Use of proton pump regulators in the preparation of reagents

Abstract

The present invention relates to a proton pump regulator and its use in the preparation of a reagent for use in improving learning ability, treating cognitive impairment, and / or treating a neurodegenerative disease. It also relates to an ATP6V1B2 regulator and its use in the preparation of a reagent for use in improving learning ability, treating cognitive impairment, and / or treating a neurodegenerative disease.

Description

[Technical field]

[0001] The present application relates to the biopharmaceutical field, and in particular to the use of proton pump modulators in the preparation of reagents. [Background technology]

[0002] Alzheimer's disease (AD) is generally caused by synapse and neuron damage, and abnormalities in the structure and function of neural circuits. Currently, the mechanisms of synapse and neuron damage are still unclear, so progress in effective prevention and treatment methods for AD is very slow.

[0003] The ATP6V1B2 protein, encoded by the ATP6V1B2 gene, is a key structural protein of the proton pump driven by ATP hydrolysis. It is widely distributed in human tissues, but is more abundant in brain tissue, kidney, osteoclasts, and other parts. It plays an important function in processes such as synaptic transmission and lysosomal acidification. Its gene mutations can cause DOORS syndrome (susceptible autosomal dominant congenital deafness syndrome with nail hypoplasia). Reduced expression of ATP6V1B2 protein may be associated with the development of AD. Summary of the Invention

[0004] The present application provides a method for improving the learning ability of a subject, and by discovering that ATP6V1B2 is a potential target for improving the learning ability of a subject, the present application further discovers a reagent that can increase the expression level and / or activity of ATP6V1B2 and / or a proton pump regulator that can be used as a potential drug for improving the learning ability of a subject.

[0005] In one aspect, the present application provides the use of a proton pump modulator in the preparation of a reagent for use in improving learning ability.

[0006] In another aspect, the present application provides the use of a proton pump modulator in the preparation of a reagent for use in the treatment of a cognitive disorder.

[0007] In another aspect, the present application provides the use of a proton pump modulating agent in the preparation of a reagent for use in the treatment of a neurodegenerative disease.

[0008] In some embodiments, the reagent increases the expression level and / or activity of a proton pump associated protein in a subject.

[0009] In some embodiments, the proton pump modulating agent increases the expression level and / or activity of a proton pump associated protein in a subject.

[0010] In some embodiments, the expression level of the proton pump associated protein comprises the expression level of a gene encoding a proton pump associated protein, the transcription level of a gene encoding a proton pump associated protein, and / or the expression level of a proton pump associated protein.

[0011] In some embodiments, the improvement comprises an increase in the expression level and / or activity of the proton pump associated protein of at least about 10% compared to the expression level and / or activity of the proton pump associated protein native to the subject.

[0012] In some embodiments, the expression level of the proton pump-associated protein is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining, and mass spectrometry.

[0013] In some embodiments, the expression level of the proton pump associated protein is measured by using a substance selected from the group consisting of primers capable of specifically amplifying a gene encoding a proton pump associated protein, a nucleic acid molecule that specifically binds to a gene encoding a proton pump associated protein, a nucleic acid molecule that specifically binds to a proton pump associated protein, a small molecule that specifically binds to a proton pump associated protein, a probe that specifically binds to a proton pump associated protein, and a polypeptide that specifically binds to a proton pump associated protein.

[0014] In some embodiments, the reagent increases the expression level and / or activity of ATP6V1B2 in the subject.

[0015] In some embodiments, the proton pump modulating agent increases the expression level and / or activity of ATP6V1B2 in a subject.

[0016] In some embodiments, the expression level of ATP6V1B2 comprises the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene and / or the expression level of the ATP6V1B2 protein.

[0017] In some embodiments, the improvement comprises an increase in the expression level and / or activity of ATP6V1B2 of at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject.

[0018] In some embodiments, the expression level of ATP6V1B2 is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining, and mass spectrometry.

[0019] In some embodiments, the expression level of ATP6V1B2 is measured by using a substance selected from the group consisting of primers capable of specifically amplifying the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 protein, a small molecule that specifically binds to the ATP6V1B2 protein, a probe that specifically binds to the ATP6V1B2 protein, and a polypeptide that specifically binds to the ATP6V1B2 protein.

[0020] In some embodiments, the proton pump modulating agent comprises a proton pump agonist.

[0021] In some embodiments, the proton pump modulating agent comprises a vesicular proton pump agonist.

[0022] In some embodiments, the proton pump modulating agent comprises an agonist of the ATP6V1B2 protein.

[0023] In some embodiments, the proton pump modulating agent is capable of binding to the ATP6V1B2 protein.

[0024] In some embodiments, the proton pump regulating agent is capable of binding to at least a portion of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein.

[0025] In some embodiments, the proton pump regulating agent is capable of binding to at least a portion of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein.

[0026] In some embodiments, the proton pump modulating agent comprises a nucleic acid or variant thereof, a polypeptide or variant thereof, and / or a small molecule.

[0027] In some embodiments, the proton pump modulating agent is capable of increasing synaptic neurotransmitter release.

[0028] In some embodiments, the increase comprises an increase of at least about 10% compared to the natural synaptic neurotransmitter release level in the subject.

[0029] In some embodiments, the proton pump modulating agent is capable of increasing the discharge frequency of excitatory postsynaptic currents.

[0030] In some embodiments, the increase comprises an increase of at least about 10% compared to the native level of excitatory postsynaptic current discharge frequency in the subject.

[0031] In some embodiments, the learning ability comprises a cognitive ability, a motor ability, a memory ability, and / or a spatial exploration ability.

[0032] In some embodiments, the improvement in learning ability comprises an improvement in the subject's improved learning ability score by at least about 50% compared to the subject's original learning ability score.

[0033] In some embodiments, the learning ability score is measured by performing a test selected from the group consisting of a novel object recognition test and a water maze test.

[0034] In some embodiments, the novel object recognition test assesses the cognitive, motor and / or spatial exploration abilities.

[0035] In some embodiments, the water maze test assesses the memory ability, motor ability, and / or spatial exploration ability.

[0036] In some embodiments, the subject comprises a mammal.

[0037] In some embodiments, the subject comprises a human.

[0038] In some embodiments, the subject comprises a non-cognitively impaired patient and / or a non-neurodegenerative disease patient.

[0039] In some embodiments, the subject comprises a neurodegenerative disease patient and / or a cognitive impairment patient.

[0040] In some embodiments, the subject comprises an Alzheimer's disease patient.

[0041] In some embodiments, the subject is in the elderly stage.

[0042] In some embodiments, the cognitive impairment comprises mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment.

[0043] In some embodiments, the cognitive disorder comprises cognitive disorder caused by normal aging, Lewis dementia (LBD), frontotemporal dementia, and / or vascular dementia.

[0044] In some embodiments, the cognitive impairment-inducing disease comprises Alzheimer's disease, multi-infarct Parkinson's disease, AIDS, and / or Creutzfeldt-Jakob disease (CJD).

[0045] In some embodiments, the neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

[0046] In some embodiments, the neurodegenerative disease comprises a neurodegenerative disease caused by neuronal death and glial cell homeostasis, a neurodegenerative disease caused by aging, a neurodegenerative disease caused by effects on CNS cell function, a neurodegenerative disease caused by abnormal communication between cells, and / or a neurodegenerative disease caused by impaired cell motility.

[0047] In some embodiments, the neurodegenerative disease comprises Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD).

[0048] In some embodiments, the reagent is formulated to be suitable for oral and / or injectable administration.

[0049] In some embodiments, the reagent is formulated to be suitable for intravenous injection.

[0050] In another aspect, the present application provides the use of ATP6V1B2, a functionally active fragment thereof or a nucleic acid molecule encoding same in improving learning ability.

[0051] In another aspect, the present application provides the use of ATP6V1B2, a functionally active fragment thereof or a nucleic acid molecule encoding same in the treatment of a cognitive disorder.

[0052] In another aspect, the present application provides the use of ATP6V1B2, a functionally active fragment thereof or a nucleic acid molecule encoding same in the treatment of a neurodegenerative disease.

[0053] In some embodiments, the ATP6V1B2 is from a human.

[0054] In some embodiments, the ATP6V1B2 comprises the amino acid sequence set forth in SEQ ID NO:8 or 16.

[0055] In some embodiments, the functionally active fragment of ATP6V1B2 has the ability to specifically bind to the amino acid sequence set forth in SEQ ID NO:5.

[0056] In some embodiments, the functionally active fragment of ATP6V1B2 comprises a truncation of ATP6V1B2.

[0057] In some embodiments, the functionally active fragment of ATP6V1B2 comprises at least a portion of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein.

[0058] In some embodiments, the functionally active fragment of ATP6V1B2 comprises at least a portion of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein.

[0059] In some embodiments, the functionally active fragment of ATP6V1B2 comprises the amino acid sequence set forth in SEQ ID NO:10 or SEQ ID NO:11.

[0060] In some embodiments, the nucleic acid molecule comprises the nucleotide sequence set forth in SEQ ID NO:9 or 17.

[0061] In some embodiments, the learning ability comprises a cognitive ability, a motor ability, a memory ability, and / or a spatial exploration ability.

[0062] In some embodiments, the improvement in learning ability comprises an improvement in the subject's improved learning ability score by at least about 50% compared to the subject's original learning ability score.

[0063] In some embodiments, the learning ability score is measured by performing a test selected from the group consisting of a novel object recognition test and a water maze test.

[0064] In some embodiments, the novel object recognition test assesses the cognitive, motor and / or spatial exploration abilities.

[0065] In some embodiments, the water maze test assesses the memory ability, motor ability, and / or spatial exploration ability.

[0066] In some embodiments, the subject comprises a mammal.

[0067] In some embodiments, the subject comprises a human.

[0068] In some embodiments, the subject comprises a non-cognitively impaired patient and / or a non-neurodegenerative disease patient.

[0069] In some embodiments, the subject comprises a neurodegenerative disease patient and / or a cognitive impairment patient.

[0070] In some embodiments, the subject comprises an Alzheimer's disease patient.

[0071] In some embodiments, the subject is in the elderly stage.

[0072] In some embodiments, the cognitive impairment comprises mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment.

[0073] In some embodiments, the cognitive disorder comprises cognitive disorder caused by normal aging, Lewis dementia (LBD), frontotemporal dementia, and / or vascular dementia.

[0074] In some embodiments, the cognitive impairment-inducing disease comprises Alzheimer's disease, multi-infarct Parkinson's disease, AIDS, and / or Creutzfeldt-Jakob disease (CJD).

[0075] In some embodiments, the neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

[0076] In some embodiments, the neurodegenerative disease comprises a neurodegenerative disease caused by neuronal death and glial cell homeostasis, a neurodegenerative disease caused by aging, a neurodegenerative disease caused by effects on CNS cell function, a neurodegenerative disease caused by abnormal communication between cells, and / or a neurodegenerative disease caused by impaired cell motility.

[0077] In some embodiments, the neurodegenerative disease comprises Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD).

[0078] In another aspect, the present application provides the use of an ATP6V1B2 modulator in the preparation of a reagent for use in improving learning ability.

[0079] In another aspect, the present application provides the use of an ATP6V1B2 modulator in the preparation of a reagent for use in the treatment of a cognitive disorder.

[0080] In another aspect, the present application provides the use of an ATP6V1B2 modulating agent in the preparation of a reagent for use in the treatment of a neurodegenerative disease.

[0081] In some embodiments, the reagent increases the expression level and / or activity of ATP6V1B2 in the subject.

[0082] In some embodiments, the ATP6V1B2 modulating agent increases the expression level and / or activity of ATP6V1B2 in a subject.

[0083] In some embodiments, the expression level comprises the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene and / or the expression level of the ATP6V1B2 protein.

[0084] In some embodiments, the increasing the activity comprises increasing the expression level and / or activity of a proton pump-associated protein.

[0085] In some embodiments, the improvement comprises an increase in the expression level and / or activity of ATP6V1B2 of at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject.

[0086] In some embodiments, the expression level of ATP6V1B2 is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining, and mass spectrometry.

[0087] In some embodiments, the expression level of ATP6V1B2 is measured by using a substance selected from the group consisting of primers capable of specifically amplifying the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 protein, a small molecule that specifically binds to the ATP6V1B2 protein, a probe that specifically binds to the ATP6V1B2 protein, and a polypeptide that specifically binds to the ATP6V1B2 protein.

[0088] In some embodiments, the ATP6V1B2 modulating agent comprises an agonist of the ATP6V1B2 protein.

[0089] In some embodiments, the ATP6V1B2 modulating agent is capable of binding to the ATP6V1B2 protein.

[0090] In some embodiments, the ATP6V1B2 regulator is capable of binding to at least a portion of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein.

[0091] In some embodiments, the ATP6V1B2 modulator is capable of binding to at least a portion of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein.

[0092] In some embodiments, the ATP6V1B2 modulating agent comprises a proton pump agonist.

[0093] In some embodiments, the ATP6V1B2 modulating agent comprises a vesicular proton pump agonist.

[0094] In some embodiments, the ATP6V1B2 modulating agent comprises a nucleic acid or variant thereof, a polypeptide or variant thereof, and / or a small molecule.

[0095] In some embodiments, the ATP6V1B2 modulating agent comprises a polypeptide or a variant thereof.

[0096] In some embodiments, the polypeptide or variant thereof specificity is capable of binding to the ATP6V1B2 protein.

[0097] In some embodiments, the polypeptide or variant thereof is capable of increasing synaptic neurotransmitter release.

[0098] In some embodiments, the increase comprises an increase of at least about 10% compared to the natural synaptic neurotransmitter release level in the subject.

[0099] In some embodiments, the polypeptide or variant thereof is capable of increasing the discharge frequency of excitatory postsynaptic currents.

[0100] In some embodiments, the increase comprises an increase of at least about 10% compared to the native level of excitatory postsynaptic current discharge frequency in the subject.

[0101] In some embodiments, the polypeptide or variant thereof comprises the amino acid sequence set forth in SEQ ID NO:5.

[0102] In some embodiments, the polypeptide or variant thereof comprises the amino acid sequence set forth in SEQ ID NO:1 or 3.

[0103] In some embodiments, the learning ability comprises a cognitive ability, a motor ability, a memory ability, and / or a spatial exploration ability.

[0104] In some embodiments, the improvement in learning ability comprises an improvement in the subject's improved learning ability score by at least about 50% compared to the subject's original learning ability score.

[0105] In some embodiments, the learning ability score is measured by performing a test selected from the group consisting of a novel object recognition test and a water maze test.

[0106] In some embodiments, the novel object recognition test assesses the cognitive, motor and / or spatial exploration abilities.

[0107] In some embodiments, the water maze test assesses the memory ability, motor ability, and / or spatial exploration ability.

[0108] In some embodiments, the subject comprises a mammal.

[0109] In some embodiments, the subject comprises a human.

[0110] In some embodiments, the subject comprises a non-cognitively impaired patient and / or a non-neurodegenerative disease patient.

[0111] In some embodiments, the subject comprises a neurodegenerative disease patient and / or a cognitive impairment patient.

[0112] In some embodiments, the subject comprises an Alzheimer's disease patient.

[0113] In some embodiments, the subject is in the elderly stage.

[0114] In some embodiments, the cognitive impairment comprises mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment.

[0115] In some embodiments, the cognitive disorder comprises cognitive disorder caused by normal aging, Lewis dementia (LBD), frontotemporal dementia, and / or vascular dementia.

[0116] In some embodiments, the cognitive impairment-inducing disease comprises Alzheimer's disease, multi-infarct Parkinson's disease, AIDS, and / or Creutzfeldt-Jakob disease (CJD).

[0117] In some embodiments, the neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

[0118] In some embodiments, the neurodegenerative disease comprises a neurodegenerative disease caused by neuronal death and glial cell homeostasis, a neurodegenerative disease caused by aging, a neurodegenerative disease caused by effects on CNS cell function, a neurodegenerative disease caused by abnormal communication between cells, and / or a neurodegenerative disease caused by impaired cell motility.

[0119] In some embodiments, the neurodegenerative disease comprises Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD).

[0120] In some embodiments, the reagent is formulated to be suitable for oral and / or injectable administration.

