Method for treating or alleviating Alzheimer's disease

Abstract

The present application relates to a method of treating or ameliorating Alzheimer's disease by reducing or inhibiting the production of amyloid beta (Abeta), alleviating anxiety, rescuing cognitive function and memory impairment, and ameliorating or ameliorating amyloid-related diseases by administering to a subject a polypeptide drug.

Description

Technical Field

[0001] This application relates to a method for treating or alleviating Alzheimer's disease and provides a corresponding drug or preparation, belonging to the field of biomedicine. Background Technology

[0002] Amyloidosis is not a single disease, but a diverse and progressive disease process characterized by the extracellular deposition of waxy, amyloid proteins called amyloids, which accumulate in one or more organs or systems. Current research suggests that the accumulation of β-amyloid (Aβ) is an early and key driver of the disease, triggering subsequent tau protein hyperphosphorylation, neuroinflammation, synaptic dysfunction, and neuronal death, ultimately leading to cognitive decline and psychosocial symptoms. For example, as Aβ accumulates, it begins to interfere with the normal function of organs or systems, having various effects on brain neural structure and function, including neuroinflammation, synaptic loss, and vascular lesions. These diseases, characterized by Aβ accumulation, include, but are not limited to, Alzheimer's disease (AD), CAA-APP (applied cerebral amyloid angiopathy), Down syndrome, and other neurological disorders. Therefore, drugs that reduce Aβ deposition are currently a hot research topic and an important treatment strategy for these diseases.

[0003] However, current drugs have two drawbacks: first, they can only delay the decline but cannot significantly reverse existing cognitive impairment; second, they have relatively serious side effects. For example, the development of drugs targeting two key enzymes in the Aβ deposition process, namely BACE and γ-secretase, has failed due to side effects. JNJ-54861911 is a BACE inhibitor that can reduce Aβ levels in cerebrospinal fluid, but it causes a significant increase in liver enzyme levels in patients (an important marker of liver function impairment), leading to the termination of its clinical research. Compared with BACE inhibitors, γ-secretase inhibitors have more pronounced drug toxicity, and most exhibit extremely strong drug toxicity, making it almost impossible to achieve inhibition in a safe manner. Some studies suggest that γ-secretase also catalyzes other transmembrane proteins, including the Notch receptor, and inhibition of this receptor can lead to severe liver and spleen side effects, posing a great challenge to the development of such drugs (see Mu Yao et al., Recent Advances in Alzheimer's Disease Drug Development, Journal of China Pharmaceutical University, 2024, 55(6): 816 − 825). Therefore, there is an urgent need for a safe (low-toxicity) drug that can effectively reduce Aβ deposition (especially in vivo experiments), effectively improve or salvage cognitive function and memory impairment, and treat amyloid-related diseases or such neurological disorders. Summary of the Invention

[0004] To address the aforementioned problems, this application first provides an isolated polypeptide, characterized in that the amino acid sequence of the polypeptide is as shown in SEQ ID NO.1 (peptide core of AG11) or SEQ ID NO.2 (peptide core of AB11). In some embodiments, the polypeptide is further prepared as a peptide mimic, fusion peptide, and / or conjugated peptide, for example, by fusing and / or conjugating the polypeptide with a label that facilitates its entry into the target site. The label may be a fluorescent tag, a cell-penetrating peptide, or a carrier peptide or ligand mediating the polypeptide's passage across the blood-brain barrier, for example, by linking the N-terminus or C-terminus of the polypeptide to a TAT peptide.

[0005] This application also provides cultured cells, formulations, compositions, or kits containing the above-described peptides.

[0006] This application also provides uses of the above-mentioned polypeptide, cultured cells of the above-mentioned polypeptide, formulations, compositions, or kits; characterized in that the uses are one of the following or two or more of the following:

[0007] ① Interfere with or inhibit the APP amyloid pathway in cells or tissues (e.g., affect the process by which APP is cleaved by amyloid protein).

[0008] ② Used to reduce or inhibit the production or accumulation of β-amyloid protein in cells or tissues;

[0009] ③ Used to treat, alleviate, inhibit, delay, or prevent amyloid-related diseases characterized by high levels of β-amyloid protein production and accumulation;

[0010] ④ Used to inhibit or reduce amyloid plaques in individuals who have or are at risk of developing neurological diseases or conditions;

[0011] ⑤ To improve or salvage cognitive function and memory impairment in individuals who have or are at risk of developing neurological diseases or conditions;

[0012] ⑥ Alleviate individual anxiety;

[0013] ⑦ For use in the preparation of drugs, formulations, compositions or devices, wherein the drugs, formulations, compositions or devices are used to achieve at least one of the uses in ①-⑥ above.

[0014] The polypeptides, drugs, formulations, or compositions of this application can be administered systemically, topically, parenterally, or non-invasively. The polypeptides, drugs, formulations, or compositions can be prepared as injections, oral preparations, patches, sprays, liniments, or lotions. The polypeptides, drugs, formulations, or compositions can be administered alone or in combination with at least one other pharmaceutical agent, for example, alone, simultaneously, or alternately with other pharmaceutical agents. Other pharmaceutical agents include fusion peptides, antibodies, cholinesterase inhibitors, glutamate modulators, vaccines, gene therapy formulations, or neurotransmitter modulators, etc. The polypeptides, cultured cells of polypeptides, formulations, compositions, or kits of this application can be provided in effective amounts. The methods of this application relate to therapeutic or non-therapeutic methods for diseases and can be performed in vivo or in vitro.

[0015] Amyloid-related diseases or neurological disorders or conditions include Alzheimer's disease (AD), traumatic brain injury, stroke, glaucoma, dementia, muscular dystrophy (MD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), cystic fibrosis, Angelman syndrome, Liddell syndrome, Paget's disease, traumatic brain injury, Reveille's body disease, post-poliomyelitis syndrome, Shay-Dregs syndrome, olivopontocerebellar atrophy, Parkinson's disease, multiple system atrophy, striatal substantia nigra degeneration, supranuclear palsy, bovine spongiform encephalopathy, scrapie, Creutzfeldt-Jakob disease, kuru, G.S.-Shaq disease, chronic wasting diseases, familial fatal insomnia, bulbar palsy, and motor neuron disease. Diseases including Carnafan disease, Huntington's disease, neuronal ceroid lipofuscin deposition disease, Alexander disease, Tourette syndrome, Menkes knot syndrome, Cockayne syndrome, Hallewarden-Schpattz syndrome, Lafra disease, Rett syndrome, Wilson's disease, Lesch-Nair syndrome and Ong-Lon syndrome, Pick's disease, spinocerebellar ataxia, Down syndrome, hereditary cerebral hemorrhage with amyloid degeneration, Guam Parkinson's disease-dementia complications, progressive supranuclear palsy, adult-onset diabetes mellitus, senile cardiac amyloid degeneration, endocrine tumors, macular degeneration, HIV-related neurocognitive impairment, Lewy body dementia, cerebral amyloid angiopathy, inclusion body myositis or mild cognitive impairment.

