Traditional Chinese medicine composition for resisting Alzheimer disease as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention provides a traditional Chinese medicine composition for resisting Alzheimer's disease as well as a preparation method and application of the traditional Chinese medicine composition. The traditional Chinese medicine composition is prepared from the following active ingredients in parts by weight: 6-50 parts of gastrodia elata, 6-50 parts of uncaria rhynchophylla, 9-60 parts of concha haliotidis, 6-50 parts of prepared rehmannia root, 6-50 parts of radix cyathulae, 3-30 parts of semen raphani, 3-30 parts of eucommia ulmoides, 3-30 parts of parasitic loranthus, 3-30 parts of scutellaria baicalensis, 3-30 parts of herba lycopi, 3-30 parts of vine of multiflower knotweed and 3-30 parts of cinnabar root poria. The raw materials of the medicine are common, the preparation method is simple and easy to operate, the medicine has wide application value, after oral administration of the medicine, cognitive impairment of a transgenic mouse model of the Alzheimer's disease can be obviously improved, deposition of age pigment in brain tissue is reduced, meanwhile, the number of activated microglia and astroglia in the brain tissue is reduced, and the treatment effect of the Alzheimer's disease is improved. The compound inhibits the formation of A beta in brain tissues, and has a new application of anti-Alzheimer's disease drugs.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a traditional Chinese medicine composition for treating Alzheimer's disease, its preparation method, and its application in the preparation of drugs for treating or preventing Alzheimer's disease. Background Technology

[0002] Alzheimer's disease (AD) is a neurodegenerative disease characterized by progressive cognitive impairment and memory loss, seriously endangering the physical and mental health of the elderly and imposing a heavy economic burden on families and society. A recent national study report shows that 15.07 million people over the age of 60 in my country suffer from dementia, of which 9.83 million suffer from AD. Acetylcholinesterase inhibitors are commonly used clinically to improve cognitive impairment in AD patients, but these drugs can only alleviate some symptoms and cannot slow or stop the progression of AD. They also have limitations such as easy drug resistance and significant side effects. Traditional Chinese medicine has gained attention in the medical community due to its higher safety profile. In traditional Chinese medicine, AD falls under the categories of "dullness syndrome," "forgetfulness," "literary dementia," "healing syndrome," and "depression syndrome." Zhang Jingyue of the Ming Dynasty first proposed dementia as an independent disease in his book *Jingyue Quanshu*, and elaborated on the etiology of dementia as mainly "liver stagnation, phlegm obstruction, and stomach deficiency." Physicians throughout history have believed that this disease is closely related to kidney deficiency and marrow depletion, liver qi stagnation, and phlegm and blood stasis obstructing the orifices. Therefore, tonifying the kidney and replenishing the marrow, soothing the liver and relieving depression, calming the mind and opening the orifices, strengthening the spleen and replenishing qi, and resolving phlegm and removing blood stasis are the basic methods for treating this disease.

[0003] Tianma Gouteng Decoction, derived from Hu Guangci's *New Interpretation of the Treatment of Miscellaneous Internal Diseases in Traditional Chinese Medicine*, consists of eleven Chinese herbs: Gastrodia elata, Uncaria rhynchophylla, Abalone shell, Achyranthes bidentata, Eucommia ulmoides, Taxillus chinensis, Gardenia jasminoides, Scutellaria baicalensis, Leonurus japonicus, Polygonum multiflorum, and Poria cocos. It has the effects of calming the liver and extinguishing wind, tonifying the liver and kidneys, clearing heat and promoting blood circulation. In this formula, Gastrodia elata and Uncaria rhynchophylla are the principal herbs, calming the liver and extinguishing wind. Abalone shell, being salty, cold, and heavy, functions to calm the liver and subdue yang, and can also clear heat and improve eyesight; combined with the principal herbs, it strengthens the effect of calming the liver and extinguishing wind. Achyranthes bidentata guides blood downwards and can promote blood circulation and diuresis; these are the assistant herbs. Eucommia ulmoides and Taxillus chinensis tonify the liver and kidneys to treat the root cause; Gardenia jasminoides and Scutellaria baicalensis clear the liver and reduce fire to subdue excessive yang; Leonurus japonicus and Achyranthes bidentata promote blood circulation and diuresis, which helps to calm liver yang; Polygonum multiflorum and Poria cocos calm the mind and soothe the nerves; these are the adjuvant herbs. This formula is primarily used to treat headaches, dizziness, and insomnia caused by hypertension, and in recent years it has also been frequently used to treat Alzheimer's disease (AD). Modern pharmacological studies have confirmed that this formula has a good neuroprotective effect. However, in recent years, there have been numerous clinical reports of adverse reactions and organ damage caused by motherwort, and experimental studies have also shown that motherwort can cause toxicity to organs such as the liver and kidneys. In addition, studies have found that long-term use (≥6 months) of gardenia and its compound preparations can cause drug accumulation in the liver and kidneys, resulting in potential liver and kidney toxicity.

[0004] Since Alzheimer's disease (AD) is one of the most common neurodegenerative diseases, patients need to take medication for a long time. Developing safer and more effective traditional Chinese medicine compound formulas for the treatment of AD is an urgent problem to be solved. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, this invention provides a traditional Chinese medicine composition for treating Alzheimer's disease, its preparation method, and its application.

[0006] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating Alzheimer's disease, wherein the effective components of the traditional Chinese medicine composition are made from raw materials comprising the following parts by weight: Gastrodia elata 6-50, Uncaria rhynchophylla 6-50, Haliotis diversicolor 9-60, Rehmannia glutinosa 6-50, Achyranthes bidentata 6-50, Raphanus sativus 3-30, Eucommia ulmoides 3-30, Taxillus chinensis 3-30, Scutellaria baicalensis 3-30, Lycopus lucidus 3-30, Polygonum multiflorum vine 3-30, and Poria cocos 3-30.