[0121] In some embodiments, the reagent is formulated to be suitable for intravenous injection.

[0122] In another aspect, the present application provides a method for screening for drugs that can improve a subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease, comprising detecting an effect of a candidate drug on the expression level and / or activity of a proton pump associated protein in a subject, wherein if the expression level and / or activity of the proton pump associated protein is improved after administration of the candidate drug, the candidate drug can improve the subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease.

[0123] In another aspect, the present application provides a method for screening for drugs that can improve a subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease, comprising a step of detecting the effect of a candidate drug on the expression level and / or activity of ATP6V1B2 in a subject, wherein if the expression level and / or activity of ATP6V1B2 is improved after administration of the candidate drug, the candidate drug can improve the subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease.

[0124] In some embodiments, the expression level of the proton pump associated protein comprises the expression level of a gene encoding a proton pump associated protein, the transcription level of a gene encoding a proton pump associated protein, and / or the expression level of a proton pump associated protein.

[0125] In some embodiments, the improvement comprises an increase in the expression level and / or activity of the proton pump associated protein of at least about 10% compared to the expression level and / or activity of the proton pump associated protein native to the subject.

[0126] In some embodiments, the expression level of the proton pump-associated protein is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining, and mass spectrometry.

[0127] In some embodiments, the expression level of the proton pump associated protein is measured by using a substance selected from the group consisting of primers capable of specifically amplifying a gene encoding a proton pump associated protein, a nucleic acid molecule that specifically binds to a gene encoding a proton pump associated protein, a nucleic acid molecule that specifically binds to a proton pump associated protein, a small molecule that specifically binds to a proton pump associated protein, a probe that specifically binds to a proton pump associated protein, and a polypeptide that specifically binds to a proton pump associated protein.

[0128] In some embodiments, the expression level of ATP6V1B2 comprises the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene and / or the expression level of the ATP6V1B2 protein.

[0129] In some embodiments, wherein said improving comprises improving the expression level and / or activity of ATP6V1B2 by at least about 10% compared to the original expression level and / or activity of ATP6V1B2 in said subject.

[0130] In some embodiments, the expression level of ATP6V1B2 is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining, and mass spectrometry.

[0131] In some embodiments, the expression level of ATP6V1B2 is measured by using a substance selected from the group consisting of primers capable of specifically amplifying the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 protein, a small molecule that specifically binds to the ATP6V1B2 protein, a probe that specifically binds to the ATP6V1B2 protein, and a polypeptide that specifically binds to the ATP6V1B2 protein.

[0132] In some embodiments, the learning ability comprises a cognitive ability, a motor ability, a memory ability, and / or a spatial exploration ability.

[0133] In some embodiments, the administration comprises oral and / or injection.

[0134] In some embodiments, the administration comprises intravenous injection.

[0135] In another aspect, the present application provides a polypeptide or a variant thereof capable of regulating the expression level and / or activity of ATP6V1B2.

[0136] In some embodiments, the polypeptide or variant thereof is capable of increasing synaptic neurotransmitter release.

[0137] In some embodiments, the increase comprises an increase of at least about 10% compared to the natural synaptic neurotransmitter release level in the subject.

[0138] In some embodiments, the polypeptide or variant thereof is capable of increasing the discharge frequency of excitatory postsynaptic currents.

[0139] In some embodiments, the increase comprises an increase of at least about 10% compared to the native level of excitatory postsynaptic current discharge frequency in the subject.

[0140] In some embodiments, the polypeptide or variant thereof is capable of binding to an ATP6V1B2 protein.

[0141] In some embodiments, the polypeptide or variant thereof is capable of increasing the expression level and / or activity of ATP6V1B2 in a subject.

[0142] In some embodiments, the expression level of ATP6V1B2 comprises the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene and / or the expression level of the ATP6V1B2 protein.

[0143] In some embodiments, the improvement comprises an increase in the expression level and / or activity of ATP6V1B2 of at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject.

[0144] In some embodiments, the polypeptide or a variant thereof is capable of binding to at least a portion of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein.

[0145] In some embodiments, the polypeptide or a variant thereof is capable of binding to at least a portion of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein.

[0146] In some embodiments, the polypeptide or variant thereof comprises the amino acid sequence set forth in SEQ ID NO:5.

[0147] In some embodiments, the polypeptide or variant thereof comprises the amino acid sequence set forth in SEQ ID NO:1 or 3.

[0148] In some embodiments, the polypeptide or variant thereof comprises a multimer.

[0149] In some embodiments, the multimer comprises a homodimer.

[0150] In some embodiments, there are no sulfhydryl blocking modifications to cysteines in the amino acid sequence of the polypeptide or variant thereof.

[0151] In some embodiments, there are no phosphorylation modifications at serine in the amino acid sequence of the polypeptide or variant thereof.

[0152] In another aspect, the present application provides a drug combination comprising a reagent described herein and / or comprising a polypeptide or variant thereof described herein.

[0153] In another aspect, the present application provides a pharmaceutical composition comprising a reagent described herein and / or comprising a polypeptide or variant thereof described herein and a pharma- ceutically acceptable carrier.

[0154] In another aspect, the present application provides the use of a polypeptide or variant thereof described herein, a reagent described herein, a drug combination described herein, and / or a pharmaceutical composition described herein in the preparation of a reagent, wherein said reagent is used for improving learning ability, treating cognitive disorders, and / or treating neurodegenerative diseases.

[0155] In some embodiments, the learning ability comprises a cognitive ability, a motor ability, a memory ability, and / or a spatial exploration ability.

[0156] In some embodiments, the cognitive impairment comprises mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment.

[0157] In some embodiments, the cognitive disorder comprises cognitive disorder caused by normal aging, Lewis dementia (LBD), frontotemporal dementia, and / or vascular dementia.

[0158] In some embodiments, the cognitive impairment-inducing disease comprises Alzheimer's disease, multi-infarct Parkinson's disease, AIDS, and / or Creutzfeldt-Jakob disease (CJD).

[0159] In some embodiments, the neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

[0160] In some embodiments, the neurodegenerative disease comprises a neurodegenerative disease caused by neuronal death and glial cell homeostasis, a neurodegenerative disease caused by aging, a neurodegenerative disease caused by effects on CNS cell function, a neurodegenerative disease caused by abnormal communication between cells, and / or a neurodegenerative disease caused by impaired cell motility.

[0161] In some embodiments, the neurodegenerative disease comprises Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD).

[0162] In some embodiments, the polypeptide, the drug combination and / or the pharmaceutical composition are formulated to be suitable for oral and / or injectable administration.

[0163] In some embodiments, the polypeptide, the drug combination and / or the pharmaceutical composition is formulated to be suitable for intravenous injection.

[0164] In another aspect, the present application provides a non-human animal or part thereof that comprises and / or expresses ATP6V1B2, functionally active fragments thereof and / or nucleic acid molecules encoding them.

[0165] In some embodiments, the ATP6V1B2 is from a human.

[0166] In some embodiments, the ATP6V1B2 comprises the amino acid sequence set forth in SEQ ID NO:8 or 16.

[0167] In some embodiments, the functionally active fragment of ATP6V1B2 has the ability to specifically bind to the amino acid sequence set forth in SEQ ID NO:5.

[0168] In some embodiments, the functionally active fragment of ATP6V1B2 comprises a truncation of ATP6V1B2.

[0169] In some embodiments, the functionally active fragment of ATP6V1B2 comprises at least a portion of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein.

[0170] In some embodiments, the functionally active fragment of ATP6V1B2 comprises at least a portion of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein.

[0171] In some embodiments, the functionally active fragment of ATP6V1B2 comprises the amino acid sequence set forth in SEQ ID NO:10 or SEQ ID NO:11.

[0172] In some embodiments, the nucleic acid molecule comprises the nucleotide sequence set forth in SEQ ID NO:9 or 17.

[0173] In some embodiments, the non-human animal is a gene-edited non-human animal, or is derived from a gene-edited non-human animal.

[0174] In some embodiments, the non-human animal is a gene knock-in non-human animal, or is derived from a gene knock-in non-human animal.

[0175] In some embodiments, expression of the ATP6V1B2 and / or functionally active fragments thereof is regulated by corresponding endogenous regulatory elements in the genome of the non-human animal.

[0176] In some embodiments, the non-human animal is a non-human mammal.

[0177] In some embodiments, the non-human animal is selected from a mouse, a rat, a rabbit, an ape, a monkey, a pig, a cow, a sheep, a horse, a chicken, a dog, an alpaca, and a cat.

[0178] In some embodiments, the non-human animal is a rodent or a primate.

[0179] In some embodiments, the portion comprises an organ, tissue and / or cell of the non-human animal.

[0180] In some embodiments, the portion comprises a bodily fluid.

[0181] In some embodiments, the bodily fluid is selected from the group consisting of blood, plasma, serum, urine, sweat, tears, saliva, semen, and cerebrospinal fluid.

[0182] In some embodiments, the plasma comprises exosomes.

[0183] In some embodiments, the portion comprises a brain or a portion thereof.

[0184] In some embodiments, the part of the brain that is included is selected from the group consisting of the olfactory bulb, amygdala, basal ganglia, hippocampus, thalamus, hypothalamus, cerebral cortex, medulla oblongata, and cerebellum.

[0185] In some embodiments, the portion includes the olfactory mucosa.

[0186] In some embodiments, the portion comprises a portion of the central nervous system, a portion of the peripheral nervous system, skin tissue, muscle tissue, and / or an internal organ.

[0187] In some embodiments, the central nervous system comprises the spinal cord.

[0188] In some embodiments, the portion comprises a cell, and the cell comprises a cell selected from the group consisting of a primary neuron, a small glial cell, an astrocyte glial cell, an oligoglial cell, a macrophage, a perivascular epithelioid cell, a B cell, a T cell, an adult stem cell, a NK cell, a totipotent stem cell, a unipotent stem cell, an embryonic stem cell, an induced pluripotent stem cell, and a gamete.

[0189] In some embodiments, the gametes include sperm and / or oocytes.

[0190] In some embodiments, the portion is incapable of developing into a complete non-human animal.

[0191] In some embodiments, offspring are obtained by hybridizing the non-human animal with another non-human animal, which may or may not express the ATP6V1B2 and / or functionally active fragments thereof.

[0192] In some embodiments, the progeny or portions thereof comprise and / or express the ATP6V1B2 and / or functionally active fragments thereof.

[0193] In another aspect, the present application provides an isolated cell comprising and / or expressing ATP6V1B2, a functionally active fragment thereof and / or the nucleic acid molecules encoding them.

[0194] In some embodiments, the ATP6V1B2 is from a human.

[0195] In some embodiments, the ATP6V1B2 and / or functionally active fragments thereof comprise an amino acid sequence set forth in any one of SEQ ID NOs: 8, 10, 11, and 16.

[0196] In some embodiments, the gene encoding ATP6V1B2 and / or a functionally active fragment thereof is homozygous or heterozygous.

[0197] In some embodiments, the nucleic acid molecule encoding ATP6V1B2 and / or functionally active fragments thereof comprises the nucleic acid sequence set forth in SEQ ID NO:9 or 17.

[0198] In some embodiments, expression of the ATP6V1B2 and / or functionally active fragments thereof is regulated by corresponding endogenous regulatory elements in the cell genome.

[0199] In some embodiments, the target cell is a non-human animal cell.

[0200] In some embodiments, the non-human animal is a non-human mammal.

[0201] In some embodiments, the non-human animal is selected from a mouse, a rat, a rabbit, an ape, a monkey, a pig, a cow, a sheep, a horse, a chicken, a dog, an alpaca, and a cat.

[0202] In some embodiments, the non-human animal is a rodent or a primate.

[0203] In some embodiments, the cells comprise cells selected from the group consisting of neuroblastoma cells, brain glioma cells, primary neurons, small glial cells, astrocytes, oligoglial cells, macrophages, perivascular epithelioid cells, B cells, T cells, adult stem cells, NK cells, totipotent stem cells, unipotent stem cells, embryonic stem cells, induced pluripotent stem cells, and gametes.

[0204] In some embodiments, the gametes include sperm and / or oocytes.

[0205] In some embodiments, the cells are not capable of developing into an entire non-human animal.

[0206] In another aspect, the present application provides tissues that contain and / or express ATP6V1B2, functionally active fragments thereof and / or nucleic acid molecules encoding them.

[0207] In some embodiments, the ATP6V1B2 is from a human.

[0208] In some embodiments, the ATP6V1B2 and / or functionally active fragments thereof comprise an amino acid sequence set forth in any one of SEQ ID NOs: 8, 10, 11, and 16.

[0209] In some embodiments, the gene encoding ATP6V1B2 and / or a functionally active fragment thereof is homozygous or heterozygous.

[0210] In some embodiments, the nucleic acid molecule encoding ATP6V1B2 and / or functionally active fragments thereof comprises the nucleic acid sequence set forth in SEQ ID NO:9 or 17.

[0211] In some embodiments, the tissue is a gene-edited tissue or is derived from a gene-edited tissue.

[0212] In some embodiments, the tissue is a gene knock-in tissue or is derived from a gene knock-in tissue.

[0213] In some embodiments, expression of the ATP6V1B2 and / or functionally active fragments thereof is regulated by corresponding endogenous regulatory elements in the tissue genome.

[0214] In some embodiments, the tissue is a non-human animal tissue.

[0215] In some embodiments, the non-human animal is a non-human mammal.

[0216] In some embodiments, the non-human animal is selected from a mouse, a rat, a rabbit, an ape, a monkey, a pig, a cow, a sheep, a horse, a chicken, a dog, an alpaca, and a cat.

[0217] In some embodiments, the non-human animal is a rodent or a primate.

[0218] In some embodiments, the tissue comprises a bodily fluid.

[0219] In some embodiments, the bodily fluid is selected from the group consisting of blood, plasma, serum, urine, sweat, tears, saliva, semen, and cerebrospinal fluid.

[0220] In some embodiments, the plasma comprises exosomes.

[0221] In some embodiments, the tissue comprises the brain or a portion thereof.

[0222] In some embodiments, the part of the brain that is included is selected from the group consisting of the olfactory bulb, amygdala, basal ganglia, hippocampus, thalamus, hypothalamus, cerebral cortex, medulla oblongata, and cerebellum.

[0223] In some embodiments, the tissue comprises the olfactory mucosa.

[0224] In some embodiments, the tissue comprises a portion of the central nervous system, a portion of the peripheral nervous system, skin tissue, muscle tissue, and / or an internal organ.

[0225] In some embodiments, the central nervous system comprises the spinal cord.

[0226] In some embodiments, the tissue is not capable of developing into an entire non-human animal.

[0227] In another aspect, the present application provides a cell line or cell culture derived from a non-human animal or part thereof described herein, derived from a progeny described herein, derived from a cell described herein, and / or derived from a tissue described herein.

[0228] In some embodiments, the cell culture is a primary cell culture.

[0229] In some embodiments, the cell culture comprises an organoid.

[0230] In another aspect, the present application provides a composition comprising a non-human animal or part thereof described herein, a progeny thereof described herein, a cell described herein, a tissue described herein, and / or a cell line or primary cell culture described herein.

[0231] In another aspect, the present application provides a reagent kit comprising a non-human animal described herein or a part thereof, a progeny described herein, a cell described herein, a tissue described herein, and / or a cell line or primary cell culture described herein, and one or more additional components selected from the group consisting of assay buffers, controls, substrates, standards, detection materials, laboratory supplies, equipment, devices, cells, organs, tissues, and user manuals or instructions.

[0232] In another aspect, the present application provides a method for screening for drugs that can improve learning ability, treat cognitive impairment, and / or treat a neurodegenerative disease in a subject, comprising administering a candidate drug to a non-human animal or part thereof described herein, a progeny described herein, a cell described herein, a tissue described herein, and / or a cell line or primary cell culture described herein, and measuring the effect of the candidate drug on the expression level and / or activity of ATP6V1B2 in the subject.

[0233] In another aspect, the present application provides a method for screening for a drug that can improve learning ability, treat cognitive impairment, and / or treat a neurodegenerative disease in a subject, the method comprising: (i) administering a candidate drug to a non-human animal or part thereof described herein, a progeny described herein, a cell described herein, a tissue described herein, and / or a cell line or primary cell culture described herein; (ii) measuring the effect of the candidate drug on the expression level and / or activity of ATP6V1B2 in the subject; and (iii) if the expression level and / or activity of ATP6V1B2 is improved after administration of the candidate drug, the candidate drug can improve learning ability, treat cognitive impairment, and / or treat a neurodegenerative disease in the subject.

