Application of sarsasapogenin structure derivative in preparation of medicine for treating Alzheimer's disease

Abstract

The invention relates to application of a sarsasapogenin structure derivative in preparation of a medicine for treating Alzheimer's disease. The structural formula of the sarsasapogenin structure derivative is shown in the specification. According to the application of the sarsasapogenin structure derivative in preparation of the medicine for treating the Alzheimer's disease, a double-transgenic Alzheimer's disease model mouse with APP protein overexpression and PS1 premature senile gene mutation can prove that the small molecule compound has a good effect of treating the Alzheimer's disease; the compound has a potential huge value for treating Alzheimer's disease, and also has important scientific and commercial application values.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to Alzheimer's disease, specifically the use of a scutellarin saponin derivative in the preparation of drugs for treating Alzheimer's disease. Background Technology

[0002] Anemarrhena asphodeloides Bunge is a common traditional Chinese medicine. It is the dried tuber of the plant, belonging to the Liliaceae family. It is used to relieve thirst and internal heat, dispel pathogenic factors, and reduce edema of the limbs, and is a commonly used yin-nourishing herb. Its extracts have been shown to possess diuretic, anti-diabetic, anti-platelet aggregation, antifungal, and metabolic regulatory activities, and also exhibit inhibitory effects on cyclic adenosine monophosphate phosphodiesterase. The main chemical components of the extract include steroidal saponins, biphenylpyranones, polysaccharides, and lignins. Among these, steroidal saponins include anemarrhena saponins AI, A-II, A-III, A-IV, BI, B-II, and B-III, as well as malcosanoside 3-O-β-D-glucopyranosyl(1→2)-β-D-galactopyranoside B, degalactoside, F-glycine saponin, and isosmilax saponin. In addition, it also contains Anemarrhena asphodeloides polysaccharides A / B / C / D, cis-cylindrical resin phenol, monomethyl-cis-cylindrical resin phenol, oxidized-cis-cylindrical resin phenol, 2,6,4'-trihydroxy-4-methoxybenzophenone, p-hydroxyphenyl crotonol, vinyl pentadecanoate, β-sitosterol, mangiferin, nicotinic acid, nicotinamide and pantothenic acid, etc.

[0003] Alzheimer's disease (AD) ranks 7th among the top 10 causes of death published by the WHO. AD is the most common type of dementia and the most expensive, deadliest, and socially burdensome neurodegenerative disease of this century [2]. The incidence of AD increases with age. In my country, the overall prevalence of AD is 5.3%, which is a huge challenge for my country, where the aging problem is becoming increasingly serious. The clinical features of AD are amyloid plaques and intracellular neurofibrillary tangles, leading to neuronal dysfunction and cell death. Currently, the FDA has approved only two types of drugs for the treatment of AD: acetylcholinesterase inhibitors, such as donepezil, galantamine, and rivastigmine; and NMDA antagonists, such as memantine. However, these drugs cannot slow down the course of AD, prevent AD, or cure AD. Their main function is to control symptoms and slow down the rate of decline in patients' thinking and memory within one year of taking them, thereby reducing the workload of caregivers. In addition, the side effects of the drugs are more serious, which also brings new suffering to AD patients.

[0004] With advancements in medicine, some terminal illnesses, including cancer, have seen the light of hope for a cure. However, Alzheimer's disease (AD) remains incurable. The most researched targets are amyloid-β (Aβ) and tau protein, which are key to understanding the disease's etiology. To date, all antibody drugs targeting Aβ have failed. While most of these drugs can reduce Aβ deposition in the brain, they offer little improvement in cognition. The tau protein hypothesis, on par with the Aβ hypothesis, is also a hot topic in AD treatment, but its clinical trials have either been suspended or declared failures, equally disappointing. Summary of the Invention

[0005] The purpose of this invention is to overcome the defects in the prior art described above. This invention provides the use of a scutellarin saponin derivative in the preparation of a drug for treating Alzheimer's disease.

