Application of Anshenbunao solution in the preparation of drugs for the prevention and treatment of Alzheimer's disease

Abstract

The present invention provides an application of Anshenbu'nao liquid in the preparation of drugs for preventing and treating Alzheimer's disease, and relates to the technical field of traditional Chinese medicine. The present invention is based on Anshenbu'nao liquid, and through research on its preparation method, it is found that Anshenbu'nao liquid can significantly improve the cognitive recognition ability, learning and memory ability, increase the number of neurons, reverse the excessive oxidative stress caused by Alzheimer's disease, and significantly improve neuronal damage and neuroinflammation of rats with Alzheimer's disease; it is shown that Anshenbu'nao liquid has a significant effect on the treatment and improvement of Alzheimer's disease, and is applied to the preparation of drugs for preventing and treating Alzheimer's disease.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to the application of anesthesia-shenbunao liquid in the preparation of drugs for preventing and treating Alzheimer's disease. Background Art

[0002] Alzheimer's disease (AD) is a progressive neurodegenerative disease with an insidious onset. It is clinically characterized by global dementia, including memory impairment, aphasia, apraxia, agnosia, visual-spatial skill impairment, executive dysfunction, and personality and behavioral changes.

[0003] Modern medical research shows that the pathogenesis of Alzheimer's disease is extremely complex, involving multiple gene defects, environmental influences, neurotoxins, cell apoptosis, oxidative stress, β-AP (β-amyloid protein) aggregation, inflammatory response, Ca 2+ Overload, cholinergic and other multi-system degeneration and other aspects. In view of the pathogenesis of Alzheimer's disease, the main therapeutic drugs on the market now include cholinesterase inhibitors, glutamate receptor antagonists, calcium ion antagonists, free radical scavengers and antioxidants, estrogen, etc. The above drugs are mainly chemical drugs with large toxic side effects.

[0004] In view of the current limitations of Western medicine in treating Alzheimer's disease, as well as the adverse reactions caused by Western medicine and the complexity of the causes of AD, it is imperative and urgent to give full play to the characteristics and advantages of traditional Chinese medicine, carry out holistic and comprehensive treatment, and exert the effects of multiple links and multiple targets to develop safe and effective drugs for the prevention and treatment of Alzheimer's disease.

[0005] With the continuous deepening of research, many Chinese medicine formulas for the prevention and treatment of Alzheimer's disease have been published. For example, Chinese invention patent 201610227537.0 discloses a Chinese medicine composition for the prevention and treatment of Alzheimer's disease and its preparation method. The composition contains 1-12 parts by weight of phosphatidylserine, and also contains the following Chinese medicine components by weight: 10-180 parts of ginseng, 10-200 parts of gastrodia, 30-240 parts of ginkgo leaves, 10-200 parts of polygala, and 10-120 parts of galangal; or 10-180 parts of ginseng and the following Chinese medicine extracts: 10-200 parts of gastrodia extract, 30-240 parts of ginkgo leaves, 10-200 parts of polygala extract, and 10-120 parts of galangal extract; or 10-180 parts of ginseng, 10-200 parts of gastrodia, 30-240 parts of ginkgo leaves, 10-200 parts of polygala, and 10-120 parts of galangal extract. The Chinese medicine composition of the invention has the effects of nourishing the liver and marrow, unblocking the meridians and resolving phlegm, improving intelligence and strengthening the brain, etc., can significantly improve memory and mild cognitive dysfunction clinically, and has no toxic side effects when taken for a long time.

[0006] Chinese invention patent 201910371018.5 discloses a Chinese medicine composition for treating Alzheimer's disease and its preparation method, wherein the Chinese medicine composition is composed of the following raw materials in parts by weight: 20-30 parts of Alpinia oxyphylla, 10-20 parts of Astragalus, 10-20 parts of Cuscuta, 5-15 parts of Chuanxiong, 10-20 parts of Rehmannia glutinosa, 10-20 parts of Morus alba, 10-20 parts of Cassia Cassia, 10-20 parts of Eucommia, 5-15 parts of Pheretima, 1-3 parts of Leech, 5-15 parts of Gastrodia, and 5-15 parts of Uncaria rhynchophylla. The invention is based on the principle of combining disease differentiation with syndrome differentiation in traditional Chinese medicine, and screens out natural medicines with the effects of tonifying the kidney, promoting blood circulation, and resolving phlegm, and is prepared according to a certain weight ratio, providing a Chinese medicine composition for treating Alzheimer's disease with significant efficacy and low cost.

[0007] Related products used to nourish qi and blood, treat amnesia and insomnia, such as Anshenbunao liquid, the main ingredients of which include velvet antler, processed Polygonum multiflorum, epimedium, dried ginger, licorice, jujube, and vitamin B1; its functions and indications: generating essence and nourishing marrow, nourishing qi and blood, strengthening brain and calming mind. It is used for dizziness, fatigue, amnesia, and insomnia caused by insufficient kidney essence and deficiency of both qi and blood; neurasthenia with the above symptoms. Related patent 200610017255.4 discloses a drug preparation for enhancing memory, which is prepared from Chinese medicines such as velvet antler, processed Polygonum multiflorum, epimedium, dried ginger, licorice, jujube, and vitamin B1, and is suitable for insomnia, amnesia, trance, forgetfulness, palpitations, dark eyes and tinnitus.

[0008] However, the currently available Chinese medicine compositions and drugs cannot effectively improve cognitive recognition ability, learning and memory ability, reverse the excessive oxidative stress caused by Alzheimer's disease, and improve neuronal damage and neuroinflammation. Therefore, a new use of Anshen Bu Nao Liquid has been developed in calming the mind and nourishing the brain, enhancing memory, and preventing and treating Alzheimer's disease. Summary of the invention

[0009] The present invention aims at the problems existing in the prior art and provides an application of an Anshen Bu Nao liquid in the preparation of drugs for preventing and treating Alzheimer's disease. Based on the Anshen Bu Nao liquid, a preparation method thereof is studied. During the preparation process, different solvents, raw materials with different particle sizes and extraction methods are used to extract Chinese medicinal ingredients. The obtained Anshen Bu Nao liquid can significantly improve cognitive recognition ability, learning and memory ability, increase the number of neurons, reverse excessive oxidative stress caused by Alzheimer's disease, and improve the significant effects of neuronal damage and neuroinflammation.

