Application of PRAS40 protein as a therapeutic target for Alzheimer's disease

Abstract

This invention relates to the field of molecular biology, specifically to the application of PRAS40 protein as a therapeutic target for Alzheimer's disease. This invention combines Western blotting, computer-generated virtual docking, and surface plasmon resonance (SPR) techniques to confirm that PRAS40 protein is a key target influencing the pathological progression of AD. Experimental results show that specific intervention (elimination or reduction) of PRAS40 protein levels is sufficient to significantly improve cognitive deficits and pathological characteristics in AD model mice, providing a clear molecular target for subsequent screening and development of novel anti-AD drugs.

Description

Technical Field

[0001] This invention relates to the field of molecular biology, specifically to the application of PRAS40 protein as a therapeutic target for Alzheimer's disease. Background Technology

[0002] Alzheimer's disease (AD) is a neurodegenerative disease characterized primarily by memory impairment, and its prevalence is increasing annually with the aging population. The main pathological features of AD include senile plaques formed by extraneuronal β-amyloid (Aβ) deposition, neurofibrillary tangles formed by abnormal phosphorylation of tau protein within neurons, neuroinflammation, and progressive loss of neurons / synapses and brain atrophy. Currently, the pathogenesis of AD is not fully understood, and there are no effective intervention drugs in clinical practice to improve the course of the disease.

[0003] Microglia, as innate immune cells of the central nervous system, play a dual role in the pathogenesis of Alzheimer's disease (AD). Current research indicates that the functional state of microglia is closely related to disease progression in AD: under normal conditions, they are responsible for engulfing metabolic waste and pathological deposits in the brain; however, in the pathological microenvironment of AD, microglia often experience functional impairment, leading to a significant decrease in their ability to phagocytose pathological proteins such as Aβ, resulting in the continuous accumulation of pathological products in the brain and further exacerbating cognitive impairment. Therefore, identifying key molecular targets that can specifically regulate the functional state of microglia and improve their ability to phagocytose and degrade pathological products such as Aβ is an important direction for developing novel AD therapeutics.

[0004] However, effective strategies for precisely intervening in microglia to restore their phagocytic function through molecular means are currently lacking. In particular, the absence of key proteins that can serve as specific therapeutic targets limits the development and application of related drugs. Therefore, screening and confirming novel targets and modulators that can improve the pathological features and cognitive deficits of Alzheimer's disease (AD) has significant clinical implications and promising application prospects.

[0005] In view of this, the present invention is hereby proposed. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention provides the application of PRAS40 protein as a therapeutic target for Alzheimer's disease.

[0007] Specifically, the technical solution of the present invention is as follows: In a first aspect, the present invention provides the application of PRAS40 protein as a therapeutic target for Alzheimer's disease for non-disease diagnostic and therapeutic purposes, including: the use of a modulator of PRAS40 protein in the preparation of a medicament for the prevention and / or treatment of Alzheimer's disease, wherein the modulator has the effect of eliminating or reducing the content, expression or function of PRAS40 protein in the human body.

[0008] Preferably, the drug comprises 4,5-bis(4-methoxyphenyl)-5-oxovalerate or 3-(4-hydroxyphenyl)-4-oxo-10-oxa-3-azatricyclo[5.2.1.0~1,5~]dec-8-ene-6-carboxylic acid.

[0009] Preferably, the drug exerts a therapeutic effect by regulating the expression or function of PRAS40 protein, reducing the deposition of β-amyloid protein and the abnormal phosphorylation level of Tau protein in the brain, and improving cognitive and memory functions.

[0010] Preferably, the drug enhances the phagocytic function of microglia in response to β-amyloid protein.

[0011] Preferably, the drug can improve cognitive impairment associated with Alzheimer's disease and improve the pathological morphological changes of microglia in hippocampal tissue.

[0012] In a second aspect, the present invention provides a method for screening drugs for treating Alzheimer's disease, comprising: determining the active binding pocket based on the three-dimensional structure of the PRAS40 protein; and performing high-throughput virtual screening using a compound database to obtain candidate compounds that can form stable interactions with the active binding pocket.