[0234] In some embodiments, the expression level of ATP6V1B2 comprises the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene and / or the expression level of the ATP6V1B2 protein.

[0235] In some embodiments, the improvement comprises an increase in the expression level and / or activity of ATP6V1B2 of at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject.

[0236] In some embodiments, the learning ability comprises a cognitive ability, a motor ability, a memory ability, and / or a spatial exploration ability.

[0237] In some embodiments, the cognitive impairment comprises mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment.

[0238] In some embodiments, the cognitive disorder comprises cognitive disorder caused by normal aging, Lewis dementia (LBD), frontotemporal dementia, and / or vascular dementia.

[0239] In some embodiments, the cognitive impairment-inducing disease comprises Alzheimer's disease, multi-infarct Parkinson's disease, AIDS, and / or Creutzfeldt-Jakob disease (CJD).

[0240] In some embodiments, the neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

[0241] In some embodiments, the neurodegenerative disease comprises a neurodegenerative disease caused by neuronal death and glial cell homeostasis, a neurodegenerative disease caused by aging, a neurodegenerative disease caused by effects on CNS cell function, a neurodegenerative disease caused by abnormal communication between cells, and / or a neurodegenerative disease caused by impaired cell motility.

[0242] In some embodiments, the neurodegenerative disease comprises Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD).

[0243] In another aspect, the present application provides the use of a non-human animal described herein or a part thereof, a progeny described herein, a cell described herein, a tissue described herein, and / or a cell line or primary cell culture described herein in the manufacture of an identification and / or screening system, wherein said system is used to screen for drugs and / or biomarkers that allow for improving learning ability, treating cognitive disorders, and / or treating neurodegenerative diseases in a subject.

[0244] In another aspect, the present application provides a non-human animal or part thereof described herein, a progeny described herein, a cell described herein, a tissue described herein, and / or a cell line or primary cell culture described herein for use in screening for drugs and / or biomarkers that can improve learning ability, treat a cognitive disorder, and / or treat a neurodegenerative disease in a subject.

[0245] In another aspect, the present application provides a method for preparing a non-human animal or part thereof as described herein, a cell as described herein, or a tissue as described herein, comprising causing the non-human animal or part thereof, cell or tissue to contain and / or express said ATP6V1B2 and / or a functionally active fragment thereof.

[0246] In another aspect, the present application provides a method of producing a disease model, the method comprising comprising comprising containing and / or expressing ATP6V1B2 and / or a functionally active fragment thereof in a non-human animal or a part, cell or tissue thereof.

[0247] In some embodiments, the ATP6V1B2 and / or functionally active fragments thereof comprise an amino acid sequence set forth in any one of SEQ ID NOs: 8, 10, 11, and 16.

[0248] In some embodiments, the method comprises transfecting a nucleic acid sequence encoding the ATP6V1B2 and / or a functionally active fragment thereof.

[0249] In some embodiments, the nucleic acid sequence encoding ATP6V1B2 and / or functionally active fragments thereof comprises the nucleic acid sequence set forth in SEQ ID NO:9 or SEQ ID NO:17.

[0250] In some embodiments, the transfecting comprises transfecting an adeno-associated virus comprising the nucleic acid sequence.

[0251] Those skilled in the art can easily discern other aspects and advantages of the present application from the following detailed description. The following detailed description shows and describes only exemplary embodiments of the present application. As will be understood by those skilled in the art, the contents of the present application allow those skilled in the art to make changes to the specific embodiments disclosed without departing from the spirit and scope of the invention covered by the present application. Correspondingly, the drawings and descriptions in the present application are illustrative only and not limiting. [Brief description of the drawings]

[0252] Particular features of the invention are set forth in the appended claims. A better understanding of the features and advantages of the invention can be obtained by reference to the exemplary embodiments and drawings described in detail below. A brief description of the drawings is given below. [Figures 1a-1i] 1 shows the effect of the polypeptides described herein on excitatory synaptic transmission. [Diagram 2] It has been shown that the polypeptides described in this application can improve cognitive behavior in a novel object discrimination test in AD mice by oral gavage. [Diagram 3] It has been shown that the polypeptides described in the present application can improve the learning ability in the water maze test in AD mice by oral gavage. [Figure 4]It has been shown that the polypeptides described in this application can improve cognitive behavior in a novel object discrimination test in AD mice by intravenous administration. [Diagram 5] It is shown that the polypeptide described in the present application can improve the learning ability in the water maze test in AD mice by intravenous administration. [Figure 6a-6b] 1 shows the results of co-immunoprecipitation of a polypeptide described in the present application with ATP6V1B2 protein. [Figure 7a-7e] We show that overexpression of ATP6V1B2 protein in the hippocampus of aged animals can improve learning and memory function. [Figure 8] 1 shows the effect of a dimer of the polypeptide described herein on excitatory synaptic transmission. [Figure 9] 1 shows the effect of the cysteine-modified polypeptides described herein on excitatory synaptic transmission. [Figure 10] 1 shows the effect of the serine-modified polypeptides described herein on excitatory synaptic transmission. [Figure 11] 1 shows the results of co-immunoprecipitation of a polypeptide described in the present application with ATP6V1B2 protein or a truncated fragment thereof. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0253] Hereinafter, the embodiments of the present invention will be described with reference to specific examples, and those skilled in the art will easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0254] Definition of Terms The term "ATP6V1B2" is usually used to refer to the ATPase H +It refers to ATPase H+transporting V1 subunit B2 protein (also called ATP6B2, DOOD, HO57, VATB, VPP3, Vma2 or ZLS2) and the gene encoding the protein. The ATP6V1B2 may be a multi-subunit enzyme that mediates the acidification of organelles in eukaryotic cells. The ATP6V1B2 may be involved in processes such as protein sorting, zymogen activation, receptor-mediated endocytosis and synaptic vesicle proton gradient generation. The ATP6V1B2 protein may include a cytoplasmic V1 domain and a transmembrane V0 domain. The accession number of human ATP6V1B2 in GenBank is 526. The accession number of human ATP6V1B2 in UniProt may be P21281.

[0255] The term "expression level" generally refers to the protein, RNA or mRNA level of a particular relevant gene. Any method known in the art can be used to measure the expression level of a relevant gene (e.g., human ATP6V1B2 gene). In this application, the "expression" generally refers to the process of converting the information encoded by a gene into a structure that exists and functions in a cell. For example, it can include reverse transcription and amplification analysis (e.g., PCR, RT-PCR linkage or RT-PCR quantification), hybridization analysis, RNA blotting (Northern blotting), dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining or mass spectrometry. It can be analyzed directly on a biological sample or on proteins / nucleic acids isolated from the sample.

[0256] The term "activity" generally refers to any activity associated with a particular protein. In the present application, the activity can include, for example, any activity associated with the ATP6V1B2 protein. The activity can include enzymatic activity associated with a protease. In some cases, the activity can include biological activity. In some cases, the activity can include binding of a protein to a receptor, for example, where the binding can result in a measurable downstream effector. In the present application, the activity can include any activity that can be attributed to the protein by a person skilled in the art.

[0257] The term "proton pump modulator" generally refers to an agent capable of modulating proton pump activity and / or function.

[0258] The term "proton pump agonist" generally refers to an agent that activates the proton pump. For example, the proton pump agonist can increase the activity of the hydrogen ion pump.

[0259] The term "vesicular proton pump agonist" generally refers to a reagent that can improve the activity of vesicular proton pumps. Vesicular or vacuolar adenosine triphosphatase (V-ATPase) is an ATP-driven proton pump, composed of a cytosolic V1 complex used for ATP hydrolysis and a membrane-embedded Vo complex used for proton translocation. The vesicular proton pump can play an important role in the acidification of vesicles, organelles and extracellular environments in eukaryotic cells.

[0260] The term "proton pump-related protein" generally refers to a related protein that codes for and / or expresses a proton pump. The proton pump may be a protein that actively transports hydrogen ions by the electrochemical potential difference of hydrogen ions across the opposite membrane in a biological membrane. The proton pump may be a Na-K pump, a Ca-K pump, or a Ca-K pump. 2+ Pump, H + -ATP pump and H +It may include a pyrophosphate pump.

[0261] The term "learning ability" generally refers to any ability related to or necessary for learning / cognitive processes. The learning ability may include the ability to acquire new information, knowledge and / or skills by processes including experience, learning or training. The learning ability may include imagination, attention, perceptual observation, reading comprehension, analytical ability, operational ability, adaptive ability, summarizing ability, problem-solving ability or a combination thereof.

[0262] The term "cognitive ability" generally refers to the ability to process information through perception. The cognitive ability may include the ability to grasp the structure of things, the relationship between performance and other things, the motivation for development, the direction of development, and basic laws.

[0263] The term "athletic ability" generally refers to the ability to participate in exercise and training. The athletic ability may include aerobic ability, muscle strength, physical flexibility, balance ability, and reaction ability. The athletic ability may be a comprehensive expression of various elements such as body type, endowment, technique, skill, and mental ability.

[0264] The term "memory ability" generally refers to the ability to remember, retain, re-understand, and reproduce content or experiences reflected in objective things. The memory ability can include sensory memory ability, short-term memory ability, and long-term memory ability.

[0265] The term "spatial exploration ability" generally refers to the ability to explore the shape and / or position of an object. The spatial exploration ability refers to observing, thinking, imagining, recognizing and / or exploring the shape and / or position of an object.

[0266] The term "learning ability assessment score" generally refers to a quantitative assessment score of the learning ability of a subject. The learning ability assessment score can be obtained by the score obtained from attention / executive function assessment (e.g., Wechsler memory test), language ability assessment (e.g., the language screening test (LAST)), visual-spatial and structural ability assessment (e.g., visual-motor integration test, Hooper visual organization test, object assembly test, figure arrangement test, clock drawing test), operational ability assessment, daily function assessment (e.g., disability assessment for dementia (DAD)) and / or neuropsychological scale. In some cases, the learning ability assessment score can be obtained by the score obtained from the mini-mental state examination (MMSE), Montreal cognitive assessment (MoCA), Alzheimer disease assessment scale-cognitive part (ADAS-cog) and clinical dementia rating scale (CDR).

[0267] The term "novel object recognition test" usually refers to a test that detects the time it takes for an animal (e.g., a mouse) to learn to distinguish a new object by having the animal distinguish the object in a specific space. In some cases, the novel object recognition test can refer to the test method described in Ennaceur et al., Behav Brain Res 80 9-25, 1996. For example, the novel object recognition test can include the steps of putting two identical objects into a container with a certain volume, putting a mouse into the container to distinguish the two objects, and replacing one of the two objects in the same container with a new object with a different shape the next day, and then measuring the time it takes the mouse to search for the new object.

[0268] The term "water maze test" generally refers to a test in which an animal (e.g., a mouse) is forced to swim to learn how to find a platform hidden in water. The water maze test (e.g., Morris water maze) can test the learning ability and / or memory ability of a mouse with respect to spatial position sense and direction sense. The water maze test can also include acquired training, exploration training, contrapositional training, or contrapositional exploration training. The shorter the time it takes an animal (e.g., a mouse) to find the platform after entering the water and the shorter the distance it travels during this time, the higher the evaluation score of the learning ability of the animal (e.g., a mouse). The water maze evaluation test can be an important experiment to evaluate learning ability.

[0269] The term "neurodegenerative disease" generally refers to cognitive impairments such as dementia resulting from the progressive loss of neuronal structure and function, including neuronal death and glial cell balance. In some cases, age (e.g., Alzheimer's disease (AD), Parkinson's disease (PD)) or genetic mutations affecting CNS cell function (e.g., Huntington's disease, early-onset AD or PD, amyotrophic lateral sclerosis (ALS)) can cause the neurodegenerative disease. The neurodegenerative disease can have changes and / or symptoms selected from protein misfolding and aggregation, neuroinflammation (e.g., CNS inflammation due to signaling stimuli such as toxic stimuli (e.g., protein aggregation), infection, traumatic injury, or autoimmunity), cell signaling changes, acquired aging / cell death (e.g., disruption of apoptotic signaling, mitochondrial dysfunction, impaired autophagy, and necrosome stress / activation of inflammation), motor cell injury, and epigenetic changes.

[0270] The term "Alzheimer's disease" usually refers to early-onset dementia or senile dementia, which is a neurodegenerative disease that develops slowly and worsens over time. The most common early symptom is short-term memory loss (difficulty remembering recent events), and as the disease progresses, at least one of the following symptoms may appear: speech impairment, disorientation (e.g., getting lost easily), mood lability, loss of motivation, inability to take care of oneself, and behavioral problems. The true cause of Alzheimer's disease remains unknown, and its progression may be related to the deposition of fibrillar amyloid protein plaques and Tau protein in the brain. Currently, there is no treatment that can stop or reverse the course of the disease, but there are some treatments that can temporarily alleviate or improve symptoms.

[0271] The term "mild cognitive impairment (MCI)" generally refers to an intermediate clinical state between normal cognition and cognitive impairment. In some cases, the MCI can include cognitive impairment that meets the criteria for dementia but is more advanced than normal aging. MCI is diverse in clinical symptoms, etiology, prognosis and prevalence. In some cases, MCI may be a pathological stage of Alzheimer's disease. Some forms of cognitive impairment are considered early symptoms of neurodegenerative diseases that ultimately lead to dementia.

[0272] The term "cognitive impairment caused by normal aging" generally refers to cognitive damage caused by normal aging. For example, said cognitive impairment caused by normal aging can be manifested as memory loss, confusion regarding the location of familiar places, taking longer than usual to complete everyday tasks, or changes in mood or personality.

[0273] The term "Lewy Body Dementia (LBD)" usually refers to Lewy Body Dementia. Lewy body dementia is characterized by an abnormal accumulation of proteins that form clumps called Lewy bodies. Lewy body dementia causes a gradual decline in mental abilities. Patients with Lewy body dementia may experience hallucinations and changes in alertness and attention. Other effects include muscle stiffness, slowness of movement, difficulty walking, and tremors. Patients with Lewy bodies in the brain may also develop plaques and tangles associated with Alzheimer's disease.

[0274] The term "frontotemporal dementia" usually refers to Pick's disease, a rare progressive disorder in which the tau protein affects only the frontal and temporal lobes of the brain. People with frontotemporal dementia have difficulty with higher order reasoning, expressive language, recognizing language, and forming memories. The frontal and temporal lobes of the brain in people with frontotemporal dementia may shrink over time.

[0275] The term "vascular dementia" generally refers to problems with reasoning, judgment and memory caused by impaired blood flow to the brain. For example, vascular dementia can include dementia due to risk factors for heart disease and stroke, such as high blood pressure and high cholesterol.

[0276] The term "multiple infarct type" usually refers to small non-cortical infarcts caused by the occlusion of a single perforator branch of a large cerebral artery.The multiple infarct type may be a special type of cerebral infarction, also called ischemic stroke.The multiple infarct type may manifest as hemisensory disturbance, aphasia, dysarthria, slowness of movement, clumsiness (especially difficulty with fine movements such as writing).

[0277] The term "Parkinson's disease" generally refers to a progressive neurodegenerative disease. The clinical features of Parkinson's disease (PD) can include motor symptoms (e.g., tremor, bradykinesia, muscle rigidity and postural instability), as well as neuropsychiatric and other non-motor symptoms. For example, the non-motor symptoms can include cognitive impairment and dementia, mood disorders (e.g., depression, anxiety, apathy) and sleep disorders.

[0278] The term "AIDS" usually refers to Acquired Immune Deficiency Syndrome (AIDS). Clinical symptoms of AIDS include changes in memory, concentration, attention, and motor skills. In some cases, cognitive impairment occurs in AIDS patients, for example, approximately 50% of infected individuals may also develop HIV-associated neurocognitive disorders (HAND).