[0010] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0011] In one aspect, the present invention provides the use of Anshenbunao liquid in the preparation of drugs for preventing and treating Alzheimer's disease.

[0012] Preferably, the drug is a drug that can improve the ability to recognize old objects and new objects.

[0013] Preferably, the drug is a drug that can improve the learning and memory ability of spatial position and direction sense.

[0014] Preferably, the drug is a drug that can increase the number of neurons in the cerebral cortex.

[0015] Preferably, the drug is a drug capable of protecting neurons.

[0016] Preferably, the drug is a drug that can inhibit the content and activity of neuroinflammatory factors.

[0017] Further preferably, the neuroinflammatory factors include at least one of Aβ42, p-Tau, AChE, NF-light and GFAP.

[0018] Preferably, the drug is a drug capable of inhibiting oxidative stress levels.

[0019] Further preferably, the drug is a drug that can significantly reduce the levels of CRP, TNF-α, IL-1β and IL-6, and increase the levels of IL-10 and GSH-Px.

[0020] The tranquilizing and brain-nourishing liquid is a product prepared by the methods described in Examples 1 and 2 of the prior patent CN1943774A, or is a tranquilizing and brain-nourishing liquid prepared by the following preparation method.

[0021] The preparation method is as follows:

[0022] (1) Mixing velvet antler with water, decocting for 4-6 times, with the solid-liquid ratio of each decoction being 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the decoction after concentration, standing until the wax layer solidifies, and adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, using a gradually decreasing ethanol concentration during the alcohol precipitation process, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate;

[0023] (2) crushing the dried ginger, mixing it with 6-10 times the amount of water, extracting the distillate by steam distillation, the steam distillation time is 5-12 hours, obtaining the dried ginger volatile oil, and recovering the dried ginger residue;

[0024] (3) Mixing the dried ginger residue with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the amount of the dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to a concentration of 70-85% in the system, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a dried ginger water extract;

[0025] (4) grinding the prepared Polygonum multiflorum, mixing it with water, and decocting it 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the prepared Polygonum multiflorum; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the prepared Polygonum multiflorum water extract;

[0026] (5) crushing epimedium, mixing with water, and decocting for 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain an epimedium water extract;

[0027] (6) Grinding licorice, mixing with water, and decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a licorice water extract;

[0028] (7) Mixing jujube with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a jujube water extract;

[0029] (8) The extract components prepared in steps (1) to (7) are mixed with vitamin B1 to obtain a tranquilizing and brain-tonifying liquid.

[0030] Preferably, step (1) is specifically as follows: mixing velvet antler with water, decocting for 5 times, with the solid-liquid ratio of each decoction being 7 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the concentrated decoction, standing until the wax layer solidifies, adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate.

[0031] Preferably, in step (1), the alcohol precipitation process specifically comprises the steps of:

[0032] S1, adding ethanol to the remaining part after removing the wax layer until the ethanol concentration in the system is 80-85%, filtering after standing, collecting the filtrate, concentrating to remove ethanol, and obtaining pilose antler alcohol precipitate extract 1;

[0033] S2, continue to add ethanol to the antler alcohol precipitate extract 1 until the ethanol concentration in the system is 70-75%, let it stand and then filter, collect the filtrate, and concentrate to remove ethanol to obtain the antler alcohol precipitate extract 2;

[0034] S3. Add ethanol to the antler alcohol precipitate extract 2 until the ethanol concentration in the system is 20-30%, let it stand and then filter, collect the filtrate, concentrate to remove the ethanol, and dry to obtain the antler water-extracted alcohol precipitate.

[0035] Further preferably, in the alcohol precipitation process, the ethanol concentration in step S1 is 80%, the ethanol concentration in step S2 is 75%, and the ethanol concentration in step S3 is 25%.

[0036] Preferably, in step (2), the dried ginger is crushed to a particle size of 50-100 mesh, the amount of water added is 8-9 times the amount of water, and the steam distillation time is 7-8 hours.

[0037] Further preferably, the dried ginger is crushed to a particle size of 50-100 mesh, the amount of water added is 9 times the amount of water, and the steam distillation time is 8 hours.

[0038] Preferably, step (3) is specifically as follows: mixing the dried ginger residue with water, decocting for 3 times, each decocting time is 2 hours, and the amount of water added for each decoction is 8 times the amount of the dried ginger residue; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain a dried ginger water extract.

[0039] Preferably, step (4) is specifically as follows: taking the prepared Polygonum multiflorum, crushing it, mixing it with water, and decocting it for 3 times, each decocting time is 2 hours, and the amount of water added for each decocting is 8 times the weight of the prepared Polygonum multiflorum; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the prepared Polygonum multiflorum water extract.

[0040] Preferably, step (5) is specifically as follows: crushing epimedium, mixing with water, and decocting for 4 times, each decocting time is 2 hours, and the amount of water added for each decoction is 8 times the weight of epimedium; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 75%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain epimedium water extract.

[0041] Preferably, step (6) is specifically as follows: grind licorice, mix with water, and decoct twice: the first decoction is 1 hour, and the second decoction is 2 hours; the amount of water added in the first decoction is 5 times the weight of the licorice powder, and the amount of water added in the second decoction is 8 times the weight of the licorice powder; filter after each decoction, combine the decoctions, concentrate, add ethanol to the system until the ethanol concentration is 80%, let stand for 48 hours, filter, collect the filtrate, concentrate and dry to obtain a licorice water extract.

[0042] Further preferably, the dry grass is crushed, and after the crushing, the licorice with a particle size of 50-100 mesh is sieved and boiled.

[0043] Preferably, step (7) is specifically as follows: taking jujube and mixing with water and decocting for 3 times, each decocting time is 2 hours, and the amount of water added for each decocting is 5 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain a jujube water extract.