[0016] The advantages of this application are: ① It inhibits the normal shearing of APP; ② It effectively reduces Aβ plaque deposition; ③ It improves or salvages cognitive function and memory impairment; ④ It is safe and has low toxicity. Attached Figure Description

[0017] Figure 1 AB11 peptide reduces the amyloid pathway in 2EB2 and BAW cells.

[0018] Figure 2 AG11 peptide reduces the amyloid pathway in 2EB2 and BAW cells.

[0019] Figure 3Safety experiment of AB11 and AG11 peptides targeting the brain of APP23 / PS45 mice.

[0020] Figure 4 AB11 and AG11 peptides effectively improved the behavioral characteristics of APP23 / PS45 mice.

[0021] Figure 5 AB11 and AG11 peptides effectively improve amyloid protein load in the brains of APP23 / PS45 mice.

[0022] Figure 6 AB11 and AG11 peptides significantly reduced Aβ levels in the cerebral cortex and hippocampus of APP23 / PS45 mice.

[0023] Figure 7 Effects of AB11 and AG11 peptides on APP cleavage in the brain of APP23 / PS45 mice. Detailed Implementation

[0024] Embodiments of this application will be illustrated exemplarily below. Although this application has been described in conjunction with these specific embodiments, it should be understood that it is not intended to limit this application to such specific embodiments. Rather, this application is intended to cover substitutions, modifications, replacements, variations, and equivalents that may be included within the spirit and scope of this application as defined by the appended claims.

[0025] This application provides a method for treating amyloid-related diseases, reducing their risk, lessening their severity, preventing or delaying their onset, the method comprising administering a therapeutically effective amount of a polypeptide, peptide nucleus, drug, formulation, or composition of this application to an individual suffering from or at risk of the disease. In some embodiments, the polypeptide is a fusion peptide, peptide mimic, or conjugated peptide.

[0026] Amyloid precursor protein (APP) is a single-transmembrane protein widely distributed in cells throughout the body. APP is mainly metabolized through two (cleavage) pathways: amyloid (β pathway) and non-amyloid (α pathway).

[0027] The APP amyloid pathway (i.e., the β pathway) refers to the cleavage of APP by β-secretase into sAPPβ and a C-terminal fragment containing 99 amino acids; the latter is further cleaved by γ-secretase into Aβ and ACID. Aβ generated through this pathway accounts for 90%–95% of the total Aβ. Therefore, interfering with or affecting the APP amyloid cleavage process can alleviate the APP amyloid pathway, thereby reducing Aβ formation or accumulation, and achieving the therapeutic effect of treating amyloid-related diseases (characterized by Aβ accumulation).

[0028] Amyloid-related diseases include, but are not limited to, Alzheimer's disease (AD), CAA-APP, Down syndrome, anxiety disorders, traumatic brain injury, stroke, glaucoma, dementia, muscular dystrophy (MD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), cystic fibrosis, Angelman's syndrome, Liddle syndrome, Paget's disease, Lewy body disease, postpoliomyelitis syndrome, Shy-Draeger syndrome, and olivopontocerebellar atrophy. atrophy, Parkinson's disease, multiple system atrophy, striatonigral degeneration, supranuclear palsy, bovine spongiform encephalopathy, scrapie, Creutzfeldt-Jakob syndrome, kuru, Gerstmann-Straussler-Scheinker disease, chronic wasting disease, fatal familial insomnia, bulbar palsy, motor neuron disease, Canavan disease, Huntington's disease, neuronal lipofuscin deposition disease. ceroid-lipofuscinosis, Alexander's disease, Tourette's syndrome, Menkes kinky hair syndrome,Cockayne syndrome, Hallervorden-Spatz syndrome, Lafora disease, Rett syndrome, hepatolenticular degeneration, Lesch-Nyhan syndrome, Unverricht-Lundborg syndrome, dementia (including but not limited to Pick's disease and spinocerebellar ataxia), hereditary cerebral hemorrhage with amyloidosis (Dutch type), Guam Parkinson-Dementia complex, progressive supranuclear palsy, HIV-related dementia. Diseases include Alzheimer's disease, HIV-related neurocognitive impairment, Lewy body dementia, cerebral amyloid angiopathy, inclusion body myositis, and mild cognitive impairment. In some preferred embodiments, the disease is a neurodegenerative disease such as Alzheimer's disease, CAA-APP (APP-related cerebral amyloid angiopathy), or Down syndrome dementia.

[0029] The polypeptides (peptide mimics containing a peptide core, fusion peptides, and / or conjugated peptides), peptide cores, pharmaceuticals, formulations, or compositions of this application may be administered alone or in combination with at least one other pharmaceutical agent (e.g., another pharmaceutical agent for treating Aβ-amyloid-related diseases). In some embodiments, the other pharmaceutical agent includes acetylcholinesterase inhibitors, particularly tetrahydroaminoacridine. The polypeptides, peptide cores, pharmaceuticals, formulations, or compositions of this application may be used simultaneously, in combination, alternately, or intermittently with other pharmaceutical agents.

[0030] In some embodiments, the polypeptide (peptide mimicry containing a peptide core, fusion peptide, and / or conjugated peptide) or peptide core of this application serves as the direct, sole, or primary active ingredient in a pharmaceutical, formulation, or composition.

[0031] In some embodiments, the terms "formulation" and "composition" are used interchangeably and refer to a mixture of two or more ingredients. In some aspects, "formulation" and "composition" can refer to a mixture of one or more active ingredients with a carrier or other excipient. A "formulation" or "composition" can take almost any physical form, including solid, liquid (e.g., solution), or gas. In some embodiments, the composition of this application is a pharmaceutical composition.

[0032] While the polypeptides of this application may be administered in the form of crude chemicals, they may also be provided as pharmaceutical formulations. Pharmaceutical formulations (or pharmaceutical compositions) comprise one or more of the polypeptides, peptide nuclei, or analogs thereof described herein, together with one or more pharmaceutically acceptable carriers; or pharmaceutical formulations (or pharmaceutical compositions) comprise one or more of the polypeptides, peptide nuclei, or analogs thereof described herein and optionally one or more active ingredients for treating other diseases. The appropriate formulation form depends on the chosen route of administration, and any suitable and well-known technology, carrier, and excipients may be used. In some embodiments, the pharmaceuticals, formulations, and compositions of this application are injections, lyophilized powders for injection, aerosols, creams, drops, ointments, or pastes; topical formulations or oral preparations; liniments, lotions, granules, sprays, or patches; or emulsions. In some embodiments, the pharmaceuticals, formulations, compositions, and devices of this application also include carriers to facilitate the delivery of the active ingredient in the polypeptides, pharmaceuticals, formulations, or compositions of this application to the brain. The pharmaceuticals, formulations, compositions, and devices of this application also include carriers or additional reagents suitable for facilitating the delivery of the active ingredient to the brain. Carriers that facilitate the delivery of active ingredients to the brain include any carrier that facilitates the crossing of the blood-brain barrier and any carrier that facilitates the uptake of active ingredients by nerve cells.