[0006] To achieve the above objectives, the present invention provides the use of a squalene saponin derivative in the preparation of a drug for treating Alzheimer's disease, characterized in that the structural formula of the squalene saponin derivative is shown below: .

[0007] The use of the sarcosine saponin derivative of the present invention in the preparation of drugs for treating Alzheimer's disease can be demonstrated by using double transgenic Alzheimer's disease model mice with APP protein overexpression and PS1 progerin gene mutation. This small molecule compound of the present invention has a good therapeutic effect on Alzheimer's disease and has great potential value for the treatment of Alzheimer's disease. At the same time, it has important scientific and commercial application value. Attached Figure Description

[0008] Figure 1 The results of the water maze test before drug administration are shown. Figure 1 A represents the incubation period for animals to find a platform during their training phase; Figure 1 B represents the proportion of time spent in the target quadrant during the animal testing period; Figure 1 C represents the number of times a platform was found during the animal testing period.

[0009] Figure 2 The image shows the results of the water maze test after 6 months of drug administration. Figure 2 A represents the incubation period during which the animal finds its platform during the training phase; Figure 2 B represents the percentage of time spent in the target quadrant during the animal testing period (%). Figure 2 C represents the number of times (N) a platform was found during the animal testing period; Figure 2 D represents the latency period (s) during which the animal first finds the platform in the testing period.

[0010] Figure 3 The results of the Barnes maze trial after 3 months of drug administration, in which Figure 3A represents the time it takes for the animal to enter the dark box (escape latency, s); Figure 3 B represents the time (in seconds) it takes for the animal to find the target hole during the training and testing periods. Figure 3 C represents the accuracy (%) of the animals in finding the target hole during the test period.

[0011] Figure 4 The results of the experiment to actively avoid the time of entering the open box (escape latency, s).

[0012] Figure 5 This is an image showing the results of new object recognition before drug administration, where... Figure 5 A represents the number of times the old object was explored; Figure 5 B represents the preference for new objects.

[0013] Figure 6 The images show the results of new object recognition after 3 months and 6 months of drug administration. Figure 6 A and 6B represent the number of times old objects were explored and the preference for new objects during the new object recognition test 3 months after drug administration; Figure 6 C and Figure 6 D represents the number of times old objects were explored and the preference for new objects during the new object recognition test 6 months after drug administration.

[0014] Figure 7 This is a diagram showing the results of a nest-building experiment. Figure 7 A represents the nesting score before drug administration; Figure 7 B represents the nesting score after 3 months of drug administration; Figure 7 C represents the nesting score after 6 months of drug administration.

[0015] Figure 8 This is a graph showing the results of the open field test data, where... Figure 8 A and 8B represent the distance traveled in the open field and the number of times the center of the open field was crossed before the drug was administered. Figure 8 C and Figure 8 D represents the distance traveled in the open field and the number of times the center of the open field was crossed during the 3 months following drug administration. Figure 8 E and Figure 8 F represents the distance traveled in the open field and the number of times the patient crossed the center of the open field 6 months after administration.

[0016] Figure 9 The image shows the quantitative detection results of Aβ1-40 and Aβ1-42 in brain tissue homogenate. Figure 9 A represents the quantitative analysis of Aβ1-40 in brain tissue homogenate; Figure 9 B represents the quantitative analysis of Aβ1-42 in brain tissue homogenate.

[0017] Figure 10 The image shows the results of ELISA detection of tau, ptau-181, and ROS content in brain tissue homogenate. Figure 10 A represents the total tau content in brain tissue; Figure 10B represents the ptau-181 content in brain tissue; Figure 10 C represents the ROS content in brain tissue.

[0018] Figure 11 The image shows the detection results of pro-inflammatory factors (IL-6, TNF-α, and IL-1β) and SOD. Figure 11 A, Figure 11 B, Figure 11 C represents the detection of serum IL-6, TNF-α, and IL-1β levels using a liquid phase chip. Figure 11 D represents the level of active SOD enzyme in serum as detected by ELISA.