[0044] The tranquilizing and brain-tonifying liquid comprises the following ingredients, measured by weight: 2-5 parts of pilose antler, 60-65 parts of processed polygonum multiflorum, 45-55 parts of epimedium, 10-15 parts of dried ginger, 5-7 parts of liquorice, 10-15 parts of jujube, and 0.3-0.8 parts of vitamin B1.

[0045] Further preferably, the tranquilizing and brain-nourishing liquid comprises the following ingredients, measured by weight: 3 parts of pilose antler, 62.5 parts of processed Polygonum multiflorum, 50 parts of epimedium, 12.5 parts of dried ginger, 6.25 parts of liquorice, 12.5 parts of jujube, and 0.5 parts of vitamin B1.

[0046] Preferably, the weight ratio of the pilose antler, dried ginger and processed Polygonum multiflorum is 1.5-6:8-18:55-70; the weight ratio is preferably 2-5:10-15:60-65, and the weight ratio is further preferably 3:12.5:62.5.

[0047] Preferably, the tranquilizing and brain-nourishing liquid further comprises, by weight: 100-300 parts of sucrose, 1-3 parts of benzoic acid, 0.3-2.3 parts of ethylparaben, and 1-3 parts of sodium benzoate.

[0048] Preferably, the Anshenbunao solution in the application can be administered alone or in combination with one or more other therapeutic agents.

[0049] Compared with the prior art, the present invention has the following beneficial effects:

[0050] 1. The present invention is based on the Anshen Bu Nao Liquid, and studies its preparation method. In the preparation process, different solvents, raw materials with different particle sizes and extraction methods are used to extract the Chinese medicinal ingredients. The obtained Anshen Bu Nao Liquid can significantly improve the cognitive recognition ability, learning and memory ability of Alzheimer's disease rats, increase the number of neurons, reverse the excessive oxidative stress caused by Alzheimer's disease, and significantly improve neuronal damage and neuroinflammation; the Anshen Bu Nao Liquid of the present invention has a significant effect on the treatment and improvement of Alzheimer's disease.

[0051] 2. The tranquilizing and brain-nourishing liquid provided by the present invention utilizes various extract components through a specific extraction method, and the various components are reasonably matched. The obtained tranquilizing and brain-nourishing liquid has high activity and good effect, and is significantly effective in treating Alzheimer's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 This is a bar graph of the novel object recognition index of rats in each group in the novel object recognition experiment.

[0053] Figure 2 It is a bar graph of the platform latency and platform crossing times of each group of rats in the Morris experiment.

[0054] Figure 3 are Nissl staining images of the cerebral cortex of rats in each group.

[0055] Figure 4 This is a graph showing the changes in Aβ42, p-Tau, AChE in the hippocampus and NF-light, GFAP in plasma of each group of rats.

[0056] Figure 5 This is a graph showing the changes in CRP, TNF-α, IL-1β, IL-6, IL-10, and GSH-Px in the cerebral cortex of rats in each group. DETAILED DESCRIPTION

[0057] The following non-limiting examples can make those of ordinary skill in the art understand the present invention more comprehensively, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention according to the disclosed content, and it should also belong to the scope of the present invention. When the embodiment gives a numerical range, it should be understood that, unless otherwise specified in the present invention, the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those of ordinary skill in the technical field to which the present invention belongs.

[0058] The present invention is further described below by way of specific examples. The various chemical reagents and Chinese medicines used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Products from different manufacturers have no significant effect on the effects. In the following examples, "parts" and "g" have the same meaning, i.e., 1 part = 1 g.

[0059] Basic Example:

[0060] The preparation method of Anshen Bu Nao Liquid is as follows:

[0061] (1) Mixing velvet antler with water, decocting for 4-6 times, with the solid-liquid ratio of each decoction being 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the decoction after concentration, standing until the wax layer solidifies, and adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, using a gradually decreasing ethanol concentration during the alcohol precipitation process, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate;

[0062] (2) crushing the dried ginger, mixing it with 6-10 times the amount of water, extracting the distillate by steam distillation, the steam distillation time is 5-12 hours, obtaining the dried ginger volatile oil, and recovering the dried ginger residue;

[0063] (3) Mixing the dried ginger residue with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the amount of the dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to a concentration of 70-85% in the system, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a dried ginger water extract;

[0064] (4) grinding the prepared Polygonum multiflorum, mixing it with water, and decocting it 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the prepared Polygonum multiflorum; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the prepared Polygonum multiflorum water extract;

[0065] (5) crushing epimedium, mixing with water, and decocting for 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain an epimedium water extract;

[0066] (6) Grinding licorice, mixing with water, and decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a licorice water extract;

[0067] (7) Mixing jujube with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a jujube water extract;

[0068] (8) Mixing the extract components prepared in steps (1) to (7) with vitamin B1, benzoic acid, sodium benzoate, ethylparaben and sucrose to obtain the tranquilizing and brain-tonifying liquid.

[0069] Basic Example 1: The preparation method of pilose antler water-extract alcohol precipitate comprises the steps of: mixing pilose antler with water, decocting for 4-6 times, with the solid-liquid ratio of each decoction being 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the concentrated decoction, standing until the wax layer solidifies, adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, collecting the extract after the alcohol precipitation, and obtaining pilose antler water-extract alcohol precipitate.

[0070] In an embodiment, the method comprises the following steps: mixing velvet antler with water, decocting for 5 times, wherein the solid-liquid ratio of each decoction is 7 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the concentrated decoction, standing until the wax layer solidifies, adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate.

[0071] The alcohol precipitation process specifically comprises the following steps: S1, adding ethanol to the remaining part after removing the wax layer until the ethanol concentration in the system is 80%, filtering after standing, collecting the filtrate, concentrating to remove the ethanol, and obtaining the antler alcohol precipitation extract 1; S2, continuing to add ethanol to the antler alcohol precipitation extract 1 until the ethanol concentration in the system is 75%, filtering after standing, collecting the filtrate, concentrating to remove the ethanol, and obtaining the antler alcohol precipitation extract 2; S3, adding ethanol to the antler alcohol precipitation extract 2 until the ethanol concentration in the system is 25%, filtering after standing, collecting the filtrate, concentrating to remove the ethanol, and drying to obtain the antler water-extracted alcohol precipitation.