[0007] In a second aspect, the present invention provides a method for preparing the traditional Chinese medicine composition described in the first aspect, comprising the following steps:

[0008] (1) Weigh each raw material according to the ratio and soak it in distilled water at room temperature for 12-24 hours;

[0009] (2) Take the soaked abalone shell and decoct it in distilled water with 10-20 times the weight of all the medicinal materials for 20-30 minutes. Then combine it with the other medicinal materials except for Uncaria rhynchophylla and decoct for 45-60 minutes. Add Uncaria rhynchophylla and decoct for 10-15 minutes. Filter and repeat the above extraction steps twice.

[0010] (3) Combine the three filtrates, remove all water under reduced pressure, and freeze dry to obtain a fine powder of the active ingredients of the traditional Chinese medicine composition.

[0011] Thirdly, the present invention provides the application of the traditional Chinese medicine composition described in the first aspect in the preparation of a drug for treating Alzheimer's disease.

[0012] The beneficial effects of this invention include: replacing Leonurus japonicus in the existing "Gastrodia and Uncaria Decoction" formula with Eupatorium fortunei, which has similar efficacy to Leonurus japonicus, removing Gardenia jasminoides, retaining Scutellaria baicalensis, which has similar efficacy, and adding Rehmannia glutinosa, a holy medicine for nourishing essence and marrow, and Raphanus sativus, a medicine for promoting digestion, lowering qi, and resolving phlegm, and optimizing their content, to obtain a traditional Chinese medicine composition for anti-Alzheimer's disease. Animal in vivo experiments have shown that the traditional Chinese medicine composition of this invention can improve cognitive impairment in AD transgenic mice, and has good prospects for development into anti-Alzheimer's disease drugs or functional foods, providing a new option for the treatment of Alzheimer's disease.

[0013] In the preferred technical solution, the following traditional Chinese medicine composition is prepared according to the following weight parts: Gastrodia elata 9 parts, Uncaria rhynchophylla 15 parts, Haliotis diversicolor 18 parts, Rehmannia glutinosa 15 parts, Achyranthes bidentata 12 parts, Raphanus sativus 12 parts, Eucommia ulmoides 9 parts, Taxillus chinensis 9 parts, Scutellaria baicalensis 9 parts, Lycopus lucidus 9 parts, Polygonum multiflorum vine 9 parts, and Poria cocos 9 parts. Using an Alzheimer's disease (AD) transgenic mouse model, the therapeutic effect of the active ingredient (MTGY) in the preferred traditional Chinese medicine composition on cognitive impairment in AD transgenic mice was observed, and its possible mechanism of action was analyzed. By evaluating various behavioral indicators of the mice, it was found that MTGY can significantly improve cognitive impairment in AD transgenic mice. Its mechanism of action may be related to inhibiting inflammatory responses and reducing the deposition of β-amyloid protein (Aβ) in the brain. Currently, there are no literature reports on the efficacy of MTGY in alleviating cognitive impairment or its application in the treatment of Alzheimer's disease. Attached Figure Description

[0014] Figure 1A , 1B 1A and 1C represent the effects of MTGY on the following in 3×Tg-AD mice: (1A) the movement path of mice in the recognition memory test; (1B) the time mice spend exploring new objects; and (1C) the NORT index of mice (n=10). *p<0.05,**p<0.01,***p<0.001,****p<0.0001.

[0015] Figure 2A , 2B 2A and 2C represent the effects of MTGY on the spatial memory of 3×Tg-AD mice: (2A) swimming path during the test period; (2B) escape latency during the training period; and (2C) time spent in the target quadrant during the test period (n=10). * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

[0016] Figure 3 The effect of MTGY on Aβ42 content in brain tissue of 3×Tg-AD mice (n=6); ****p<0.0001.

[0017] Figure 4 This study describes the effects of MTGY on Aβ plaque deposition and astrocytes in the cerebral cortex and hippocampus of 3×Tg-AD mice (n=3). GFAP: glial fibrillary acidic protein, an astrocyte-specific marker; DAPI: 4',6-diamidinyl-2-phenylindole, a nuclear-specific marker; Merge: integration map. * p<0.05, *** p<0.001.

[0018] Figure 5 This study describes the effects of MTGY on amyloid plaques and neuritis in the brain tissue of 3×Tg-AD mice (n=3). *p<0.05, **p<0.01 compared with the 3×Tg-AD (3×Tg) control group. GFAP: glial fibrillary acidic protein, astrocyte-specific marker; DAPI: 4',6-diamidinyl-2-phenylindole, nuclear-specific marker; Merge: integrated map.

[0019] Figure 6 These are the expression protein blot bands of APH-1, p-APP, CTFs, ADAM-10, NEP, and IDE proteins in mouse brain tissue.

[0020] Figure 7A , 7B 7C, 7D, 7E, and 7F represent the effects of MTGY on the expression levels of Aβ plaque deposition-related proteins APH-1, p-APP, CTFs, ADAM-10, NEP, and IDE in the brain tissue of 3×Tg-AD mice (n=6). * p<0.05, ** p<0.01, *** p<0.001. Detailed Implementation

[0021] In this article, "room temperature" and "normal temperature" both refer to a temperature between 23-25℃.

[0022] 1. The therapeutic effect of the traditional Chinese medicine composition of the present invention on Alzheimer's disease.

[0023] 1.1 Experimental Materials

[0024] 1.1.1 Laboratory animals and drugs

[0025] SPF-grade 3×Tg-AD transgenic mice, female, 24-26g, were purchased from the Laboratory Animal Centre of the Chinese University of Hong Kong. The animals were housed separately, with bedding changed every two days, and underwent a 7-day acclimatization period. During this period, they had free access to water and food. The ambient temperature was 22±2℃, the relative humidity was 60%, and the environment featured a 12-hour light and 12-hour darkness cycle.

[0026] 1.1.2 Preparation of test reagents

[0027] Source: Extracted from the laboratory.