[0279] The term "Creutzfeldt-Jakob disease (CJD)" usually refers to a type of transmissible spongiform encephalopathy that occurs in humans. CJD is a disease caused by a prion infection. CJD patients may develop paranoid behavior, confusion, loss of appetite and weight, and depression; a small number of patients present with visual or hearing abnormalities, and in the progressive stages, progressive neurological deterioration (e.g., paresthesia, speech impairment, aphasia).

[0280] The term "multiple sclerosis (MS)" generally refers to demyelinated neurological lesions. When the insulating material (i.e., myelin sheath) on the surface of nerve cells in the brain or spinal cord of MS patients is destroyed and signaling in the nervous system is impaired, it causes a series of possible symptoms that affect the patient's activity, psyche, and even mental state. These symptoms can include double vision, loss of vision on one side, muscle weakness, reduced sensation, or impaired coordination.

[0281] The term "amyotrophic lateral sclerosis (ALS)" generally refers to amyotrophic lateral sclerosis, a motor neuron disease that is a progressive and fatal neurodegenerative disease. A small number of ALS patients may develop frontotemporal dementia. Some ALS patients experience a decrease in sensation, vision, touch, smell and taste, and a very small number of amyotrophic lateral sclerosis patients develop dementia at the same time.

[0282] The term "Huntington's disease (HD)" or Huntington's chorea usually refers to a genetic disorder that causes brain cell death. As the disease progresses, HD patients experience a gradual decline in abilities as the coordination of their body movements becomes more apparent, eventually making movement difficult and making speech impossible. Mental abilities often decline and lead to dementia.

[0283] The term "elderly stage" generally refers to the aging stage of a subject. For example, in the case of a human, the elderly stage may be 60 years or older, 70 years or older, or 75 years or older, and in the case of a mouse, the elderly stage may be 10 months or older, such as 13 months or older, or 18 months or older. In some cases, the elderly stage subject may suffer from one or more symptoms of learning deficiency, memory impairment, memory deficiency, and / or brain dysfunction.

[0284] The term "variant" generally refers to a polypeptide comprising an amino acid sequence that differs from the amino acid sequence of a parent or reference polypeptide (e.g., a wild-type polypeptide) by at least one amino acid residue. In the present application, the variant can have a relatively high (e.g., at least 80%) homology to the parent or reference polypeptide. The homology can include sequence similarity or identity. In the present application, the homology can be determined by standard techniques known in the art (see, e.g., Smith and Waterman, Adv. Appl. Math. Advances in Applied Mathematics), and the percentage of identity shared by polynucleotide or polypeptide sequences is measured by direct comparison of intermolecular sequence information, which is determined by sequence alignment, using methods known in the art. An example of an algorithm suitable for determining sequence similarity is the BLAST algorithm (see, e.g., Altschul et al., J. Mol. Biol., 215:403-410

[1990] ). Software used for BLAST analyses is publicly available from the National Center for Biotechnology Information (NCBI).

[0285] The term "transmitter release" generally refers to the release of neurotransmitters, that is, the process in which a neuron releases vesicles-encapsulated neurotransmitters into the synaptic gap to act on another neuron, thereby transmitting information. The transmitter release process may involve synapses, which are the basic structures of neural circuits. In some cases, the transmitter release may be referred to as synaptic transmission. The transmitter release method may include synchronous release, asynchronous release, and spontaneous release.

[0286] The term "discharge frequency" usually refers to the frequency of action potential discharge. Action potentials can refer to the process of rapid and reversible reversal and recovery of the potential across the membrane generated from the resting potential when an excitable cell is stimulated. Action potentials consist of a peak potential and an afterpotential, which correspond to the processes of depolarisation and hyperpolarisation, respectively. The firing of action potentials can have a pulsed character. In some cases, the frequency of pulse firing is the ratio of the number of pulses fired to time.

[0287] In this application, the term "neuron" generally refers to a nerve cell, which is the primary functional unit of the nervous system. A neuron consists of a cell body and its neurites, an axon, and one or more dendrites. Neurons can send messages to other neurons or cells by releasing neurotransmitters at synapses.

[0288] The term "multimer" generally refers to a molecule having two or more polypeptide chains associated by covalent bonds, non-covalent bonds, or both covalent and non-covalent interactions. The multimer can include a dimer.

[0289] The term "homodimer" generally refers to a molecule formed from two identical monomers that are capable of aggregating, complexing, or associating with one another through covalent and / or non-covalent interactions.

[0290] The term "mercapto blocking" generally refers to blocking of free mercapto so that intramolecular and / or intermolecular disulfide bonds are less likely to form. In the present application, the mercapto blocking can occur at cysteine ​​residues. The mercapto blocking prevents disulfide bonds from forming between cysteine ​​residues in proteins, thus preventing cross-linking or modification of proteins. The mercapto blocking can be achieved by using a blocking agent that is a reducing agent. The blocking agent can include dithiothreitol (DTT), β-mercaptoethanol (BME), and tris(2-carboxyethyl)phosphine hydrochloride (TCEP·HCl).

[0291] The term "serine phosphorylation" generally refers to a phosphorylation modification that occurs at a serine residue. The serine phosphorylation may be a process in which a phosphate group of a donor (e.g., ATP or GTP) is transferred to a serine residue. The serine phosphorylation may be mediated by a protein kinase. The serine phosphorylation may cause a change in protein activity.

[0292] The term "about" generally refers to a numerical range of 20% more or less than a particular value. For example, "about X" includes numerical ranges of ±20%, ±10%, ±5%, ±2%, ±1%, ±0.5%, ±0.2%, or ±0.1% of X, where X is the numerical value.

[0293] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT In one aspect, the present application provides the use of a proton pump modulator in the preparation of a reagent for use in improving learning ability.

[0294] In another aspect, the present application provides the use of a proton pump modulator in the preparation of a reagent for use in the treatment of a cognitive disorder.

[0295] In another aspect, the present application provides the use of a proton pump modulating agent in the preparation of a reagent for use in the treatment of a neurodegenerative disease.

[0296] In another aspect, the present application provides the use of an ATP6V1B2 modulator in the preparation of a reagent for use in improving learning ability.

[0297] In another aspect, the present application provides the use of an ATP6V1B2 modulator in the preparation of a reagent for use in the treatment of a cognitive disorder.

[0298] In another aspect, the present application provides the use of an ATP6V1B2 modulating agent in the preparation of a reagent for use in the treatment of a neurodegenerative disease.

[0299] In the present application, when the expression level and / or activity of ATP6V1B2 in the subject is improved, the learning ability of the subject can be significantly improved (e.g., the cognitive ability, motor ability, memory ability and / or spatial exploration ability can be significantly improved compared to before the expression level and / or activity of ATP6V1B2 in the subject is improved). Therefore, ATP6V1B2 can be a potential target for improving learning ability. For example, ATP6V1B2 can be a potential target for treating neurodegenerative diseases (e.g., Alzheimer's disease) and / or cognitive disorders.

[0300] Increased expression level and / or activity of proton pump-related protein, ATP6V1B2 In the present application, the reagent is capable of increasing the expression level and / or activity of ATP6V1B2 in a subject.

[0301] In the present application, the proton pump regulator can increase the expression level and / or activity of ATP6V1B2 in a subject.

[0302] In the present application, the ATP6V1B2 regulator is capable of increasing the expression level and / or activity of ATP6V1B2 in a subject.

[0303] In the present application, the expression level of ATP6V1B2 and / or its functionally active fragments can include the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene, and / or the expression level of the ATP6V1B2 protein and / or its functionally active fragments. For example, the expression level can include the amount of a specific gene (e.g., human ATP6V1B2 gene) polynucleotide, mRNA, or amino acid product or protein. The expression level can include the amount of a polynucleotide transcribed by a specific gene (e.g., human ATP6V1B2 gene), a translated protein, or a fragment of a post-translationally modified protein.

[0304] In the present application, the improvement can include an improvement in the expression level of ATP6V1B2 by at least about 10% compared to the original expression level of ATP6V1B2 in the subject, for example, an improvement of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0305] In the present application, the expression level of ATP6V1B2 can be measured by using a substance selected from the group consisting of primers that specifically amplify the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 protein, a small molecule that specifically binds to the ATP6V1B2 protein, a probe that specifically binds to the ATP6V1B2 protein, and a polypeptide that specifically binds to the ATP6V1B2 protein.

[0306] In the present application, the expression level of the ATP6V1B2 can be measured by performing a test selected from the group consisting of reverse transcription and amplification analysis (e.g., PCR, RT-PCR linked or RT-PCR quantification), hybridization analysis, RNA blotting (Northern blotting), dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining or mass spectrometry. For example, the expression level of the ATP6V1B2 described in the present application can be measured by qPCR, qRT-PCR, northern hybridization, western hybridization and / or ELISA detection. The expression level of the ATP6V1B2 can also be measured by direct analysis of a biological sample or a protein / nucleic acid isolated from the sample.

[0307] In the present application, the ATP6V1B2 may be derived from a human or a mouse.

[0308] In the present application, the ATP6V1B2 may comprise the amino acid sequence shown in SEQ ID NO:8 or SEQ ID NO:16.

[0309] In the present application, the functionally active fragment of ATP6V1B2 can have the ability to specifically bind to the amino acid sequence shown in SEQ ID NO: 5. For example, the functionally active fragment of ATP6V1B2 can include a truncated form of ATP6V1B2. For example, the functionally active fragment of ATP6V1B2 can include at least a portion of the sequence of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein. For example, the functionally active fragment of ATP6V1B2 can include at least a portion of the sequence of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein.

[0310] In the present application, the ATP6V1B2 and / or a functionally active fragment thereof may comprise an amino acid sequence as shown in any one of SEQ ID NOs: 8, 10, 11, and 16.

[0311] In the present application, the nucleic acid sequence encoding the ATP6V1B2 and / or a functionally active fragment thereof may comprise the nucleic acid sequence shown in SEQ ID NO:9 or SEQ ID NO:17.

[0312] In the present application, the activity of ATP6V1B2 can include the biological activity of the ATP6V1B2 protein and / or its functionally active fragment (e.g., can include a measurable downstream effector caused thereby). For example, the activity of ATP6V1B2 can include an increase in the expression level and / or activity of the proton pump-related protein.

[0313] In the present application, the improvement can include an improvement in the activity of the ATP6V1B2 by at least about 10% compared to the original activity of ATP6V1B2 in the subject, such as an improvement of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0314] In the present application, the reagent is capable of increasing the expression level and / or activity of a proton pump-associated protein in a subject.

[0315] In the present application, the proton pump regulator can increase the expression level and / or activity of a proton pump-associated protein in a subject.

[0316] In the present application, the ATP6V1B2 regulator can increase the expression level and / or activity of a proton pump-related protein in a subject.

[0317] In the present application, the expression level of the proton pump-associated protein can include the expression level of a gene encoding a proton pump-associated protein, the transcription level of a gene encoding a proton pump-associated protein, and / or the expression level of a proton pump-associated protein. For example, the expression level can include the amount of a polynucleotide, mRNA, or amino acid product or protein of a particular gene (e.g., a gene encoding a proton pump-associated protein). The expression level can include the amount of a polynucleotide, a translated protein, or a post-translationally modified protein fragment transcribed by a particular gene (e.g., a gene encoding a proton pump-associated protein).

[0318] In the present application, the improvement can include an improvement of the expression level of the proton pump associated protein by at least about 10% compared to the expression level of the original proton pump associated protein in the subject, for example, an improvement of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0319] In the present application, the expression level of the proton pump-associated protein can be measured by using a substance selected from the group consisting of primers capable of specifically amplifying a gene encoding the proton pump-associated protein, a nucleic acid molecule that specifically binds to a gene encoding the proton pump-associated protein, a nucleic acid molecule that specifically binds to the proton pump-associated protein, a small molecule that specifically binds to the proton pump-associated protein, a probe that specifically binds to the proton pump-associated protein, and a polypeptide that specifically binds to the proton pump-associated protein.

[0320] In the present application, the expression level of the proton pump-associated protein can be measured by performing a test selected from the group consisting of reverse transcription and amplification analysis (e.g., PCR, RT-PCR linked or RT-PCR quantification), hybridization analysis, RNA blotting (Northern blotting), dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining or mass spectrometry. For example, the expression level of the proton pump-associated protein described in the present application can be measured by qPCR, qRT-PCR, northern hybridization, western hybridization and / or ELISA detection. The expression level of the proton pump-associated protein can also be measured by direct analysis of a biological sample or a protein / nucleic acid isolated from the sample.

[0321] In the present application, the proton pump-related proteins may include NADH dehydrogenase, coenzyme Q, succinate-coenzyme Q reductase, cytochrome c, and / or coenzyme Q-cytochrome c reductase.

[0322] In the present application, the activity of the proton pump-associated protein may include the biological activity of the proton pump-associated protein.

[0323] The activity of the proton pump-related proteins is controlled by the hydrogen / potassium adenosine triphosphate enzyme system (also called hydrogen / potassium ion ATP enzyme, i.e. H + / K +ATPase) activity level and / or H2 receptor activity level. In the present application, the improvement of the activity of the proton pump-associated protein may include an improvement of the activity of the proton pump-associated protein by at least about 10% compared to the activity of the original proton pump-associated protein in the subject. For example, the improvement may include an improvement of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0324] Learning Ability In the present application, the learning ability may include any ability related to or requiring a learning / cognitive process. For example, the learning ability may include cognitive ability, motor ability, memory ability, and / or spatial exploration ability.

[0325] In the present application, the improvement of the learning ability may include an improvement of at least about 50% in the evaluation score of the improved learning ability of the subject compared to the evaluation score of the original learning ability of the subject, for example, an improvement of at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0326] In the present application, the assessment score of the learning ability can be obtained by a score obtained in an attention / executive function assessment (e.g., Wechsler Memory Test), a language ability assessment (e.g., the language screening test (LAST)), a visuospatial and structural ability assessment (e.g., the visual-motor integration test, the Hooper visual organization test, the object binding test, the figure arrangement test, the clock drawing test), an operational ability assessment, an everyday function assessment (e.g., the disability assessment for dementia (DAD)), and / or a neuropsychological scale. For example, the assessment score of the learning ability can be measured by performing a test selected from the group consisting of a new object recognition test and a water maze test.

[0327] In the present application, the new object recognition test can evaluate the cognitive ability, motor ability and / or spatial exploration ability. In the present application, in the new object recognition test, a subject (e.g., a mouse) can explore and learn a specific shape of object in a fixed container, and can distinguish a new object that is different in shape from the previous object that is newly displayed in the fixed container after a few days based on the memory obtained by learning. The shorter the time that the mouse takes to distinguish the new object with a different shape after a few days, the higher the learning ability of the mouse is correspondingly. The water maze evaluation test can be an important experiment to evaluate the learning ability of a subject.

[0328] In the present application, the water maze test can evaluate the memory ability, motor ability and / or spatial exploration ability. In the present application, the water maze test (e.g., Morris water maze) is forced to swim so that the subject (e.g., mouse) learns how to find a platform hidden in water, and further tests the spatial exploration ability and / or memory ability of the mouse's sense of spatial position and sense of direction. The water maze test can include acquired training, exploration training, contrapositional training or contrapositional exploration training. The shorter the time from when the mouse enters the water to when it finds the platform, and the shorter the distance it moves during this period, the higher the learning ability of the mouse is correspondingly. The water maze evaluation test can be an important experiment to evaluate the learning ability of a subject.

[0329] Target and indications In the present application, the subject can include a mammal. For example, the subject can include a rodent and / or a primate. For example, the subject can include a human.

[0330] In the present application, the subject can include non-cognitive disorder patients and / or non-neurodegenerative disease patients.For example, the subject can be a normal human and / or a healthy human.For example, the subject can have a need and / or a desire to further improve its learning ability.

[0331] In the present application, the neurodegenerative disease can include acute neurodegenerative disease and chronic neurodegenerative disease.For example, the neurodegenerative disease can include neurodegenerative disease caused by neuronal death and glial cell homeostasis, neurodegenerative disease caused by aging, neurodegenerative disease caused by influence on CNS cell function, neurodegenerative disease caused by abnormal communication between cells and / or neurodegenerative disease caused by impaired cell movement.

[0332] In the present application, the subject may include a patient with a neurodegenerative disease. For example, the subject may include a patient with Alzheimer's disease. For example, the patient with Alzheimer's disease may be in the early, early, middle or late stage of Alzheimer's disease.