[0072] Basic Example 2: The preparation method of dried ginger volatile oil and dried ginger water extract comprises the steps of: crushing dried ginger, mixing dried ginger with a mesh size of 50-100 with 9 times the amount of water, extracting the distillate by steam distillation, the steam distillation time is 8 hours, and obtaining dried ginger volatile oil; recovering the dried ginger residue.

[0073] The above-mentioned dried ginger residue was mixed with water and decocted for 3 times, each decoction time was 2 hours, and the amount of water added for each decoction was 8 times the amount of the dried ginger residue; after each decoction, the decoctions were combined, concentrated, and ethanol was added to the system until the ethanol concentration was 80%, allowed to stand for 48 hours, filtered, the filtrate was collected, and concentrated and dried to obtain the dried ginger water extract.

[0074] Basic Example 3: The preparation method of the water extract of Polygonum multiflorum comprises the steps of: taking processed Polygonum multiflorum, crushing it, mixing it with water, decocting it 3 times, each decocting time is 2 hours, and the amount of water added for each decoction is 8 times the weight of the processed Polygonum multiflorum; filtering after each decoction, combining the decoctions, concentrating them, adding ethanol to the system until the concentration of ethanol is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the water extract of processed Polygonum multiflorum.

[0075] Basic Example 4: The preparation method of epimedium water extract comprises the steps of: taking epimedium, crushing it, mixing it with water, decocting it 4 times, each decoction time is 2 hours, and the amount of water added for each decoction is 8 times the weight of epimedium; filtering after each decoction, combining the decoctions, concentrating them, adding ethanol to the system until the concentration of ethanol is 75%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain epimedium water extract.

[0076] Basic Example 5: The preparation method of licorice water extract comprises the steps of: crushing and sieving licorice to a particle size of 50-100 mesh, mixing licorice with water, and decocting twice: the first decoction is 1 hour, and the amount of water added is 5 times the weight of the licorice powder; the second decoction is 2 hours, and the amount of water added is 8 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the concentration of ethanol is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the licorice water extract.

[0077] Basic Example 6: The preparation method of jujube water extract comprises the steps of: mixing jujube with water and decocting for 3 times, each decocting time is 2 hours, and the amount of water added for each decoction is 5 times the weight of the jujube; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the jujube water extract.

[0078] Example 1

[0079] The product prepared by the method described in Example 1 of patent CN1943774A.

[0080] Example 2

[0081] The product prepared by the method described in Example 2 of patent CN1943774A.

[0082] Embodiment 3-5

[0083] The tranquilizing and brain-nourishing liquid for preventing and treating Alzheimer's disease is shown in Table 1, and includes the following raw material components in parts by weight:

[0084] Table 1

[0085] Weight Example 3 Example 4 Example 5 Deer antler 2 5 3 Processed Polygonum multiflorum 65 60 62.5 Epimedium 55 45 50 Dried ginger 10 15 12.5 Licorice 5 7 6.25 Jujube 15 10 12.5 <![CDATA[Vitamin B1]]> 0.3 0.8 0.5

[0086] Example 3 further comprises the following components, calculated by weight: 100 parts of sucrose, 1 part of benzoic acid, 0.3 parts of ethylparaben and 1 part of sodium benzoate.

[0087] Example 4 further comprises the following components, in parts by weight: 300 parts of sucrose, 2.5 parts of benzoic acid, 2.0 parts of ethylparaben and 3 parts of sodium benzoate.

[0088] Example 5, in parts by weight, further comprises the following components: 200 parts of sucrose, 2.2 parts of benzoic acid, 1.8 parts of ethylparaben and 1.5 parts of

[0089] Sodium benzoate.

[0090] Preparation method: Weigh the raw materials according to the weight of each raw material described in Table 1, then mix the various extracts prepared according to Basic Examples 1-6 with vitamin B1, benzoic acid, sodium benzoate, ethylparaben and sucrose, add water to make the volume to 1000 mL, and obtain a tranquilizing and brain-tonifying solution.

[0091] Comparative Example 1

[0092] The difference from Example 5 is that 3.5 parts of Epimedium are replaced by 3.5 parts of Poria.

[0093] The preparation method of the water extract of Poria cocos is the same as that of the water extract of Epimedium: grind Poria cocos, mix with water, and decoct for 4 times, each decoction time is 2 hours, and the amount of water added for each decoction is 8 times the weight of Epimedium; filter after each decoction, combine the decoctions, concentrate, add ethanol to the system until the concentration of ethanol is 75%, let stand for 48 hours, filter, collect the filtrate, concentrate and dry to obtain the water extract of Poria cocos. The rest is the same as Example 5, and the product of Comparative Example 1 is obtained.

[0094] Comparative Example 2

[0095] Different from Example 5, the Anshenbunao liquid does not contain the component Polygonum multiflorum.

[0096] Comparative Example 3

[0097] This comparative example contains only a single ingredient, prepared Polygonum multiflorum, and the weight portion of prepared Polygonum multiflorum is 147.25 parts, which is the same as the total weight portions of the ingredients in the Anshen Bu Nao Liquid described in Example 5.

[0098] The preparation method is as follows: grind the processed Polygonum multiflorum, mix it with water, and decoct it for 3 times, each decoction time is 2 hours, and the amount of water added for each decoction is 8 times the weight of the processed Polygonum multiflorum; filter after each decoction, combine the decoctions, concentrate, add ethanol to the system until the concentration of ethanol is 80%, let it stand for 48 hours, filter, collect the filtrate, concentrate and dry to obtain the product.

[0099] Then it was mixed with sodium benzoate and fructose syrup to obtain the product of Comparative Example 3.