[0013] Preferably, the method further includes: verifying the binding affinity of candidate compounds to recombinant PRAS40 protein using surface plasmon resonance technology, and screening for target compounds with dissociation constants in the micromolar range.

[0014] Preferably, the ameliorative effect of the target compound on AD pathological features is verified in an Alzheimer's disease cell model or a 3×Tg-AD transgenic mouse model.

[0015] Preferably, verifying the ameliorative effect of the target compound on AD pathological features specifically includes verifying whether it can reduce β-amyloid protein levels or reduce Tau protein phosphorylation levels.

[0016] Beneficial effects: This invention combines Western blotting, computer-generated virtual docking, and surface plasmon resonance (SPR) techniques to confirm that the PRAS40 protein is a key target influencing the pathological progression of Alzheimer's disease (AD). Experimental results show that specific intervention (elimination or reduction) of PRAS40 protein levels is sufficient to significantly improve cognitive deficits and pathological characteristics in AD model mice, providing a clear molecular target for subsequent screening and development of novel anti-AD drugs. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in this invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be described below.

[0018] Figure 1 Figure 1 shows the experimental results of targeting and knocking down PRAS40 in microglia to improve cognitive deficits and pathological features in 3×Tg-AD mice. Figure A shows the experimental procedure and grouping of 3×Tg-AD mice after receiving recombinant adeno-associated virus (rAAV) injection and behavioral tests; Figure B shows the statistical graph of the discrimination index of mice in the Novel Object Recognition (NOR) test; Figure C shows the representative swimming trajectory of mice in the Morris Water Maze (MWM) exploration test; Figure D shows the statistical graph of the number of times mice crossed the platform in the exploration test; Figure E shows the statistical graph of the time mice spent in the target quadrant; Figure F shows the statistical graph of the latency period for mice to first enter the platform position; Figure G shows the representative bands of PRAS40 and AD-related proteins (Aβ, phosphorylated Tau protein AT8, and total Tau protein HT7) in the hippocampus; Figure H shows the quantitative statistical analysis of the relative expression levels of the above proteins.

[0019] Figure 2Figure 1 shows the experimental results of virtual screening of novel inhibitors based on PRAS40 structure and their improvement of microglia phagocytic function. Figure A is a schematic diagram of the virtual screening process for identifying PRAS40-binding small molecules based on the SPECS database; Figure B is a two-dimensional interaction diagram of molecular docking between compound F1 and PRAS40 protein; Figure C is a two-dimensional interaction diagram of molecular docking between compound F2 and PRAS40 protein; Figure D is a sensor map and fitted kinetic curve of surface plasmon resonance (SPR) detection of the binding affinity between compound F1 and PRAS40 protein; Figure E is a sensor map and fitted kinetic curve of surface plasmon resonance (SPR) detection of the binding affinity between compound F2 and PRAS40 protein; Figure F shows representative immunofluorescence micrographs of BV2 microglia phagocytosis of pHrodo-labeled synaptosomes under different treatment conditions (control, Aβ model, Aβ combined with compounds F1-F4, or DDS treatment); Figure G is a quantitative statistical analysis of the synaptic phagocytic efficiency (phagocytic index) of microglia in each experimental group. Detailed Implementation

[0020] This invention provides the application of a 40kDa proline-rich Akt substrate (PRAS40) as a therapeutic target for Alzheimer's disease.

[0021] Based on this, the present invention further provides the use of inhibitors or modulators of PRAS40 protein in the preparation of medicaments for the prevention and / or treatment of Alzheimer's disease.

[0022] As a preferred embodiment of the application described in this invention, the drug improves cognitive dysfunction by reducing the expression of PRAS40 protein or inhibiting its function, thereby reducing the deposition of β-amyloid protein (Aβ) and abnormal phosphorylation of Tau protein in the brain.

