleep all night. The onset time can be long or short. The shorter one can get better in a few days, and the longer one can last for several days and be difficult to recover. It is easy to be awakened, some are sensitive to sound, and some are sensitive to light. Many people with insomnia like to have wild thoughts. Insomnia can cause fatigue, uneasiness, general discomfort, listlessness, slow reaction, headache, and poor memory. Its biggest impact is mental, and a serious one can lead to schizophrenia.

[0003] Insomnia is one of the common clinical diseases. Although it is not a critical disease, it hinders people's normal life, work, study and health, and can aggravate or induce palpitations, chest pain, dizziness, headache, stroke and other symptoms. Insomnia brings long-term pain to patients. The drugs for treating insomnia mainly include drugs for calming the nerves and nourishing the brain, sedatives and sleeping pills. The more common drugs for treating insomnia in clinical practice mainly include non-benzodiazepines and benzodiazepines. Long-term use of the above-mentioned sleeping pills is prone to dependence and addiction. If the drug is stopped suddenly, rebound symptoms will occur, insomnia will become more difficult to fall asleep, and long-term use can also lead to slow reaction and memory loss. Sleeping pills have a central muscle relaxant effect. Some people may experience dizziness, general weakness, unsteady walking and other symptoms after taking them. There is a risk of accidents such as falls, especially in the elderly population, which affects life and needs special attention. Sleeping pills can also cause patients with respiratory insufficiency. After taking them, they may suppress breathing and endanger their lives. Therefore, such people should not take them. There is also the possibility of liver and kidney damage. Taking a large amount of sleeping pills at one time can lead to overdose poisoning, respiratory depression, and endanger life. At the same time, long-term use of sleeping pills can also cause other iatrogenic diseases. At the same time, some insomnia patients take melatonin tablets. Melatonin is an endogenous hormone-like drug that can mainly regulate the endocrine function of the human body. Melatonin is a health product. Health products refer to foods with specific health functions. They are suitable for specific groups of people to take and have the function of regulating body functions, but are not intended to treat diseases. Excessive use of melatonin may lead to a decrease in the secretion capacity of melatonin and cause endocrine disorders. Men who take melatonin frequently may suffer from problems such as decreased libido and sexual dysfunction.

[0004] Based on the drawbacks of the above-mentioned sleeping pills and health products, the present invention develops a series of pharmaceutical compositions for insomnia (heart and spleen deficiency type) and memory loss. It has an excellent therapeutic effect on patients with heart and spleen deficiency type insomnia, fundamentally solves the pain of patients with heart and spleen deficiency type insomnia, and patients who maintain physical and mental pleasure can completely return to a normal sleep state, and can also improve the memory of insomnia patients. It also has the effect of improving the memory of patients with severe memory loss. The pharmaceutical composition of the present application is a drug that brings the best treatment and improvement effect to patients with heart and spleen deficiency type insomnia and patients with severe memory loss.

[0005] At present, the drugs for treating insomnia are mainly sedatives and sleeping pills, which are all Western medicines. Although they can produce immediate therapeutic effects, taking sedatives and hypnotic drugs may cause adverse symptoms such as short-term memory damage, hallucinations, coordination disorders, dizziness and vertigo, which may still persist the next day. Such drugs are addictive. Long-term medication will increase the patient's tolerance to the drug and cause dependence on the drug. Compared with Western medicine, Chinese herbal medicines have unique advantages in treating insomnia, such as significant efficacy, few side effects, low recurrence rate, and no addiction. However, most of the Chinese patent medicines for treating insomnia on the market are not effective and cannot achieve therapeutic effects. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides a pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss and a preparation method thereof.

[0007] The invention discloses a medicine composition for insomnia (heart and spleen deficiency type) and memory loss. The medicine composition comprises 5 to 100 parts of Acanthopanax senticosus, 1 to 20 parts of Ginseng leaves, 1 to 20 parts of Schisandra chinensis, 2 to 50 parts of Astragalus, 2 to 60 parts of Ganoderma lucidum and 1 to 20 parts of Gastrodia elata in parts by weight.

[0008] Furthermore, the pharmaceutical composition is composed of 10-80 parts of Acanthopanax senticosus, 5-15 parts of Ginseng Leaves, 5-15 parts of Schisandrae Chinensis, 5-40 parts of Astragalus, 5-50 parts of Ganoderma Lucidum and 5-15 parts of Gastrodia elata in parts by weight.

[0009] Furthermore, the pharmaceutical composition is composed of 20-60 parts of Acanthopanax senticosus, 10-15 parts of Ginseng Leaves, 10-15 parts of Schisandrae Chinensis, 10-30 parts of Astragalus, 10-40 parts of Ganoderma Lucidum and 10-15 parts of Gastrodia elata in parts by weight.

[0010] Furthermore, the pharmaceutical composition is composed of 40-50 parts of Acanthopanax senticosus, 10-15 parts of Ginseng Leaves, 10-15 parts of Schisandrae Chinensis, 10-20 parts of Astragalus, 10-30 parts of Ganoderma Lucidum and 10-15 parts of Gastrodia elata in parts by weight.

[0011] Furthermore, the pharmaceutical composition is composed of 25 parts of Acanthopanax senticosus, 5 parts of Ginseng Leaves, 3 parts of Schisandrae Chinensis, 15 parts of Astragalus, 10 parts of Ganoderma Lucidum and 5 parts of Gastrodia elata in parts by weight.

[0012] The method for preparing the granules of the Bujuexiao series of pharmaceutical compositions for insomnia and memory loss of heart and spleen deficiency type of the present invention is carried out in the following manner:

[0013] 1) Weigh 5-100 parts of Acanthopanax senticosus, 1-20 parts of Ginseng Leaves, 1-20 parts of Schisandrae Chinensis, 2-50 parts of Astragalus, 2-60 parts of Ganoderma Lucidum and 1-20 parts of Gastrodia elata according to weight;

[0014] 2) mixing the medicinal materials weighed in step 1), adding 8 times the amount of water, reflux extraction, filtering, and obtaining filtrate 1; ultrasonic-assisted extraction of the medicinal residue, filtering, and obtaining filtrate 2; adding 8 times the amount of water to dissolve the residue, reflux extraction, filtering, and obtaining filtrate 3; combining filtrate 1, filtrate 2, and filtrate 3 for later use;

[0015] 3) concentrating the combined filtrate from step 2) by a reduced pressure concentration method;

[0016] 4) Add auxiliary materials to make tablets, granules, capsules or oral liquid.

[0017] Further, in step 2), 8 times the amount of water was added, refluxed for 2 hours, and filtered to obtain filtrate 1; the residue was ultrasonically extracted for 10 minutes at 40°C, and filtered to obtain filtrate 2; the residue was dissolved in 8 times the amount of water, refluxed for 1.5 hours, and filtered to obtain filtrate 3.

[0018] Furthermore, the concentration in step 3) is concentrated to an extract having a relative density of 1.10 to 1.30 at 60°C.

[0019] Furthermore, the granules are prepared as follows:

[0020] After drying the concentrated extract in step 3), measuring its water content ≤ 3%, taking it out and cooling it; adding dextrin and citric acid as auxiliary materials to mix with the dried extract in step 4), then adding 10% of water based on the total amount of the original auxiliary materials, stirring and mixing to obtain a soft material, granulating, drying, granulating, and packaging.

[0021] Furthermore, the oral liquid is prepared as follows:

[0022] 1) Weigh 5-100 parts of Acanthopanax senticosus, 1-20 parts of Ginseng Leaves, 1-20 parts of Schisandrae Chinensis, 2-50 parts of Astragalus, 2-60 parts of Ganoderma Lucidum and 1-20 parts of Gastrodia elata according to weight;

[0023] 2) mixing the medicinal materials weighed in step 1), adding 8 times the amount of water, reflux extraction, filtering, and obtaining filtrate 1; ultrasonic-assisted extraction of the medicinal residue, filtering, and obtaining filtrate 2; adding 8 times the amount of water to dissolve the residue, reflux extraction, filtering, and obtaining filtrate 3; combining filtrate 1, filtrate 2, and filtrate 3 for later use;

[0024] 3) concentrating the combined filtrate from step 2) by a reduced pressure concentration method;

[0025] 4) Add the concentrated extract to water at 70-85°C and stir evenly, then add potassium sorbate, sodium citrate and stevioside excipients, add water to make up to volume, centrifuge, filter, fill and sterilize.

[0026] Acanthopanax contains many chemical components, mainly glycosides, polysaccharides, flavonoids, lignin, coumarins, and a variety of trace elements and amino acids, among which glycosides, flavonoids and polysaccharides are the main active ingredients of Acanthopanax. Acanthopanax has the functions of regulating the central nervous system, improving blood supply to brain tissue, regulating immunity, resisting fatigue, lowering blood sugar, resisting ultraviolet radiation, and resisting tumors.