[0033] In some embodiments, the polypeptides, drugs, formulations, or compositions of this application are contained within packaging, containers, or devices. Packaging may be a soft bag, blister pack, etc.; containers may be vials, ampoules, etc. In some embodiments, the polypeptides, drugs, formulations, or compositions of this application are substantially uniformly distributed within packaging, containers, or devices. In some embodiments, the polypeptides, drugs, formulations, or compositions of this application are provided in single-dose or multi-dose packaging, containers, or devices. That is, the polypeptides, drugs, formulations, or compositions of this application may be provided in single-dose or multi-dose form. In one embodiment, multi-dose refers to containing at least two doses.

[0034] The polypeptides, pharmaceuticals, formulations, compositions, devices, or treatments of this application control, slow, alleviate, reduce, delay, and / or reverse the symptoms of a disease in the treated individual within the following ranges relative to untreated: approximately 5%-95%, approximately 5%-90%, approximately 5%-80%, approximately 5%-70%, approximately 5%-60%, approximately 5%-50%, approximately 5%-40%, approximately 5%-30%, approximately 5%-20%, approximately 10%-100%, approximately 20%-90%, approximately 30%-90%, approximately 40%-90%, approximately 50%-90%, or approximately 75%-90%.

[0035] The polypeptides, peptide cores, formulations, compositions, devices, or methods of this application may be used in individuals or subjects with respect to amyloid-related diseases. Individuals or subjects (such as patients) may be mammals or non-mammals, typically humans. However, this application is not limited to treating humans and may also be used for veterinary purposes, such as for farm animals (e.g., sheep, pigs, cattle, or horses); pet animals (e.g., dogs and cats); or laboratory animals (e.g., mice, rabbits, or monkeys).

[0036] The cells or tissues in this application can be any cells or tissues of an individual or subject, and can be nerve cells or tissues, preferably brain cells or tissues, such as hippocampus, cerebral cortex, etc.; they can also be model cells for in vitro experiments, such as HEK293 cells or SH-SY5Y cells, etc.

[0037] Terms and Definitions

[0038] In this specification and the appended claims, unless the context explicitly specifies otherwise, the singular forms, including the singular forms “a,” “an,” and “the,” specifically cover the plural indicators of the terms they refer to. Additionally, as used herein, unless otherwise specifically indicated, the word “or” is used in its inclusive meaning of “and / or” rather than its exclusive meaning of “or / or.”

[0039] As used herein, references to the numerical range of variables are intended to convey that this application can be implemented with variables equal to any value within that range. Thus, for intrinsically discontinuous variables, the variable may be equal to any integer value within the numerical range, including the endpoints of the range. Similarly, for intrinsically continuous variables, the variable may be equal to any real value within the numerical range, including the endpoints of the range. For example, a variable described as having a value between 0 and 2 may be 0, 1, or 2 for intrinsically discontinuous variables, and may be 0.0, 0.1, 0.01, 0.001, or any other real value for intrinsically continuous variables.

[0040] As used herein, the term “about” will be understood by those skilled in the art and will vary to some extent depending on the context in which it is used. If the use of this term is unclear to those skilled in the art, then in the context of its use, “about” will mean a value within the range of the listed value plus or minus 10%.

[0041] "Prevention" refers to reducing the occurrence of disorder or condition in a treated sample relative to an untreated control sample, or delaying the occurrence of one or more symptoms of disorder or condition relative to an untreated control sample, or reducing the severity of one or more symptoms of disorder or condition.

[0042] "Treatment," "mitigation," "delay," "inhibition," or "control" refers to therapeutic measures aimed at alleviating (reducing) or terminating a target symptom or disorder. In some implementations, "treatment" does not necessarily require the complete elimination of disease symptoms, but those skilled in the art will understand that "treatment," "mitigation," "delay," "inhibition," "control," or "prevention" all refer to direct effects and not to indirect means of disease healing such as health maintenance or adjunctive effects, like enhancing blood circulation, flavoring, or improving comfort.

[0043] "Effective dose" refers to an amount sufficient to achieve the desired therapeutic and / or preventive effect, such as causing prevention or relief of symptoms associated with the disease. The amount of the composition administered to the recipient will depend on the type and severity of the disease and the individual's characteristics, such as general health, age, sex, weight, and tolerance to the drug. The amount also depends on the degree, severity, and type of the disease. A skilled technician will be able to determine the appropriate dosage based on these and other factors. For example, "effective dose" may refer to the average level of pharmacological action that minimizes the symptoms.

[0044] "Peptide analogs": Peptide analogs that mimic the key structural features and biological activities of the original peptide (or natural peptide).

[0045] "Fusion peptides" are formed by linking peptides with other molecules (mostly via peptide bonds). Applications include adding affinity tags (such as His-tag, GST, MBP) for protein purification; improving peptide solubility / stability (by fusing with highly soluble proteins); facilitating detection / visualization (by fusing with reporter proteins such as GFP, luciferase, etc.); or serving as vaccine antigens (by fusing antigenic peptides to carrier proteins to enhance immunogenicity).

[0046] "Conjugated peptides": These are formed by linking peptides with other molecules (mostly through chemical bonds). For example, in labeling and detection, they are linked with fluorescent dyes (such as FITC), enzymes (for ELISA / WB), or biotin (for affinity capture); in drug delivery, drug molecules are linked to target peptides or antibodies (such as antibody-drug conjugates).

[0047] "Carrier peptide": A peptide expressed on a therapeutic polypeptide through bioengineering fusion.

[0048] "Ligand": A substance that is chemically coupled to a drug carrier (such as nanoparticles) or a drug molecule.

[0049] "Cell-penetrating peptides": A type of functional peptide that enhances the ability of molecules (such as polypeptides, proteins, nucleic acids, or drug carriers) to cross the cell membrane. These peptides have an inherent ability to penetrate the cell membrane and often carry macromolecules or particles linked to them into the cell, such as TAT ​​peptide (fragment YGRKKRRQRRR of the HIV TAT protein transduction domain), penetrantin, or polyarginine peptides.