[0028] Preparation of the active ingredient (hereinafter referred to as MTGY) in the traditional Chinese medicine composition: All medicinal materials (Gastrodia elata 9g, Uncaria rhynchophylla 15g (added later), Haliotis diversicolor 18g (decocted first), Rehmannia glutinosa 15g, Achyranthes bidentata 12g, Raphanus sativus 12g, Eucommia ulmoides 9g, Taxillus chinensis 9g, Scutellaria baicalensis 9g, Lycopus lucidus 9g, Polygonum multiflorum vine 9g, Poria cocos 9g) were soaked in 10 times the amount of water for 24 hours. Haliotis diversicolor was decocted for 20 minutes first, then combined with the other medicinal materials except Uncaria rhynchophylla and decocted for another 45 minutes. Uncaria rhynchophylla was then added and decocted for another 15 minutes. The mixture was filtered, and the above extraction steps were repeated twice. The filtrates from the three extractions were combined, recovered under reduced pressure, and freeze-dried to obtain the optimized extract of Gastrodia elata and Uncaria rhynchophylla decoction (i.e., the active ingredient, MTGY), with an extraction rate of 26.5% (w / w). The above extract was prepared into a test solution sample with a concentration of 0.5g raw material / mL using distilled water.

[0029] Positive control group test solution (PC): Accurately weigh 20 mg donepezil hydrochloride and dissolve it in 40 mL of double-distilled water to prepare a test solution sample with a concentration of 0.5 mg / mL. After preparation, store the above-mentioned drugs at 4°C for later use.

[0030] 1.1.3 Experimental Reagents

[0031] Dosing reagents: Donepezil hydrochloride, Sigma-Aldrich, USA; Coomassie Brilliant Blue Quantitative Reagent, Bio-Rad, Inc.

[0032] Reagents for frozen section preparation and staining:

[0033] Western blot assay reagents: RIPA lysis buffer, protease inhibitor cocktail, PMSF (100mM), phosphorylated protease inhibitor, protein loading buffer, SDS-PAGE gel preparation reagent, TRIS buffer, glycine, SDS, Tween-20 (Sigma-Aldrich, USA); PVDF membrane (0.22μm) (Bio-Rad); BSA (Thermo Scientific); β-actin (Santa Cruz); filaggrin (FLG) and lobelin (LOR) antibodies (Cell Signal Technology); protein markers, HRP-labeled goat anti-rabbit antibody, HRP-labeled goat anti-mouse antibody (Thermo Scientific).

[0034] 1.1.4 Experimental Apparatus

[0035] 752-P UV spectrophotometer, Shanghai Xianke Instrument Co., Ltd.; SHIMADZU analytical balance, Guangzhou Xiangyi Electromechanical Equipment Co., Ltd.; PL-203 electronic balance, Mettler Toledo Instruments (Shanghai) Co., Ltd.; IKA10 tissue homogenizer, IKA GmbH, Germany; BH22 optical microscope, Olympus GmbH, Japan; FLUO star Optima microplate reader, BMG Labtec GmbH, Germany; ABI-7500 real-time PCR instrument, ABI, USA; FBZ2001-up-p standard reagent pure water system, Qingdao Fullerm Technology Co., Ltd.; benchtop high-speed refrigerated centrifuge, Heal Force; DYY-6C electrophoresis apparatus, Beijing Liuyi Instrument Factory.

[0036] 1.2 Animal grouping and administration

[0037] Six-month-old female wild-type mice served as the normal control group (WT), and age-matched 3×Tg-AD transgenic mice were randomly divided into three groups: a 3×Tg-AD model control group, a positive control group, and MTGY groups (high-dose, medium-dose, and low-dose). Ten animals were assigned to each group. The normal control and model control groups received the same volume of solvent; the positive control group received donepezil (5 mg / kg), and the MTGY groups received low-dose MTGY (5 g / kg), medium-dose MTGY (10 g / kg), and high-dose MTGY (20 g / kg). The 3×Tg-AD transgenic mice were administered the drug via gavage once daily for four months, starting at six months of age. At ten months of age, the mice underwent a new object recognition test and a water maze test to investigate the effects of MTGY on cognitive impairment in the 3×Tg-AD transgenic mice.

[0038] 1.3 Novel Object Recognition Test (NORT) and Evaluation in Mice

[0039] Experimental equipment includes a square open field experimental chamber and camera equipment; three types of objects, A, B, and C, where objects A and B are exactly the same, and object C is very different from objects A and B.

[0040] The experiment consisted of three phases. Phase 1, the adaptation period, involved mice freely moving within the experimental setup (without objects) for 10 minutes. Phase 2, the familiarization period, involved placing two identical objects (AB, ensuring the objects were odorless and could not be moved) into the setup. The mice were placed with their backs to the objects from an equal distance, and explored the setup for 5 minutes. Phase 3, the testing period, involved replacing one of the two objects with a new object (AC or BC). The mice were again placed with their backs to the object from an equal distance. A camera and software recorded the time the mice spent exploring each object (exploration was defined as the mice's mouths or noses touching the object or approaching it within approximately 2-3 cm). The number of times and the duration of each object's exploration were measured within 5 minutes.

[0041] 1.4 Mouse Water Maze Test (MWMT)

[0042] Experimental equipment includes a circular swimming pool and camera equipment. A circular pool (120 cm in diameter and 30 cm in height) is placed in the behavioral laboratory, maintaining a water temperature of 25°C. The pool maze is divided into four equal quadrants, and four different shapes are placed on the pool walls to mark the different quadrants. A hidden platform (5 cm in diameter) is placed 0.5 cm below the water surface at the midpoint of one of the quadrants.

[0043] The MWMT exercise lasted five days. The first four days were a spatial training period, during which mice were trained to find hidden platforms in a water maze. On day 1 of spatial training, each mouse swam three times from different directions, each time for a maximum of 60 seconds. Then, each mouse was guided to a platform and held there for 30 seconds. From day 2 to day 4, each mouse was trained to swim for a maximum of 60 seconds each time. Escape latency was recorded during the training. On day 5, the hidden platforms were removed, and the mice were allowed to swim freely in the water for 60 seconds without the hidden platforms. Swimming parameters were recorded and analyzed using the video tracking software SuperMaze V2.0 (Shanghai Xinruan, China).