[0027] Ginseng leaves contain many chemical components, including saponins, ginseng polysaccharides, flavonoids, amino acids, organic acids, volatile oils, etc. Saponins are the main active ingredients. Ginseng leaves have the functions of protecting nerves, resisting fatigue, delaying aging, inhibiting tumor cell growth, improving body immunity, and improving cardiovascular and cerebrovascular blood supply insufficiency.

[0028] Schisandra contains many chemical components, including lignans, volatile oils, organic acids, terpenes, flavonoids, polysaccharides and inorganic elements. Among them, lignans and polysaccharides are the main components of Schisandra, which are the basic substances of various pharmacological activities. Schisandra has the functions of protecting cardiovascular and liver, anti-oxidation, and immunomodulation.

[0029] Astragalus contains many chemical components, mainly polysaccharides, flavonoids, saponins, in addition to amino acids, alkaloids, trace elements and other effective ingredients. Among them, polysaccharides, saponins and flavonoids are the main active ingredients of Astragalus. Astragalus has the effects of improving immunity, protecting cardiovascular and cerebrovascular, neuroprotection, anti-tumor effect, hypoglycemic effect, anti-aging effect, and protecting internal organs (liver, kidney, lung).

[0030] Ganoderma contains many chemical components, mainly Ganoderma polysaccharides, Ganoderma triterpenes, nucleosides, sterols and proteins, among which Ganoderma triterpenes and Ganoderma polysaccharides are the main active ingredients of Acanthopanax senticosus. Ganoderma has immune regulation, nervous system regulation, anti-oxidation, anti-tumor, liver protection and anti-inflammatory, blood sugar and blood lipid lowering, and antibacterial effects.

[0031] Gastrodia elata contains many chemical components, mainly including phenols and their glycosides, organic acids, polysaccharides, sterols, various amino acids and trace elements required by the human body, etc. Among them, gastrodin, gastrodin aglycone and gastrodia polysaccharide are the main active ingredients of Gastrodia elata. Gastrodia elata has the effects of improving memory and anti-dementia, regulating the central nervous system, enhancing immunity, protecting the liver, anti-tumor, anti-oxidation, lowering blood pressure, lowering blood lipids, anti-dizziness, brain protection, and intelligence.

[0032] Solution:

[0033] Insomnia is also called insomnia. According to traditional Chinese medicine, although there are many causes of insomnia, its pathological changes are always due to the excess of yang and the decline of yin, and the mismatch of yin and yang. The main location of the disease is the heart, which is closely related to the liver, spleen, and kidney. Because the heart governs the mind, one can sleep when the mind is at peace, and one cannot sleep when the mind is not at peace. The source of yin and yang, qi and blood, is transformed from the essence of water and grain, and is presented to the heart, so that the mind can be nourished. If the heart blood is sufficient and the essence is nourished in the brain, the brain function will be normal and the mind will be at peace.

[0034] Modern people generally experience irregular daily work and life, like to eat fatty and greasy food, have little activity, high work pressure, and overthinking. Over time, this will lead to imbalance between work and rest. Excessive fatigue will damage the spleen, and too much rest and little movement will also lead to spleen deficiency and weak qi, poor transportation and transformation, insufficient source of qi and blood, and inability to serve the heart, causing the heart to lose its nourishment, and then the heart blood and essence cannot be nourished to the brain, and the spirit is not nourished and suffers from insomnia; or because of excessive worry, the heart and spleen are damaged, and the heart damages the yin blood and the spirit is not in the house; the spleen is damaged, and the appetite is poor, the source of biochemical is insufficient, and the blood is deficient, and it cannot be served to the heart, causing restlessness. It can be seen that insufficient heart and spleen will lead to blood deficiency, which will lead to insomnia.

[0035] This prescription is mainly used to treat insomnia caused by deficiency of both heart and spleen. Acanthopanax can not only replenish the Qi of the heart and spleen, but also calm the mind and benefit the will. Astragalus can both replenish the spleen and help produce blood. The two are the main medicines to replenish the spleen, replenish Qi and produce blood; Ginseng leaves replenish Qi, Ganoderma lucidum replenishes heart blood, benefits the heart Qi and calms the mind; Schisandra chinensis replenishes the heart and kidneys, calms the mind and calms the mind. The three are the minister medicines to assist the main medicine in replenishing Qi and blood, nourishing the heart and calming the mind; Gastrodia elata is used as an adjuvant medicine to calm liver yang, which is used to improve symptoms such as dizziness and headache caused by insufficient liver yin and hyperactivity of liver yang disturbing the brain.

[0036] The above drugs cooperate with each other to treat the heart and spleen together, and play the role of replenishing qi and blood, nourishing the heart and calming the mind. By replenishing the heart and spleen, the essence of water and grain is transported normally, the biochemical transportation of qi and blood is promoted, and sufficient nourishment is provided to the heart, so that the mind is nourished, the heart blood is sufficient, the essence and qi are nourished to the brain, and the brain function is normal, thereby improving insomnia. The addition of Gastrodia elata relieves the symptoms of dizziness, headache and other symptoms associated with insomnia, and improves the overall treatment effect.

[0037] To sum up, the prescription composed of Acanthopanax, Astragalus, Ganoderma lucidum, Ginseng leaf, Schisandra chinensis, and Gastrodia elata is mainly aimed at insomnia caused by deficiency of both heart and spleen, and exerts synergistic effects from multiple aspects. It is mainly used to treat difficulty falling asleep and frequent dreams and easy awakening. At the same time, it also has a certain effect on symptoms such as palpitations, forgetfulness, fatigue, loss of appetite, dizziness, and fatigue in the limbs associated with insomnia caused by deficiency of both heart and spleen.

[0038] The present invention has the following beneficial effects:

[0039] 1. The pharmaceutical composition of the present invention is composed of Acanthopanax senticosus, Ginseng Leaf, Schisandra, Astragalus, Ganoderma Lucidum, and Gastrodia. There are no toxic medicinal materials in the prescription, and the prescription composition does not have 18 anti-incompatibility and 19 fear incompatibility taboos. The Chinese medicinal materials in the prescription are cheap, the production cost is low, and it is suitable for a wide range of people. At the same time, a variety of dosage forms can be prepared, including but not limited to tablets, granules, capsules, oral liquids and other dosage forms.

[0040] 2. The pharmaceutical composition of the present invention has a significant effect in treating insomnia, which is better than the therapeutic effect of western medicines such as sezpiclone tablets and estazolam tablets. It shortens the time of inducing sleep, and can effectively shorten the time of falling asleep for patients with difficulty falling asleep. At the same time, it is the best to prolong the sleep time. It has a significant effect in treating insomnia, and will not cause drowsiness or sleepiness. At the same time, there is no sense of dependence, and insomnia is fundamentally treated.

[0041] 3. In the long-term toxicity experiment, the physical signs of mice after taking the medicine were detected. The results of routine tests on organ coefficients and blood and urine showed that the mice taking the pharmaceutical composition of the present invention for a long time had normal physical signs, no organic changes were induced, and long-term use of the medicine had no toxic side effects.

[0042] 4. The results of the tolerance experiment show that the pharmaceutical composition of the present invention is still effective after continuous administration for 15 days. It also proves that the pharmaceutical composition of the present invention has no tolerance performance of decreased effect after long-term administration, and the pharmaceutical composition can be used for a long time to treat insomnia, whether it is to reduce the time to fall asleep or to prolong the sleep time, and has a long-term effective effect. Insomnia patients can take the pharmaceutical composition of the present invention as needed.

[0043] 5. Clinical trials have shown that the pharmaceutical composition of the present invention can be used to treat insomnia of the type of deficiency of both heart and spleen, and its therapeutic effect is more significant than that of traditional Chinese medicine and sedative hypnotic drugs for treating insomnia on the market; at the same time, it also has a certain alleviating effect on possible symptoms such as cough and asthma; the Gastrodia elata in the composition has the effect of calming liver yang, promoting fluid production and nourishing blood, and can relieve the accompanying symptoms such as dizziness and headache. The drug combination of the present invention did not cause side effects such as dizziness and headache during the administration of the drug in the clinical trial, and the side effects were smaller than those of the control group drugs in the experiment.