[0100] Comparative Example 4

[0101] The difference from Example 5 is that the weight parts of pilose antler, dried ginger and processed Polygonum multiflorum are different, specifically: 9.5 parts pilose antler, 49.5 parts

[0102] Processed Polygonum multiflorum, 50 parts of Epimedium, 19 parts of dried ginger, 6.25 parts of licorice, 12.5 parts of jujube, and 0.5 part of vitamin B1.

[0103] The rest is the same as in Example 5.

[0104] Comparative Example 5

[0105] The difference from Example 5 is that the preparation method is different: the concentration of ethanol in Basic Example 3-Basic Example 6 is different, and ethanol is added until the concentration of ethanol is 20%:

[0106] The decoctions were combined and filtered after each decoction. After concentration, ethanol was added until the concentration of ethanol in the system was 20%. The mixture was allowed to stand for 48 hours, filtered, the filtrate was collected, concentrated and dried to obtain processed Polygonum multiflorum water extract, Epimedium water extract, Licorice water extract and Jujube water extract, respectively.

[0107] Specifically:

[0108] (1) The preparation method of antler water-extraction alcohol precipitation is the same as that of basic example 1;

[0109] (2) The preparation method of dried ginger volatile oil and dried ginger water extract is the same as that of basic example 2;

[0110] (3) Preparation method of Polygonum multiflorum water extract: grind prepared Polygonum multiflorum, mix with water, and decoct for 3 times, each decoction time is 2 hours, and the amount of water added for each decoction is 8 times the weight of prepared Polygonum multiflorum; filter after each decoction, combine the decoctions, concentrate, add ethanol to the system until the concentration of ethanol in the system is 20%, let stand for 48 hours, filter, collect the filtrate, concentrate and dry to obtain prepared Polygonum multiflorum water extract;

[0111] (4) Preparation method of epimedium water extract: crush epimedium, mix with water, and boil for 4 times, each boiling time is 2 hours, and the amount of water added for each boiling is 8 times the weight of epimedium; filter after each boiling, combine the decoctions, concentrate, add ethanol to the system until the concentration of ethanol in the system is 20%, let stand for 48 hours, filter, collect the filtrate, concentrate and dry to obtain epimedium water extract;

[0112] (5) Preparation method of liquorice aqueous extract: licorice is crushed and sieved to a particle size of 50-100 mesh. The liquorice is mixed with water and decocted twice: the first decoction is 1 hour, and the amount of water added is 5 times the weight of the liquorice powder; the second decoction is 2 hours, and the amount of water added is 8 times the weight of the liquorice powder; after each decoction, the decoction is filtered, the decoction is combined, concentrated, and ethanol is added to the system until the concentration of ethanol is 20%, and the system is allowed to stand for 48 hours, filtered, and the filtrate is collected, concentrated and dried to obtain the liquorice aqueous extract.

[0113] (6) Preparation method of jujube water extract: jujube and water are mixed and decocted for 3 times, each decocting time is 2 hours, and the amount of water added for each decocting is 5 times the weight of the jujube; after each decocting, the decoction is filtered, the decoction is combined, and after concentration, ethanol is added to the system until the ethanol concentration in the system is 80%, and the system is allowed to stand for 48 hours, filtered, the filtrate is collected, and concentrated and dried to obtain the jujube water extract.

[0114] The prepared extract component is mixed with vitamin B1, benzoic acid, sodium benzoate, ethylparaben and sucrose, and water is added to make the volume to 1000 mL to obtain a tranquilizing and brain-tonifying liquid.

[0115] Comparative Example 6

[0116] The difference from Example 5 is that the particle size of the dried ginger powder in step (2) is different. In this comparative example, the particle size of the dried ginger powder used in preparing the dried ginger volatile oil is between 10 and 20 meshes. The rest is the same as Example 5.

[0117] Comparative Example 7

[0118] The difference from Example 5 is that the particle size of the licorice after crushing in step (6) is different. In this comparative example, licorice powder with a particle size of 300-400 mesh is used to prepare the licorice water extract. The rest is the same as Example 5.

[0119] Technical effect comparison test

[0120] 1. Experimental Animals

[0121] SD rats, male, weighing (220±20) g, were provided by Beijing Huafukang Biotechnology Co., Ltd., animal quality certificate number: 44829700017323. The animals were kept in an SPF environment, with a room temperature of 18-23°C and a relative humidity of 45%-55%, free access to water and food, and adaptive feeding for 3 days. All animal experiments were approved by the Animal Ethics Committee of the experimental unit (2023067).

[0122] 2. Reagents and Materials

[0123] Anshenbunao liquid, donepezil hydrochloride tablets (batch number: 022200701, Chongqing Zhien Pharmaceutical Co., Ltd., China), scopolamine hydrobromide (batch number: S302239, Shanghai Aladdin Biochemical Technology Co., Ltd.), and isoflurane obtained in the examples and comparative examples of the present invention.

[0124] 3. Experimental instruments

[0125] The animal behavior and pain measurement system (model: Panlabharved, Huaibei Zhenghua Biological Instrument Equipment Co., Ltd.) includes Morris water maze constant temperature swimming pool and open field ZH-ZFT, animal behavior software SMART 3.0 (Panlab, Spain), fully automatic intelligent dehydrator (model: HP300, Dakoway Biotechnology Co., Ltd.), tissue embedding machine (model: HistoStar, Thermo Fisher Scientific (China) Co., Ltd.), fully automatic staining and sealing integrated workstation (model: ST5020, Leica Instrument Co., Ltd., Germany), paraffin sectioner (model: RM2245, Leica Instrument Co., Ltd., Germany), upright fluorescence microscope (model: BX53, Olympus (China) Co., Ltd.), and medical centrifuge (model: DT5-3, Beijing Times Beili Centrifuge Co., Ltd.).