[0050] "Blood-brain barrier penetrating peptide": A carrier peptide or ligand that mediates the passage of polypeptides across the blood-brain barrier, and a functional peptide that is parallel to cell-penetrating peptide.

[0051] “Peptide core”: the shortest active sequence or the smallest functional fragment. In this application, it refers to a polypeptide with an amino acid sequence as shown in SEQ ID NO.1 or SEQ ID NO.2.

[0052] "Polypeptide": In this application, it includes a peptide core, as well as peptide mimics, fusion peptides, and / or conjugated peptides containing a peptide core.

[0053] "Topical application" refers to the direct application of a drug to the body part to be affected or a method of administration that can achieve local efficacy, including but not limited to: epidermal administration, inhalation administration, enema administration, ocular administration, and nasal administration.

[0054] "Systemic administration," also known as systemic drug delivery, includes methods such as intravenous administration, oral administration, intramuscular injection, and subcutaneous injection. Drugs can be transported throughout the body via the bloodstream.

[0055] "Administration" includes any route by which it is introduced into or delivered to an individual to perform its intended function. It can be performed by any suitable route, including systemic administration, local administration, parenteral (via intravenous, intramuscular, intraperitoneal, or subcutaneous) administration, or non-invasive administration.

[0056] "Device" refers to instruments, equipment, consumables, systems, medical devices, and healthcare products that can release drugs, have drug delivery functions, or have the potential to deliver drugs. For example, a device delivers drugs in a sustained-release manner.

[0057] "Active ingredient," also known as "direct active ingredient" or "active pharmaceutical ingredient," refers to a substance that has a direct therapeutic effect on a disease. It is a necessary condition for treating a disease, or its dosage, administration method, and other parameters directly affect the therapeutic effect. There is a clear causal relationship between the application of this ingredient and the achievement of the therapeutic effect. For example, in a compound preparation, if the efficacy of the preparation after its removal is significantly worse than that of the original formulation, then the removed ingredient is considered to be the direct active ingredient.

[0058] "Sole active ingredient" refers to a final product (such as a peptide or drug) in which only one ingredient exerts its medicinal effect, such as directly inhibiting, preventing, and / or treating the symptoms of a disease.

[0059] "Main active ingredient" refers to an active ingredient whose added amount, proportion, concentration, or efficacy is no less than that of other active ingredients in the final product (such as peptides or drugs); or whose content accounts for more than 50%, 60%, 70%, 80%, or 90% of all active ingredients, wherein the percentage is a mass ratio or molar ratio.

[0060] "Acceptable" means compatible with other components of the formulation and harmless to its recipients. "Pharmaceutically acceptable" refers to substances approved by regulatory authorities such as the CFDA (China), EMEA (Europe), and / or FDA (US) and / or any other national regulatory authority for use in animals, preferably for humans, such as pharmaceutical carriers, drug concentrations, or forms of drug presence. Pharmaceutical carriers include, for example, buffers, excipients, stabilizers, or preservatives.

[0061] "Simultaneous administration" means administering at least two active ingredients to an individual via the same route and at the same or substantially at the same time.

[0062] "Single administration" refers to administering one or more active ingredients to an individual at the same time or substantially at the same time through different routes.

[0063] "Alternating administration" refers to administering at least two active ingredients to an individual at different times, via the same or different routes.

[0064] Example 1, Experimental Design

[0065] 1. Peptide design

[0066] The peptides were treated with the core sequences GVVIATVIVIT (SEQ ID NO.1) and SEVKMDVEFRH (SEQ ID NO.2), respectively, with the unintended sequence VEDFKMREVHS (SEQ ID NO.3) as a control. Further, the TAT peptide YGRKKRRQRRR (SEQ ID NO.4) was bound to the amino terminus of both the treated and control peptides. Further still, a FITC fluorescent tag was coupled to the amino terminus of the TAT peptide. The final peptides are shown below (see...). Figure 3 B):

[0067] SB11:YGRKKRRQRRR-VEDFKMREVHS.

[0068] AB11:YGRKKRRQRRR-SEVKMDVEFRH.

[0069] AG11:YGRKKRRQRRR-GVVIATVIVIT.

[0070] FITC-SB11:FITC-YGRKKRRQRRR-VEDFKMREVHS.

[0071] FITC-AB11FITC-YGRKKRRQRRR-SEVKMDVEFRH.

[0072] FITC-AG11FITC-YGRKKRRQRRR-GVVIATVIVIT.

[0073] The above-mentioned peptide was synthesized by GL Biochem (Shanghai). The peptide was dissolved in PBS. Eight-week-old male APP23 / PS45 mice were intraperitoneally injected with the peptide at a dose of 0.5 mg / kg / day for 8 consecutive weeks. Behavioral and biochemical analyses were then performed on the mice.

[0074] 2. Cell Culture

[0075] All cell lines were cultured in 85% Dulbecco Modified Eagle Medium (DMEM) (Cat#C11995500BT, Gibco, USA) supplemented with 15% fetal bovine serum (FBS) (Cat#10270-106, Gibco, USA) and maintained in a 37% CO2 incubator. The 2EB2 cell line consisted of human embryonic kidney (HEK293) cells stably expressing the human Swedish mutant APP695 and human BACE1, and was maintained in 85% DMEM supplemented with 15% FBS containing 100 µg / mL zeocin (Cat#R25001, Invitrogen, USA) and 50 µg / mL G418 (Cat#11811031, Gibco, USA). The BAW cell line consisted of human embryonic kidney (HEK293) cells stably expressing human wild-type APP695 and human BACE1, maintained in 85% DMEM and 15% FBS containing 100 µg / mL zeocin (Cat#R25001, Invitrogen, USA) and 50 µg / mL G418 (Cat#11811031, Gibco, USA). 2EB2 and BAW cells are not included in cell lines typically identified by the International Committee on Cell Line Identification. The treated cell lines were identified by Western blotting to determine overexpression bands. Stable cell lines (2EB2 and BAW) were treated with AB11 or AG11 peptides at 0, 1, 2, 4, or 10 μM for 24 h, and the expression of various proteases along the APP cleavage pathway was characterized by Western blotting.

[0076] 3. Mouse strains

[0077] We used the APP23 / PS45 mouse model because they developed visible senile plaques at 8 weeks of age and exhibited behavioral differences at 16 weeks of age. APP23 / PS45 transgenic mice were obtained by crossing APP23 mice carrying the Swedish APP751 (KM→NL) mutant transgene with PS45 mice overexpressing the human G384A mutant presenilin-1 (PS1). The mice were housed at the Institute of Gerontology, Wenzhou Medical University, in individually ventilated cages (IVCs) with constant temperature and humidity, under a 12-hour light-dark cycle (light on from 08:00 to 20:00), with free access to food and water. DNA was extracted from tail tissue using polymerase chain reaction (PCR) for genotyping. All animal experiments were conducted in accordance with the "Guidelines for the Care and Use of Laboratory Animals" formulated by the Ethics Committee of Wenzhou Medical University.