[0044] 6. The pharmaceutical composition of the present invention has the effect of improving memory in the experiment of improving memory in PCPA insomnia mice. The water maze navigation experiment and water maze space exploration experiment of PCPA insomnia mice proved that the pharmaceutical composition of the present application has a significant effect on improving memory. Therefore, the pharmaceutical composition of the present invention can not only be used to treat insomnia of heart and spleen deficiency type, but also improve the symptoms of patients with memory loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 This is a graph showing the effect of drugs on the number of activities;

[0046] Figure 2This is a diagram showing the effects of drugs on GABA;

[0047] Figure 3 This is a graph showing the effects of drugs on 5-HT;

[0048] Figure 4 For space exploration experiment-blank group picture;

[0049] Figure 5 It is a space exploration experiment-model group picture;

[0050] Figure 6 For space exploration experiments - Estazolam tablets group picture;

[0051] Figure 7 For space exploration experiments - right zopiclone tablets group picture;

[0052] Figure 8 This is a set of pictures of space exploration experiment-test. DETAILED DESCRIPTION

[0053] In order to make the objectives, technical solutions and advantages of the embodiments of the present invention more clearly understood, the spirit of the contents disclosed by the present invention will be described in detail below. After understanding the embodiments of the contents of the present invention, any technician in the relevant technical field can change and modify them according to the techniques taught by the contents of the present invention without departing from the spirit and scope of the contents of the present invention.

[0054] The exemplary embodiments of the present invention and the description thereof are used to explain the present invention but are not intended to limit the present invention.

[0055] Example 1

[0056] Granules prepared from the Bujuexiao series of pharmaceutical compositions for treating insomnia (heart and spleen deficiency type) and improving memory

[0057] Prescription: Acanthopanax 25g, Ginseng Leaf 5g, Schisandra 3g, Astragalus 15g, Ganoderma Lucidum 10g, Gastrodia elata 5g.

[0058] 1. Weighing

[0059] Weigh each medicinal material according to the prescription, pre-treat them separately and set aside.

[0060] 2. Extraction

[0061] Take the medicinal materials in the formula amount, add 8 times the amount of water, reflux extract for 2 hours, filter, and obtain filtrate 1; extract the medicinal residue with ultrasound assistance for 10 minutes at 40°C, filter, and obtain filtrate 2; dissolve the residue in 8 times the amount of water, reflux extract for 1.5 hours, filter, and obtain filtrate 3. Combine filtrate 1, filtrate 2, and filtrate 3 for later use.

[0062] 3. Concentration

[0063] The reduced pressure concentration temperature is 60°C, and the extract is concentrated to about 1.10-1.30 (50°C).

[0064] 4. Drying

[0065] The extract was dried by microwave drying method, and its moisture content was measured to be ≤3%, then taken out and cooled.

[0066] 5. Granulation

[0067] Mix 3g dextrin and 1g citric acid with the extract, add 10% water of the total amount of raw materials and auxiliary materials, stir for 20 minutes to mix the materials evenly to obtain a soft material, granulate with 20 mesh, dry at 40℃ until the moisture content is ≤2%, then take out and cool.

[0068] 6. Whole grain and subpackaging

[0069] Use an oscillating screen to granulate and pack the particles according to the required filling quantity.

[0070] Example 2

[0071] Oral liquid prepared from the Bujuexiao series of drug compositions for treating insomnia (heart and spleen deficiency type) and improving memory

[0072] Prescription: Acanthopanax 27g, Ginseng Leaf 6g, Schisandra 2g, Astragalus 15g, Ganoderma Lucidum 12g, Gastrodia elata 3g.

[0073] 1. Weighing

[0074] Weigh each medicinal material according to the prescription, pre-treat them separately and set aside.

[0075] 2. Extraction

[0076] Take the medicinal materials in the formula amount, add 8 times the amount of water, reflux extract for 2 hours, filter, and obtain filtrate 1; extract the medicinal residue with ultrasound assistance for 10 minutes at 40°C, filter, and obtain filtrate 2; dissolve the residue in 8 times the amount of water, reflux extract for 1.5 hours, filter, and obtain filtrate 3. Combine filtrate 1, filtrate 2, and filtrate 3 for later use.

[0077] 3. Concentration

[0078] The concentrated solution was concentrated by a reduced pressure concentration method, the temperature was controlled at 60°C, and the concentrate was concentrated to an extract of about 1.10 to 1.30 (50°C).

[0079] 4. Liquid preparation

[0080] Add 200 ml of water (80° C.) to the above extract and stir evenly, then add 2 g of potassium sorbate, 2 g of citric acid and 3 g of stevioside, and add water to make up to the prepared amount.

[0081] 5. Centrifugation

[0082] Set the centrifuge to 2500r / min and centrifuge for 10 minutes. Remove the supernatant and filter it with 20-mesh silk cloth for later use.

[0083] 6. Filling

[0084] Oral solution filling equipment is used for filling and capping.

[0085] 7. Sterilization

[0086] Check that the sterilizer is in good condition, set the parameters, the sterilization temperature is 100-130℃, the sterilization time is 15-30min, and cool.

[0087] Experimental Example 1

[0088] Effect of drug combination on hypnotic effect of PCPA insomnia mice

[0089] The pharmaceutical composition of the present invention has the effect of shortening the latency period of falling asleep and prolonging the sleeping time. In order to verify that the pharmaceutical composition of the present invention has the above-mentioned effect of treating insomnia, a sleep experiment was conducted using PCPA insomnia mice, and the sleeping time of the mice after taking the medicine was recorded. The sleeping conditions of the insomnia mice improved by each group of drugs were compared. The therapeutic drugs include the pharmaceutical composition of the present invention, commonly used sedative hypnotic drugs (eszolam tablets, eszopiclone tablets) and health foods (melatonin tablets). The experimental contents are as follows.

[0090] 1. Experimental Materials

[0091] 1.1 Experimental drugs and reagents

[0092] Pharmaceutical composition: Prepare the preparation according to Example 2;

[0093] p-Chlorophenylalanine (PCPA): Shanghai Yuanye Biotechnology Co., Ltd., batch number: A07GS157185;

[0094] Pentobarbital sodium: Sinopharm Chemical Reagent Co., Ltd., batch number WS20050411;

[0095] Sodium bicarbonate: Wuhan Pronosai Life Science Technology Co., Ltd., batch number: WH01112107SP;

[0096] Estazolam: Shandong Xinyi Pharmaceutical Co., Ltd., national medicine standard H37023047;

[0097] R-zopiclone tablets: Jiangsu Tasly Diyi Pharmaceutical Co., Ltd., national medicine standard H20090209;

[0098] Melatonin tablets: By-Health Co., Ltd., National Food and Health Note G20150889;

[0099] Physiological saline: Harbin Sanlian Pharmaceutical Co., Ltd., national medicine standard H23020611

[0100] 1.2 Experimental animals

[0101] Kunming mice: SPF grade, male, purchased from Liaoning Changsheng Biotechnology Co., Ltd. (Liaoning Provincial Experimental Animal Resources Center, NO.SCXK(Liao)2020-0001), body weight 18-22 g, free access to drinking water and food, room temperature 18-24 ° C, humidity 40%-50%, adaptive feeding for seven days before the start of the experiment.

[0102] 2. Experimental Grouping and Drug Administration

[0103] 2.1 Experimental Animal Grouping

[0104] The mice were randomly divided into 11 groups according to the suprathreshold experiment: blank control group, model control group, 2 mg estazolam tablets positive drug group, 2 mg eszopiclone tablets positive drug group, 0.4 g melatonin tablets positive drug group, and 6 groups of suprathreshold insomnia treatment drug combination groups, with 10 mice in each group. They were kept in an experimental environment with constant temperature and humidity, and had free access to water and food.

[0105] 2.2 Preparation of PCPA insomnia mouse model

[0106] According to the preliminary test of the research group and reference to the literature, PCPA suspension was prepared: 35 mg / mL PCPA suspension was prepared with weak alkaline saline (PH10.1), and the suspension was placed in a water bath at 80°C for 20 minutes and mixed thoroughly. Except for the blank control group, each group of mice was intraperitoneally injected with 350 mg / kg PCPA suspension at 0.1 mL / 10 g body weight, once a day for three consecutive days, and the blank control group was given an equal amount of weak alkaline saline. Model evaluation: After modeling, the mice showed signs of disappearance of circadian rhythm, messy hair, less food intake, easy irritation and fighting, damp litter, and abnormal gray feces, indicating that the model was successfully prepared.

[0107] 2.3 Administration

[0108] The mice were gavaged and given the drug at 10 am every day. The gavage dose of each group was converted according to the equivalent dose of humans and mice. The weight of the mice was measured to be 20±2g. The conversion formula D from standard weight to non-standard weight was used. b ’ =D 艾司唑仑片2mg ×R 艾司唑仑片2mgb ×Sb(R 艾司唑仑片2mgb =12.33, Sb=0.874), the dosage of the experimental drug was obtained and controlled by gavage at 0.1mL / 10g. The blank control group and the model control group were gavaged with an equal volume of normal saline once a day for seven consecutive days.