[0023] Furthermore, the present invention also provides a method for screening drugs for treating Alzheimer's disease, the method comprising the following steps: Based on the three-dimensional structure of the PRAS40 protein, its active binding pocket was determined. High-throughput virtual screening was performed using a compound database to obtain candidate compounds that could form stable interactions (such as hydrogen bonds and hydrophobic interactions) with the active binding pocket. The direct binding affinity of the candidate compound to the recombinant PRAS40 protein was verified using surface plasmon resonance (SPR) technology. Validate the effectiveness of the candidate compounds in alleviating AD pathological features such as Aβ deposition or Tau protein phosphorylation in Alzheimer's disease models such as 3×Tg-AD transgenic mouse models.

[0024] Furthermore, the present invention also provides a medicament for treating Alzheimer's disease, wherein the medicament specifically targets the PRAS40 protein.

[0025] As a preferred embodiment of the drug described in this invention, the drug can promote the phagocytic capacity of microglia for AD pathological products (especially β-amyloid protein), thereby alleviating neurodegenerative diseases.

[0026] As one of the contributions of this invention, it is the first to discover and confirm a novel potential therapeutic target for Alzheimer's disease (AD). This invention combines Western blotting, computer-generated virtual docking, and surface plasmon resonance (SPR) techniques to confirm that the PRAS40 protein is a key target influencing the pathological progression of AD. Experimental results show that specific intervention in PRAS40 protein levels is sufficient to significantly improve cognitive deficits and pathological characteristics in AD model mice, providing a clear molecular target for subsequent screening and development of novel anti-AD drugs.

[0027] As one of the contributions of this invention, it reveals the crucial role of the PRAS40 target in restoring the phagocytic capacity of microglia in Alzheimer's disease (AD) pathology. This invention demonstrates that, under AD pathological conditions, reducing PRAS40 protein levels significantly enhances the phagocytic efficiency of microglia for Aβ-amyloid (Aβ). Through this mechanism, it effectively reduces Aβ deposition and abnormal phosphorylation levels of Tau protein in the hippocampus of AD mice, thereby alleviating the core pathological changes of AD.

[0028] As one of the contributions of this invention, it significantly improves AD-related cognitive and memory impairments. Animal behavioral experiments (novel object recognition test and Morris water maze) confirmed that specific interventions targeting PRAS40 (such as gene knockdown or pharmacological regulation) can significantly enhance short-term discriminative memory and spatial learning memory abilities in 3×Tg-AD model mice. More importantly, this invention also demonstrated that when PRAS40 is artificially overexpressed, it blocks the therapeutic benefits of related drugs (such as dapsone), further confirming the specificity and necessity of this target.

[0029] As one of the contributions of this invention, a novel drug screening strategy based on the PRAS40 structure is constructed. This invention establishes a screening process of "virtual screening-physical binding (SPR)-in vivo / in vitro functional validation." Using this strategy, this invention successfully screened several small molecule compounds from a compound library that specifically bind to the active pocket of the PRAS40 protein. These compounds exhibited pathological amelioration activities comparable to positive control drugs in validation experiments, confirming the good druggability of this target and the practical value of this screening method.

[0030] Furthermore, studies have shown that inhibiting Pim1 to reduce phosphorylated PRAS40 levels can alleviate Aβ and Tau pathological manifestations and salvage cognitive deficits by enhancing proteasome function. This finding seems contradictory to the findings of this invention, but the analysis suggests it may be related to the dynamic changes in microglia during AD progression. When faced with oxidative stress or pathological protein challenges, microglia activate the mTOR signaling pathway to meet increased energy demands and protein synthesis. Acute mTOR activation can transiently enhance glycolysis to meet immediate energy needs, while sustained activation may lead to mitochondrial dysfunction, causing microglia to transform into a pro-inflammatory phenotype dominated by glycolysis. In the early stages of AD, this mechanism may enhance the phagocytic capacity of microglia, effectively clearing misfolded tau proteins; however, in the later stages of the disease, sustained mTOR activation may lead to microglia dysfunction, thereby promoting pathological progression.

[0031] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification.

[0032] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0033] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "specific implementation," or "some specific implementations," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which 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. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0034] The endpoints and any values ​​of the ranges disclosed in this specification are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0035] In the embodiments provided in this specification, unless specific techniques or conditions are specified, the techniques or conditions described in the literature in this field, or the product instructions, shall be followed. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased from legitimate channels.