[0126] 4. Experimental methods

[0127] 4.1 Animal grouping and drug administration

[0128] After 3 days of adaptive feeding, SD male rats were randomly divided into normal control group (CON), Alzheimer's disease model group (AD), low-dose group (ASL) in Example 1, medium-dose group (ASM) in Example 1, high-dose group (ASH) in Example 1, medium-dose group in Example 2-5, medium-dose group in Comparative Examples 1-7, and positive drug donepezil hydrochloride group (DON), with 10 rats in each group for experiment. The examples, comparative examples and positive drug groups were all drug administration groups. The normal control group and the model group were given distilled water by gavage at 1.05 mL / 100 g, the low, medium and high dose groups were given Anshenbunao liquid by gavage at a dose of 1.05 mL / kg / d (human dose 10 mL / day), 2.1 mL / kg / d (human dose 20 mL / day), and 4.2 mL / kg / d (human dose 40 mL / day), and the donepezil hydrochloride group was given donepezil hydrochloride tablets by gavage at 0.52 g / kg / d, and the administration was continued for 4 weeks. The model was established 19 days after administration. The model group and all the treatment groups were given scopolamine at 3 mg / kg / d by intraperitoneal injection, and the normal control group was given an equal amount of saline by subcutaneous injection, and the model was established for 9 consecutive days.

[0129] 4.2 Indicator Observation

[0130] 4.2.1 New object recognition experiment

[0131] The novel object recognition test was tested in an opaque plastic chamber (35 cm × 41.5 cm × 50 cm). A camera mounted on the wall just above the test chamber was used to record the test process for offline analysis. Before starting the test, all rats were placed in the plastic chamber to adapt for 5 minutes. The test began 24 hours later, and two identical objects were placed in the corner, and each mouse stayed in the plastic chamber for 5 minutes. After 24 hours, object recognition was tested by replacing the habitual object with a new object, and the exploratory behavior of the mice was recorded for 5 minutes.

[0132] 4.2.2 Morris water maze

[0133] Hidden platform test: At the beginning of each day, the mouse is placed in the water facing the pool wall from any of the four quadrants (northeast, northwest, southeast, southwest), and the platform is placed in the southeast quadrant. Each time the mouse swims for a total of 60 seconds to find the hidden platform. If the platform is successfully found, the mouse can get a 10-second rest time on the platform; if the platform is not successfully found within 60 seconds, the experimenter will manually place the mouse on the platform and give it a 10-second rest time. Sometimes the mouse may fall off the platform or jump into the water to continue swimming before the 10-second interval is reached. Once this happens, the mouse is put back on the platform and the time is reset so that the time interval reaches 10 seconds. This ensures that each mouse has equal time to observe and obtain spatial information after each experiment.

[0134] Spatial exploration test: 24 hours after the hidden platform test, the platform was removed. Then, the mouse was placed in the water at the same entry point, and the swimming path of the mouse within 60 seconds was recorded. The mouse's stay time in the original platform quadrant and the number of times it crossed the original platform were recorded. The spatial positioning ability of the test mouse and the changing rules during the spatial exploration process were observed. The original location of the platform was marked with a circular ring on the computer screen, so that the number of times the original platform location was crossed could be recorded.

[0135] 4.3 Sample collection

[0136] 12 hours after the last injection of scopolamine, all rats were anesthetized with isoflurane. Blood was collected from the abdominal aorta of the rats, and the blood in the procoagulant blood collection tube was centrifuged at 3500rpm for 10 minutes at 4°C. The upper serum was separated and stored in a -80°C refrigerator. After the blood was collected, the brain, heart, liver, lung, kidney, colon, and feces of the rats were collected by dissection, quenched with liquid nitrogen, and stored in a -80°C refrigerator. The removed brain tissue was cut about 3mm along the transverse coronal plane and placed in an embedding box and immersed in 4% tissue cell fixative.

[0137] 4.4 Histopathology

[0138] The brain and colon were immersed in fixative and then dehydrated, embedded, dewaxed, sectioned, and Nissl-stained according to standard procedures.

[0139] 4.5 Elisa test

[0140] According to the instructions, the levels of interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 10 (IL-10), tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), and glutathione peroxidase (GSH-Px) in the rat cerebral cortex; amyloidβ-42 (Aβ42), phosphorylated Tau protein (phospho-microtubule-associated protein tau, p-Tau), acetylcholinesterase (AChE) in the hippocampus; and neurofilament light chain (NF-light) and glial fibrillary acidic protein (GFAP) in plasma were detected.

[0141] 4.6 Statistical analysis

[0142] The data were imported into GraphPad Prism 8 software for graphing and statistical analysis. Welch's t test was used for comparison analysis between two groups. P < 0.05 indicated that the difference was statistically significant.

[0143] 5. Technical Effect

[0144] 5.1 New Object Recognition

[0145] The new object recognition experiment mainly tests the rats' ability to recognize old and new objects, including remembering old things and becoming familiar with new things. This experiment evaluates the animals' cognitive memory ability through behavioral methods, such as the length of time they explore familiar objects and new unfamiliar objects. If the rats have poor cognitive ability, there will be no difference in the exploration of new and old objects; if the rats have normal cognitive ability, they will explore new objects longer than old objects.

[0146] The formula for calculating the cognitive index is: cognitive index = new object / (new object + old object) × 100%. Figure 1As shown in the figure, compared with the normal control group, the exploration time of the model group rats for the new object showed a decreasing trend (P=0.475). Compared with the model group, the new object recognition index of the rats in the positive drug group and the embodiment group increased (ASL vs AD P=0.738, ASM vs AD P=0.595, ASH vs AD P=0.624, DON vs AD P=0.306).

[0147] The novel object recognition experiment showed that: ① the exploration desire of scopolamine-induced AD model rats for new objects was significantly reduced; ② both the example group and the positive drug (donepezil hydrochloride tablets) could improve the learning ability of AD rats, but there was no statistical difference compared with the model group; ③ different doses of the drugs prepared by Anshenbu Nao Liquid could improve the learning ability of AD rats, among which the high-dose group of Example 1 had better efficacy than the low- and medium-dose groups, showing a certain dose-effect relationship.