[0109] 3. Testing of hypnotic test indexes of sodium pentobarbital above threshold dose

[0110] The grouping modeling and drug administration were carried out according to the above method. 30 minutes after the last drug administration, each group of mice was intraperitoneally injected with 46 mg / kg of sodium pentobarbital (the dose was obtained by preliminary test, the minimum dose for 100% loss of righting reflex in mice), and the mice were observed for sleep. The loss of righting reflex was taken as a sign of falling asleep. When the mice were placed in a dorsal position, if they could not right themselves for more than 30 seconds, it was considered that the righting reflex had disappeared and they fell asleep. The recovery of righting reflex was considered as animal awakening. The time from the start of sodium pentobarbital injection to the disappearance of the righting reflex of the mice was the animal's sleep latency, and the time from the disappearance to the recovery of the righting reflex was the animal's sleep time. The sleep latency and sleep time of each group of mice were recorded respectively.

[0111] 4. Effects of drug combination on sleep time of mice induced by suprathreshold dose of sodium pentobarbital

[0112] Compared with the normal control group, the sleep latency of mice in the model control group was prolonged (P<0.01) and the sleep time was shortened (P<0.01). After the treatment of insomnia drugs in each group, the sleep condition of PCPA insomnia mice was significantly improved. See Tables 1 and 2. The results of the sleep latency test showed that the drug-treated groups were better than the positive drug group, and the induced sleep time was also better than the positive drug group.

[0113] Table 1 Effects of suprathreshold test on sleep onset latency

[0114]

[0115] Note: Compared with the blank control group, the model control group has significant differences. # indicates P<0.05; compared with the model control group, each drug-treated group has significant differences. * indicates P<0.05, and ** indicates P<0.01.

[0116] Table 2 Effects of suprathreshold experiments on sleep time

[0117]

[0118] Note: Compared with the blank control group, the model control group has significant differences. # indicates P<0.05; compared with the model control group, each drug-treated group has significant differences. * indicates P<0.05.

[0119] Conclusion: Compared with the normal control group, the sleep latency of mice in the model control group was prolonged (P<0.05) and the sleep time was shortened (P<0.05). After each group was treated with drugs for insomnia, the sleep condition of PCPA insomnia mice was significantly improved. In the experiment, the sleep latency of the group treated with the pharmaceutical composition of the present invention was only 4.89min, which was shortened by 17.3% compared with the model group, while the sleep time was prolonged from 40.8min to 100.72min, which was prolonged by 146.9% compared with the model group. The effect of the pharmaceutical composition of the present invention on insomnia treatment was better than that of the positive drug group of 2mg of sezpiclone tablets, 2mg of estazolam tablets and 0.4g of melatonin tablets, and the induction of sleep time and the extension of sleep time were the best.

[0120] Experimental Example 2

[0121] Effect of drug combination on the sleep rate of mice administrated with subthreshold dose of sodium pentobarbital

[0122] In order to verify whether the pharmaceutical composition of the present invention can improve the effect of subthreshold dose sodium pentobarbital, a subthreshold dose sodium pentobarbital mouse sleep rate experiment was conducted to investigate the difference between the pharmaceutical composition of the present invention and the blank control group to determine its effect on the sleep rate. The detailed experimental content is as follows.

[0123] Choose 150 male mice of Kunming species (body weight 18-22g), be randomly divided into 5 groups (30 in every group), group as shown in the table below.Administer by gavage (medicine is embodiment 2 preparation preparation), once a day, 5 consecutive days.After the last administration in 1 hour, intraperitoneal injection of sodium pentobarbital (25mg / Kg), in 30 minutes, animal righting reflex disappears and reaches 1 minute above person, shows that it has entered sleep.Relatively each group animal falls asleep number, experimental result is shown in Table 3.

[0124] Table 3 Effect of the drug combination on the sleep rate of mice with subthreshold dose of sodium pentobarbital

[0125]

[0126] According to the above experimental results, the sleeping rate of the mice in the high-dose group of the pharmaceutical composition with a subthreshold dose of sodium pentobarbital is significantly higher than that in the blank control group (P<0.05), and the sleeping rate can reach 73.3%; at the same time, the sleeping rate of the low-dose group is also higher than that of the blank control group. It shows that the pharmaceutical composition of the present invention can reduce the number of autonomous activities of mice and significantly increase the number of sleeping mice. At the same time, the pharmaceutical composition has the effect of significantly improving the sleeping rate of mice with a subthreshold dose of sodium pentobarbital. The combination of the composition of the present invention and sodium pentobarbital has a synergistic sedative and hypnotic effect. After taking the pharmaceutical composition of the present invention, the dosage of barbiturates can be reduced, and the incidence of adverse reactions and side effects of barbiturates can be reduced.

[0127] Experimental Example 3

[0128] Hypnotic effects of different drug combinations on PCPA-induced insomnia mice

[0129] Based on the composition of the pharmaceutical composition of the present invention, the effect of the prescription after adding or reducing the corresponding medicinal materials on insomnia was investigated, and the effect of each composition on insomnia was compared with the currently commonly used commercial sleeping pills, eszopiclone tablets and estazolam tablets. The above experimental method of Experimental Example 1 was used to conduct an experiment on insomnia mice, and the test results are shown in Table 4 below.

[0130] Prescription composition and dosage:

[0131] Prescription 1: 5g ginseng, 15g astragalus;

[0132] Prescription 2: 5g ginseng, 15g astragalus, 3g schisandra;

[0133] Prescription 3: 5g ginseng, 15g astragalus, 9g wolfberry, 10g ganoderma;

[0134] Prescription 4: 5g ginseng, 15g astragalus, 9g wolfberry, 10g ganoderma, 5g gastrodia;

[0135] Prescription 5: Acanthopanax senticosus 25g, Ginseng 5g, Schisandra 3g, Astragalus 15g, Ganoderma lucidum 10g, Gastrodia elata 5g;

[0136] Prescription 6: Acanthopanax senticosus 25g, Ginseng 5g, Schisandra 3g, Astragalus 15g, Ganoderma lucidum 10g, Gastrodia elata 5g, Lycium barbarum 9g, Ligustrum lucidum 10g;

[0137] Prescription 7: Acanthopanax senticosus 25g, Ginseng 5g, Schisandra 3g, Astragalus 15g, Ganoderma lucidum 10g, Gastrodia elata 5g, Lycium barbarum 9g, Ligustrum lucidum 10g, Polygonatum 12g, White Peony 12g;

[0138] Prescription 8: 5g ginseng, 15g astragalus, 9g wolfberry, 10g Ligustrum lucidum, 10g ganoderma lucidum;

[0139] Prescription 9: 5g ginseng, 15g astragalus, 9g wolfberry, 10g Ligustrum lucidum, 8g golden cherry, 9g dodder seed;

[0140] Prescription 10: 5g ginseng, 15g astragalus, 9g wolfberry, 10g glossy privet fruit, 8g rose hip, 9g dodder seed, 12g polygonatum, 12g white peony root, 8g licorice, 12g malt, and 8g epimedium.

[0141] Table 4 Comparison of the effects of different compositions in treating insomnia

[0142]

[0143]

[0144] Conclusion: By increasing and decreasing the ingredients of the composition, the results showed that prescription 5 had the best effect in treating insomnia, with a tendency to shorten the sleep latency, and the sleep time was as high as 82.42±5.14min. Compared with s-zopiclone tablets and estazolam tablets, the sleep time was further prolonged. This shows that the composition has the therapeutic effect of shortening the sleep latency and prolonging the sleep time.

[0145] Experimental Example 4

[0146] Effects of different dosages of drug combinations on the efficacy of treating insomnia

[0147] When different drug doses are combined together, the effects produced are not the same. The therapeutic effect is not positively increasing. Therefore, it is very necessary to investigate the effects of different doses of corresponding medicinal materials on insomnia according to the composition of the drug composition. The experimental method of Experimental Example 1 was used to conduct an experiment on insomnia mice, and the test results are shown in Table 5 below.

[0148] Table 5 Effects of different dosages of insomnia combination drugs on the sleep of insomnia mice

[0149]

[0150]

[0151] Conclusion: According to the above experimental results, Acanthopanax senticosus 27g, Ginseng Leaf 6g, Schisandra 2g, Astragalus 15g, Ganoderma Lucidum 12g, Gastrodia 3g are the best curative combination. Its sleep latency is shortened by 3.75min compared with the model group, and the sleep time is reduced by more than 50%. At the same time, the sleep time is longer than that of R-zopiclone tablets and Estazolam tablets, and the effect of treating insomnia is more significant than sedative drugs. After replacing ginseng with ginseng leaf in the formula, the therapeutic effect is equivalent. The cost of ginseng leaf is lower than that of ginseng. The cost of 10-day prescription is 4.37 yuan, and the cost of 11-day prescription is 2.51 yuan. After using ginseng leaf, the daily dosage cost is reduced by 1.86 yuan, which can reduce the daily dosage cost by 42.6%, and the production cost is controlled to the minimum, providing insomnia patients with a highly cost-effective therapeutic drug.