[0078] 4. Y-maze experiment

[0079] Experimental mice were placed in the center of a Y-maze, with the three arms spaced 120° apart, and allowed to freely explore the arms for 5 minutes. Video was recorded using Anymaze video tracking software, and the spontaneous alternation rate was then calculated manually.

[0080] 5. Open field experiment

[0081] Open field experiments were conducted in an open field chamber. Mice were allowed to explore freely for 10 minutes, and the total distance traveled and the percentage of time spent were observed. Anymaze was used to automatically calculate the time and distance spent by the mice in the central and surrounding areas.

[0082] 6. New Object Recognition Experiment

[0083] Twenty-four hours prior to the test, each mouse was acclimatized in a 40 cm × 40 cm × 40 cm box for 15 minutes. During the test, the mice were exposed to a set of identical objects for 6 minutes. The mice were then removed from the box, the chamber was cleaned with 70% ethanol, and one of the objects was replaced with a new one. The mice were then returned to the laboratory, and the interaction time between the mice and the familiar or unfamiliar objects was recorded. The total interaction time was determined as the sum of the number of interactions (old object + new object). The discrimination index (%) was defined as the time spent exploring the new object / total interaction time × 100. Interaction was defined as actively observing the object when the mouse was pointing at it and within 1 cm of it.

[0084] 7. Morris's Water Maze Experiment

[0085] The MWM test was conducted in a circular tank filled with opaque water, with four reference shapes of different colors and patterns affixed to the walls around the tank. During the hidden platform training phase, mice were placed at one of four random points and allowed 60 seconds to find the hidden platform. If a mouse did not find the platform within 60 seconds, it was guided to the platform and rested for 10 seconds. The time spent finding the hidden platform was recorded using Anymaze. During the platform testing phase, the hidden platform was removed. The time spent in each quadrant and the number of times the mouse traversed the platform were recorded.

[0086] 8. Statistical Analysis

[0087] GraphPad Prism 8.0 was used for statistical analysis and the creation of quantified figures. All results are expressed as mean ± standard error (SEM). Comparisons between two groups were analyzed using t-tests, and multiple comparisons were analyzed using one-way or two-way ANOVA followed by post-hoc Bonferroni's, Dunnett's, or Tukey's multiple comparison tests; P < 0.05 was considered statistically significant.

[0088] It should be noted that, in order to illustrate the therapeutic principle and effect of the peptide of this application on amyloid-related diseases, this application only uses AD as the experimental symptom design (see [link to application]). Figure 3 A). Since the accumulation of amyloid Aβ is a key pathological feature of amyloid-related diseases, those skilled in the art can expect that the peptides of this application can also produce similar therapeutic effects on other amyloid-related diseases (such as CAA-APP, Down syndrome, etc.) through the same treatment principle (reducing the accumulation of Aβ).

[0089] Example 2, In vitro experiment: Safe and significantly improves the pathology of AD

[0090] 1. AB11 peptide has no significant cytotoxic effects.

[0091] To evaluate the effects of AB11 peptide on cell viability and potential cytotoxicity, we conducted tests in HEK293 and SH-SY5Y cells using the CCK8 assay and LDH release assay, respectively. After treating cells with 0, 2, 4, 10, 20, and 40 μM AB11 peptide for 24 hours, the CCK8 assay showed no significant changes in cell viability at any concentration. Similarly, the LDH release assay in SH-SY5Y cells also indicated that treatment with different concentrations of AB11 peptide did not induce significant cytotoxic effects. In conclusion, AB11 peptide has no significant cytotoxic effect on cells within the above concentration range. Figure 1 A).

[0092] 2. AB11 peptide affects the cleavage of amyloid precursor protein (APP) in 2EB2 and BAW cells.

[0093] To determine whether AB11 treatment affected the amyloidosis process and APP processing, we first treated stable cell lines (2EB2 and BAW) with AB11 peptide solutions at concentrations of 0, 1, 2, 4, or 10 μM for 24 hours. Figure 1 B, Figure 1 G). We characterized the expression of various proteins and APP hydrolysates in the APP processing pathway using immunoblotting analysis. We used the C-terminal antibody C20 immunoblotting method to detect CTF (CTF-C99, C89) and APP protein levels. We found that compared with the control group SB11, treatment with the AB11 peptide significantly reduced the protein level of CTF-C99 in 2EB2 cells. Figure 1 C, 4 μM P = 0.0355, 10 μM P = 0.0183), while reducing the level of CTF-C99 / APP ( Figure 1C, 4 μM P = 0.1978, 10 μM P = 0.1331). In BAW cells, treatment with the AB11 peptide reduced the protein level of CTF-C89 (C, 4 μM P = 0.1978, 10 μM P = 0.1331). Figure 1 H, 2 μM P = 0.1490, 4 μM P = 0.3655, 10 μM P = 0.2000), while AB11 peptide significantly reduced the level of CTF-C89 / APP (H, 2 μM P = 0.1490, 4 μM P = 0.3655, 10 μM P = 0.2000), and AB11 peptide significantly reduced the level of CTF-C89 / APP (H, 2 μM P = 0.1490, 4 μM P = 0.3655, 10 μM P = 0.2000). Figure 1 H, 2 μM *P = 0.0272, 4 μM *P = 0.0223, 10 μM *P = 0.0126), and AB11 treatment did not significantly affect the protein levels of APP or BACE1. Figure 1 D, Figure 1 I). Furthermore, consistent with these in vitro results, we found that AB11 peptide treatment significantly reduced Aβ40 secretion in 2EB2 cells (I). Figure 1 E, 10 μMP = 0.1059) and the level of Aβ42 ( Figure 1 F, 10 μM *P = 0.0186), while significantly reducing the level of Aβ42 secreted in BAW cells (F, 10 μM *P = 0.0186), Figure 1 K, 4 μM **P = 0.0081, 10 μM **P = 0.0054), but the level of Aβ40 did not change ( Figure 1 In summary, we have determined that the AB11 peptide plays a mitigating role in the APP amyloid pathway in 2EB2 and BAW cells.

[0094] 3. AG11 peptide has no significant cytotoxic effects.