[0019] Figure 12 This is a graph showing the statistical results of amyloid plaque area, where... Figure 12 A represents the percentage of amyloid plaques in the hippocampus; Figure 12 B represents the area of ​​the cortical amyloid plaque.

[0020] Figure 13 For Aβ 1-40 and Aβ 1-42 Statistical data of staining, in which Figure 13 A and Figure 13 B represents the hippocampus and cortical Aβ, respectively. 1-40 Positive area; Figure 13 C and 13D represent hippocampal and cortical Aβ, respectively. 1-42 Positive area. Detailed Implementation

[0021] To better understand the technical content of this invention, the specific implementation method of this invention will be further described below.

[0022] The small molecule compound S1 used in the following experiments has the following structural formula: It is derived from Beijing Qingbo Huineng Pharmaceutical Technology Co., Ltd., and is a white powder. The preparation method can be found in CN116621912A. Donepezil hydrochloride was used as a positive control.

[0023] This experiment used 100 male APP / PS1 SPF mice (source: Beijing Huafukang Biotechnology Co., Ltd.) and 20 male C57BL / 6J SPF mice (source: Beijing Huafukang Biotechnology Co., Ltd.) aged 18 weeks and weighing 25.00-33.00 g. The background strain of the 100 male APP / PS1 mice was the C57BL / 6J APP / PS1 mice, which were bred by crossing PrP-hAPPK595N / M596L dementia model mice and PrP-hPS1dE9 dementia mice. They have stable gene expression, show cognitive and behavioral changes at 3-6 months of age, develop senile plaques at about 5 months of age, and have a large number of senile plaques at about 12 months of age. They are the model mice commonly used in Alzheimer's drug efficacy trials.

[0024] Experimental mice were divided into six groups: a normal control group (C57BL / 6J mice), a solvent control group (Vehicle, model control group, APP / PS1 mice), a positive control group (Donepezil, donepezil hydrochloride 1.5 mg / kg, APP / PS1 mice), a low-dose group of small molecule compound S1 (12 mg / kg, APP / PS1 mice), a medium-dose group of small molecule compound S1 (36 mg / kg, APP / PS1 mice), and a high-dose group of small molecule compound S1 (72 mg / kg, APP / PS1 mice), with 20 animals in each group. Animals with scores close to the mean within ±50% of the target quadrant in the water maze were randomly assigned to groups based on their time spent in the maze. After grouping, the mice were administered the medication orally via gavage once daily for six months, and their condition was observed.

[0025] Normal control group and model control group: administered physiological saline and solvent, respectively; positive control group: administered donepezil hydrochloride; small molecule compound S1 administration groups: administered different doses of small molecule compound S1. The solvent was sulfobutyl-β-cyclodextrin (SBβCD). Donepezil hydrochloride was prepared using ultrapure water, and compound S1 was prepared using the solvent.

[0026] The day of administration is defined as day 1 of the trial.

[0027] The specific dosage design is shown in Table 1 below.

[0028] Behavioral tests were conducted 3 months after drug administration, including open field test, nesting test, Barnes maze test, and new object recognition test; behavioral tests were conducted again 6 months after drug administration, including open field test, nesting test, water maze, active avoidance test, and new object recognition test.

[0029] The Morris water maze primarily assesses spatial and reference memory in animals. Since mice retain long-term spatial and reference memories of the Morris water maze, repeated testing may interfere with the results. Therefore, only two water maze tests were conducted throughout the experiment, one before drug administration and one 6 months after administration. The water maze test consisted of two phases: orienteering (training phase) and spatial exploration (testing phase). The orienteering phase primarily assessed the latency period for mice to locate the hidden underwater platform. Comparison of latency periods over multiple training days reflected the learning process. The spatial exploration phase primarily assessed the number of loop crossings, latency period in exploring the target area, and percentage of time spent in the target quadrant, reflecting the memory outcome.