[0333] In the present application, the cognitive impairment can include mild cognitive impairment (MCI), intermediate cognitive impairment and late cognitive impairment. For example, the cognitive impairment can include cognitive impairment caused by normal aging, Lewis' physical dementia (LBD), frontotemporal dementia and / or vascular dementia. For example, the cognitive impairment-inducing disease can include Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and / or Creutzfeldt-Jakob disease (CJD).

[0334] In the present application, the subject may include a patient with cognitive impairment. For example, the subject may suffer from mild cognitive impairment (MCI) (e.g., loss of short-term memory, difficulty expressing or understanding abstract things, unstable mood or behavior, difficulty learning new things or following complex instructions, impaired judgment and / or needing reminders from others for basic self-care), mid-stage cognitive impairment (e.g., confusion between long-term memory and memory of real-life situations, lack of ability to communicate clearly, behavioral or personality changes or unstable mood and / or needing help from others for self-care), or late-stage cognitive impairment (e.g., memory impairment, reduced physical activity or mental state, inability to express or communicate effectively, inability to take care of oneself, and / or abnormalities in the body clock). In the present application, the subject may suffer from a disease that can induce the cognitive impairment. For example, the subject may suffer from Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS, and / or Creutzfeldt-Jakob disease (CJD).

[0335] In the present application, the subject can be in the elderly stage.For example, the subject shows cognitive impairment caused by normal aging.For example, the subject shows symptoms of mild cognitive impairment (MCI).For example, the subject shows symptoms of neurodegenerative disease (for example, Alzheimer's disease).

[0336] Polypeptides or variants thereof, reagents and pharmaceutical compositions and uses thereof In the present application, the proton pump regulator may include a proton pump agonist.

[0337] In the present application, the proton pump regulator may include a vesicular proton pump agonist.

[0338] In the present application, the proton pump regulator may include an agonist of the ATP6V1B2 protein.

[0339] In the present application, the proton pump regulator can bind to ATP6V1B2 protein. For example, the proton pump regulator can include a ligand of ATP6V1B2 protein. For example, the proton pump regulator can include an agonist of ATP6V1B2 protein. For example, the proton pump regulator can specifically bind to a specific position (e.g., a specific epitope) in the ATP6V1B2 protein.

[0340] In the present application, the proton pump modulating agent can include a nucleic acid or a variant thereof, a polypeptide or a variant thereof, and / or a small molecule.

[0341] In the present application, the proton pump modulator is capable of increasing the release of synaptic neurotransmitters.

[0342] As used herein, the increase includes an increase of at least about 10% compared to the original synaptic neurotransmitter release level in the subject, for example, an increase of at least about 20%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0343] In the present application, the proton pump regulator is capable of increasing the discharge frequency of excitatory postsynaptic currents.

[0344] In the present application, the increase includes an increase of at least about 10% compared to the original level of excitatory postsynaptic current discharge frequency in the subject, for example, an increase of at least about 20%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0345] In the present application, the ATP6V1B2 regulator may include an ATP6V1B2 agonist.

[0346] In the present application, the ATP6V1B2 can bind to the ATP6V1B2 protein. For example, the ATP6V1B2 regulator can include a ligand of the ATP6V1B2 protein. For example, the ATP6V1B2 regulator can include an agonist of the ATP6V1B2 protein. For example, the ATP6V1B2 regulator can specifically bind to a specific position (e.g., a specific epitope) in the ATP6V1B2 protein.

[0347] In the present application, the ATP6V1B2 regulator may include a proton pump agonist, for example, a vesicular proton pump agonist.

[0348] In the present application, the ATP6V1B2 modulating agent may include a nucleic acid or a variant thereof, a polypeptide or a variant thereof, and / or a small molecule.

[0349] In the present application, the ATP6V1B2 modulator is capable of increasing synaptic neurotransmitter release.

[0350] As used herein, the increase includes an increase of at least about 10% compared to the original synaptic neurotransmitter release level in the subject, for example, an increase of at least about 20%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0351] In the present application, the ATP6V1B2 regulator can increase the discharge frequency of excitatory postsynaptic currents.

[0352] In the present application, the increase includes an increase of at least about 10% compared to the original level of excitatory postsynaptic current discharge frequency in the subject, for example, an increase of at least about 20%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0353] In another aspect, the present application provides polypeptides capable of regulating the expression level and / or activity of ATP6V1B2.

[0354] In the present application, the polypeptide can increase the release of synaptic neurotransmitters. In the present application, the increase includes being able to increase by at least about 10% compared to the original release level of synaptic neurotransmitters in the subject. For example, it can be improved by at least about 20%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0355] In the present application, the polypeptide can increase the discharge frequency of excitatory postsynaptic current. In the present application, the increase includes being able to increase by at least about 10% compared to the original level of the discharge frequency of excitatory postsynaptic current in the subject. For example, it can be increased by at least about 20%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0356] In the present application, the polypeptide or variant thereof can specifically bind to the ATP6V1B2 protein. For example, the polypeptide or variant thereof can have a "ligand-receptor" relationship with the ATP6V1B2 protein.

[0357] In the present application, the polypeptide or its variant can bind to at least a portion of the amino acid sequence of mouse ATP6V1B2 protein from positions 288 to 512. In the present application, the polypeptide or its variant can bind to at least a portion of the amino acid sequence of human ATP6V1B2 protein from positions 288 to 512.

[0358] As used herein, the polypeptide or variants thereof can include molecules made up of monomers (amino acids) linearly linked by amide bonds (also called peptide bonds). The polypeptide can include any chain having two or more amino acids, and no specific length of the product is intended. The variants can include polypeptides that contain one or more amino acid substitutions, additions, or deletions in addition to the polypeptide. The variants can include products of modifications to the polypeptide, for example, the modifications can include glycosylation, acetylation, phosphorylation, acylation, derivatization with known protecting / blocking groups, proteolytic cleavage, and / or modification with non-naturally occurring amino acids.

[0359] In the present application, the polypeptide or its variant can improve the expression level and / or activity of ATP6V1B2 in a subject. Here, the expression level of ATP6V1B2 can include the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene, and / or the expression level of the ATP6V1B2 protein. In the present application, the improvement can include an improvement in the expression level and / or activity of ATP6V1B2 by at least about 10% compared to the original expression level and / or activity of ATP6V1B2 in the subject. For example, the expression level and / or activity of ATP6V1B2 can be improved by at least about 20%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 500% or more.

[0360] In the present application, the polypeptide can comprise the amino acid sequence shown in SEQ ID NO:5.

[0361] In the present application, the polypeptide can comprise the amino acid sequence shown in SEQ ID NO:1 or SEQ ID NO:3.

[0362] In the present application, the polypeptide or its variant can form a multimer, for example, a homodimer. In the present application, the homodimer can still have the ability to bind to the ATP6V1B2 protein.

[0363] The homodimer may be a tandem structure of two completely identical polypeptides. For example, the homodimer may comprise the amino acid sequence shown in SEQ ID NO:18.

[0364] In the present application, the cysteine ​​in the polypeptide or its variant amino acid sequence may not have a sulfhydryl blocking modification. For example, if the cysteine ​​residue in the polypeptide or its variant amino acid sequence is blocked by a sulfhydryl group, it may lose the ability to bind to the ATP6V1B2 protein.

[0365] In the present application, the serine in the polypeptide or its variant amino acid sequence may not have a phosphorylation modification. For example, if the serine residue in the polypeptide or its variant amino acid sequence is modified by phosphorylation, it may lose the ability to bind to the ATP6V1B2 protein.

[0366] In the present application, the formulation (e.g., the polypeptide or its variant) can improve the learning ability of the subject (e.g., significantly improve the evaluation score of the learning ability of the subject compared to before administration of the polypeptide or its variant).

[0367] In the present application, the preparation (e.g., the polypeptide or a variant thereof) can regulate the expression level and / or activity of the learning ability-related gene in the subject. In the present application, the learning ability-related gene can include the APP gene. The APP gene is an amyloid precursor protein gene, and the gene can be associated with Alzheimer's disease, for example, familial Alzheimer's disease (FAD). The learning ability-related gene can also include the presenilin 1 gene (PSEN1) and the presenilin 2 gene (PSEN2). For example, the learning ability-related gene can include a gene encoding a glutamate receptor. For example, the glutamate receptor can include an N-methyl-D-aspartate receptor (NMDAR), a kainate receptor (KAR), and an α-amino-3-hydroxy-5-methyl-4-isoxazole receptor (AMPAR). For example, the learning ability-related gene can include a gene encoding an agonist receptor for memantine (e.g., 1-amino-3,5-dimethylamantadine hydrochloride). For example, the agonist receptor for memantine can include an NMDA receptor. For example, the polypeptide can reduce the expression level and / or activity of the gene described above in this paragraph.

[0368] In the present application, the formulations described herein, the ATP6V1B2 regulators, the proton pump regulators, and / or the polypeptides described herein may be potential drugs for improving learning ability. The formulations described herein, the ATP6V1B2 regulators, the proton pump regulators, and / or the polypeptides described herein may be potential drugs for treating cognitive disorders and / or treating neurodegenerative diseases.

[0369] In another aspect, the present application provides a combination of drugs that are reagents as described herein.

[0370] The drug combination may include drug products suitable for drug use (eg, improving learning ability, treating cognitive disorders, and / or treating neurodegenerative diseases (eg, Alzheimer's disease)).

[0371] In another aspect, the present application provides a pharmaceutical composition comprising a reagent described herein and a pharma- ceutically acceptable carrier.

[0372] The pharmaceutical composition may be a composition comprising one or more active ingredients (e.g., a formulation described herein) and one or more inactive ingredients, as well as any product obtained directly or indirectly by combination, complexation or aggregation of any two or more ingredients, or by dissociation of one or more ingredients, or by any other type of reaction or interaction of one or more ingredients.

[0373] The pharma- ceutically acceptable carriers can include sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstituting sterile injectable solutions or dispersions immediately before use. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (e.g., propylene glycol, propylene glycol, polyethylene glycol, etc.), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic solvents such as ethyl oleate.

[0374] In another aspect, the present application provides the use of a reagent described herein, a drug combination described herein, and / or a pharmaceutical composition described herein in the preparation of a medicament for increasing synaptic neurotransmitter release.

[0375] In another aspect, the present application provides the use of a reagent described herein, a drug combination described herein, and / or a pharmaceutical composition described herein in the preparation of a medicament for improving learning ability.

[0376] In another aspect, the present application provides the use of a reagent described herein, a drug combination described herein, and / or a pharmaceutical composition described herein in the preparation of a medicament for treating a cognitive disorder and / or for treating a neurodegenerative disease.

[0377] The polypeptides described herein, the reagents described herein, the drug combinations described herein, and / or the pharmaceutical compositions described herein can be formulated to be suitable for oral and / or injectable administration. The polypeptides described herein, the reagents described herein, the drug combinations described herein, and / or the pharmaceutical compositions described herein can be formulated to be suitable for intravenous administration.

[0378] Drug Screening Methods In another aspect, the present application provides a method for screening for drugs that can improve a subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease, comprising detecting an effect of a candidate drug on the expression level and / or activity of a proton pump associated protein in a subject, wherein if the expression level and / or activity of the proton pump associated protein is improved after administration of the candidate drug, the candidate drug can improve the subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease.

[0379] In another aspect, the present application provides a method for screening for drugs that can improve a subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease, comprising a step of detecting the effect of a candidate drug on the expression level and / or activity of ATP6V1B2 in a subject, wherein if the expression level and / or activity of ATP6V1B2 is improved after administration of the candidate drug, the candidate drug can improve the subject's learning ability, treat a cognitive impairment, and / or treat a neurodegenerative disease.

[0380] In some cases, the candidate drug may include a reagent capable of modulating the expression level and / or activity of ATP6V1B2, and in some cases, the candidate drug may include a reagent capable of modulating the expression level and / or activity of a proton pump-associated protein. In some cases, the candidate drug may include a reagent capable of binding to the ATP6V1B2 protein.

[0381] In this application, the administration can include oral administration and / or injection.

[0382] In the present application, the administration can include intravenous injection.

[0383] The present application further provides a detection reagent kit capable of detecting the expression level and / or activity of ATP6V1B2. The detection reagent kit can include an instruction manual describing specific steps for detecting the expression level and / or activity of proton pump-associated protein and / or ATP6V1B2 using the detection reagent kit, and / or specific steps for determining whether the candidate drug can improve the learning ability of a subject using the detection result.

[0384] In the present application, the detection reagent kit may further include a reagent capable of detecting other targets to determine whether the candidate drug can improve the learning ability of a subject.

[0385] Non-human animals, their descendants or parts thereof In any embodiment of the present application, the non-human animal can be a gene-edited non-human animal or can be derived from a gene-edited non-human animal. For example, the non-human animal can be a gene knock-in non-human animal or can be derived from a gene knock-in non-human animal.

[0386] For example, the non-human animal can be a non-human mammal. In some embodiments, the non-human animal is a rodent or a primate. In some embodiments, the non-human animal is selected from a mouse, a rat, a rabbit, an ape, a monkey, a pig, a cow, a sheep, a horse, a chicken, a dog, an alpaca, and a cat.

[0387] In any aspect of the present application, the part of the non-human animal or its progeny may include organs, tissues and / or cells thereof.

[0388] In some embodiments, the portion can include a bodily fluid. For example, the bodily fluid can be selected from the group consisting of blood, plasma, serum, urine, sweat, tears, saliva, semen, and cerebrospinal fluid. In some embodiments, the plasma includes exosomes. In some embodiments, the portion includes a brain (e.g., brain tissue) or a portion thereof. The portion of the brain can include a portion selected from the group consisting of the olfactory bulb, amygdala, basal ganglia, hippocampus, thalamus, hypothalamus, cerebral cortex, medulla oblongata, and cerebellum. In some embodiments, the portion includes an olfactory mucosa. In some embodiments, the portion includes a portion of the central nervous system, a portion of the peripheral nervous system, skin tissue, muscle tissue, and / or visceral organs. For example, the central nervous system can include a spinal cord.

[0389] In some embodiments, the portion can include cells. The cells can include cells selected from the group consisting of primary neurons, small glial cells, astrocytes, oligoglial cells, macrophages, perivascular epithelioid cells, B cells, T cells, adult stem cells, NK cells, totipotent stem cells, unipotent stem cells, embryonic stem cells, induced pluripotent stem cells, and gametes. For example, the gametes can include sperm and / or oocytes.

[0390] In some embodiments, the part (eg, the organ, tissue, or cell) is not capable of developing into a complete non-human animal.

[0391] In some embodiments, the non-human animal, its progeny, or a portion thereof, comprises the ATP6V1B2 and / or a functionally active fragment thereof. For example, the ATP6V1B2 and / or a functionally active fragment thereof may be expressed by the animal, its progeny, or its cells, or may be derived from another source and then introduced (e.g., injected) into the animal, its progeny, or its cells, thereby causing them to comprise the ATP6V1B2 and / or a functionally active fragment thereof.

[0392] In some embodiments, the non-human animal, its progeny, or part thereof expresses the ATP6V1B2 and / or a functionally active fragment thereof.

[0393] The expression of ATP6V1B2 can include expression of the ATP6V1B2 gene, expression of a transcript of the ATP6V1B2 gene, and / or expression of the ATP6V1B2 protein. For example, the expression can include expression of a polynucleotide, mRNA, or amino acid product or protein of a particular gene (e.g., human ATP6V1B2 gene). The expression can include expression of a fragment of a polynucleotide, a translated protein, or a post-translationally modified protein transcribed by a particular gene (e.g., human ATP6V1B2 gene).

[0394] The expression of the functionally active fragment of ATP6V1B2 can include expression of a gene encoding a functionally active fragment of ATP6V1B2, a transcript of a gene encoding a functionally active fragment of ATP6V1B2, and / or expression of a functionally active fragment protein of ATP6V1B2. For example, the expression can include expression of a polynucleotide, mRNA, or amino acid product or protein of a specific gene (e.g., encoding a functionally active fragment of human ATP6V1B2 (e.g., an amino acid sequence from positions 288 to 512)). The expression can include expression of a fragment of a polynucleotide transcribed by a specific gene, a translated protein, or a post-translationally modified protein.