[0095] To evaluate the effects of AG11 peptide on cell viability and potential cytotoxicity, we performed CCK8 assays and LDH release assays in HEK293 and SH-SY5Y cells. After treating cells with 0, 2, 4, 10, 20, and 40 μM AG11 peptide for 24 hours, the CCK8 assay showed no significant changes in cell viability at any concentration. Similarly, the LDH release assay in SH-SY5Y cells also indicated that treatment with different concentrations of AG11 peptide did not induce significant cytotoxic effects. In conclusion, AG11 peptide has no significant cytotoxic effect on cells within the above concentration range. Figure 2 A).

[0096] 4. AG11 peptide affects the cleavage of amyloid precursor protein (APP) in 2EB2 and BAW cells.

[0097] Subsequently, to determine whether AG11 treatment affected the amyloidosis process and APP processing, we first treated stable cell lines (2EB2 and BAW) with AG11 peptide solution at concentrations of 0, 1, 2, 4, or 10 μM for 24 hours. Figure 2 B. Figure 2 G). We found that treatment with the AG11 peptide increased the protein level of CTF-C99 in 2EB2 cells compared to the control group SB11. Figure 2 C, 10 μM P = 0.4710), while increasing the level of CTF-C99 / APP ( Figure 2 C, 10 μM P = 0.2931). In BAW cells, treatment with the AG11 peptide increased the protein level of CTF-C89 (C, 10 μM P = 0.2931). Figure 2 H, 10 μM P = 0.1665), but we then found that AG11 significantly increased the level of CTF-C89 / APP (H ...). Figure 2 H, 2 μM P = 0.0705, 4 μM * P = 0.0448, 10 μM P = 0.0650), while AG11 treatment did not significantly affect the protein levels of APP or BACE1. Figure 2 D, Figure 2 I). Furthermore, consistent with these in vitro results, we found that AG11 peptide treatment significantly reduced Aβ40 secretion in 2EB2 cells (I). Figure 2 E, 10 μM *P = 0.0477) and the level of Aβ42 ( Figure 2 F, 10 μM P = 0.0824), while significantly reducing the level of Aβ40 secreted in BAW cells (F, 10 μM P = 0.0824), Figure 2 J, 4 μM P = 0.0524) and the level of Aβ42 ( Figure 2 K, P = 0.0969). In summary, we confirm that AG11 also plays a mitigating role in the APP amyloid pathway in 2EB2 and BAW cells.

[0098] The above results indicate that treatment with AB11 and AG11 peptides alone can significantly improve AD pathology in vitro, and both have low toxicity. The in vivo experiments described below show even more significant effects.

[0099] Example 3: In vivo experiment: Effects on APP shearing in the brain of APP23 / PS45 mice

[0100] 1. AB11 and AG11 have good ability to cross the blood-brain barrier.

[0101] To assess the permeability of transmembrane peptides across the blood-brain barrier via intraperitoneal injection, we intraperitoneally injected FITC-conjugated peptides FITC-SB11, FITC-AB11, and FITC-AG11 into APP23 / PS45 mice. We found that FITC fluorescence signals were detectable in both the cortex and hippocampus of immunofluorescent brain slices from the FITC-injected group. Figure 5 A). Furthermore, after peripherally injecting the transmembrane peptide via the tail vein, we used in vivo two-photon imaging to assess its permeability to the blood-brain barrier. We found that FITC fluorescence signals could be detected in cerebral blood vessels. Figure 3 D). The results showed that AB11 and AG11 peptides could cross the blood-brain barrier (BBB).

[0102] 2. AB11 and AG11 peptides affect APP cleavage in the brains of APP23 / PS45 mice.

[0103] To determine whether the reduction in amyloid deposition after treatment with AB11 or AG11 peptides was due to changes in APP processing, we characterized the expression of various proteins in the APP processing pathway using Western blot analysis. Figure 7 A, Figure 7 C Figure 7 E, Figure 7 G). We found that, compared with the SB11 control group, AB11 peptide treatment significantly reduced APP (G) levels in the cerebral cortex of APP23 / PS45 mice. Figure 7 B, **P = 0.0038) and its cleavage product (CTFβ) ( Figure 7 B, P = 0.0508) protein levels, while reducing APP (B, P = 0.0508) in the hippocampus Figure 7 F, *P = 0.0182) and its cleavage product (CTFβ) ( Figure 7 The protein levels (F, **P = 0.0022) were observed. AG11 peptide treatment did not affect the levels of PS1 in the mouse cerebral cortex (F, **P = 0.0022). Figure 7 D) and the PS1 level in the hippocampus ( Figure 7 H). Simultaneously, CTFβ in the cerebral cortex ( Figure 7 D, P = 0.2351) and CTFα ( Figure 7 Both D and P = 0.1374 showed varying degrees of increase; CTFβ in the hippocampus ( Figure 7 H, P = 0.2990) and CTFα ( Figure 7 H (P = 0.1437) also showed varying degrees of increase. These results indicate that treatment with AB11 and AG11 peptides reduces the amyloid protein pathway by affecting various components of the amyloid degeneration processing pathway (such as APP, BACE1, or PS1), thereby reducing the generation and deposition of Aβ.

[0104] Example 4, In vivo experiment: Intraperitoneal injection of AB11 and AG11 peptides significantly improved amyloid pathology in APP23 / PS45 mice.

[0105] First, we assessed the ability of AB11 and AG11 to cross the blood-brain barrier. To evaluate the permeability of the peptides across the blood-brain barrier via intraperitoneal injection, we intraperitoneally injected FITC-conjugated peptides FITC-SB11, FITC-AB11, and FITC-AG11 into APP23 / PS45 mice. We found that FITC fluorescence signals were detectable in the cerebral cortex and hippocampus of immunofluorescent brain slices from the FITC-injected group. Figure 5 A) The results showed that AB11 and AG11 peptides can penetrate the blood-brain barrier (BBB).

[0106] Then, we performed histological analysis on amyloid plaques from APP23 / PS45 mouse cerebral cortex and hippocampus sections treated with SB11, AB11, and AG11 peptides. Staining with Aβ antibody (6E10) and thiamine-S showed ( Figure 5 B, Figure 5 E). Compared with mice treated with SB11 peptide, the total area of ​​Aβ plaques in the cerebral cortex of APP23 / PS45 mice treated with AB11 peptide ( Figure 5 C, **P = 0.0053) and the number of plaques ( Figure 5 C, **P = 0.0093) significantly reduced compared to the SB11 peptide control group. Simultaneously, the area and number of plaques in the hippocampus showed a decreasing trend, but no statistically significant difference was observed. Figure 5 D), these results demonstrate the therapeutic effect of AB11 peptide on brain plaques in AD mice. Compared to the SB11 peptide control group, the total area of ​​Aβ plaques in the cerebral cortex of APP23 / PS45 mice treated with AG11 peptide (D) Figure 5 C, **P = 0.0034) and the number of plaques ( Figure 5 C, **P = 0.0049) was also significantly reduced. Meanwhile, the area and number of plaques in the hippocampus of the AG11 mouse treatment group showed a decreasing trend, but there was no significant statistical difference. Figure 5 (D) These results indicate that treatment with the AG11 peptide can delay the progression of brain plaques in AD mice. Although both the area and number of plaques in the cortex were significantly reduced, the specific differences in the hippocampus region of AD mice may be attributed to inherent regional differences in Aβ deposition and clearance dynamics. The main role of AB11 and AG11 peptides may be to limit the growth and maturation of existing plaques by inhibiting the addition of new Aβ peptides, thereby significantly reducing the overall plaque burden, even if the initial seed number in specific regions does not change drastically.