[0148] 5.2 Morris water maze

[0149] The water maze experiment is mainly used to evaluate the spatial learning and memory ability of rats. The experiment forces the experimental animals to swim and learn how to find a platform hidden in the water. The learning and memory ability of the experimental animals' sense of spatial position and direction (spatial positioning) is tested by measuring the number of platform crossings, the latency to reach the platform, and the residence time in the target quadrant.

[0150] Figure 2 A in the figure shows (platform latency), the latency of the model rats to reach the platform was significantly prolonged (P=0.046); compared with the model group, the latency of the rats in the positive drug control group and the embodiment group to reach the platform was shortened, among which the positive group and the high-dose group (ASH) of Example 1 had significant effects (ASL vs AD P=0.014, ASM vs AD P=0.072, ASH vs AD P=0.002, DON vs AD P=0.048). Figure 2 As shown in B (number of platform crossings), compared with the normal control group, the number of times the model group rats crossed the platform was reduced (P=0.042). Both the positive drug and the drugs in the examples can increase the number of times the rats crossed the platform, and there is a statistical difference between the positive drug group and the high-dose group (ASH) of Example 1 (ASL vs AD P=0.081, ASM vs AD P=0.174, ASH vs AD P=0.012, DON vs AD P=0.021).

[0151] Figure 2 In the table, #normal control group was compared with model group, P<0.05; *drug administration group was compared with model group, P<0.05; **drug administration group was compared with model group, P<0.01.

[0152] The water maze experiment showed that: ① the learning and memory ability of scopolamine-induced AD model rats was significantly reduced, and they were unable to quickly and accurately locate the position of the platform, which was manifested by a significant prolongation of the latency to reach the platform (P<0.05) and a decrease in the number of platform crossings (P<0.05); ② the Anshenbunao liquid of the embodiment and the positive drug (donepezil hydrochloride tablets) can effectively improve the learning and memory ability of AD rats, among which donepezil hydrochloride and the high-dose group (ASH) of Example 1 were statistically different from the model group (P<0.05); ③ different doses of the Anshenbunao liquid of the embodiment can improve the learning and memory ability of AD rats, among which the high-dose group had the best therapeutic effect.

[0153] 5.3 Histopathology

[0154] From the statistical results, compared with the normal control group, the number of neurons in the cerebral cortex of the AD model rats decreased, but there was no statistical difference. Different doses of the example Anshenbu Nao liquid can increase the number of neurons in AD rats, and the high dose group (ASH) of Example 1 has better efficacy than the low and medium doses. Donepezil hydrochloride can also increase the number of neurons.

[0155] Figure 3 The staining histopathological results showed that: ① the number of neurons in the cerebral cortex of AD model rats induced by scopolamine decreased; ② both the example group and the positive drug (donepezil hydrochloride tablets) could increase the number of neurons in AD rats, but there was no statistical difference compared with the model group; ③ different doses of the tranquilizing and brain-nourishing solution of the present invention could increase the number of neurons in AD rats, among which the high-dose group (ASH) of Example 1 had better therapeutic effect than the low- and medium-dose groups.

[0156] 5.4 Elisa test

[0157] like Figure 4 As shown in the figure, compared with the normal control group, the content of Aβ42 and p-Tau in the brain of the AD model rats increased significantly, and the AChE activity was significantly upregulated. Different doses of the example Anshenbu Nao liquid can downregulate the levels of Aβ42 and p-Tau and inhibit AChE activity. The positive drug donepezil hydrochloride tablets can also reduce the content of Aβ42 and p-Tau and inhibit AChE activity.

[0158] Compared with the normal control group, the content of NF-light in the brain of rats in the AD model group increased significantly, indicating neuronal damage. Different doses of the example Anshenbunao liquid medicine and positive medicine (donepezil hydrochloride tablets) can reduce the content of NF-light, indicating that the Anshenbunao liquid and positive medicine of the present invention have the effect of protecting neurons.

[0159] Compared with the normal control group, the content of GFAP in the brain of the rats in the AD model group increased significantly, indicating the occurrence of neuroinflammation. Different doses of the Example Anshenbunao solution and the positive drug (donepezil hydrochloride tablets) can reduce the content of GFAP, and the Example Anshenbunao solution and the positive drug can inhibit neuroinflammation.

[0160] Compared with the example groups (ASH and ASM groups), the neuroinflammation (p-Tau, Aβ42, AChE, NF-light, GFAP content) of rats in the comparative example 1-7 groups were significantly different (P<005), indicating that the tranquilizing brain nourishing solution of the present invention has a better effect of inhibiting neuroinflammation than the drugs provided in comparative examples 1-7.

[0161] Figure 4 middle, ### CON vs AD, P < 0.001; ** AD vs drug-treated group, P < 0.01; *** AD vs drug administration group, P<0.001. Table 2 shows the contents of Aβ42, p-Tau, AChE in the hippocampus and NF-light and GFAP in the plasma of rats in each group ( ### CON vs AD, P < 0.001; ** AD vs drug-treated group, P < 0.01; *** AD vs drug-treated group, P < 0.001; & The other example groups were compared with Example 1 ASM, P < 0.05; && The other example groups were compared with Example 1 ASM, P<0.01).

[0162] Table 2

[0163]

[0164] In addition, from the inflammatory factor indicators ( Figure 5 ), compared with the normal control group, the inflammatory response in the rats of the AD model group increased significantly, as shown by the significant increase in the content of pro-inflammatory factors CRP, TNF-α, IL-1β, and IL-6, and the significant decrease in the content of the anti-inflammatory factor IL-10. Different doses of Example 1 Anshenbu Nao Liquid can reverse the excessive inflammatory response caused by AD. The positive drug (Donepezil Hydrochloride Tablets) can also inhibit the inflammatory response in AD rats.

[0165] From the oxidative stress index, compared with the normal control group, the oxidative stress in the AD model group rats increased significantly, which was manifested by a significant decrease in the enzyme activity of GSH-Px. Different doses of Example 1 Anshenbu Nao Liquid can reverse the excessive oxidative stress caused by AD. Positive drugs (Donepezil Hydrochloride Tablets) can also inhibit oxidative stress in AD rats.