[0395] Preparation method The present application further provides a method for preparing a non-human animal or part thereof described herein, a cell described herein, or a tissue described herein, comprising comprising causing the non-human animal or part thereof, cell or tissue to contain and / or express ATP6V1B2 and / or a functionally active fragment thereof.

[0396] In another aspect, the present application further provides a method of producing a disease model, the method comprising comprising comprising comprising and / or expressing ATP6V1B2 and / or a functionally active fragment thereof in a non-human animal or a part, cell or tissue thereof.

[0397] For example, the method can include introducing the ATP6V1B2, a functionally active fragment thereof, a nucleic acid molecule encoding the ATP6V1B2, and / or a nucleic acid molecule encoding a functionally active fragment of ATP6V1B2 into the non-human animal or part thereof, the cell, tissue, cell line, and / or cell culture. The introduction can include, for example, injecting the non-human animal, part thereof, tissue, or cell (e.g., injecting a protein or a nucleic acid molecule). For example, the introduction can further include other means of introducing an exogenous protein, an exogenous nucleic acid molecule, etc. into a cell, tissue, or animal body, such as nucleic acid molecule transfection, viral transduction, etc. For example, the transfection can include transfection using an adeno-associated virus.

[0398] In some embodiments, the method may include knocking in a nucleic acid sequence encoding the ATP6V1B2 and / or a functionally active fragment thereof into the non-human animal or part thereof, the cell, tissue, cell line and / or cell culture.

[0399] In some embodiments, the method can further include identifying the modified cell or non-human animal that contains the knocked-in heterologous nucleic acid sequence.

[0400] The donor nucleic acid molecule comprising a heterologous nucleic acid sequence may also comprise a 5' homologous arm and a 3' homologous arm. The 5' homologous arm and the 3' homologous arm may be located on the flank of the heterologous nucleic acid sequence encoding the ATP6V1B2 and / or its functionally active fragment. The homologous arm (e.g., the 5' homologous arm or the 3' homologous arm) in the donor nucleic acid molecule may have sufficient ability to promote interaction with an endogenous target site.

[0401] In the present application, the homologous arms and the target gene site (i.e., the homologous gene region or corresponding region in the endogenous ATP6V1B2 gene site) match each other or correspond to each other. In this case, the mutual identity of the two local sequences is sufficient to serve as the basis for the homologous recombination reaction. "Homology" refers to a DNA sequence being the same as or having a certain sequence identity with the corresponding or matching sequence. The sequence identity between a particular target site and the corresponding homologous arm found in the donor nucleic acid molecule can be any degree of sequence identity that allows the occurrence of homologous recombination. For example, to facilitate homologous recombination of sequences, the degree of sequence identity shared by the homology arms of the donor nucleic acid molecule (or fragments thereof) and the target site (or fragment) can be at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83% at least about 84%, at least about 85%, at least about 86%%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%%, at least about 97%, at least about 98%, at least about 99%, or at least about 100%.

[0402] Screening / Identification / Design Methods For example, the device may include a medical device, such as a medical device that is claimed to be effective in improving a subject's learning ability, treating a cognitive disorder, and / or treating a neurodegenerative disease.

[0403] The identification, screening or design methods described herein can be in vitro methods, ex vivo methods or in vivo methods.

[0404] In the present application, a candidate substance may be a synthetic compound, a peptide, a protein, a DNA library or a nucleic acid molecule within such a library, an animal (e.g., a mammal such as mouse, rat, pig, cow, sheep, monkey or human) tissue extract or cell culture supernatant, a plant or microbial extract or culture product, or a mixture of any of the above substances.

[0405] Cognitive, motor, memory and / or spatial exploration abilities can also be compared between a treatment group administered with a candidate drug and a control group, for example, by performing animal behavior analysis. For example, if significant improvements in cognitive, motor, memory and / or spatial exploration abilities are observed after treatment with a candidate drug compared to the control group, the candidate drug can be selected for further study (e.g., as a potential therapeutic drug for improving the subject's learning ability, treating cognitive disorders, and / or treating neurodegenerative diseases).

[0406] In some cases, the screening method can also be performed by using non-human animal tissues and / or cells according to the present application, such as neuronal cells, brain regions, other tissues containing / corresponding to AD lesions, or tissue parts thereof (e.g., olfactory bulb, amygdala, basal ganglia, hippocampus, thalamus, hypothalamus, cerebral cortex, medulla oblongata, central nervous system, peripheral nervous system, spinal cord, cerebellum, skin tissue, muscle tissue, and / or visceral organs). For example, after culturing the tissues and / or cells in vitro or ex vivo and incubating for a suitable period of time (e.g., hours, days, weeks, or months), a candidate drug is administered to the cultured tissues and / or cells, and the treated brain tissue / cells are examined using the above-mentioned method.

[0407] In some cases, for example, in a method for screening biomarkers, samples (e.g., cells, tissues, or other samples containing DNA or RNA, samples containing proteins, and / or samples containing metabolites) can be collected from a non-human animal derived from the present application, a part thereof, or a progeny thereof before and after the non-human animal shows signs related to a nervous system disease or condition (e.g., Aβ deposition, neuronal fiber tangles, morphological or functional abnormalities (atrophy) synapses, neuronal cell death, or memory and learning impairment), and then gene transcription products (transcripts), gene translation products (proteins), lipids (lipids), or metabolites (metabolites) from the samples can be analyzed comprehensively to identify substances that change before and after the non-human animal shows signs related to the nervous system disease or condition. In some embodiments, the sample can include bodily fluids of the non-human animal, such as blood, plasma (e.g., plasma containing exosomes), serum, urine, sweat, tears, saliva, semen, and / or cerebrospinal fluid of a rat.

[0408] Nucleic acid microarray chips (microarrays), such as DNA microarray chips, can be used to analyze gene transcription products (e.g., transcripts). Gel electrophoresis (e.g., two-dimensional gel electrophoresis) or mass spectrometry (e.g., time-of-flight mass spectrometry, electrospray ionization mass spectrometry), capillary HPLC / MS and LC / MS / MS can be used to analyze gene translation products (e.g., proteins). NMR, capillary electrophoresis, LC / MS and LC / MS / MS can be used to analyze metabolites (e.g., metabolites).

[0409] If the presence / content of a substance shows a significant difference before and after the onset of relevant symptoms, the substance can be considered a biomarker for improving a subject's learning ability, treating a cognitive disorder, and / or treating a neurodegenerative disease.

[0410] Without intending to be bound by any theory, the following examples are intended only to illustrate the fusion proteins, preparation methods and uses of the present application, and are not intended to limit the scope of the present invention.

[0411] Working Example Example 1 Overexpression of ATP6V1B2 protein can improve learning and memory function Test steps: 15-month-old aged mice were randomly divided into a control group and an experimental group, and an adeno-associated virus carrying ATP6V1B2-mCherry fusion protein (pAAV2 / 9-hSyn-ATP6V1B2-Myc-2A-mCherry, where the amino acid sequence of the ATP6V1B2-mCherry fusion protein is shown in SEQ ID NO:6) or a control adeno-associated virus carrying mCherry was injected into the dorsal hippocampus of both sides of the mouse brain in the experimental group and the control group, respectively, to overexpress ATP6V1B2-mCherry protein or mCherry protein in neurons in the dorsal hippocampus of both sides of the mouse brain in the experimental group and the control group, respectively. One month after surgery, the mice were subjected to a water maze experiment. (See Example 4 for the test steps) The results showed that overexpressing ATP6V1B2 protein in the hippocampus of 15-month-old aged mice (see Fig. 7a, b) could obviously improve the learning and memory abilities of the aged mice in the water maze behavioral experiment (see Fig. 7c, d, e).

[0412] Example 2 Polypeptide targeting ATP6V1B2 protein Test steps (see E. Harlow D. Ryan, American E. Harlow D. Ryan, Shen Qin et al., Antibody Technology Experimental Guidelines [J]. Science Publishers, 2002): (1) Polypeptide QD202 was synthesized according to the amino acid sequence shown in SEQ ID NO:1, a biotin label was added to the amino C' terminus of the polypeptide, the amino acid sequence of QD202 was randomly shuffled, and a shuffled control QD201 (the amino acid sequence is shown in SEQ ID NO:7) was synthesized, and a biotin label was added to the amino C' terminus of the polypeptide. Polypeptides QD202 and QD201 were dissolved in PBS to prepare a stock solution with a concentration of 500 μM. (2) Biotin-labeled QD202 and QD201 were intravenously administered to two 8-month-old C57BL / 6 mice at a dose of 6.25 mg / kg, respectively. After 4 hours, the mice were anesthetized, brain hippocampal tissue was harvested, lysis buffer was added, the tissue was crushed by mechanical polishing, and the supernatant was collected after centrifugation. Biotin antibody and Protein G agarose beads were added to the supernatant to perform immunoprecipitation, and the test steps were referred to the "Antibody Technology Experimental Guidelines" published by Science Publishing Co., Ltd. in 2002. Proteins were eluted using protein injection buffer. (3) SDS-PAGE protein electrophoresis gel was prepared, the concentration of the lower separation gel was 10% and the concentration of the upper concentration gel was 4%, the sample was added to the injection well, and SDS-PAGE electrophoresis was performed. After the electrophoresis was completed, the separation gel was cut out and stained using Coomassie Brilliant Blue, and the results were shown in Figure 6a. (4) A specific band with a molecular weight of approximately 55 kDa, as indicated by Coomassie Brilliant Blue staining, was excised, subjected to pre-mass spectrometry treatment, and the protein was enzymatically degraded into peptide fragments and desalted. The sample was separated into ion fragments according to mass number using a mass analyzer, and a mass spectrum was obtained using a detector. The identified peptide fragment sequences were aligned with a Western database using search software to identify the type of protein. (5) By comparison with the shuffled control QD201, we detected ATP6V1B2, a protein that specifically binds to QD202. (6) A mouse Atp6v1b2 expression plasmid with a myc tag added to the 3' end was constructed, and the plasmid was transfected into HEK 293 cells. After 2 days, the cells were harvested and the cellular proteins were extracted. The cell lysate was then co-immunoprecipitation with biotin antibody and Protein G agarose beads. [1] and the protein was eluted with protein injection buffer. (7) SDS-PAGE protein electrophoresis gel was prepared, the concentration of the lower separation gel was 10% and the concentration of the upper concentration gel was 4%, the sample was added to the injection well, and the protein was detected by immunoblotting, the results are shown in Figure 6b. By the method of co-immunoprecipitation, it was found that QD202 specifically binds to ATP6V1B2 protein.

[0413] The results showed that QD202 bound to ATP6V1B2 protein by protein profiling and co-immunoprecipitation methods, and the results are shown in Figure 6a, b.

[0414] Example 3 Biological activity test of polypeptides Test steps: The polypeptide QD202 was synthesized according to the amino acid sequence shown in SEQ ID NO:1.

[0415] Polypeptide QD202 N'-1 was synthesized according to the amino acid sequence shown in SEQ ID NO:2.

[0416] Polypeptide QD202 C'-1 was synthesized according to the amino acid sequence shown in SEQ ID NO:3.

[0417] Polypeptide QD202 C'-2 was synthesized according to the amino acid sequence shown in SEQ ID NO:4.

[0418] C57 mice aged 2-3 months were anesthetized by intraperitoneal injection of 0.1-0.15 mL of 20% uracilin, then decapitated and brain tissue was excised and placed in an ice-water mixture of artificial cerebrospinal fluid (ACSF) that had been previously gassed with a mixture of 95% O2 and 5% CO2, and left for 2 min. The ACSF formulation was 11.7 mM NaCl, 0.36 mM KCl, 0.12 mM NaH2PO4, 0.25 mM CaCl2, 0.12 mM MgCl2, 25 mM NaHCO3, and 11 mM glucose. 350 μm coronal hippocampal brain slices were cut using a vibrating microtome. Hippocampal brain slices were left in room temperature ACSF for more than 30 min to recover, then placed in a recording tank and continuously perfused with ACSF through the mixed gas at a rate of 2-2.5 mL / min. Hippocampal CA1 pyramidal neurons were blind sealed under an Olympus BX51 upright microscope, and whole-cell patch clamp recordings were performed using an Axon700B amplifier and a 1550B digital-to-analog converter. Spontaneous excitatory postsynaptic currents (sEPSCs) of pyramidal neurons were recorded in voltage clamp at a clamp voltage of -70 mV. After 5 min of sEPSCs, 5 μM QD202 or QD202 N'-1, C'-1, C'-2, etc. were administered to synthesize peptides for 5 min, and then perfused and washed with ACSF for 10 min. Recorded data were analyzed using Minianalysis software.

[0419] The results showed that QD202 could increase the spontaneous excitatory synaptic transmitter release in hippocampal CA1 neurons, mainly reflected by an increase in the release frequency of excitatory postsynaptic currents, but had no effect on the amplitude of excitatory postsynaptic currents (see Fig. 1a, b, c).

[0420] In comparison with the effect of QD202 on excitatory synaptic transmission, when one amino acid at the N-terminus was removed, no similar effect to that of QD202 was observed (see Fig. 1d, e); when one amino acid at the C-terminus was removed, a similar effect to that of QD202 was still observed (see Fig. 1f, g); and when two amino acids at the C-terminus were removed, no similar effect to that of QD202 was observed (see Fig. 1h, i).

[0421] Example 4: Improvement of learning ability by polypeptides New Object Identification Test The novel object discrimination test was performed using an open field box (40 × 40 × 35 cm, made of blue opaque plastic) with reference to the behavioral test plan described in Leger, M., et al. Object recognition test in mice. Nat Protoc. 8, 2531-2537 (2013). On the first day, mice were placed in the open field box and allowed to adapt for 10 min. The next day, each mouse was carefully placed in the center of the box, two similar objects (AA batteries) were placed in the central area, and the mice were allowed to explore freely for 10 min, and then the mice were returned to their breeding cages. After 3 h, the mice were placed back in the box (one battery was replaced with a 10 cm tall doll toy) again for 10 min to perform the memory retention test. The recordings were analyzed using Etho Vision XT 14 software, and the time it took the mice to explore the new / old objects was recorded. The discrimination index was calculated as (Tnovel-Tfamliar) / (Tnovel+Tfamiliar).

[0422] Morris Water Maze Test The Morris water maze was performed according to the test plan described in Qing-Feng Wu et al., Fibroblast growth factor 13 is a microtubule-stabilizing protein regulating neuronal polarization and migration. Cell. 149, 1549-1564 (2012). The test was performed in a circular pool filled with water (120 cm in diameter, 30 cm deep, made opaque by adding titanium dioxide, and maintained at 21 ± 1 °C) in a fixed environment room. The experiment was divided into an adaptation period (1 day), a training period (5-6 days), and a test period (1 day). Adaptation period: The platform was placed 0.5 cm away from the water surface, and the mouse was guided to the platform. Training day: The platform was placed 0.5 cm underwater, and the mouse was trained to find the platform. The timing was stopped when the mouse reached the platform, and if the mouse did not reach the platform within 1 min, it was guided to the platform and held on the platform for 30 s. Mice were trained four times a day (between 8:00 and 16:00), with a different water entry point each time, with at least 30 min between training sessions, and the order of water entry points was changed each day for a total of 6 days. Experimental day: Experiments were performed 24 hours after the end of the training day. The platform was removed and the mouse entered the water from an untrained water entry point and was recorded for 1 min. Recordings were analyzed using Etho Vision XT 14 software, and recording parameters included avoidance latency to enter the goal quadrant, number of times the platform was crossed, and dwell time in the goal quadrant.

[0423] 4.1 Oral Administration of Polypeptides Ten-month-old APP / PS1 transgenic mice (purchased from Minami-Momo Bio) were selected and divided into three groups. These mice were orally administered with PBS buffer (group 2), QD202 (group 3, the dose of QD202 was 6.25 mg / kg (wherein the concentration of QD202 was 500 μM)) and memantine (group 4, the dose of memantine was 1.25 mg / kg, wherein the concentration of memantine was 1 mM) by gavage, respectively, and 10-month-old C57 BL / 6 mice that were not given any reagent by gavage served as control (group 1).

[0424] The four groups of mice were subjected to the above-mentioned behavioral tests.