[0044] 1.5. Serum and skin sampling

[0045] After the experiment, blood was collected from mice via orbital blood sampling. The blood samples were allowed to stand at room temperature for 2 hours, then centrifuged at 3000 rpm for 15 minutes, and the supernatant was separated to obtain serum samples. Mice were euthanized by cervical dislocation after blood collection, and brain tissue was immediately removed. The cortex and hippocampus were extracted and frozen at -80°C for subsequent experiments. Four mice were randomly selected from each group for cardiac perfusion, and the brains were then fixed in 4% (v / v) paraformaldehyde for 24 hours for preparing pathological tissue sections.

[0046] 1.6. Western Blot detection of related protein expression

[0047] 1.6.1 Total Protein Extraction

[0048] Wash tissue blocks 2-3 times with cold PBS to remove blood contamination, cut into small pieces, and place in a homogenizer. Add 10 times the tissue volume of lysis buffer (add protease inhibitor a few minutes before use) and homogenize thoroughly on ice. Transfer the homogenate to a 1.5 mL centrifuge tube and vortex. Incubate on ice for 30 min, repeatedly pipetting during this time to ensure complete cell lysis. Then centrifuge at 12000g for 10 min and collect the supernatant, which is the total protein solution.

[0049] 1.6.2 Protein concentration determination: Protein concentration was determined using the BCA protein concentration kit.

[0050] 1.6.3 SDS-PAGE electrophoresis

[0051] Prepare the SDS-PAGE gel according to the instructions of the SDS-PAGE gel preparation kit. After adding sufficient electrophoresis buffer, load the sample into the electrophoresis wells for electrophoresis (stacking gel voltage 70V, 1h; separating gel voltage 120V, 30min). Stop electrophoresis when bromophenol blue just runs out, and then proceed with the membrane transfer.

[0052] 1.6.4 Transfer of film

[0053] Prepare six 7×9cm filter papers and one appropriately sized PVDF membrane. Activate the PVDF membrane with methanol for 15 seconds before use. Place the transfer clamp, two sponge pads, filter paper, and the activated PVDF membrane in a bowl containing transfer buffer. Open the clamp so the black side is horizontal. Place the sponge pads and three layers of filter paper on the mats. Carefully peel off the separating gel and place it on the filter paper. Place the membrane on the gel, removing any air bubbles. Then cover the membrane with three more layers of filter paper, removing any air bubbles. Finally, cover with the remaining sponge pad. Transfer conditions (wet transfer): fast transfer, 250mA constant current transfer for 45 minutes.

[0054] 1.6.5 Immune Response

[0055] The transferred membrane was blocked for 2 hours on a decolorizing shaker at room temperature with 5% (w / v) skim milk (prepared with 0.5% (w / v) TBST). The primary antibody was diluted (5% (w / v) skim milk dissolved in TBST; phosphorylated proteins were treated with 5% (w / v) BSA dissolved in TBST), and incubated overnight at 4°C. The membrane was then washed three times with TBST on a decolorizing shaker at room temperature for 5 minutes each time. The secondary antibody was diluted 2000-fold with TBST and incubated for 2 hours at room temperature. The membrane was then washed three times with TBST on a decolorizing shaker at room temperature for 5 minutes each time.

[0056] 1.6.6 Chemiluminescence

[0057] Mix equal volumes of ECLA and ECLB reagents in a centrifuge tube. Place the PVDF membrane, protein side up, into the mixture for 1-2 minutes. Remove any remaining liquid, wrap the membrane, and place it in a dark box for exposure. Finally, develop and fix the membrane using developing and fixing reagents, adjusting the exposure conditions according to different light intensities.

[0058] 1.7 Pathological and Histological Observation

[0059] Brain tissue was fixed in 4% (v / v) paraformaldehyde for 24 h, then transferred to 30% (w / v) sucrose solution for dehydration, followed by OCT embedding and then frozen sectioning. Sections were blocked in 5% (w / v) BSA at room temperature for 20 min. Sections were incubated overnight at 4°C with primary antibodies (anti-Iba-1 antibody (ab178846, Abcam), anti-GFAP antibody (ab68428, Abcam), and anti-Aβ antibody (A5213, Sigma). The next day, sections were incubated with donkey anti-rabbit IgG H&L (Alexa). 488, 647 (Abcam) secondary antibody was incubated at room temperature in the dark for 1 hour. Finally, it was incubated with ProLong (containing DAPI, Invitrogen) TM Diamond antifade mountant was used for mounting, which also allows for staining of cell nuclei. Images were taken using a Zeiss AxioScope A1 microscope.

[0060] 1.8 Determination of Aβ42 content in brain tissue

[0061] The procedure was performed according to the ELISA kit instructions. The brain tissue was accurately weighed, and after adding cell lysis buffer, a tissue homogenate was prepared using a high-speed homogenizer. The homogenate was centrifuged at 12,000×g and 4℃ for 15 min, and the supernatant was separated to determine the content of Aβ42 in the brain tissue.

[0062] 1.9 Statistical Analysis

[0063] All data are expressed as mean ± standard error (x ± SEM). GraphPad Prism 9 software was used for graphing and statistical analysis. One-way ANOVA was used to compare differences between groups, and Dunnett's method was used for pairwise multiple comparisons between groups. A p < 0.05 was considered statistically significant, *p < 0.05, **p < 0.01.

[0064] 2. Experimental results of the therapeutic effect of the traditional Chinese medicine composition of the present invention on Alzheimer's disease.

[0065] 2.1 Effects of MTGY on cognitive impairment in 3×Tg-AD transgenic mice

[0066] To assess whether MTGY treatment affects cognitive impairment in 3×Tg-AD transgenic mice, this invention used two behavioral tests, including NORT (as described in 1.3 above) and MWMT (as described in 1.4 above), and the results are as follows:

[0067] 2.1.1 Effects of MTGY on recognition and memory in 3×Tg-AD transgenic mice

[0068] like Figure 1A , 1B As shown in Figure 1C, the activity pathways of the mice revealed that wild-type WT mice, MTGY-treated mice, and donepezil mice (positive control group) may have preferred exploring new objects, while 3×Tg-AD transgenic mice preferred to spend more time exploring familiar objects. By comparing the NORT index, 3×Tg-AD transgenic mice showed reduced recognition memory compared to wild-type mice, while MTGY (5 g / kg, 10 g / kg, and 20 g / kg) significantly improved cognitive impairment in 3×Tg-AD transgenic mice (p<0.01).