[0030] Pre-drug water maze test results are an important indicator for determining whether to proceed with the next step (treatment). When APP / PS1 mice exhibited spatial memory cognitive impairment, treatment was considered feasible, and they were grouped for treatment based on their pre-drug water maze test results (selecting the percentage of time spent in the target quadrant). The 6-month water maze test is a powerful tool for verifying the drug's effect on improving the cognitive function of APP / PS1 mice. Based on the water maze test results, it was determined whether the drug treatment for APP / PS1 mice achieved the expected results.

[0031] Before drug administration, 5-month-old WT and APP / PS1 animals underwent a 6-day water maze test (WT, n = 20, APP / PS1, n = 100), with days 1-5 as the training period and spatial cognition testing on day 6. Figure 1 As shown in Figure A, with the increase of training days, the time spent by both APP / PS1 and WT mice to find the platform during the training period decreased (D1 vs. D5). P <0.001), but the latency during the APP / PS1 training period was significantly longer than that during WT ( P <0.001); such as Figure 1 B and Figure 1 As shown in C, during the testing period, the number of times the APP / PS1 searched for the platform and the percentage of time spent in the target quadrant were significantly lower than WT ( P <0.001). Pre-drug test data showed that APP / PS1 mice had reduced learning and memory and cognitive impairment, so subsequent drug treatment could proceed.

[0032] Six months after administration, all animals underwent a water maze test again. Figure 2 A to Figure 2 As shown in D, analysis of the latency during the training period reveals that the latency for the Vehicle group to find the platform is significantly longer compared to the WT group. P<0.001), low, medium and high doses of S1 can significantly shorten the latency period in APP / PS1 animals, but the medium and high doses have the most significant effect. P = 0.031, P =0.020). Analysis of the animals' time spent in the target quadrant, the number of times they found the platform, and the latency period for platform finding during the test period showed that, compared to the WT group, the Vehicle group had a shorter time spent in the target quadrant, a longer latency period for platform finding, and fewer platform finding attempts. P< 0.001, P = 0.127, P< 0.001); Compared with the Vehicle group, the treatment group spent more time in the target quadrant, found more platforms, and had a shorter latency to find platforms, but the Donepezil group spent even longer in the target quadrant. P = 0.014); Low, medium, and high doses of S1 significantly shortened the latency period for animals to find the platform ( P = 0.024, P =0.014, P = 0.006), while the S1 36 mg / kg treatment group significantly increased the number of times APP / PS1 found a platform ( P = 0.027).

[0033] Based on the water maze results after 6 months of drug administration, the training group showed shorter latency to find the platform and stronger spatial memory compared to the Vehicle group, with the S1 36 mg / kg and S1 72 mg / kg groups performing best. During the testing period, analysis was conducted based on the percentage of time spent in the target quadrant, the number of times the platform was found, and the latency to find the platform. Low, medium, and high doses of S1 all performed well in all three tests, with S1 36 mg / kg and S1 72 mg / kg showing the best results. In summary, after 6 months of drug administration, low, medium, and high doses of S1 showed good improvement in spatial memory and cognitive abilities in APP / PS1 mice, with the medium dose group showing the best effect.

[0034] Barnes Maze Mice retain memories acquired in the Morris water maze test for a long time. To avoid the influence of repeated testing in a short period, the Barnes maze was used for the second round of spatial memory testing three months after drug administration. The Barnes maze is designed based on the light-avoiding, dark-loving, and exploratory characteristics of rodents. The animal escapes from a brightly lit, open platform to a dark, small target box located below the platform. After seven days of training, the animal learns and remembers the location of the target box. On the eighth day, its learning and memory abilities are tested by removing the target box.