[0152] Experimental Example 5

[0153] Effects of drug combination on neurotransmitters in PCPA insomnia mice

[0154] In order to verify the therapeutic effect of the pharmaceutical composition for treating insomnia of the present invention on insomnia from a physiological perspective, an experiment was conducted using PCPA insomnia mice to detect the changes in various transmitter indicators before and after using the pharmaceutical composition of the present invention, and then to compare whether the pharmaceutical composition of the present invention has a better effect on insomnia. The specific experimental steps are as follows.

[0155] 1 Materials

[0156] 1.1 Experimental drugs and reagents

[0157] Bujuexiao series pharmaceutical compositions: prepare the preparation according to Example 2;

[0158] p-Chlorophenylalanine (PCPA): Shanghai Yuanye Biotechnology Co., Ltd., batch number: A07GS157185;

[0159] Pentobarbital sodium: Sinopharm Chemical Reagent Co., Ltd., batch number WS20050411;

[0160] Sodium bicarbonate: Wuhan Pronosai Life Science Technology Co., Ltd., batch number: WH01112107SP;

[0161] Estazolam: Shandong Xinyi Pharmaceutical Co., Ltd., national medicine standard H37023047;

[0162] R-zopiclone tablets: Jiangsu Tasly Diyi Pharmaceutical Co., Ltd., national medicine standard H20090209;

[0163] Normal saline: Harbin Sanlian Pharmaceutical Co., Ltd., national medicine approval number H23020611.

[0164] 1.2 Experimental animals

[0165] Kunming mice: SPF grade, male, purchased from Liaoning Changsheng Biotechnology Co., Ltd. (Liaoning Provincial Experimental Animal Resource Center), body weight 18-22g, free access to water and food, room temperature 18-24°C, humidity 40%-50%, adaptive feeding for seven days before the start of the experiment.

[0166] 1.3 Main instruments and equipment

[0167] Electronic balance, Shanghai Jingqi Instrument Co., Ltd.;

[0168] Constant temperature water bath, Tianjin Test Instrument Co., Ltd.;

[0169] Microplate reader, Bio-Rad, USA;

[0170] ZZ-6 mouse locomotor activity tester and small animal behavior system (Chengdu Taimeng Technology Co., Ltd.).

[0171] 2 Experimental methods

[0172] 2.1 Experimental Animal Grouping

[0173] The mice were randomly divided into 11 groups: a blank control group, a model control group, an estazolam positive control group, and 8 groups of insomnia treatment drug compositions (the compositions were prepared according to Example 2), as shown in Table 6. Each group had 15 mice, which were kept in an experimental environment with constant temperature and humidity and had free access to water and food.

[0174] Table 6 Experimental groups

[0175]

[0176]

[0177] 2.2 Mouse modeling

[0178] After the mice were adapted to feeding for 7 days, they were randomly divided into blank control group, insomnia model group, positive drug group, and drug administration group on the 8th day, with 15 mice in each group. PCPA [400 mg / (kg·d)] was intraperitoneally injected into the mice in the insomnia model group and drug administration group for 3 consecutive days, and the blank control group mice were injected with an equal volume of weak alkaline saline in parallel. 24 hours after the last intraperitoneal injection of PCPA suspension, the activities of the mice were monitored to find evidence of the successful establishment of the insomnia model. The insomnia model includes loss of circadian rhythm, increased excitability, increased aggression, increased urine and feces, messy and dark hair, and gray feces. The mice were weighed with an electronic balance before and after the model was established. The drug administration treatment was carried out by gavage at 9:00-10:00 am every day. The blank control group and the insomnia model group were gavaged with an equal amount of deionized water for 7 consecutive days. After gavage treatment every day, the mice were returned to the cage, and mouse food and water were placed in the cage to ensure that the mice in each group could eat freely normally.

[0179] 2.3 Direct sleep test:

[0180] After the last administration, the mice were observed to see if they fell asleep. The disappearance of the righting reflex was used as an indicator of sleep. When the mice were placed in a dorsal recumbent position, they could turn right immediately. If they could not turn right for more than 30 to 60 seconds, it was considered that the righting reflex disappeared and they fell asleep. The recovery of the righting reflex was considered as the awakening of the animals, and the period from the disappearance of the righting reflex to the recovery was considered as the sleeping time of the animals. The differences in the number of sleeping animals and the sleeping time were compared between the blank control group and the model group, and between the model group and the test sample group.

[0181] 2.4 Autonomic activity instrument test experiment:

[0182] 30 minutes after the last intragastric administration, mice in each group were placed in the autonomic activity recorder in turn and adapted for 5 minutes. After the instrument was turned on, the number of autonomic activities and standing times within 5 minutes were measured.

[0183] 2.5 Elisa method for neurotransmitter measurement

[0184] The hypothalamus samples of mice in each group were weighed, and 10 times the amount of normal saline was added. The samples were homogenized at 4°C for 180 s, centrifuged at 3000 r / min for 15 min, and the supernatant was taken for detection according to the instructions of the Elisa kit.

[0185] 3 Experimental results

[0186] 3.1 Direct sleep test

[0187] The drug was administered continuously for 7 days, and no sleep phenomenon was observed in the mice after the last administration.

[0188] 3.2 Autonomic activity instrument test experiment

[0189] After PCPA modeling, the excitability of mice increased and the number of activities increased. After 7 days of administration, all drugs could reduce the number of activities of mice after modeling excitement. The strength of the effect is shown in Table 7 and Figure 1 .

[0190] Table 7 Effect of the composition on the number of spontaneous activities of mice

[0191]

[0192] Note: Compared with the blank control group, the different drug-treated groups have significant differences. ## indicates P<0.01; compared with the model control group, the different drug-treated groups have significant differences. ** indicates P<0.01.

[0193] 3.3 Results of Elisa assay for neurotransmitters

[0194] 3.3.1 GABA content

[0195] γ-Aminobutyric acid (GABA) is an inhibitory neurotransmitter. The GABA content of mice decreased after PCPA modeling. After 7 days of administration, the GABA content increased significantly. The test results are shown in Table 8 and Figure 2 shown.

[0196] Table 8 Effects of drugs on GABA content in mice

[0197]

[0198]

[0199] 3.3.2 5-HT Content

[0200] 5-hydroxytryptamine (5-HT) is an inhibitory neurotransmitter. After PCPA modeling, the inhibitory neurotransmitter was significantly reduced compared with the blank group. After 7 days of administration, the content of 5-HT in each group was significantly increased. The test results are as follows: Figure 3 shown.

[0201] Table 9 Effects of drugs on 5-HT content in mice

[0202]

[0203] Conclusion: Through the detection of the autonomic activity meter, the number of activities reflects the sedative effect of the drug. This indicator shows that the drug has a significant sedative effect. Among the number of activities, the effect of composition 8 (2 times the medium dose for adults per day) is the most obvious, and the number of activities when the dosage reaches 1 / 2 of the adult's dose once is equivalent to that of the positive control group. When the dosage is normal (adults once full dose), the effect of reducing the number of activities is significantly better than that of the positive administration group. Combined with the results of the sodium pentobarbital sleep induction experiment, it can be seen that the pharmaceutical composition of the present invention is very effective in inducing sleep and shortening the time to fall asleep.

[0204] It can be seen from the results of the Elisa method for detecting neurotransmitters that the pharmaceutical composition of the present invention has a significant effect on inhibitory neurotransmitters, and as the dosage increases, the concentration of inhibitory neurotransmitters also increases. The low-dose pharmaceutical composition has an effect equivalent to that of the positive control group, and the effects of the medium and high-dose pharmaceutical preparations are much higher than those of the blank group and the positive control group. The normal-dose composition can fully achieve the effects of sedation and hypnosis. For occasional difficulty falling asleep caused by stress and anxiety, those with poor sleep quality or weak physical fitness can choose to take a low-dose pharmaceutical composition; for those with staged difficulty falling asleep, induced difficulty falling asleep, and those who are easily awakened due to life stress and anxiety, they can choose to take a medium-dose pharmaceutical composition; for patients with intractable difficulty falling asleep and long-term sleep disorders, high-dose pharmaceutical compositions can be selected for treatment in stages.

[0205] Experimental Example 6

[0206] Safety test of drug combination (long-term toxicity test)

[0207] In order to verify whether there are any potential safety hazards after long-term use of the insomnia treatment drug composition of the present invention, the toxic effects of the Bujuexiao series of insomnia drug compositions on experimental animals were observed through long-term administration for several weeks, including organ damage, changes in urine routine and blood routine indicators, abnormal biochemical indicators, etc. Ensuring the safety of the drug and identifying potential adverse reactions, long-term toxicity experiments are of great significance to ensuring the safety of patients' medication.