[0069] 2.1.3. Effects of MTGY on spatial recognition memory in 3×Tg-AD transgenic mice.

[0070] like Figure 2A , 2B As shown in 2C, in the same spatial memory and learning function tests conducted in the MWMT, all mice exhibited shortened platform escape latency during training. There were differences in mean latency between the MTGY (5, 10, and 20 g / kg) treatment groups and 3×Tg-AD transgenic mice by day 4 of training. During the test period, 3×Tg-AD transgenic mice spent significantly less time in the target quadrant than the wild-type WT group (p<0.01), while MTGY (5, 10, and 20 g / kg) treatment effectively improved the time spent in the target quadrant in 3×Tg-AD transgenic mice (both p<0.01). These results indicate that MTGY can improve learning and memory impairments in 3×Tg-AD transgenic mice.

[0071] 2.2. Effects of MTGY on Aβ plaque content and related protein expression in the brain tissue of 3×Tg-AD transgenic mice.

[0072] 2.2.1 Effect of MTGY on Aβ42 content in brain tissue of 3×Tg-AD transgenic mice

[0073] like Figure 3As shown, this paper illustrates the effect of MTGY on the Aβ42 content in the brain of 3×Tg-AD transgenic mice. The results indicate that the Aβ42 content in the brain tissue of 3×Tg-AD transgenic mice was significantly reduced in the MTGY treatment group.

[0074] 2.2.2 Effects of MTGY on amyloid plaques and neuritis in the brain tissue of 3×Tg-AD transgenic mice

[0075] Frozen sections of mouse brain were used for antigen retrieval with sodium citrate buffer (pH 6.0). After blocking for 30 min, primary antibodies against Aβ1-42, 4G8, GFAP, and Iba-1 (1:1000, purchased from CST) were added, and the sections were incubated overnight at 4°C. The sections were then washed with PBS for 5 min × 3 times. Secondary antibodies (1:1000, Invitrogen, USA) were added, and the sections were incubated at 37°C for 1 h. The sections were then washed with PBS for 5 min × 3 times. DAPI staining was performed at room temperature for 5 min, followed by PBS washing for 5 min × 3 times before mounting. Fluorescence microscopy was used for observation and image acquisition, and ImageJ was used to analyze the expression levels of Aβ, GFAP, and Iba-1 positive proteins.

[0076] like Figure 4 The effects of MTGY on Aβ plaque deposition and astrocytes in the cerebral cortex (A) and hippocampus (B) of 3×Tg-AD mice (C) (n=3). In the figure, GFAP: glial fibrillary acidic protein, astrocyte-specific marker; DAPI: 4',6-diamidinyl-2-phenylindole, nuclear-specific marker; Merge: integrated diagram. * p<0.05, *** p<0.001. Figure 5 This study investigated the effects of MTGY on amyloid plaques and neuritis in the brain tissue of 3×Tg-AD mice (n=3). *p<0.05, **p<0.01 compared with the 3×Tg-AD (3×Tg) control group. Results showed that MTGY (5g / kg or 20g / kg) significantly reduced the expression levels of Aβ, GFAP, and Iba-1 positive proteins in the hippocampus and cortex of 3×Tg-AD mouse brain tissue.

[0077] 2.2.3. Effect of MTGY on the expression level of Aβ-related protein in the brain tissue of 3×Tg-AD transgenic mice

[0078] like Figure 6 The image shows the expression bands of proteins APH-1, p-APP, CTFs, ADAM-10, NEP, and IDE in mouse brain tissue. Figure 7A , 7B7C, 7D, 7E, and 7F represent the effects of MTGY on the expression levels of Aβ plaque deposition-related proteins APH-1, p-APP, CTFs, ADAM-10, NEP, and IDE in the brain tissue of transgenic mice in 3×Tg-AD mice (n=6). * p<0.05, ** p<0.01, *** p<0.001. The results showed that the expression levels of proteins related to Aβ deposition, such as p-APP, APH-1, CTFs, and ADAM-10, were increased in the brain tissue of 3×Tg-AD transgenic mice. After MTGY treatment, the expression of these proteins was significantly reduced (p<0.05 compared with the model group). Conversely, the expression levels of proteins related to Aβ hydrolysis, such as IDE and NEP, were decreased in the brain tissue of 3×Tg-AD transgenic mice. After MTGY treatment, the expression of these proteins was significantly increased, indicating that MTGY has an inhibitory effect on the protein pathways related to Aβ plaque deposition in the brain of 3×Tg-AD transgenic mice.

[0079] 3. Discussion

[0080] Alzheimer's disease (AD) is a progressive neurodegenerative disease, primarily characterized by cognitive impairment. AD symptoms worsen over time. Generally, AD progression is defined as three stages: mild, moderate, and severe. The mild stage is the early stage of AD, typically characterized by cognitive impairment symptoms that do not significantly impact daily life, such as forgetting familiar words or the location of everyday objects, and short-term memory loss. In addition, patients in the mild stage may experience neuropsychiatric symptoms such as anxiety, apathy, irritability, and depression. Alleviating cognitive impairment symptoms in the mild stage of AD and slowing disease progression to reduce its impact on the patient's quality of life are crucial aspects of AD treatment.

[0081] Currently, the two generally accepted hypotheses related to the pathogenesis of Alzheimer's disease (AD) are: senile plaques formed by the deposition of β-amyloid protein (Aβ) and ganglion intractability caused by excessive phosphorylation of tau protein. Previous studies have reported that Aβ binds to receptors on the surface of nerve cells and alters synaptic structure, thereby disrupting neurotransmitter transmission between neurons and ultimately leading to neuronal death. Therefore, based on the Aβ hypothesis, reducing the phosphorylation of β-amyloid precursor protein (APP) and increasing APP hydrolysis can reduce Aβ protein deposition in the brain, thereby slowing the progression of AD and improving the cognitive impairment caused by AD.