[0035] Using strong light stimulation as the driving force for the Barnes maze, animals were trained for 7 days. The time it took for the animals to escape from the strong light and enter the dark box (escape latency, s) was analyzed. Figure 3 A to Figure 3 As shown in C, the time to enter the dark chamber was significantly shorter in the S1 12 mg / kg, S1 36 mg / kg, and Donepezil groups after seven days of training ( P <0.05), while there was no statistically significant difference in the time to enter the dark chamber between the Vehicle group and the S1 72mg / kg group after seven days of training ( P> 0.05); and repeated measures analysis comparing the time animals took to enter the dark chamber with S1 12 mg / kg, S1 36 mg / kg, and Donepezil with the Vehicle group showed statistically significant differences ( P = 0.035, P = 0.008, P = 0.037), of which the S1 36 mg / kg group was able to enter the dark chamber in a shorter time ( P = 0.008). Analysis of the time it took for animals to first discover the target hole during the training and testing periods revealed that the time was shorter in each treatment group compared to the Vehicle group, but the difference was not statistically significant. P >0.05). Analysis of the animals' accuracy in exploring the target hole during the test period showed that the Donepezil group had the highest accuracy rate in exploring the target hole. P = 0.031), there was no significant difference between the S1 test sample group and the Vehicle group.

[0036] Based on the Barnes maze results after 3 months of drug administration, during the training period, S1 12 mg / kg, S1 36 mg / kg, and Donepezil showed faster ability to find the dark box and escape bright light compared to the Vehicle group, with S1 36 mg / kg showing the best performance. During the test period, the animals' accuracy in finding the target hole was lower than that of the Donepezil group. In summary, after 3 months of drug administration, the low and medium doses of S1 showed a good improvement in spatial memory and cognitive ability in APP / PS1 mice during the training period, with the medium dose showing the best effect. During the test period, the test product improved the spatial cognition of APP / PS1 mice, but the difference was not statistically significant.

[0037] Active avoidance test The active avoidance test utilizes the rodents' tendency to seek darkness and avoid light. The animal is placed in a dark box, and during training, a single foot shock is given from the dark box. The animal then enters the bright box through the middle door to escape the shock, allowing the animal to develop a fear memory of the aversive stimulus. After 24 hours, the animal is returned to the dark box, and the latency period for the animal to escape from the dark box and enter the bright box is recorded to test the animal's memory ability.

[0038] like Figure 4 As shown, the latency period for animals in the model group to enter the open box during the test period was significantly longer than that in the control group. P = 0.015). A one-way ANOVA was performed to analyze the latency of animals escaping the dark box and entering the bright box during the test period. The S1 group (36 mg / kg) was able to escape the dark box and enter the bright box much faster than the Vehicle group. P = 0.016), the escape latency of Donepezil tended to be shorter compared to the Vehicle group ( P = 0.068).

[0039] New Object Recognition The novel object recognition test is a learning and memory test method established based on the principle that animals have an innate tendency to explore new objects, used to test the animal's episodic memory. By analyzing the number of times the animal explores old objects, we can observe the animal's desire to explore the environment and its curiosity about things. The novel object preference index (DI) = new object / (new object + old object) × 100% is used to examine the animal's episodic memory ability or working memory. This test was conducted before drug administration, 3 months after drug administration, and 6 months after drug administration.

[0040] like Figure 5 As shown, before drug administration, there was no difference between APP / PS1 mice and WT mice in their exploration of old objects during the training period and their preference for new objects during the testing period. P = 0.161, P = 0.358), indicating that the APP / PS1 episodic memory ability has not changed; after 3 months of drug administration, the Vehicle group explored old objects less frequently than the WT group ( P = 0.058), compared with the Vehicle group, the S1 12 mg / kg group explored more old objects ( P = 0.012), but while each treatment group showed an improvement in new object preference compared to the Vehicle group, the difference was not statistically significant. P >0.05).

[0041] like Figure 6 A to Figure 6 As shown in D, after 6 months of administration, the WT and S1 36 mg / kg groups showed a significant increase in exploration of old objects. P = 0.041, P = 0.032), after treatment with 13B 72 mg / kg, the animals' preference for new objects was improved compared with the Vehicle group. P = 0.063).