[0166] Compared with the Example 1 group (ASH and ASM groups), the oxidative stress proinflammatory factors (CRP, TNF-α, IL-1β, IL-6 content, anti-inflammatory factor IL-10, GSH-Px content) of the rats in the comparative examples 1-7 groups were significantly different (P<0.05), indicating that the tranquilizing brain nourishing solution of the present invention has a better effect of inhibiting oxidative stress in AD rats than the drugs provided in comparative examples 1-7.

[0167] Figure 5 middle, ### CON vs AD, P < 0.001; ** AD vs drug-treated group, P < 0.01; *** AD vs drug-treated group, P<0.001.

[0168] Table 3 shows the contents of CRP, TNF-α, IL-1β, IL-6, IL-10, and GSH-Px in the cerebral cortex of rats in each group ( ### CON vs AD, P < 0.001; ** AD vs drug-treated group, P < 0.01; *** AD vs drug-treated group, P < 0.001; & The other example groups were compared with Example 1 ASM, P < 0.05; && The other example groups were compared with Example 1 ASM, P<0.01).

[0169] Table 3

[0170]

[0171]

[0172] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. The use of Anshenbunao liquid in the preparation of drugs for preventing and treating Alzheimer's disease, characterized in that: The ingredients of the tranquilizing and brain-nourishing liquid are as follows, by weight: 2-5 parts of pilose antler, 60-65 parts of processed polygonum multiflorum, 45-55 parts of epimedium, 10-15 parts of dried ginger, 5-7 parts of liquorice, 10-15 parts of jujube, and 0.3-0.8 parts of vitamin B1; The preparation method of Anshenbunao liquid is as follows: (1) Mix velvet antler with water, and boil for 4-6 times, with the solid-liquid ratio of each boiling being 3-8 g / mL, filter, and combine the filtrate to obtain a decoction; after the decoction is concentrated, add beeswax, and let stand until the wax layer solidifies, and add ethanol to the remaining part after removing the wax layer for alcohol precipitation, using a gradually decreasing ethanol concentration during the alcohol precipitation process, and collect the extract after the alcohol precipitation to obtain a velvet antler water-extracted alcohol precipitate; The alcohol precipitation process specifically comprises the steps of: S1, adding ethanol to the remaining part after removing the wax layer until the ethanol concentration in the system is 80-85%, filtering after standing, collecting the filtrate, concentrating to remove ethanol, and obtaining pilose antler alcohol precipitate extract 1; S2, continue to add ethanol to the antler alcohol precipitate extract 1 until the ethanol concentration in the system is 70-75%, let it stand and then filter, collect the filtrate, and concentrate to remove ethanol to obtain the antler alcohol precipitate extract 2; S3, add ethanol to the antler alcohol precipitate extract 2 until the ethanol concentration in the system is 20-30%, let it stand and then filter, collect the filtrate, concentrate to remove ethanol, and dry to obtain the antler water-extracted alcohol precipitate; (2) Grind the dried ginger into 50-100 mesh, mix it with 6-10 times the amount of water, and extract the distillate by steam distillation. The steam distillation time is 5-12 hours to obtain the dried ginger volatile oil, and recover the dried ginger residue; (3) Mix the dried ginger residue with water and boil for 2-5 times, with each boiling time being 1-3 hours and the amount of water added being 4-10 times the amount of the dried ginger residue; filter after each boiling, combine the decoctions, concentrate, add ethanol until the concentration of ethanol in the system is 70-85%, let stand for 40-68 hours, filter, collect the filtrate, concentrate and dry to obtain the dried ginger water extract; (4) Grind the prepared Polygonum multiflorum, mix with water, and boil for 2-5 times, each boiling time is 1-3 hours, and the amount of water added is 4-10 times the weight of the prepared Polygonum multiflorum; filter after each boiling, combine the decoctions, concentrate, add ethanol until the concentration of ethanol in the system is 70-85%, let stand for 40-68 hours, filter, collect the filtrate, concentrate and dry to obtain the prepared Polygonum multiflorum water extract; (5) crushing epimedium, mixing with water, and decocting for 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain epimedium water extract; (6) Grind and sieve licorice to a particle size of 50-100 mesh, mix with water, and boil for 2-5 times, each boiling time is 1-3 hours, and the amount of water added is 4-10 times the weight of the licorice powder; filter after each boiling, combine the decoctions, concentrate, add ethanol to the system until the ethanol concentration is 70-85%, let stand for 40-68 hours, filter, collect the filtrate, concentrate and dry to obtain a licorice water extract; (7) Mixing jujube with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a jujube water extract; (8) The extract components prepared in steps (1) to (7), vitamin B1, benzoic acid, sodium benzoate, ethylparaben and sucrose are mixed to obtain the tranquilizing and brain-tonifying solution.

2. The use according to claim 1, characterized in that: The drug can improve the ability to recognize old and new objects.

3. The use according to claim 1, characterized in that: The medicine can improve the learning and memory ability of spatial position sense and direction sense.

4. The use according to claim 1, characterized in that: The drug can increase the number of neurons in the cerebral cortex.

5. The use according to claim 1, characterized in that: The drug is capable of protecting neurons.

6. The use according to claim 1, characterized in that: The medicine can inhibit the content and activity of neuroinflammatory factors.

7. The use according to claim 6, characterized in that: The neuroinflammatory factors in neuroinflammation include at least one of Aβ42, p-Tau, AChE, NF-light and GFAP.

8. The use according to claim 1, characterized in that: The drug is able to inhibit oxidative stress levels.

9. The use according to claim 8, characterized in that: The inhibition of oxidative stress level specifically includes: significantly reducing the contents of CRP, TNF-α, IL-1β and IL-6, and increasing the content of IL-10.

10. The use according to claim 1, characterized in that: During the alcohol precipitation process of step (1), the ethanol concentration in step S1 is 80%, the ethanol concentration in step S2 is 75%, and the ethanol concentration in step S3 is 25%.

11. The use according to claim 1, characterized in that: The Anshen Bu Nao Liquid can be administered alone or in combination with one or more other therapeutic agents.

Images