[0425] The results showed that after continuous oral gavage of QD202 for 1 week, APP / PS1 transgenic AD mice showed obvious improvement of learning cognitive function in detection of novel object discrimination (see Figure 2 ) and water maze behavior (see Figure 3 ).

[0426] 4.2 Intravenous injection of polypeptides Ten-month-old APP / PS1 transgenic mice (purchased from Minami-Momo Bio) were selected and divided into two groups. These mice were intravenously injected with PBS buffer (group 2, PBS concentration 500 μM), QD202 (group 3, QD202 dosage 1.25 mg / kg, where QD202 concentration 500 μM), respectively. Ten-month-old C57 BL / 6 mice that did not receive any reagents by intravenous injection served as control (group 1).

[0427] The four groups of mice were subjected to the above-mentioned behavioral tests.

[0428] The results showed that after continuous intravenous injection of QD202 for one week, APP / PS1 transgenic AD mice showed obvious improvement of learning cognitive function in detection of novel object discrimination (see Figure 4 ) and water maze behavior (see Figure 5 ).

[0429] As can be seen, QD202 had a significant improving effect on learning ability, particularly memory ability and cognitive ability.

[0430] Example 5 Modified Polypeptides and Their Improving Effect on Learning Ability 5.1 Dimeric forms of polypeptides Preparation of QD202 dimer: QD202 was taken and dissolved in water at a concentration of 10 mg / mL, and diluted ammonia water was added to adjust the pH value to between 8 and 9, and the mixture was oxidized with stirring for 24 hours, during which the reaction end point was monitored by HPLC. After the oxidation was completed, the mixture was further purified by HPLC to obtain QD202 dimer. The QD202 dimer contained the amino acid sequence shown in SEQ ID NO:18.

[0431] According to the method described in Example 4, QD202 dimers were used to detect spontaneous excitatory postsynaptic currents in mouse pyramidal neurons, where the control refers to the spontaneous excitatory postsynaptic current results recorded during normal perfusion of artificial cerebrospinal fluid.

[0432] The results are shown in Figure 8. The results show that QD202 dimer can increase the spontaneous excitatory synaptic transmitter release of hippocampal CA1 neurons, which is mainly reflected in the increase in the release frequency of excitatory postsynaptic currents and the increase in the amplitude of excitatory postsynaptic currents.

[0433] 5.2 Cysteine-modified polypeptides Preparation of cysteine-modified QD202: Fmoc-Cys(CAm)-OH was substituted for Fmoc-Cys(Trt)-OH, which was directly synthesized according to the amino acid sequence of QD202 to obtain QD202 containing cysteine ​​modification.

[0434] Cysteine-modified QD202 was used to detect spontaneous excitatory postsynaptic currents in mouse pyramidal neurons according to the method described in Example 4. Here, the control refers to the spontaneous excitatory postsynaptic current results recorded during normal perfusion of artificial cerebrospinal fluid.

[0435] The results are shown in Figure 9. The results showed that cysteine-modified QD202 lost the activity of increasing spontaneous excitatory synaptic transmitter release in hippocampal CA1 neurons, which was mainly reflected in a decrease in the release frequency of excitatory postsynaptic currents and / or a decrease in the amplitude of excitatory postsynaptic currents.

[0436] 5.3 Serine-modified polypeptides Preparation of serine-modified QD202: Fmoc-Ser(HPO3Bzl)-OH was substituted for Fmoc-Ser(tbu)-OH, and the QD202 was directly synthesized according to the amino acid sequence of QD202 to obtain serine-modified QD202 in which the serine (S) at position 12 from the N-terminus was phosphorylated.

[0437] According to the method described in Example 4, serine-modified QD202 was used to detect spontaneous excitatory postsynaptic currents in mouse pyramidal neurons, where the control refers to the spontaneous excitatory postsynaptic current results recorded during normal perfusion of artificial cerebrospinal fluid.

[0438] The results are shown in Figure 10. The results showed that serine-modified QD202 lost the activity of increasing spontaneous excitatory synaptic transmitter release in hippocampal CA1 neurons, which was mainly reflected in a decrease in the release frequency of excitatory postsynaptic currents and / or a decrease in the amplitude of excitatory postsynaptic currents.

[0439] Example 6 Binding site between polypeptide and ATP6V1B2 subunit Construct expression plasmids containing 1-286aa truncated sequence or 1-172aa truncated sequence of mouse ATP6V1B2 full length with Myc tag, where the amino acid sequence of mouse ATP6V1B2 full length is shown in SEQ ID NO: 8, the amino acid sequence of mouse ATP6V1B2 1-286aa is shown in SEQ ID NO: 10, and the amino acid sequence of mouse ATP6V1B2 1-172aa is shown in SEQ ID NO: 11, and these plasmids are called ATP6V1B2 full length expression plasmid, ATP6V1B2 1-286aa expression plasmid, and ATP6V1B2 1-172aa expression plasmid, respectively. At the same time, construct QD202 expression plasmid with EGFP tag.

[0440] 293 cells (purchased from the Cell Bank / Stem Cell Bank of the Chinese Academy of Sciences) were co-transfected with QD202 expression plasmid, ATP6V1B2 full-length expression plasmid, ATP6V1B2 1-286aa expression plasmid, or ATP6V1B2 1-172aa expression plasmid. After 48 hours, cell lysates were collected and Myc antibody (purchased from Sigma-Aldrich) was added to the lysates to enrich for ATP6V1B2 full-length protein, 1-286aa cleavage sequence, or 1-172aa cleavage sequence.

[0441] Then, Protein G agarose beads combined with Myc antibody were added to co-precipitate the ATP6V1B2 full-length protein, the 1-286aa cleavage sequence or the 1-172aa cleavage sequence, and proteins that bind to each other, and finally the co-immunoprecipitated proteins were detected by immunoblotting.

[0442] The results are shown in Figure 11, where Atp6v1b2-Myc, b2-286Δ-Myc and b2-172Δ-Myc correspond to the ATP6V1B2 full-length protein, the 1-286aa truncated sequence and the 1-172aa truncated sequence, respectively. The results showed that only the full-length ATP6V1B2 protein could bind to the QD202 polypeptide, indicating that the binding site between ATP6V1B2 and QD202 is located within the region of amino acids 288 to 512 from the N-terminus of ATP6V1B2.

[0443] The above detailed description is provided by way of interpretation and enumeration, and is not intended to limit the scope of the appended claims. Various modifications of the embodiments recited herein will be apparent to those skilled in the art, and are within the scope of the appended claims and their equivalents.

Claims

1. Use of a proton pump regulator in the preparation of a reagent for use in improving learning ability.

2. 13. Use of a proton pump modulator in the preparation of a reagent for use in the treatment of cognitive disorders.

3. Use of a proton pump modulator in the preparation of a reagent for use in the treatment of a neurodegenerative disease.

4. The reagent increases the expression level and / or activity of a proton pump-associated protein in a subject. The use according to any one of claims 1 to 3.

5. The proton pump regulator increases the expression level and / or activity of a proton pump-associated protein in a subject. The use according to any one of claims 1 to 4.

6. The expression level of the proton pump-associated protein includes the expression level of a gene encoding the proton pump-associated protein, the transcription level of a gene encoding the proton pump-associated protein, and / or the expression level of the proton pump-associated protein; The use according to any one of claims 4 to 5.

7. The improvement comprises an improvement of at least about 10% in the expression level and / or activity of the proton pump-associated protein compared to the expression level and / or activity of the proton pump-associated protein native to the subject. The use according to any one of claims 4 to 6.

8. The expression level of the proton pump-related protein is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining, and mass spectrometry; The use according to any one of claims 4 to 7.

9. The expression level of the proton pump associated protein is measured by using a substance selected from the group consisting of a primer capable of specifically amplifying a gene encoding the proton pump associated protein, a nucleic acid molecule that specifically binds to a gene encoding the proton pump associated protein, a nucleic acid molecule that specifically binds to the proton pump associated protein, a small molecule that specifically binds to the proton pump associated protein, a probe that specifically binds to the proton pump associated protein, and a polypeptide that specifically binds to the proton pump associated protein. The use according to any one of claims 4 to 8.

10. The reagent increases the expression level and / or activity of ATP6V1B2 in a subject. Use according to any one of claims 1 to 9.

11. The proton pump regulator increases the expression level and / or activity of ATP6V1B2 in a subject. The use according to any one of claims 1 to 10.

12. The expression level of ATP6V1B2 includes the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene, and / or the expression level of the ATP6V1B2 protein; 12. Use according to any one of claims 10 and 11.

13. The improvement includes an improvement in the expression level and / or activity of ATP6V1B2 by at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject. The use according to any one of claims 10 to 12.

14. The expression level of ATP6V1B2 is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining and mass spectrometry; The use according to any one of claims 10 to 13.

15. The expression level of ATP6V1B2 is measured by using a substance selected from the group consisting of a primer capable of specifically amplifying the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 protein, a small molecule that specifically binds to the ATP6V1B2 protein, a probe that specifically binds to the ATP6V1B2 protein, and a polypeptide that specifically binds to the ATP6V1B2 protein. The use according to any one of claims 10 to 14.

16. The proton pump regulator comprises a proton pump agonist. Use according to any one of claims 1 to 15.

17. The proton pump regulator comprises a vesicular proton pump agonist. Use according to any one of claims 1 to 16.

18. Proton pump regulators include agonists of the ATP6V1B2 protein. Use according to any one of claims 1 to 17.

19. The proton pump regulator is capable of binding to the ATP6V1B2 protein. Use according to any one of claims 1 to 18.

20. The proton pump regulator is capable of binding to at least a portion of the amino acid sequence at positions 288 to 512 of mouse ATP6V1B2 protein. Use according to any one of claims 1 to 19.

21. The proton pump regulator is capable of binding to at least a portion of the amino acid sequence at positions 288 to 512 of the human ATP6V1B2 protein. Use according to any one of claims 1 to 20.

22. The proton pump regulator comprises a nucleic acid or variant thereof, a polypeptide or variant thereof, and / or a small molecule. Use according to any one of claims 1 to 21.

23. The proton pump modulator is capable of increasing synaptic neurotransmitter release. Use according to any one of claims 1 to 22.

24. The increase comprises an increase of at least about 10% compared to the original synaptic neurotransmitter release level in the subject.

24. The use according to claim 23.

25. The proton pump regulator can increase the discharge frequency of excitatory postsynaptic currents. Use according to any one of claims 1 to 24.

26. The increase comprises an increase of at least about 10% compared to the original level of excitatory postsynaptic current discharge frequency in the subject.

26. The use according to claim 25.

27. The learning ability includes cognitive ability, motor ability, memory ability and / or spatial exploration ability; Use according to any one of claims 1, 4 to 26.

28. The improvement in learning ability includes an improvement of at least about 50% in the assessment score of the improved learning ability of the subject compared to the assessment score of the subject's original learning ability. Use according to any one of claims 1, 4 to 27.

29. The learning ability evaluation score is measured by performing a test selected from the group consisting of a new object recognition test and a water maze test. Use according to any one of claims 1, 4 to 28.

30. The new object recognition test evaluates the cognitive, motor and / or spatial exploration abilities.

30. The use according to claim 29.

31. The water maze test evaluates the memory ability, motor ability and / or spatial exploration ability. Use according to any one of claims 29 to 30.

32. The subject includes a mammal. Use according to any one of claims 4 to 31.

33. The subject includes a human. Use according to any one of claims 4 to 32.

34. The subject includes a non-cognitively impaired patient and / or a non-neurodegenerative disease patient. Use according to any one of claims 4 to 33.

35. The subject includes a patient with a neurodegenerative disease and / or a patient with a cognitive impairment. Use according to any one of claims 4 to 34.

36. The subject includes an Alzheimer's disease patient. Use according to any one of claims 4 to 35.

37. The subject is in an elderly stage. Use according to any one of claims 4 to 36.

38. The cognitive impairment includes mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment; Use according to any one of claims 2, 4 to 37.

39. The cognitive impairment includes cognitive impairment caused by normal aging, Lewis' physical dementia (LBD), frontotemporal dementia and / or vascular dementia; Use according to any one of claims 2, 4 to 38.

40. The cognitive impairment-inducing disease includes Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and / or Creutzfeldt-Jakob disease (CJD); Use according to any one of claims 2, 4 to 39.

41. The neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases. Use according to any one of claims 3 to 40.

42. The neurodegenerative diseases include neurodegenerative diseases caused by neuronal death and glial cell homeostasis, neurodegenerative diseases caused by aging, neurodegenerative diseases caused by effects on CNS cell function, neurodegenerative diseases caused by abnormal communication between cells and / or neurodegenerative diseases caused by impaired cell motility. Use according to any one of claims 3 to 41.

43. The neurodegenerative disease includes Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD); Use according to any one of claims 3 to 42.

44. The reagent is formulated to be suitable for oral and / or injectable administration; 44. Use according to any one of claims 1 to 43.

45. The reagent is formulated to be suitable for intravenous injection.

45. Use according to any one of claims 1 to 44.

46. Use of ATP6V1B2, its functionally active fragments or the nucleic acid molecules encoding them, in improving learning ability.

47. Use of ATP6V1B2, its functionally active fragments or the nucleic acid molecules encoding them, in the treatment of cognitive disorders.

48. Use of ATP6V1B2, its functionally active fragments or the nucleic acid molecules encoding them, in the treatment of neurodegenerative diseases.

49. The ATP6V1B2 is derived from a human. Use according to any one of claims 46 to 48.

50. The ATP6V1B2 comprises the amino acid sequence shown in SEQ ID NO: 8 or 16.

50. The use according to any one of claims 46 to 49.

51. The functionally active fragment of ATP6V1B2 has the ability to specifically bind to the amino acid sequence set forth in SEQ ID NO:

5. Use according to any one of claims 46 to 50.

52. The functionally active fragment of ATP6V1B2 includes a truncation of ATP6V1B2.

52. Use according to any one of claims 46 to 51.

53. The functionally active fragment of ATP6V1B2 comprises at least a portion of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein.

53. The use according to any one of claims 46 to 52.

54. The functionally active fragment of ATP6V1B2 comprises at least a part of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein.

54. The use according to any one of claims 46 to 53.

55. The functionally active fragment of ATP6V1B2 comprises the amino acid sequence shown in SEQ ID NO: 10 or 11.

55. The use according to any one of claims 46 to 54.

56. The nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 9 or 17.

56. The use according to any one of claims 46 to 55.

57. The learning ability includes cognitive ability, motor ability, memory ability and / or spatial exploration ability; 57. Use according to any one of claims 46, 49 to 56.

58. The improvement in learning ability comprises an improvement of at least about 50% in the assessment score of the improved learning ability of the subject compared to the assessment score of the subject's original learning ability.

58. Use according to any one of claims 46, 49 to 57.

59. The learning ability evaluation score is measured by performing a test selected from the group consisting of a new object recognition test and a water maze test.

59. Use according to any one of claims 46, 49 to 58.

60. The new object recognition test evaluates the cognitive, motor and / or spatial exploration abilities.

60. The use according to claim 59.

61. The water maze test evaluates the memory ability, motor ability and / or spatial exploration ability. Use according to any one of claims 59 to 60.

62. The subject includes a mammal.

62. Use according to any one of claims 58 to 61.

63. The subject includes a human.

63. The use according to any one of claims 58 to 62.

64. The subject includes a non-cognitively impaired patient and / or a non-neurodegenerative disease patient.

64. The use according to any one of claims 58 to 63.

65. The subject includes a patient with a neurodegenerative disease and / or a patient with a cognitive impairment.

65. The use according to any one of claims 58 to 64.

66. The subject includes an Alzheimer's disease patient.

66. Use according to any one of claims 58 to 65.

67. The subject is in an elderly stage.

67. Use according to any one of claims 58 to 66.

68. The cognitive impairment includes mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment; 68. Use according to any one of claims 47, 49 to 67.

69. The cognitive impairment includes cognitive impairment caused by normal aging, Lewis physical dementia (LBD), frontotemporal dementia and / or vascular dementia; 69. The use according to any one of claims 47, 49 to 68.

70. The cognitive impairment-inducing disease includes Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and / or Creutzfeldt-Jakob disease (CJD); 70. The use according to any one of claims 47, 49 to 69.