[0042] According to the new object recognition experiment, different doses of S1 can enhance animals' curiosity about objects and increase their exploratory nature. S1 administration for 6 months can improve the episodic memory ability of APP / PS1 mice.

[0043] like Figure 7 A to Figure 7 As shown in C, low and medium doses of S1 improved the nesting score of APP / PS1. Figure 8 A to Figure 8 As shown in F, open field test data indicate that S1 36 mg / kg can improve APP / PS1 motor performance and mood.

[0044] After 6 months of drug administration, serum IL-1β, IL-6, and TNF-α levels were detected using a liquid chromatography-array kit, and Aβ levels in brain tissue homogenate were detected using a liquid chromatography-array kit. 1-40 Aβ 1-42 The levels of these substances were measured, and the expression of Tau, p-Tau181, and ROS in brain tissue was detected. Histopathological examination was performed on hippocampal amyloid plaques (Thioflavin S staining) and Aβ. 1-40 Aβ 1-42 Precipitation, fibrotic tangles (p-Tau181), neuronal death (NeuN staining), astrocytes (GFAP staining), microglia (Iba-1 staining, CD86, CD206 and CD68), and vascular staining (CD31).

[0045] Brain tissue Aβ 1-40 and Aβ 1-42 Liquid chip detection The main protein component of the plaque is amyloid-beta protein (Aβ), a polypeptide of 40-42 amino acids. 1-40 and Aβ 1-42 20 mg of brain tissue was weighed from each sample and homogenized with 200 μL of lysis buffer (Cell Signaling Lysis Buffer, Lot #3867843). Then, a 1:200 solution of a protease inhibitor (Protease Inhibitor Cocktail Set, EDTA-Free, 539134) was added. The sample was centrifuged at 3000 r / min for 10 minutes, and the supernatant was collected for homogenization and protein extraction. Total protein concentration was determined using the BCA method. After adjusting the protein concentration, Aβ was used for further analysis. 1-40 and Aβ 1-42 Liquid phase chip detection.

[0046] like Figure 9 A to Figure 9 As shown in Figure B, the results showed that the brain tissue homogenate in the Vehicle group showed Aβ 1-40 and Aβ 1-42The levels of Aβ in all four treatment groups (Donepezil, S1 low, medium, and high) were significantly higher than those in the WT group (P < 0.001). 1-40 and Aβ 1-42 The mean brain tissue content of S1 was lower in the AD mice than in the Vehicle group, with low, medium, and high doses of S1 showing significantly lower levels (P < 0.05). This indicates that low, medium, and high doses of S1 can reduce Aβ levels in the brains of AD mice. 1-40 and Aβ 1-42 .

[0047] ROS ELISA detection in brain tissue Reactive oxygen species (ROS)-induced oxidative stress leads to oxidative modifications of proteins and lipids, ultimately resulting in neuronal dysfunction. For example... Figure 10 A to Figure 10 As shown in C, the ROS expression level in the vehicle group was significantly higher than that in the WT group when ROS was detected in the brain tissue homogenate. P = 0.001), indicating a pathological increase in ROS in APP / PS1 mice, while ROS significantly decreased after treatment with S1 36 mg / kg. P = 0.026). This indicates that a medium dose of S1 can reduce ROS in the brains of APP / PS1 mice, thereby reducing oxidative stress and protecting neurons.

[0048] Detection of pro-inflammatory factors (IL-6, TNF-α, and IL-1β) and SOD like Figure 11 A to Figure 11 As shown in D, the levels of pro-inflammatory factors IL-6, TNFα, and IL1β in serum were detected by liquid chromatography-mass spectrometry. There were no statistically significant differences in the levels of these three factors between the treatment group and the vehicle group. P >0.05); Superoxide dismutase (C-Zn superoxide dismutase, SOD) is an antioxidant enzyme that catalyzes the decomposition of superoxide free radicals. ELISA analysis of serum SOD levels showed no statistically significant difference between the treatment group and the vehicle group. P >0.05). Serum factor detection indicated that APP / PS1 mice did not show significant changes in inflammation and SOD compared to WT mice. After treatment with 13B drug, there was no increase or decrease in inflammatory factors in the serum, nor were there any changes in SOD.