[0082] This invention experimentally confirmed that MTGY can significantly reduce the content of β-amyloid protein 1-42 in brain tissue, inhibit the activation of microglia and astrocytes in the brain, and downregulate the expression levels of proteins related to β-amyloid formation in the brain. This invention uses an in vivo model of cognitive impairment in 3×Tg-AD transgenic mice as a validation model simulating human Alzheimer's disease. 3×Tg-AD transgenic mice simultaneously overexpress three human mutant genes, including APP Swe, tau P301L, and 1Lfa PSEN1 (Jackson Laboratory Stock No.: 34830-JAX). When 3×Tg-AD mice reach 6 months of age, significant neuroinflammation, Aβ deposition, and neurofibrillary tangles appear in the hippocampus, accompanied by AD pathological features such as cognitive impairment, which worsen with age. Therefore, this model can more comprehensively and accurately simulate the pathological features of AD, and is particularly suitable for preclinical anti-AD drug research. In the study of this invention, a decline in cognitive ability was observed in 3×Tg-AD mice. Mice treated with MTGY showed significant improvements in spatial memory and cognitive abilities, suggesting that MTGY has a therapeutic effect on cognitive impairment in 3×Tg-AD mice.

[0083] This invention utilizes molecular biology techniques such as immunohistochemistry and Western blot to explore the effects of MTGY on Aβ metabolism, APP hydrolysis, and APP phosphorylation in the brains of transgenic mice, thereby investigating the anti-AD mechanism of MTGY. The research results will not only benefit the development and utilization of MTGY but also provide a basis for elucidating its anti-AD mechanism and offer new insights into the anti-AD mechanism of traditional Chinese medicine compound formulas.

[0084] The advantages of this invention include: (1) This invention found that MTGY has a therapeutic effect on cognitive impairment in 3×Tg-AD mice. After four months of treatment with MTGY, cognitive impairment in mice can be significantly improved, and the content of Aβ plaques in brain tissue can be reduced. At the same time, MTGY inhibits the expression of proteins related to Aβ plaque formation. (2) The traditional Chinese medicines in this invention are all derived from commonly used traditional Chinese medicinal materials. The effective components of the traditional Chinese medicine composition can effectively treat Alzheimer's disease and can be used to prepare drugs or functional foods for the treatment of Alzheimer's disease. (3) This invention is based on the perspective of anti-Alzheimer's disease, elucidating the pharmacodynamic material basis and mechanism of action of MTGY, providing new options for the future development of drugs and functional products to combat Alzheimer's disease.

[0085] The traditional Chinese medicine composition of the present invention also includes medically acceptable excipients, which can be formulated with the active ingredients into any pharmaceutically acceptable oral dosage form, such as granules, capsules, concentrated pills, oral liquids, or tablets. These oral dosage forms are illustrated below through some examples.

[0086] Example 1 (Tablet)

[0087] Take 1000g of this MTGY (hereinafter also referred to as "the extract"), add 480g of lactose and 500g of starch, mix well, use 300g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 20g of magnesium stearate, mix well, and compress into 10,000 tablets, each containing 100mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0088] Example 2 (tablet)

[0089] Take 800g of this MTGY, add 480g of lactose and 700g of starch, mix well, use 300g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 20g of magnesium stearate, mix well, and compress into 10,000 tablets, each containing 80mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0090] Example 3 (tablet)

[0091] Take 500g of this MTGY, add 480g of lactose and 1000g of starch, mix well, use 300g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 20g of magnesium stearate, mix well, and compress into 10,000 tablets, each containing 50mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0092] Example 4 (tablet)

[0093] Take 1000g of this MTGY, add 2984g of starch and mix well. Use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, compress into 10,000 tablets, each containing 100mg of the extract, with a net weight of 0.4g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0094] Example 5 (tablet)

[0095] Take 800g of this MTGY, add 3184g of starch and mix well. Use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, compress into 10,000 tablets, each containing 80mg of the extract, with a net weight of 0.4g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0096] Example 6 (tablet)

[0097] Take 500g of this MTGY, add 3484g of starch and mix well. Use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, compress into 10,000 tablets, each containing 50mg of the extract, with a net weight of 0.4g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0098] Example 7 (Tablet)

[0099] Take 1000g of this MTGY, add 594g of lactose and 390g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, and compress into 10,000 tablets, each containing 100mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0100] Example 8 (tablet)

[0101] Take 800g of this MTGY, add 614g of lactose and mix well, add 570g of starch and mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, compress into 10,000 tablets, each containing 80mg of extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0102] Example 9 (Tablet)

[0103] Take 500g of MTGY, add 574g of lactose and mix well. Use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, compress into 10,000 tablets, each containing 50mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of atopic dermatitis. Symptoms include: skin erythema, edema, peeling, or lichenification.

[0104] Example 10 (Tablet)

[0105] Take 1000g of this MTGY, add 480g of dextrin and 1114g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, and compress into 10,000 tablets, each containing 100mg of the extract, with a net weight of 0.1g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0106] Example 11 (Tablet)

[0107] Take 800g of MTGY, add 480g of dextrin and 1134g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, and compress into 10,000 tablets, each containing 80mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0108] Example 12 (Tablet)

[0109] Take 500 g of MTGY, add 480 g of dextrin and 1094 g of starch, mix well, use 350 g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16 g of magnesium stearate, mix well, and compress into 10,000 tablets, each containing 50 mg of the extract, with a net weight of 0.2 g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0110] Example 13 (Tablet)

[0111] Take 1000g of this MTGY, add 480g of lactose and 1114g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of talc powder, mix well, and compress into 10,000 tablets, each containing 100mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0112] Example 14 (Tablet)

[0113] Take 800g of this MTGY, add 480g of lactose and 1134g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of talc powder, mix well, and compress into 10,000 tablets, each containing 80mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of atopic dermatitis. Symptoms include: skin erythema, edema, peeling, or lichenification.