[0107] In addition, to detect whether such changes occurred in extracellular plaques, we stained with thiosulfate-S. We found that the total area of ​​Aβ plaques in the cerebral cortex of AB11 peptide-treated APP23 / PS45 mice ( Figure 5 F, *P = 0.0148) and quantity ( Figure 5 F, *P = 0.0121) was significantly reduced compared to the SB11 peptide control group. We also found that the total area of ​​Aβ plaques in the cerebral cortex of APP23 / PS45 mice treated with AG11 peptide (F, *P = 0.0121) was significantly reduced. Figure 5 F, **P = 0.0069) and quantity ( Figure 5 The F (*P = 0.0191) was significantly reduced compared to the SB11 peptide control group.

[0108] Next, we analyzed the levels of Aβ40 and Aβ42 in the lysates of the mouse cortex and hippocampus using ELISA. We found that Aβ40 levels in the cerebral cortex of APP23 / PS45 mice treated with AB11 peptide (Aβ40) were significantly higher than those in the lysates of the hippocampus. Figure 6 A, *P = 0.0336) and Aβ42 ( Figure 6 The level of Aβ40 (*P=0.0026) in the cerebral cortex of APP23 / PS45 mice treated with AG11 peptide was significantly reduced; Figure 6 A, **P = 0.0112) and Aβ42 ( Figure 6 The levels of Aβ40 (B, P = 0.0205) were significantly reduced. Similarly, in the hippocampus, AB11 peptide treatment also alleviated Aβ40 (B, P = 0.0205) levels in the cerebral cortex of APP23 / PS45 mice. Figure 6 A, *P = 0.0347) and Aβ42 ( Figure 6 B, *P = 0.0109) levels; AG11 peptide treatment also alleviated Aβ40 (B, *P = 0.0109) levels in the cerebral cortex of APP23 / PS45 mice. Figure 6 A, **P = 0.0112) and Aβ42 ( Figure 6 The level of B (P = 0.0436) was significantly reduced.

[0109] In summary, these results indicate that treatment with AB11 and AG11 peptides significantly improves amyloid pathology in APP23 / PS45 mice, regardless of whether the disease is in the cortex or hippocampus.

[0110] Example 5, In vivo experiments: Relieving anxiety and improving, rescuing and reversing individual cognitive and memory impairments.

[0111] 1. AB11 and AG11 peptides improve anxiety levels and working memory in mice.

[0112] We administered SB11, AB11, and AG11 peptides intraperitoneally to APP23 / PS45 mice at a dose of 0.5 mg / kg / day for 8 consecutive weeks, during which time the mice's body weight was measured weekly. We found that the peptide injections did not change the animals' body weight. Figure 3 C) and none of the organs were affected. Figure 3 E) indicates that these peptides are safe and non-toxic.

[0113] Following 8 consecutive weeks of intraperitoneal injections of SB11, AB11, or AG11 peptides in APP23 / PS45 mice, the following procedure was performed:

[0114] Open field test. We did not find significant differences in total distance traveled or average speed in the open field test. However, AD model mice injected with AB11 and AG11 peptides showed longer dwell times in the central region of the open field, indicating that there was no significant difference in motor function among the different groups of mice injected with the peptides, but anxiety levels were significantly reduced. Figure 4 A, AB11 *P = 0.0185, AG11 ***P = 0.0036).

[0115] Y-maze test. We found that, with the SB11 group as the control group, both AB11 and AG11 peptides could rescue the spontaneous alternation rate in mice. Figure 4 B, AB11 ***P = 0.0005, AG11 ***P = 0.0005). These results indicate that treatment with AB11 and AG11 peptides alone has a rescuing effect on working memory in APP23 / PS45 mice.

[0116] 2. AB11 and AG11 peptides improve cognition and memory in mice.

[0117] In the novel object recognition test, we found that the AB11 peptide treatment group significantly increased the percentage of time mice spent exploring novel objects during the test period. Figure 4 C, AB11 male ***P = 0.0001) and discrimination index ( Figure 4 D, AB11 male ****p < 0.0001), indicating that the AB11 peptide can reverse the discrimination index (DI) in mice and improve their cognitive function and memory. Additionally, although the AB11 peptide treatment group did not significantly increase its exploration time percentage during the testing period ( Figure 4 C), but we found that AG11 treatment significantly improved its discrimination index (DI). Figure 4D, AG11 male ***P = 0.0005), indicating that AG11 peptide can also effectively improve cognitive function and memory in mice. In conclusion, treatment with both AB11 and AG11 peptides can significantly alleviate cognitive and memory deficits in APP23 / SP45 mice.

[0118] 3. AB11 and AG11 peptides improve spatial learning and memory in mice.

[0119] Based on the findings that AB11 and AG11 peptide treatments affect working memory and cognitive function in mice, the effects of AB11 and AG11 peptide treatments on spatial learning and memory were further investigated using the Morris water maze experiment.