[0049] Histopathological examination Amyloid plaque detection Amyloid-β (Aβ) plaques, formed by fibrotic deposition of amyloid protein, are a pathological feature of Alzheimer's disease (AD). In healthy neurons, amyloid precursor proteins with three domains (intracellular, intracellular, and extracellular) are digested by α and γ secretions. This digestive reaction produces some soluble peptides that can be broken down and recycled within the cell. However, when β-secretase combines with γ-secretase, insoluble β-amyloid peptides are produced. When these β-amyloid peptides aggregate, they form harmful β-amyloid plaques. β-amyloid plaques damage surrounding neurons by disrupting signal transduction between healthy neurons and by initiating an immune response that leads to inflammation. These problems cause severe brain damage and loss of functions such as memory and learning, while the clearance of amyloid plaques helps improve cognitive function.

[0050] Thiamine S can specifically bind to mature Aβ amyloid protein and possesses its own green fluorescence. It is used to label the content and distribution of amyloid protein in brain parenchyma and is an important indicator for evaluating the pathological status of Alzheimer's disease. For example... Figure 12 As shown, thiosulfate S staining of brain tissue from AD mice revealed that, compared to WT mice, the area of ​​green fluorescence in the Vehicle group was significantly increased in both the hippocampus and cortex. P All values ​​were less than 0.001. The area of ​​amyloid plaque-positive regions in the hippocampus of the S1 36 mg / kg group was significantly smaller than that of the Vehicle group. P = 0.020), with no significant differences among the other groups.

[0051] like Figure 13 A to Figure 13 As shown in D, Aβ 1-40 Immunohistochemical staining results showed that the positive area in the hippocampus and cortex of the Vehicle group was significantly larger than that in the control group. P All values ​​were less than 0.001. Low, medium, and high doses of Donepezil and S1 significantly reduced Aβ in the hippocampus of APP / PS1 mice. 1-40 Positive area ( P = 0.014, P = 0.005, P = 0.025, P = 0.008). Low, medium and high doses of S1 all significantly reduced Aβ in the cortical region of APP / PS1 mice. 1-40 positive area ( P = 0.010, P = 0.007 and P = 0.003).

[0052] Aβ 1-42Immunohistochemical staining results showed that the positive area in the hippocampus and cortex of the Vehicle group was significantly larger than that in the control group. P All values ​​were less than 0.001. Donepezil and low-to-medium doses of 13B significantly reduced Aβ levels in the hippocampus of APP / PS1 mice. 1-42 Positive area ( P = 0.026, P = 0.025, P = 0.029). Both donepezil and low- and high-dose 13B significantly reduced Aβ in the cortical region of APP / PS1 mice. 1-42 Positive area ( P = 0.021, P = 0.005, P = 0.030), medium dose S1 reduces Aβ in the cortical region. 1-42 Trend of positive area ( P = 0.053).

[0053] Pathological examination of amyloid plaques showed that low, medium, and high doses of S1 could reduce the mean plaque size in the brains of AD mice, but a medium dose of 13B significantly reduced plaque deposition in the hippocampus. P = 0.020); and S1 at low, medium and high doses significantly improved Aβ in the cerebral cortex and hippocampus of AD mice. 1-40 and Aβ 1-42 The expression ( P <0.05).

[0054] In this specification, the invention has been described with reference to specific embodiments thereof. However, it will be apparent that various modifications and variations can be made without departing from the spirit and scope of the invention. Therefore, this specification should be considered illustrative rather than restrictive.

Claims

1. The use of sarsaponin derivatives in the preparation of drugs for treating Alzheimer's disease, characterized in that, The structural formula of the sarsaponin derivative is shown below: 。

Images