[0114] Example 15 (Tablet)

[0115] Take 500g of MTGY, add 480g of lactose and 1094g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of talc powder, mix well, and compress into 10,000 tablets, each containing 50mg of extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0116] Example 16 (Tablet)

[0117] Take 1000g of this MTGY, add 480g of dextrin and 504g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of talc powder, mix well, and compress into 10,000 tablets, each containing 100mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0118] Example 17 (Tablet)

[0119] Take 800g of this MTGY, add 480g of dextrin and 704g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of talc powder, mix well, and compress into 10,000 tablets, each containing 80mg of the extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0120] Example 18 (Tablet)

[0121] Take 500g of MTGY, add 480g of dextrin and 1004g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of talc powder, mix well, and compress into 10,000 tablets, each containing 50mg of extract, with a net weight of 0.2g per tablet. For oral administration, take 3 tablets twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0122] Example 19 (Capsule)

[0123] Take 1000g of this MTGY, add 980g of lactose and 1004g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, fill into No. 1 capsules to make 10,000 capsules, each capsule contains 100mg of extract, and each capsule has a net weight of 0.3g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0124] Example 20 (Capsule)

[0125] Take 800g of this MTGY, add 980g of lactose and 1204g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, fill into No. 1 capsules to make 10,000 capsules, each capsule contains 80mg of extract, each capsule has a net weight of 0.3g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0126] Example 21 (Capsule)

[0127] Take 500g of this MTGY, add 980g of lactose and 1504g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, fill into No. 1 capsules to make 10,000 capsules, each capsule containing 50mg of extract, each tablet net weight 0.3g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0128] Example 22 (Capsule)

[0129] Take 1000g of this MTGY, add 2984g of starch and mix well. Use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, fill into No. 1 capsules to make 10,000 capsules, each with a net weight of 0.4g and containing 100mg of extract. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include forgetfulness and irritability.

[0130] Example 23 (Capsule Formulation)

[0131] Take 800g of this MTGY, add 3184g of starch and mix well. Use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, fill into No. 1 capsules to make 10,000 capsules, each with a net weight of 0.4g and containing 80mg of extract. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include forgetfulness and irritability.

[0132] Example 24 (Capsule)

[0133] Take 500g of this MTGY, add 3484g of starch and mix well. Use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate and mix well, fill into No. 1 capsules to make 100,000 capsules, each with a net weight of 0.4g and containing 50mg of extract. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include forgetfulness and irritability.

[0134] Example 25 (Capsule)

[0135] Take 1000g of this MTGY, add 1480g of lactose and 2504g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, fill into size 0 capsules to make 10,000 capsules, each capsule contains 100mg of extract, each capsule has a net weight of 0.5g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0136] Example 26 (Capsule)

[0137] Take 800g of this MTGY, add 1480g of lactose and 2704g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, fill into size 0 capsules to make 10,000 capsules, each capsule contains 80mg of extract, and each capsule has a net weight of 0.5g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0138] Example 27 (Capsule)

[0139] Take 600g of this MTGY, add 1480g of lactose and 2904g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, fill into size 0 capsules to make 10,000 capsules, each capsule contains 50mg of extract, and each capsule has a net weight of 0.5g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0140] Example 28 (Capsule)

[0141] Take 500g of this MTGY, add 1480g of lactose and 3004g of starch, mix well, use 350g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16g of magnesium stearate, mix well, fill into No. 2 capsules to make 10,000 capsules, each capsule contains 50mg of extract, and each capsule has a net weight of 0.2g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0142] Example 29 (Capsule)

[0143] Take 400 g of MTGY, add 480 g of lactose and 1104 g of starch, mix well, use 350 g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16 g of magnesium stearate, mix well, fill into No. 2 capsules to make 10,000 capsules, each capsule contains 40 mg of extract, and each capsule has a net weight of 0.2 g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0144] Example 30 (Capsule)

[0145] Take 250 g of MTGY, add 580 g of lactose and 1154 g of starch, mix well, use 350 g of 7% (w / v) starch slurry as a binder, wet granulate, dry, add 16 g of magnesium stearate, mix well, fill into No. 2 capsules to make 10,000 capsules, each capsule contains 25 mg of extract, and each capsule has a net weight of 0.2 g. For oral administration, take 3 capsules twice daily for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0146] Example 31 (Oral Liquid)

[0147] Take 50g of MTGY and add 15% (v / v) ethanol, stirring thoroughly. Then add appropriate excipients (such as 0.1% (w / v) sorbic acid as an antibacterial agent, 0.1% (w / v) citric acid as an antioxidant, and 0.1% (w / v) mannitol as a flavoring agent), dissolve evenly, filter to clarify, and fill the contents into ampoules or easy-open bottles, then sterilize. The product is an oral solution containing 166mg of extract per bottle, with a net content of 10mL per bottle. For oral administration, once daily, one bottle each time, for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0148] Example 32 (Oral Liquid)

[0149] Take 100g of MTGY and add 15% (v / v) ethanol, stirring thoroughly. Then add appropriate excipients (such as 0.1% (w / v) sorbic acid as an antibacterial agent, 0.1% (w / v) citric acid as an antioxidant, and 0.1% (w / v) mannitol as a flavoring agent), dissolve evenly, filter to clarify, and fill the contents into ampoules or easy-open bottles, then sterilize. The product is an oral solution containing 300mg of extract per bottle, with a net content of 10mL per bottle. For oral administration, once daily, one bottle each time, for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0150] Example 33 (Oral Liquid)

[0151] Take 200g of MTGY, add 15% (v / v) ethanol, and stir thoroughly. Then add appropriate excipients (such as 0.1% (w / v) sorbic acid as an antibacterial agent, 0.1% (w / v) citric acid as an antioxidant, and 0.1% (w / v) mannitol as a flavoring agent), dissolve evenly, filter to clarify, and fill the contents into ampoules or easy-open bottles, then sterilize. The specification is an oral solution containing 600mg of extract per bottle, with a net content of 10mL per bottle. For oral administration, once daily, one bottle each time, for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0152] Example 34 (Oral Liquid)