[0208] 1. Experimental Animals

[0209] SPF-grade healthy SD mice, weighing (200±20) g, half male and half female, were purchased from Liaoning Changsheng Biotechnology Co., Ltd., with experimental animal production license number SCXK(Liao)2020-0001. The animals were kept in the animal room of the School of Pharmacy, Harbin University of Commerce, under 12h / 12h day and night cycle conditions, free access to food, room temperature of 22±2℃, and relative humidity of 45-55%. The experiment began after one week of adaptive feeding.

[0210] 2. Experimental methods

[0211] SD mouse grouping: After seven days of adaptive feeding, the mice were randomly divided into 4 groups: 2 blank groups and 2 test groups (see Table 10 for drug grouping), 20 mice in each group, half of which were male and half were female. The drug-treated groups were given drugs by gavage for 12 weeks, and the blank groups were given the same dose of normal saline by gavage. The drug grouping and dosage are shown in Table 10.

[0212] Table 10 Long-term toxicity test groups

[0213]

[0214] Mouse sampling: Each group of mice was continuously administered for 12 weeks. After the last administration, the mice were fasted and deprived of water for 12 hours. The mice were placed in metabolic cages overnight, and urine from each group of mice was collected for routine urine tests. After all mice had their urine taken, they were anesthetized by intraperitoneal injection of chloral hydrate, and blood was taken from the abdominal aorta. First, 2 mL of whole blood from the mice was collected using an anticoagulant blood collection tube containing EDTA, which was labeled and left to stand at 37°C for routine blood tests of mice; whole blood from mice was collected using ordinary disposable blood collection tubes, and after standing for 30 minutes, the heart was pumped (3000 rpm, 10 minutes), the supernatant was collected, and the supernatant was packaged. 200 μL of supernatant was used to detect the blood biochemical indicators of mice, and the remaining serum was used to detect the content of ALT, AST, AKP, and CK. The detection indicators are shown in 2; the heart, liver, spleen, lung, and kidney of the mice were weighed, and the organ coefficient of the mice was calculated. The liver, kidney, and brain tissues were fully immersed in formalin for HE staining observation.

[0215] 3. Experimental results

[0216] After the autopsy, the organs were weighed, and the results were shown in Table 11. There was no significant difference (P>0.05). Urine routine, blood routine and biochemical index tests were performed on mice in each group, and the test results are shown in Table 12 below.

[0217] Table 11 Mouse organ coefficients

[0218]

[0219] Table 12 Test results of each group of indicators

[0220]

[0221]

[0222] Conclusion: Through the detection of organs, urine routine, blood routine and biochemical indicators, it can be known that after taking the pharmaceutical composition of the present invention, there is no obvious change in the mass of various organs and no abnormal organs are found. There is no overall obvious difference in urine routine, blood routine and biochemical indicators. The results of this safety experiment show that long-term use of the pharmaceutical composition of the present invention does not lead to organic mutagenesis, has no toxic side effects, and does not affect long-term medication.

[0223] Experimental Example 7

[0224] Effect of drug combination on hypnotic tolerance in mice

[0225] Because all kinds of therapeutic drugs have tolerance problems, the effect after long-term use is not as good as the initial effect, and even worse, there is no effect after multiple uses. In order to confirm whether the series of drug compositions of the present invention produce tolerance after long-term use, a mouse hypnosis tolerance experiment is conducted to verify it. The experimental content is as follows. This experiment is of great significance to determine whether tolerance is produced after long-term use of the drug composition, which is closely related to the therapeutic effect of patients after long-term use.

[0226] 1 Materials

[0227] 1.1 Experimental drugs

[0228] Bujuexiao series of pharmaceutical compositions: Preparation of preparations in Example 2;

[0229] Estazolam: Shandong Xinyi Pharmaceutical Co., Ltd., national medicine standard H37023047;

[0230] Normal saline: Harbin Sanlian Pharmaceutical Co., Ltd., national medicine approval number H23020611.

[0231] 1.2 Experimental animals

[0232] Kunming mice: SPF grade, male, body weight 18-22 g, adaptively raised for seven days before the experiment.

[0233] 2 Methods

[0234] Animal grouping and drug administration: The mice were randomly divided into blank control, positive control, low-dose, medium-dose and high-dose groups of the drug combination. The blank group was gavaged with an equal amount of saline. Each group was continuously administered for 15 days. 60 minutes after gavage on the morning of the 7th and 15th days, the mice's sleep latency and sleep duration were measured respectively to observe the tolerance phenomenon of the hypnotic effect of the drug after repeated administration.

[0235] 3 Results

[0236] Each group administered the drug to insomniac mice for 15 days according to the above experimental method. At the same time, the mice's sleep latency and sleep time were counted at 7 and 15 days after drug administration. The results are shown in Table 13 below.

[0237] Table 13 Effect of repeated use of the drug composition on hypnotic tolerance in mice

[0238]

[0239] Conclusion: The results of repeated use of each group of drugs on the hypnotic tolerance of mice showed that the sleep latency of the positive control group, low-dose group, medium-dose group and high-dose group was significantly lower than that of the blank group on the 7th day of administration; the sleep time of each administration group was significantly higher than that of the blank group on the 7th and 15th day of administration; there was no difference in the sleep latency before and after the 7th day and the 15th day, the sleep time of the positive control group on the 15th day was significantly shorter than that of the 7th day, the sleep time of the high-dose group on the 15th day was significantly longer than that of the 7th day, and the low-dose and medium-dose groups on the 15th day had a tendency to be longer than that of the 7th day. According to the experimental data, it can be seen that the pharmaceutical composition of the present invention does not show tolerance to the decline of the effect after long-term medication.

[0240] Experimental Example 8

[0241] Effect of the drug composition on improving memory in PCPA insomnia mice

[0242] In addition to treating insomnia of deficiency of both heart and spleen, the pharmaceutical composition of the present invention is also found to have the effect of improving memory. In order to verify the memory-improving effect of the Bujuexiao series of pharmaceutical compositions, water maze positioning navigation experiments and water maze space exploration experiments were performed on PCPA insomnia mice to verify the effect of improving memory.

[0243] 1 Materials

[0244] 1.1 Experimental drugs and reagents

[0245] The experimental drugs and reagents were the same as those in Experimental Example 1.

[0246] 1.2 Experimental animals

[0247] Kunming mice: SPF grade, male, purchased from Liaoning Changsheng Biotechnology Co., Ltd., with a body weight of 18-22 g, were adaptively raised for seven days before the experiment.

[0248] 2 Experimental methods

[0249] 2.1 Experimental Animal Grouping

[0250] The mice were randomly divided into: blank control group, model control group, positive drug group of 2 mg estazolam tablets, positive drug group of 2 mg eszopiclone tablets, and test group (10 ml of the drug composition prepared in Example 2), with 10 mice in each group, and were raised in an experimental environment with constant temperature and humidity, with free access to water and food.

[0251] 2.2 Mouse modeling

[0252] After the mice were adapted to feeding for 7 days, they were randomly divided into groups on the 8th day, with 10 mice in each group. PCPA [400 mg / (kg·d)] was intraperitoneally injected into the insomnia model group and the drug group for 3 consecutive days, and the blank control group mice were injected with an equal volume of normal saline in parallel. 24 hours after the last intraperitoneal injection of PCPA suspension, the activities of the mice were monitored to find evidence of the successful establishment of the insomnia model. The insomnia model includes loss of circadian rhythm, increased excitability, increased aggression, increased urine and feces production, messy and dark hair, and gray feces. The mice were weighed with an electronic balance before and after the model was established. The treatment was given by gavage at 9:00-10:00 am every day. The blank control group and the insomnia model group were gavaged with an equal amount of deionized water for 7 consecutive days. After gavage treatment every day, the mice were returned to the cage, and mouse food and water were placed in the cage to ensure that the mice in each group could eat freely normally.

[0253] 2.3 Morris water maze test

[0254] Morris water maze test was performed on the 4th day of drug administration. The experiment was conducted in a circular pool with a black inner wall, which was equally divided into 4 quadrants (the 4 areas divided by the horizontal and vertical axes in the plane rectangular coordinate system, the upper right is called Quadrant I, the lower right is called Quadrant II, the lower left is called Quadrant III, and the upper left is called Quadrant IV). The target platform was located 1 cm below the water surface in Quadrant IV. The water temperature was constant (22±2)℃, the light was constant, and there was no influence of daylight changes. The experimental process was recorded using a video tracking and analysis system.