[0036] Unless otherwise specified, all raw materials used in the embodiments of this invention are commercially available. See Table 1 for details.

[0037] Table 1 Raw Material Information

[0038] Example 1 The therapeutic effect of specifically silencing the PRAS40 gene in microglia on Alzheimer's disease.

[0039] This embodiment aims to verify whether specifically reducing the expression level of PRAS40 in microglia is sufficient to improve AD-related pathological phenotypes, thereby confirming the effectiveness of PRAS40 as a therapeutic target.

[0040] 1. Experimental materials and construction of viral vectors.

[0041] Viral vector origin: The adeno-associated virus (AAV) vector was constructed and produced by Brain Case Technology (Wuhan, China).

[0042] Knockdown virus (rAAV-shPRAS40): To achieve microglia-specific PRAS40 knockdown, an AAV vector carrying a short hairpin RNA (shRNA) targeting mouse PRAS40, driven by the mIBA1 promoter, was constructed. Its complete construct is: rAAV-mlBA1-EGFP-5'miR30a-shRNA(mPRAS40)-3'miR30a-WPRE-4×miR9T.

[0043] Control virus (rAAV-shC.V.): The corresponding scrambled control viral vector has the following construct elements: rAAV-mlBA1-EGFP-5'miR30a-shRNA(Scramble)-3'miR30a-WPRE-4×miR9T.

[0044] Viral titer: The viral titers of all prepared viruses were greater than 1.0 × 10⁻⁶. 13 vg / mL.

[0045] 2. Animal models and stereotactic injection into the brain.

[0046] Experimental animals: 36-week-old (approximately 9 months old) male 3×Tg-AD transgenic mice were selected.

[0047] Surgical methods: such as Figure 1 As shown in Figure A, mice were anesthetized with 1.8-2% isoflurane and fixed on a stereotaxic apparatus. The virus was injected bilaterally into the CA1 region of the mouse hippocampus (0.5 μL per side).

[0048] Injection coordinates (relative to the anterior fontanelle Bregma): anteroposterior (AP) -2.0 mm; medial lateral (ML) +1.5 mm; dorsoventral (DV) -1.7 mm.

[0049] Postoperative recovery: After injection, the mice were placed on a heating pad until fully awake. They were then fed for 6 weeks to ensure that the virus reached a stable expression level in the body, after which behavioral tests and tissue samples were collected.

[0050] 3. Behavioral test results.

[0051] Six weeks after viral expression, cognitive function was assessed in mice from each group.

[0052] New Object Recognition Experiment (NOR): such as Figure 1 As shown in Figure B, compared with the control group (rAAV-shC.V.), the knockdown mice injected with rAAV-shPRAS40 showed a significantly higher discrimination index, indicating that reducing the PRAS40 level in microglia significantly improved the short-term memory ability of AD mice.

[0053] Morris Water Maze (MWM): In spatial memory tests, PRAS40 knockdown mice showed a significant increase in platform crossings during the exploration experiment. Figure 1 (Figure D), and the latency to first entry into the target area is significantly shortened ( Figure 1 (Figure F). The swimming trajectories of mice showed representative differences among the groups ( Figure 1 (Figure C), but no statistically significant difference was observed in the time spent in the target quadrant (Figure C). Figure 1 (China E map).

[0054] 4. Analysis of hippocampal tissue protein expression: After the behavioral test, hippocampal tissue proteins were extracted for Western blotting analysis.

[0055] Target knockdown validation: such as Figure 1As shown in Figures G and H, the results confirm that rAAV-shPRAS40 effectively reduces the expression of PRAS40 protein at the tissue level, verifying the effectiveness of viral interference.

[0056] Improvement in pathological markers: Along with the downregulation of PRAS40, Aβ levels in the hippocampus were significantly reduced, and the phosphorylation level of Tau protein at the AT8 site also decreased significantly. Total Tau protein (HT7) levels remained unchanged between the two groups. Figure 1 (G diagram, H diagram).

[0057] 5. Conclusion.

[0058] These results indicate that inhibiting PRAS40 expression in microglia alone is sufficient to improve cognitive deficits (especially memory discrimination and spatial search strategies) in 3×Tg-AD mice and significantly alleviate two key AD pathological features: Aβ deposition and Tau protein hyperphosphorylation. This confirms that PRAS40 is a key therapeutic target for AD.