[0153] Take 300g of MTGY, add 15% (v / v) ethanol, and stir thoroughly. Then add appropriate excipients (such as 0.1% (w / v) sorbic acid as an antibacterial agent, 0.1% (w / v) citric acid as an antioxidant, and 0.1% (w / v) mannitol as a flavoring agent), dissolve evenly, filter to clarify, and fill the contents into ampoules or easy-open bottles, then sterilize. The specification is an oral solution containing 900mg of extract per bottle, with a net content of 10mL per bottle. For oral administration, once daily, one bottle each time, for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0154] Example 35 (Oral Liquid)

[0155] Take 400 g of MTGY, add 15% (v / v) ethanol, and stir thoroughly. Then add appropriate excipients (such as 0.1% (w / v) sorbic acid as an antibacterial agent, 0.1% (w / v) citric acid as an antioxidant, and 0.1% (w / v) mannitol as a flavoring agent), dissolve evenly, filter to clarify, and fill the contents into ampoules or easy-open bottles, then sterilize. The specification is an oral solution containing 1333 mg of extract per bottle, with a net content of 10 mL per bottle. For oral administration, once daily, one bottle each time, for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0156] Example 36 (Oral Liquid)

[0157] Take 500g of MTGY, add 15% (v / v) ethanol, and stir thoroughly. Then add appropriate excipients (such as 0.1% (w / v) sorbic acid as an antibacterial agent, 0.1% (w / v) citric acid as an antioxidant, and 0.1% (w / v) mannitol as a flavoring agent), dissolve evenly, filter to clarify, and fill the contents into ampoules or easy-open bottles, then sterilize. The specification is an oral solution containing 1666mg of extract per bottle, with a net content of 10mL per bottle. For oral administration, once daily, one bottle each time, for the treatment of Alzheimer's disease and cognitive impairment. Symptoms include: forgetfulness and irritability.

[0158] The background section of this invention may include background information about the problems or environment in which the invention is being developed, and is not necessarily a description of prior art. Therefore, the content included in the background section does not constitute an admission of prior art by the applicant.

[0159] The above description provides a further detailed explanation of the present invention in conjunction with specific / preferred embodiments, and it should not be construed that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, various substitutions or modifications can be made to these described embodiments without departing from the concept of the present invention, and all such substitutions or modifications should be considered within the scope of protection of the present invention. In the description of this specification, the reference to terms such as "an embodiment," "some embodiments," "preferred embodiment," "example," "specific example," or "some examples," etc., indicates that the specific features, structures, materials, or characteristics described in connection with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any suitable manner in one or more embodiments or examples. Without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification and the features of different embodiments or examples. Although the embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions, and modifications can be made herein without departing from the scope of protection of the patent application.

Claims

1. A traditional Chinese medicine composition for treating Alzheimer's disease, characterized in that, The active ingredients in the traditional Chinese medicine composition are made from the following raw materials in parts by weight: Gastrodia elata 6-50g, Uncaria rhynchophylla 6-50g, Abalone shell 9-60g, Rehmannia glutinosa 6-50g, Achyranthes bidentata 6-50g, Radish seed 3-30g, Eucommia ulmoides 3-30g, Taxillus chinensis 3-30g, Scutellaria baicalensis 3-30g, Lycopus lucidus 3-30g, Polygonum multiflorum vine 3-30g, Poria cocos 3-30g.

2. The traditional Chinese medicine composition for treating Alzheimer's disease according to claim 1, characterized in that, The weight proportions of each raw material in the effective ingredients are as follows: Gastrodia elata 9, Uncaria rhynchophylla 15, Haliotis diversicolor 18, Rehmannia glutinosa 15, Achyranthes bidentata 12, Raphanus sativus 12, Eucommia ulmoides 9, Taxillus chinensis 9, Scutellaria baicalensis 9, Lycopus lucidus 9, Polygonum multiflorum vine 9, and Poria cocos 9.

3. The traditional Chinese medicine composition for treating Alzheimer's disease according to claim 1, characterized in that, The traditional Chinese medicine composition also includes medically acceptable excipients.

4. The traditional Chinese medicine composition for treating Alzheimer's disease according to any one of claims 1-3, characterized in that, The traditional Chinese medicine composition is any pharmaceutically acceptable oral dosage form.

5. The traditional Chinese medicine composition for treating Alzheimer's disease according to claim 4, characterized in that, The traditional Chinese medicine composition is in the form of granules, capsules, concentrated pills, oral liquids, or tablets.

6. The traditional Chinese medicine composition for treating Alzheimer's disease as described in any one of claims 1-3, characterized in that: The active ingredients in the traditional Chinese medicine composition are prepared from each raw material through the following process: Abalone shell is decocted for a first predetermined time, then combined with other medicinal materials except Uncaria rhynchophylla and decocted for a second predetermined time, and finally Uncaria rhynchophylla is added and decocted for a third predetermined time. After repeating this process multiple times, the filtrates are filtered and combined, and the water in the filtrate is removed and the mixture is dried.

7. A method for preparing a traditional Chinese medicine composition for treating Alzheimer's disease according to any one of claims 1-6, characterized in that, Includes the following steps: (1) Weigh each raw material according to the ratio and soak it in distilled water at room temperature for 12-24 hours; (2) Take the soaked abalone shell and decoct it in distilled water with 10-20 times the weight of all the medicinal materials for 20-30 minutes. Then combine it with the other medicinal materials except for Uncaria rhynchophylla and decoct for 45-60 minutes. Add Uncaria rhynchophylla and decoct for 10-15 minutes. Filter and repeat the above extraction steps twice. (3) Combine the three filtrates, remove all water under reduced pressure, and freeze dry to obtain a fine powder of the active ingredients of the traditional Chinese medicine composition.

8. The preparation method according to claim 7, characterized in that, It also includes the following steps: (4) The fine powder and a predetermined amount of medically acceptable excipients are prepared into any pharmaceutically acceptable oral dosage form.

9. The use of the traditional Chinese medicine composition according to any one of claims 1-6 in the preparation of a medicament for treating Alzheimer's disease.

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