[0255] (1) Positioning and navigation experiment: The mice were placed in the water facing the inner wall at a fixed location farthest from the target platform (quadrant II). The time taken by each group of mice from entering the water to finding and climbing onto the target platform and staying there for 15 seconds was recorded, which was the escape latency of the mice. If the mice did not find the target platform within 90 seconds, they were guided and stayed on the platform for 15 seconds. All mice were trained twice a day for 2 days.

[0256] (2) Spatial exploration experiment: The platform was removed and mice from each group were placed in the water at the same position facing the inner wall of the pool. The movement trajectory of the mice, swimming speed, and the number of times, time, and distance they crossed the target quadrant within 60 seconds were recorded.

[0257] 3 Experimental results

[0258] 3.1 Results of positioning navigation experiment

[0259] The escape latency was used as an indicator to evaluate the spatial learning ability of insomnia mice. After 4 guided trainings, the escape latency of mice in each group decreased to varying degrees. The fourth training was the final positioning navigation test result. Compared with the control group, the escape latency of mice in the model group increased significantly (P < 0.05); compared with the model group, the escape latency of mice in the drug-treated group decreased significantly (P < 0.01), among which the escape latency of the experimental group on the fourth day was shortened the most. The experimental results are shown in Table 14.

[0260] Table 14 Morris water maze behavioral experimental results

[0261]

[0262] Note: Compared with the blank control group, the model control group has significant differences. ## indicates P<0.01; compared with the model control group, different drug-treated groups have significant differences. * indicates P<0.05, ** indicates P<0.01.

[0263] 3.2 Space Exploration Experiment

[0264] The mouse water maze tester tracking imaging system was used to record the movement trajectory of each group of mice in the pool. Figure 4-8 As shown in the figure. The quadrant where the original platform was located was quadrant IV. After entering the water, the mice in the blank group were able to autonomously go to quadrant IV to find the platform, and their tracks were relatively loose; the tracks of the mice in the model group were concentrated in quadrant II of the water, and they rarely explored other quadrants; the mice in other drug-treated groups wandered in quadrant IV where the platform was located, and had a tendency to enter quadrant IV in a straight line after entering the water, indicating that after the combination of drugs was administered, the learning and memory ability of mice was adjusted back. GrAph PAd Pri software was used to analyze the four indicators of target quadrant residence time, distance into the target quadrant, average speed, and number of times entering the platform quadrant. As shown in Tables 15-18, compared with the control group, the time, distance, number of times and average speed of the model group mice entering quadrant IV after the platform was removed were significantly reduced (P < 0.01). In the experiment, it was observed that they mostly wandered around the water entry or explored aimlessly; compared with the model group, the distance, residence time, number of times and average speed of the mice in each drug-treated group entering the target quadrant were significantly prolonged (P < 0.05, 0.01), indicating that the drug-treated groups can improve the learning and memory ability of insomniac mice.

[0265] Table 15 Number of times of crossing the platform quadrant in water maze spatial exploration

[0266]

[0267] Note: Compared with the blank control group, the model control group has significant differences. ## indicates P<0.01; compared with the model control group, different drug-treated groups have significant differences. * indicates P<0.05, ** indicates P<0.01.

[0268] Table 16 The time spent in the target quadrant of the water maze spatial exploration

[0269]

[0270] Note: Compared with the blank control group, the model control group has significant differences. ## indicates P<0.01; compared with the model control group, different drug-treated groups have significant differences. * indicates P<0.05, ** indicates P<0.01.

[0271] Table 17 Water maze spatial exploration distance into the target quadrant

[0272]

[0273] Note: Compared with the blank control group, the model control group has significant differences. ## indicates P<0.01; compared with the model control group, different drug-treated groups have significant differences. * indicates P<0.05, ** indicates P<0.01.

[0274] Table 18 Average speed of spatial exploration in water maze

[0275]

[0276] Note: Compared with the blank control group, the model control group has significant differences. ## indicates P<0.01; compared with the model control group, different drug-treated groups have significant differences. * indicates P<0.05, ** indicates P<0.01.

[0277] 4 Conclusion

[0278] In the water maze navigation experiment, the experimental group showed significant differences compared with the model group, and the time to find the platform was relatively reduced compared with the 2mg sezopiclone tablets and 2mg estazolam groups. In the water maze spatial exploration experiment, the experimental group crossed the platform more times than the positive drug; the residence time was equivalent to 2mg sezopiclone tablets and 2mg estazolam; the distance into the quadrant was lower than 2mg sezopiclone tablets and 2mg estazolam tablets; the average speed was equivalent to 2mg sezopiclone tablets and 2mg estazolam. Compared with the model group, the distance, residence time, number of times and average speed of mice in each drug administration group entering the target quadrant were significantly prolonged (P < 0.05, 0.01), among which the pharmaceutical composition of the present invention had the best prolongation effect, indicating that the pharmaceutical composition of the present invention can improve the learning and memory ability of insomnia mice.

[0279] Experimental Example 9

[0280] Clinical trials

[0281] In order to verify the therapeutic effect of the Bujuexiao series of pharmaceutical compositions of the present invention on patients with insomnia of the deficiency of both heart and spleen, the experiment was conducted on patients clinically diagnosed with this disease, and the therapeutic efficacy of the pharmaceutical compositions of the present invention on patients with insomnia of the deficiency of both heart and spleen was evaluated based on the treatment of real cases, so as to determine whether it can be applied to this type of insomnia.

[0282] 1. Diagnostic criteria

[0283] 1.1 Western medicine diagnostic criteria

[0284] (1) Insomnia is the most critical symptom, including difficulty falling asleep, shallow sleep, easy awakening, difficulty falling asleep again after awakening, frequent dreams, early awakening, feeling uncomfortable after waking up, fatigue, and daytime sleepiness;

[0285] (2) The above insomnia-related symptoms occur at least three times a week and last for more than one month;

[0286] (3) Paying too much attention to insomnia and being extremely concerned about the consequences of insomnia;

[0287] (4) Severe dissatisfaction with sleep time and sleep quality, which affects mental, psychological, emotional or social functions;

[0288] (5) Exclude secondary insomnia caused by physical or mental illness.

[0289] 1.2 TCM diagnostic criteria

[0290] (1) Difficulty falling asleep, waking up easily after falling asleep, not being able to fall asleep after waking up, waking up early, and in severe cases, not being able to fall asleep all night.

[0291] (2) Often accompanied by symptoms such as headache, dizziness, palpitations, forgetfulness, and frequent dreams.

[0292] (3) No abnormalities were found in any system or laboratory examinations.

[0293] 1.3 Diagnostic criteria for insomnia due to deficiency of both heart and spleen

[0294] Heart and spleen deficiency is a concept in traditional Chinese medicine, which refers to the coexistence of insufficient heart blood and weak spleen. In this case, patients may show the following symptoms and manifestations:

[0295] (1) Palpitations: Due to insufficient blood in the heart, heart function is affected, which may cause palpitations.

[0296] (2) Insomnia and nightmares: Insufficient blood in the heart can affect sleep quality, leading to symptoms such as insomnia and nightmares.

[0297] (3) Pale or pale complexion: Insufficient blood in the heart cannot nourish the facial skin, making the complexion look bloodless.

[0298] (4) Loss of appetite and abdominal distension: A weak spleen will affect the digestion and absorption function, leading to digestive system symptoms such as loss of appetite and abdominal distension.

[0299] (5) Loose stools: Spleen deficiency cannot transport and transform water and dampness, which may lead to loose stools.

[0300] (6) Dizziness: Insufficient Qi and blood may affect blood supply to the brain, which may cause dizziness.

[0301] (7) Fatigue and weakness: Qi, blood and body fluids cannot nourish the whole body, resulting in fatigue and weakness.

[0302] (8) Menstrual abnormalities: Female patients may experience abnormal menstrual symptoms such as scanty menstrual flow, light-colored menstruation, or prolonged menstrual bleeding.

[0303] 2. Case inclusion criteria

[0304] (1) Meet the Western medical diagnostic criteria for insomnia;

[0305] (2) Meet the TCM diagnostic criteria for insomnia;

[0306] (3) Meet the diagnostic criteria for the syndrome of deficiency of both heart and spleen;

[0307] (4) Any gender, age 20-60 years old:

[0308] (5) No sedatives were used within 1 week:

[0309] (6) Voluntarily participate in this study and sign the informed consent form and be able to cooperate with follow-up.

[0310] 3. Experimental plan

[0311] 3.1 Case selection

[0312] According to the above inclusion criteria, 90 patients with new heart and spleen deficiency type insomnia were selected as the subjects of this study. They were randomly divided into a control group and an experimental group, with 30 patients in each group. There were 13 males and 17 females in the control group 1, aged 20 to 60 years old; there were 16 males and 14 females in the control group 2, aged 25 to 60 years old; there were 15 males and 15 females in the experimental group, aged 22 to 60 years old. There was no significant difference in the general data of the three groups (P>0.05), and they were comparable.