[0059] Example 2 Virtual drug screening and discovery of new compounds based on PRAS40 structure.

[0060] This embodiment aims to demonstrate the feasibility of using the PRAS40 protein structure for drug screening and to verify the biological activity of the screened PRAS40-bound small molecule compounds in an in vitro model.

[0061] 1. Structure-based virtual screening, such as... Figure 2 Figure A illustrates the drug screening workflow of this embodiment.

[0062] Database source: SPECS compound database, which contains 202,381 small molecule compounds.

[0063] Receptor model: The predicted three-dimensional structure model of the PRAS40 protein (PDB ID: AF-Q96B36-F1) was used as the receptor for molecular docking.

[0064] Screening process: Funnel-style hierarchical screening using Glide software: High-throughput virtual screening (HTVS): Performs initial screening on the entire database and retains the top 10% of molecules by score; Glide SP (Standard Precision Matching): Re-screens the molecules that were matched in the previous round, retaining the top 10%. Glide XP (Glide Precision Docking): Performs high-precision conformational search and scoring, retaining the top 5%.

[0065] Candidate selection: The XP docking score threshold was set to be less than -4.901. Finally, 15 candidate compounds (Hits) with high predicted binding affinity were selected. Their detailed list and scores are shown in Table 2.

[0066] Table 2. Top 15 candidate compounds obtained from virtual screening of the SPECS database based on receptors (using Schrödinger Maestro)

[0067] 2. Molecular docking mode analysis.

[0068] The binding mode was analyzed using representative compounds F1 and F2, which had high scores.

[0069] Compound F1: such as Figure 2 As shown in Figure B, the F1 molecule intercalates into the binding pocket of PRAS40 via hydrogen bonding and hydrophobic interactions. Its key interacting amino acid residues include CYS44, ARG70, HIE69 (a specific protonated state of histidine), and TYR46.

[0070] Compound F2: such as Figure 2 As shown in Figure C, the F2 molecule forms a stable interaction network with the key residues ALA74, ARG70, and ALA66 of PRAS40.

[0071] 3. Surface plasmon resonance (SPR) affinity verification.

[0072] The SPR technique was used to quantitatively detect the direct binding ability of candidate compounds to the PRAS40 protein.

[0073] Experimental method: The recombinant PRAS40 protein was immobilized on the surface of the sensor chip, and the test compound was injected into the mobile phase at different gradient concentrations (6.25 μM-100 μM), and the response units (RU) were recorded.

[0074] Data Analysis and Computational Methods: Biacore Evaluation Software was used to analyze the sensorgrams obtained from the experiment. A 1:1 Langmuir binding model was used for global fitting of the bound and dissociated phases.

[0075] Calculation formula: Equilibrium dissociation constant (K) D ) by the dissociation rate constant (k ddissociation rate constant and binding rate constant (k a The ratio of K to the association rate constant is calculated using the formula: K D = k d / k a ; Where, k a The rate of complex formation per unit time (M) -1 S -1 ), k d The rate of dissociation of the complex per unit time (s) -1 The software uses an iterative algorithm to minimize the residual between the theoretical curve and the experimental data, thereby obtaining the optimal k. a and k d The value is then used to calculate K. D value.

[0076] Experimental results: Compound F1: such as Figure 2 As shown in Figure D, the sensor image exhibits typical fast binding and dissociation kinetics. After software fitting, the binding rate constant k... a and dissociation rate constant k d Derive the final dissociation constant K D = 1.35μM, indicating a strong affinity between F1 and PRAS40.

[0077] Compound F2: such as Figure 2 As shown in Figure E, a specific binding signal is also displayed, K D The value is 79.9 μM.

[0078] The results confirmed that the compounds obtained by virtual screening could indeed bind directly to the PRAS40 target.

[0079] 4. Verification of phagocytic function of BV2 microglia.

[0080] To verify the biological function of the PRAS40-binding compound, this embodiment examined its effect on improving the phagocytic capacity of microglia under Aβ injury.