[0120] We found that during the visual plateau phase training on the first day of the water maze, the average swimming speed during the visual plateau phase was significantly lower in both the AB11 peptide treatment group and the AG11 peptide treatment group compared to the SB11 control group. Figure 4 E, AB11 male P = 0.7432; AB11 male P = 0.9292) and the platform incubation period ( Figure 4 There were no significant differences in E, AB11 male P = 0.8418; AB11 male P = 0.9496, excluding differences in motor ability and vision among the three groups of mice. On day 4, the latency to reach the plateau in the AB11 peptide treatment group was significantly reduced compared with that in the SB11 control group (E, AB11 male P = 0.8418; AB11 male P = 0.9496). Figure 4 F, AB11 male** P = 0.002. On day 5, the latency to reach the plateau in the AB11 peptide treatment group was significantly reduced compared to the SB11 control group (F, AB11 male **P = 0.002). Figure 4 F, AB11 male *P = 0.0195). Additionally, on day 3, the latency to reach the plateau in the AG11 peptide-treated group was significantly reduced compared to the SB11 control group (F, AB11 male *P = 0.0195). Figure 4 F, AG11 male P = 0.0693). On day 4, the latency to reach the plateau in the AG11 peptide treatment group was significantly reduced compared to the SB11 control group (F, AG11 male P = 0.0693). Figure 4 F, AG11 male *P = 0.0197). On day 5, the latency to reach the plateau in the AG11 peptide-treated group was significantly reduced compared to the SB11 control group (F, AG11 male *P = 0.0197). Figure 4 F, AG11 male, P = 0.1925). On day 6, a probe test was performed to assess the mice's learning and memory abilities. The platform was removed, allowing the mice to freely explore the pool for 60 seconds. The average trajectory heatmap and swimming path map of the three groups of mice during the testing period were obtained from day 6. Figure 4In G), we found that mice in the AB11 and AG11 peptide treatment groups spent more time remaining in the quadrant where the plateau was located compared to mice in the SB11 peptide control group. Figure 4 H, AB11 male *P = 0.0092, AG11 male *P = 0.0479), and the number of times the treated group mice crossed the platform was significantly greater than that of the control group mice. Figure 4 H, AB11 male P = 0.0502, AG11 male P = 0.0984). These results indicate that treatment with both AB11 and AG11 peptides significantly alleviated spatial memory impairment in APP23 / PS45 mice.

[0121] In summary, in cell experiments, treatment with AB11 or AG11 improved AD pathology. Figures 1-2 AC,FH), and reduced the generation of Aβ ( Figure 1-2 (DE, IJ). Subsequently, we verified the membrane-penetrating properties of the membrane-penetrating peptides using FITC-conjugated FITC-AB11 and FITC-AG11 (DE, IJ). Figure 5 A). More importantly, after treatment with AB11 or AG11, APP23 / PS45 mice performed better in the Y maze ( Figure 4 B) New object recognition Figure 4 C, Figure 4 D) and the water maze ( Figure 4 E- Figure 4 The H) results were superior to the control group, indicating that cognitive function and memory improved in mice after treatment. Correspondingly, we found that after treatment with AB11 or AG11 transmembrane peptides, immunofluorescence of mouse brain tissue sections showed fewer and smaller neural plaques (H). Figure 5 B- Figure 5 G), subsequent ELISA results showed that the treatment group mice had less Aβ40 / 42 in their cerebral cortex tissue (G), Figure 6 A- Figure 6 B). Furthermore, Western blot experiments on mouse brain tissue also demonstrated that after treatment with AB11 and AG11, the corresponding components related to APP cleavage were altered. Figure 7 ).

Claims

1. An isolated polypeptide, characterized in that, The amino acid sequence of the polypeptide is shown in SEQ ID NO.1 or SEQ ID NO.

2.

2. The polypeptide according to claim 1, characterized in that, The polypeptide was further prepared into peptide mimics, fusion peptides, and / or conjugated peptides.

3. The polypeptide according to claim 2, characterized in that, The peptide is fused and / or conjugated with a marker that helps it enter the target site.

4. The polypeptide according to claim 3, characterized in that, The label is a fluorescent tag, a cell-penetrating peptide, or a carrier peptide or ligand that mediates the passage of the polypeptide across the blood-brain barrier, for example, linking the N-terminus or C-terminus of the polypeptide to a TAT peptide.

5. Cells, formulations, compositions or kits comprising the polypeptides of any one of claims 1-4.

6. Use of the polypeptide according to any one of claims 1-4, cultured cells of the polypeptide, formulations, compositions, or kits thereof; characterized in that, The intended use is one of the following, or two or more of the following: ① Interfere with or inhibit the APP amyloid pathway in cells or tissues; ② Used to reduce or inhibit the production or accumulation of β-amyloid protein in cells or tissues; ③ Used to treat, alleviate, inhibit, delay, or prevent amyloid-related diseases characterized by high levels of β-amyloid protein production and accumulation; ④ Used to inhibit or reduce amyloid plaques in individuals who have or are at risk of developing neurological diseases or conditions; ⑤ To improve or salvage cognitive function and memory impairment in individuals who have or are at risk of developing neurological diseases or conditions; ⑥ Alleviate individual anxiety; ⑦ For use in the preparation of drugs, formulations, compositions or devices, wherein the drugs, formulations, compositions or devices are used to achieve at least one of the uses in ①-⑥ above.

7. The use as described in claim 6, characterized in that, The polypeptide, drug, formulation, or composition is administered alone or in combination with at least one other agent.

8. The use as described in claim 7, characterized in that, The other agents include fusion peptides, antibodies, cholinesterase inhibitors, glutamate modulators, vaccines, gene therapy agents, or neurotransmitter modulators.

9. The use as described in any one of claims 6-8, characterized in that, The amyloid-related diseases or neurological disorders or conditions include Alzheimer's disease, CAA-APP, Down syndrome, anxiety disorder, traumatic brain injury, stroke, glaucoma, dementia, muscular dystrophy, multiple sclerosis, amyotrophic lateral sclerosis, cystic fibrosis, Angelman syndrome, Liddell syndrome, Paget's disease, traumatic brain injury, Reveille's body disease, post-poliomyelitis syndrome, Shay-Dregs syndrome, olivopontocerebellar atrophy, Parkinson's disease, multiple system atrophy, striatal substantia nigra degeneration, supranuclear palsy, bovine spongiform encephalopathy, scrapie, Creutzfeldt-Jakob disease, kuru, G.S.-Shahar disease, chronic wasting diseases, familial fatal insomnia, medullary diseases, etc. Paralysis, motor neuron disease, Carnafan disease, Huntington's disease, neuronal ceroid lipofuscin deposition disease, Alexander disease, Tourette syndrome, Menkes knot syndrome, Cockayne syndrome, Hallewarden-Schpattz syndrome, Lafra disease, Rett syndrome, Wilson's disease, Lesch-Nair syndrome, Won-Lon syndrome, Pick's disease, spinocerebellar ataxia, hereditary cerebral hemorrhage with amyloid degeneration, Guam Parkinson's disease-dementia complications, progressive supranuclear palsy, adult-onset diabetes mellitus, senile cardiac amyloid degeneration, endocrine tumors, macular degeneration, HIV-related neurocognitive impairment, Lewy body dementia, cerebral amyloid angiopathy, inclusion body myositis or mild cognitive impairment.

10. The use as described in any one of claims 6-9, characterized in that, The polypeptide, drug, formulation, or composition is administered systemically, topically, parenterally, or non-invasively; the polypeptide, drug, formulation, or composition is prepared as an injection, oral preparation, patch, spray, liniment, or lotion; or the polypeptide, drug, formulation, or composition is administered alone, simultaneously, or alternately with at least one other agent.

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