[0313] 3.2 Treatment methods

[0314] Test group: Orally take the pharmaceutical preparation of the present invention (prepared in Example 2), 10 ml / time, twice a day;

[0315] Control group 1: Oral administration of Anshenbunao liquid, 1 bottle / time, twice a day;

[0316] Control group 2: Oral administration of estazolam tablets, 1 tablet / time, 3 times a day;

[0317] The above three groups were treated with medication for 1 month.

[0318] 4. Evaluation criteria and test results

[0319] 4.1 Evaluation criteria

[0320] Cured: Symptoms such as panic, palpitations, insomnia, and nightmares disappear, and the patient returns to normal life;

[0321] Significant effect: The number of panic and palpitations is significantly reduced, and the occasional difficulty in falling asleep and insomnia and dreaminess occur, and the daytime life is basically restored to normal;

[0322] Effective: symptoms of panic and palpitations are alleviated, difficulty falling asleep and insomnia are reduced, and normal life is restored in some days;

[0323] Ineffective: Symptoms such as panic, heart palpitations, insomnia, and nightmares were not improved, and some of them tended to get worse.

[0324] 4.2 Test results

[0325] The effects of the three groups of experiments on treating insomnia of heart and spleen deficiency type are shown in Table 19 below.

[0326] Table 19 Comparison of the effective rates of the three groups after treatment

[0327] Group Number of cases cure Significant effect efficient invalid Efficiency Cure rate Experimental Group 30 19 6 3 2 93.3％ 63.3％ Control group 1 30 7 12 4 7 76.7％ 23.3％ Control group 2 30 6 10 9 5 83.3％ 20.0％

[0328] Conclusion: According to the data analysis of Table 19, the effective rate of the test group is as high as 93.3%, which is significantly higher than 76.7% of the control group 1 and 83.3% of the control group 2. In particular, the cure rate of the test group reached 63.3%, which is much higher than 23.3% of the control group 1. This data shows that the drug composition used in the test group performs well in improving the symptoms of insomnia caused by deficiency of both heart and spleen. Acanthopanax senticosus in the composition promotes the biochemical transformation of qi and blood, replenishes the qi of the heart and spleen, and plays a key role in calming the mind and helping sleep; Astragalus membranaceus not only replenishes the spleen qi but also helps to produce blood, providing sufficient qi and blood support for the mind; Ganoderma lucidum has the effect of replenishing qi and calming the mind, and cooperates with other drugs to strengthen the nourishment of the mind and relieve insomnia symptoms. At the same time, it also has a certain relief effect on possible symptoms such as cough and asthma. Ginseng leaves assist Acanthopanax and Astragalus to enhance the power of replenishing qi, and its calming and intelligence-enhancing effects help improve insomnia; Schisandra chinensis nourishes the heart and kidneys, calms the mind and calms the nerves, and has a highly effective synergistic effect with Acanthopanax; Gastrodia elata has the effect of calming liver yang, promoting fluid production and nourishing blood, and can relieve the accompanying symptoms such as dizziness and headache. The Chinese medicine composition composed of six medicinal materials has obvious therapeutic effects on symptoms such as palpitations, palpitations, insomnia, and dreams. At the same time, the experimental group did not produce side effects such as dizziness and headache compared with the control group 1 and the control group 2, and the patients with memory loss symptoms in the experimental group were significantly improved. The reason why the experimental group showed better therapeutic effects is that it adopted a more effective and targeted treatment strategy. This strategy may act more directly on the pathological mechanism of insomnia of both heart and spleen deficiency type, thereby promoting the recovery of patients. In summary, this experiment shows that the pharmaceutical composition of the present invention shows a better therapeutic effect than the control group in the treatment of insomnia of both heart and spleen deficiency type.

Claims

1. A pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss, characterized in that The medicine composition is composed of 5-100 parts of Acanthopanax senticosus, 1-20 parts of Ginseng Leaves, 1-20 parts of Schisandrae Chinensis, 2-50 parts of Astragalus, 2-60 parts of Ganoderma Lucidum and 1-20 parts of Gastrodia elata in parts by weight.

2. A pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss according to claim 1, characterized in that The medicine composition is composed of 10-80 parts of Acanthopanax senticosus, 5-15 parts of Ginseng Leaves, 5-15 parts of Schisandrae Chinensis, 5-40 parts of Astragalus, 5-50 parts of Ganoderma Lucidum and 5-15 parts of Gastrodia elata in parts by weight.

3. A pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss according to claim 1 or 2, characterized in that The medicine composition is composed of 20-60 parts of Acanthopanax senticosus, 10-15 parts of Ginseng Leaves, 10-15 parts of Schisandrae Chinensis, 10-30 parts of Astragalus, 10-40 parts of Ganoderma Lucidum and 10-15 parts of Gastrodia elata in parts by weight.

4. A pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss according to claim 1 or 3, characterized in that The medicine composition is composed of 40-50 parts of Acanthopanax senticosus, 10-15 parts of Ginseng Leaves, 10-15 parts of Schisandrae Chinensis, 10-20 parts of Astragalus, 10-30 parts of Ganoderma Lucidum and 10-15 parts of Gastrodia elata in parts by weight.

5. A pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss according to claim 1 or 4, characterized in that The pharmaceutical composition is composed of 25 parts of Acanthopanax senticosus, 5 parts of Ginseng Leaves, 3 parts of Schisandrae Chinensis, 15 parts of Astragalus, 10 parts of Ganoderma Lucidum and 5 parts of Gastrodia elata in parts by weight.

6. A method for preparing the granules of the pharmaceutical composition for insomnia (heart and spleen deficiency type) and memory loss according to claim 1, characterized in that The preparation method is carried out as follows: 1) Weigh 5-100 parts of Acanthopanax senticosus, 1-20 parts of Ginseng Leaves, 1-20 parts of Schisandrae Chinensis, 2-50 parts of Astragalus, 2-60 parts of Ganoderma Lucidum and 1-20 parts of Gastrodia elata according to weight; 2) mixing the medicinal materials weighed in step 1), adding 8 times the amount of water, reflux extraction, filtering, and obtaining filtrate 1; ultrasonic-assisted extraction of the medicinal residue, filtering, and obtaining filtrate 2; adding 8 times the amount of water to dissolve the residue, reflux extraction, filtering, and obtaining filtrate 3; combining filtrate 1, filtrate 2, and filtrate 3 for later use; 3) concentrating the combined filtrate from step 2) by a reduced pressure concentration method; 4) Add auxiliary materials to make tablets, granules, capsules or oral liquid.

7. The preparation method according to claim 5, characterized in that In step 2), 8 times the amount of water was added, refluxed for 2 hours, and filtered to obtain filtrate 1; the residue was ultrasonically extracted for 10 minutes at 40° C., and filtered to obtain filtrate 2; the residue was dissolved in 8 times the amount of water, refluxed for 1.5 hours, and filtered to obtain filtrate 3.

8. The preparation method according to claim 5, characterized in that The concentration in step 3) is concentrated to an extract having a relative density of 1.10 to 1.30 at 60°C.

9. The preparation method according to claim 5, characterized in that The granules are prepared as follows: After drying the concentrated extract in step 3), measuring its water content ≤ 3%, taking it out and cooling it; adding dextrin and citric acid as auxiliary materials to mix with the dried extract in step 4), then adding 10% of water based on the total amount of the original auxiliary materials, stirring and mixing to obtain a soft material, granulating, drying, granulating, and packaging.

10. The preparation method according to claim 5, characterized in that The oral solution is prepared as follows: 1) Weigh 5-100 parts of Acanthopanax senticosus, 1-20 parts of Ginseng Leaves, 1-20 parts of Schisandrae Chinensis, 2-50 parts of Astragalus, 2-60 parts of Ganoderma Lucidum and 1-20 parts of Gastrodia elata according to weight; 2) mixing the medicinal materials weighed in step 1), adding 8 times the amount of water, reflux extraction, filtering, and obtaining filtrate 1; ultrasonic-assisted extraction of the medicinal residue, filtering, and obtaining filtrate 2; adding 8 times the amount of water to dissolve the residue, reflux extraction, filtering, and obtaining filtrate 3; combining filtrate 1, filtrate 2, and filtrate 3 for later use; 3) concentrating the combined filtrate from step 2) by a reduced pressure concentration method; 4) Add the concentrated extract to water at 70-85°C and stir evenly, then add potassium sorbate, sodium citrate and stevioside excipients, add water to make up to volume, centrifuge, filter, fill and sterilize.

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