[0081] Cell model: BV2 microglia were used.

[0082] Experimental grouping and treatment: Control group (CON): Normal culture. BV2 microglia were seeded into culture dishes containing Dulbecco's Modified Eagle Medium (DMEM), with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin added. Cells were cultured in a 37°C, 5% CO2 incubator using standard methods, with the medium replaced every 2–3 days until the cells reached the logarithmic growth phase for subsequent experiments.

[0083] Model group (Aβ): Aβ was added for 24 hours to simulate the pathological environment of AD.

[0084] In the treatment group, the selected compounds (F1, F2, F3, F4) or the positive control compound (DDS) were added simultaneously with Aβ, with a final concentration of 50 μM, and incubated for 24 hours.

[0085] Detection method: Add pHrodo fluorescently labeled synaptosomes, observe and calculate the phagocytic index using immunofluorescence microscopy.

[0086] Experimental results: like Figure 2 As shown in Figures F and G, Aβ treatment significantly inhibited the phagocytosis of synaptosomes by BV2 cells (P<0.05).

[0087] Compared with the model group, all PRAS40-binding compound (F1-F4) treatment groups significantly restored the phagocytic activity of microglia.

[0088] Quantitative analysis showed that ( Figure 2 (Figure G) The extent to which compounds F1, F2, F3, and F4 improved phagocytosis efficiency was comparable to that of the positive control compound DDS, and some compounds (such as F2 and F3) even showed a better trend.

[0089] 5. Conclusion.

[0090] This embodiment demonstrates that the small molecule regulators obtained based on PRAS40 structure screening can not only specifically bind to the target protein physically, but also improve Aβ-induced microglial dysfunction at the cellular level. This further indicates that PRAS40 is a therapeutic target with high development potential, and that the screening method described in this patent can effectively obtain lead compounds with therapeutic activity.

[0091] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the present invention.

Claims

1. The application of PRAS40 protein as a therapeutic target for Alzheimer's disease for non-disease diagnostic and therapeutic purposes, characterized in that, include: The use of regulators of PRAS40 protein in the preparation of drugs for the prevention and / or treatment of Alzheimer's disease, wherein the regulators have the effect of eliminating or reducing the content, expression or function of PRAS40 protein in the human body.

2. The application according to claim 1, characterized in that, The drug exerts its therapeutic effect by regulating the expression or function of PRAS40 protein, reducing the deposition of β-amyloid protein and the abnormal phosphorylation level of Tau protein in the brain, and improving cognitive and memory functions.

3. The application according to claim 1, characterized in that, The drug can enhance the phagocytic function of microglia in β-amyloid protein.

4. The application according to claim 1, characterized in that, The drug can improve cognitive impairment associated with Alzheimer's disease and improve the pathological morphological changes of microglia in hippocampal tissue.

5. The application according to claim 1, characterized in that, The drug comprises 4,5-bis(4-methoxyphenyl)-5-oxovalerate or 3-(4-hydroxyphenyl)-4-oxo-10-oxa-3-azatricyclo[5.2.1.0~1,5~]dec-8-ene-6-carboxylic acid.

6. A method for screening drugs for treating Alzheimer's disease, characterized in that: This includes determining the active binding pocket based on the three-dimensional structure of the PRAS40 protein; High-throughput virtual screening was performed using a compound database to obtain candidate compounds that could form stable interactions with the active binding pocket.

7. The method for screening drugs for treating Alzheimer's disease according to claim 6, characterized in that, Also includes: The binding affinity of candidate compounds to recombinant PRAS40 protein was verified by surface plasmon resonance technology, and target compounds with dissociation constants in the micromolar range were screened.

8. The method for screening drugs for treating Alzheimer's disease according to claim 7, characterized in that, The ameliorative effect of the target compound on AD pathological features was verified in Alzheimer's disease cell models or 3×Tg-AD transgenic mouse models.

9. The method for screening drugs for treating Alzheimer's disease according to claim 8, characterized in that: The verification of the target compound's effect on improving AD pathological features specifically includes verifying whether it can reduce β-amyloid protein levels or reduce Tau protein phosphorylation levels.

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