71. The neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases. Use according to any one of claims 48 to 70.

72. The neurodegenerative diseases include neurodegenerative diseases caused by neuronal death and glial cell homeostasis, neurodegenerative diseases caused by aging, neurodegenerative diseases caused by effects on CNS cell function, neurodegenerative diseases caused by abnormal communication between cells and / or neurodegenerative diseases caused by impaired cell motility.

72. Use according to any one of claims 48 to 71.

73. The neurodegenerative disease includes Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD); 73. The use according to any one of claims 48 to 72.

74. Use of an ATP6V1B2 modulator in the preparation of a reagent for use in improving learning ability.

75. Use of an ATP6V1B2 modulator in the preparation of a reagent for use in the treatment of cognitive disorders.

76. Use of an ATP6V1B2 modulating agent in the preparation of a reagent for use in the treatment of a neurodegenerative disease.

77. The reagent increases the expression level and / or activity of ATP6V1B2 in a subject.

77. Use according to any one of claims 74 to 76.

78. The ATP6V1B2 modulator increases the expression level and / or activity of ATP6V1B2 in a subject.

78. Use according to any one of claims 74 to 77.

79. The expression level includes the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene, and / or the expression level of the ATP6V1B2 protein; 79. Use according to any one of claims 74 to 78.

80. The improvement in activity includes improving the expression level and / or activity of a proton pump-related protein.

80. The use according to any one of claims 77 to 79.

81. The improvement includes an improvement in the expression level and / or activity of ATP6V1B2 by at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject. Use according to any one of claims 77 to 80.

82. The expression level of ATP6V1B2 is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining and mass spectrometry; 82. Use according to any one of claims 77 to 81.

83. The expression level of ATP6V1B2 is measured by using a substance selected from the group consisting of a primer capable of specifically amplifying the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 protein, a small molecule that specifically binds to the ATP6V1B2 protein, a probe that specifically binds to the ATP6V1B2 protein, and a polypeptide that specifically binds to the ATP6V1B2 protein.

83. The use according to any one of claims 77 to 82.

84. ATP6V1B2 regulators include agonists of the ATP6V1B2 protein.

84. The use according to any one of claims 74 to 83.

85. The ATP6V1B2 modulator is capable of binding to the ATP6V1B2 protein.

85. The use according to any one of claims 74 to 84.

86. The ATP6V1B2 regulator can bind to at least a portion of the amino acid sequence at positions 288 to 512 of mouse ATP6V1B2 protein.

86. Use according to any one of claims 74 to 85.

87. The ATP6V1B2 regulator can bind to at least a portion of the amino acid sequence at positions 288 to 512 of the human ATP6V1B2 protein.

87. Use according to any one of claims 74 to 86.

88. The ATP6V1B2 modulator comprises a proton pump agonist.

88. Use according to any one of claims 74 to 87.

89. The ATP6V1B2 regulator comprises a vesicular proton pump agonist.

89. Use according to any one of claims 74 to 88.

90. The ATP6V1B2 modulator comprises a nucleic acid or variant thereof, a polypeptide or variant thereof, and / or a small molecule.

90. Use according to any one of claims 74 to 89.

91. The ATP6V1B2 regulator comprises a polypeptide or a variant thereof. Use according to any one of claims 74 to 90.

92. The polypeptide or variant thereof is capable of binding to the ATP6V1B2 protein; 92. The use according to claim 91.

93. The polypeptide or variant thereof is capable of increasing synaptic neurotransmitter release.

93. The use according to any one of claims 91 to 92.

94. The increase comprises an increase of at least about 10% compared to the original synaptic neurotransmitter release level in the subject.

94. The use according to claim 93.

95. The polypeptide or variant thereof can increase the release frequency of excitatory postsynaptic currents.

95. Use according to any one of claims 91 to 94.

96. The increase comprises an increase of at least about 10% as compared to the native level of excitatory postsynaptic current discharge frequency in the subject.

96. The use according to claim 95.

97. The polypeptide or variant thereof comprises the amino acid sequence shown in SEQ ID NO:

5.

97. Use according to any one of claims 91 to 96.

98. The polypeptide or variant thereof comprises the amino acid sequence shown in SEQ ID NO: 1 or 3.

98. Use according to any one of claims 91 to 97.

99. The learning ability includes cognitive ability, motor ability, memory ability and / or spatial exploration ability; 99. Use according to any one of claims 74, 77 to 98.

100. The improvement in learning ability includes an improvement of at least about 50% in the assessment score of the improved learning ability of the subject compared to the assessment score of the subject's original learning ability.

99. Use according to any one of claims 74, 77 to 99.

101. The learning ability evaluation score is measured by performing a test selected from the group consisting of a new object recognition test and a water maze test. Use according to any one of claims 74, 77 to 100.

102. The new object recognition test evaluates the cognitive, motor and / or spatial exploration abilities.

102. The use according to claim 101.

103. The water maze test evaluates the memory ability, motor ability and / or spatial exploration ability.

103. The use according to any one of claims 101 or 102.

104. The subject includes a mammal. Use according to any one of claims 77 to 103.

105. The subject includes a human. Use according to any one of claims 77 to 104.

106. The subject includes a non-cognitively impaired patient and / or a non-neurodegenerative disease patient. Use according to any one of claims 77 to 105.

107. The subject includes a patient with a neurodegenerative disease and / or a patient with a cognitive impairment. Use according to any one of claims 77 to 106.

108. The subject includes an Alzheimer's disease patient. Use according to any one of claims 77 to 107.

109. The subject is in an elderly stage. Use according to any one of claims 77 to 108.

110. The cognitive impairment includes mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment; Use according to any one of claims 75, 77 to 109.

111. The cognitive impairment includes cognitive impairment caused by normal aging, Lewis' physical dementia (LBD), frontotemporal dementia and / or vascular dementia; 111. Use according to any one of claims 75, 77 to 110.

112. The cognitive impairment-inducing disease includes Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and / or Creutzfeldt-Jakob disease (CJD); 112. The use according to any one of claims 75, 77 to 111.

113. The neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

13. The use according to any one of claims 76 to 112.

114. The neurodegenerative diseases include neurodegenerative diseases caused by neuronal death and glial cell homeostasis, neurodegenerative diseases caused by aging, neurodegenerative diseases caused by effects on CNS cell function, neurodegenerative diseases caused by abnormal communication between cells and / or neurodegenerative diseases caused by impaired cell motility. Use according to any one of claims 76 to 113.

115. The neurodegenerative disease includes Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD); 115. The use according to any one of claims 76 to 114.

116. The reagent is formulated to be suitable for oral and / or injectable administration; 16. Use according to any one of claims 74 to 115.

117. The reagent is formulated to be suitable for intravenous injection.

117. Use according to any one of claims 74 to 116.

118. 1. A method for screening for drugs that can improve learning ability, treat cognitive disorders, and / or treat neurodegenerative diseases in a subject, comprising: The method includes detecting the effect of a candidate drug on the expression level and / or activity of a proton pump associated protein in a subject, wherein if the expression level and / or activity of the proton pump associated protein is improved after administration of the candidate drug, the candidate drug enables improvement of the subject's learning ability, treatment of a cognitive impairment, and / or treatment of a neurodegenerative disease.

119. 1. A method for screening for drugs that can improve learning ability, treat cognitive disorders, and / or treat neurodegenerative diseases in a subject, comprising: The method includes a step of detecting the effect of a candidate drug on the expression level and / or activity of ATP6V1B2 in a subject, wherein if the expression level and / or activity of ATP6V1B2 is improved after administration of the candidate drug, the candidate drug enables improvement of the subject's learning ability, treatment of cognitive impairment, and / or treatment of a neurodegenerative disease.

120. The expression level of the proton pump-associated protein includes the expression level of a gene encoding the proton pump-associated protein, the transcription level of a gene encoding the proton pump-associated protein, and / or the expression level of the proton pump-associated protein; The method of claim 118.

121. The improvement comprises an improvement of at least about 10% in the expression level and / or activity of the proton pump-associated protein compared to the expression level and / or activity of the proton pump-associated protein native to the subject.

121. The method of any one of claims 118 or 120.

122. The expression level of the proton pump-related protein is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining, and mass spectrometry; 122. The method according to any one of claims 118, 120 to 121.

123. The expression level of the proton pump associated protein is measured by using a substance selected from the group consisting of a primer capable of specifically amplifying a gene encoding the proton pump associated protein, a nucleic acid molecule that specifically binds to a gene encoding the proton pump associated protein, a nucleic acid molecule that specifically binds to the proton pump associated protein, a small molecule that specifically binds to the proton pump associated protein, a probe that specifically binds to the proton pump associated protein, and a polypeptide that specifically binds to the proton pump associated protein.

122. The method according to any one of claims 118, 120 to 121.

124. The expression level of ATP6V1B2 includes the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene, and / or the expression level of the ATP6V1B2 protein; 124. The method according to any one of claims 119 to 123.

125. The improvement includes an improvement in the expression level and / or activity of ATP6V1B2 by at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject.

125. The method of any one of claims 119 to 124.

126. The expression level of ATP6V1B2 is measured by performing a test selected from the group consisting of qPCR, qRT-PCR, hybridization analysis, RNA blotting, dot blotting, in-situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, Western blotting, immunohistochemistry, immunostaining and mass spectrometry; 126. The method of any one of claims 119 to 125.

127. The expression level of ATP6V1B2 is measured by using a substance selected from the group consisting of a primer capable of specifically amplifying the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 gene, a nucleic acid molecule that specifically binds to the ATP6V1B2 protein, a small molecule that specifically binds to the ATP6V1B2 protein, a probe that specifically binds to the ATP6V1B2 protein, and a polypeptide that specifically binds to the ATP6V1B2 protein.

127. The method of any one of claims 119 to 126.

128. The learning ability includes cognitive ability, motor ability, memory ability and / or spatial exploration ability; 128. The method of any one of claims 118 to 127.

129. The administration includes oral and / or injection.

129. The method of any one of claims 118 to 128.

130. The administration comprises intravenous injection.

130. The method of any one of claims 118 to 129.

131. A polypeptide or a variant thereof capable of regulating the expression level and / or activity of ATP6V1B2.

132. Can increase the release of synaptic neurotransmitters, A polypeptide or variant thereof according to claim 131.

133. The increase comprises an increase of at least about 10% compared to the original synaptic neurotransmitter release level in the subject. A polypeptide or variant thereof according to claim 132.

134. Increases the frequency of excitatory postsynaptic current discharge, A polypeptide or a variant thereof according to any one of claims 131 to 133.

135. The increase comprises an increase of at least about 10% compared to the original level of excitatory postsynaptic current discharge frequency in the subject. A polypeptide or variant thereof according to claim 134.

136. capable of binding to the ATP6V1B2 protein; A polypeptide or a variant thereof according to any one of claims 131 to 135.

137. The expression level and / or activity of ATP6V1B2 in a subject can be improved. A polypeptide or a variant thereof according to any one of claims 131 to 136.

138. The expression level of ATP6V1B2 includes the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene, and / or the expression level of the ATP6V1B2 protein; A polypeptide or variant thereof according to claim 137.

139. The improvement includes an improvement in the expression level and / or activity of ATP6V1B2 by at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject.

139. A polypeptide or variant thereof according to any one of claims 137 or 138.

140. It is capable of binding to at least a portion of the amino acid sequence at positions 288 to 512 of mouse ATP6V1B2 protein. A polypeptide or variant thereof according to any one of claims 131 to 139.

141. capable of binding to at least a portion of the amino acid sequence at positions 288 to 512 of the human ATP6V1B2 protein; A polypeptide or a variant thereof according to any one of claims 131 to 140.

142. Comprising the amino acid sequence shown in SEQ ID NO:5, A polypeptide or a variant thereof according to any one of claims 131 to 141.

143. Contains the amino acid sequence shown in SEQ ID NO: 1 or 3; A polypeptide or a variant thereof according to any one of claims 131 to 142.

144. Including multimers, A polypeptide or a variant thereof according to any one of claims 131 to 143.

145. The multimer comprises a homodimer. A polypeptide or variant thereof according to claim 144.

146. No sulfhydryl blocking modifications to cysteines in the amino acid sequence; A polypeptide or variant thereof according to any one of claims 131 to 145.

147. There is no phosphorylation modification of serine in the amino acid sequence; A polypeptide or variant thereof according to any one of claims 131 to 146.

148. The polypeptide according to any one of claims 131 to 147 or a variant thereof, Drug combinations.

149. A method for producing a pharmaceutical composition comprising administering to a patient a therapeutically effective amount of a polypeptide or variant thereof according to any one of claims 131 to 147, and a pharma- ceutically acceptable carrier. Pharmaceutical compositions.

150. Use of a polypeptide or variant thereof according to any one of claims 131 to 147, a drug combination according to claim 148 and / or a pharmaceutical composition according to claim 149 in the preparation of a reagent, comprising The agent is used to improve learning ability, treat cognitive disorders and / or treat neurodegenerative diseases.

151. The learning ability includes cognitive ability, motor ability, memory ability and / or spatial exploration ability; 151. The use according to claim 150.

152. The cognitive impairment includes mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment; 152. Use according to any one of claims 150 to 151.

153. The cognitive impairment includes cognitive impairment caused by normal aging, Lewis' physical dementia (LBD), frontotemporal dementia and / or vascular dementia; 153. The use according to any one of claims 150 to 152.

154. The cognitive impairment-inducing disease includes Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and / or Creutzfeldt-Jakob disease (CJD); 154. Use according to any one of claims 150 to 153.

155. The neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

155. Use according to any one of claims 150 to 154.

156. The neurodegenerative diseases include neurodegenerative diseases caused by neuronal death and glial cell homeostasis, neurodegenerative diseases caused by aging, neurodegenerative diseases caused by effects on CNS cell function, neurodegenerative diseases caused by abnormal communication between cells and / or neurodegenerative diseases caused by impaired cell motility.

156. Use according to any one of claims 150 to 155.

157. The neurodegenerative disease includes Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD); 157. Use according to any one of claims 150 to 156.

158. The polypeptide, the drug combination and / or the pharmaceutical composition are formulated to be suitable for oral and / or injectable administration.

158. Use according to any one of claims 150 to 157.

159. The polypeptide, the drug combination and / or the pharmaceutical composition are formulated to be suitable for intravenous injection.

159. Use according to any one of claims 150 to 158.

160. ATP6V1B2, functionally active fragments thereof and / or nucleic acid molecules encoding them, Non-human animals or parts thereof.

161. The ATP6V1B2 is derived from a human.

161. A non-human animal or part thereof according to claim 160.

162. The ATP6V1B2 comprises the amino acid sequence shown in SEQ ID NO: 8 or 16. A non-human animal or a part thereof according to any one of claims 160 to 161.

163. The functionally active fragment of ATP6V1B2 has the ability to specifically bind to the amino acid sequence shown in SEQ ID NO:

5. A non-human animal or a part thereof according to any one of claims 160 to 162.

164. The functionally active fragment of ATP6V1B2 includes a truncation of ATP6V1B2. A non-human animal or a part thereof according to any one of claims 160 to 163.

165. The functionally active fragment of ATP6V1B2 comprises at least a portion of the amino acid sequence from positions 288 to 512 of human ATP6V1B2 protein. A non-human animal or a part thereof according to any one of claims 160 to 164.

166. The functionally active fragment of ATP6V1B2 comprises at least a part of the amino acid sequence from positions 288 to 512 of mouse ATP6V1B2 protein. A non-human animal or a part thereof according to any one of claims 160 to 165.

167. The functionally active fragment of ATP6V1B2 comprises the amino acid sequence shown in SEQ ID NO: 10 or 11. A non-human animal or a part thereof according to any one of claims 160 to 166.

168. The nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 9 or 17. A non-human animal or a part thereof according to any one of claims 160 to 167.

169. The non-human animal is a gene-edited non-human animal or is derived from a gene-edited non-human animal. A non-human animal or a part thereof according to any one of claims 160 to 168.

170. The non-human animal is a gene knock-in non-human animal or is derived from a gene knock-in non-human animal. A non-human animal or a part thereof according to any one of claims 160 to 169.

171. The expression of the ATP6V1B2 and / or its functionally active fragment is regulated by a corresponding endogenous regulatory element in the genome of the non-human animal. A non-human animal or a part thereof according to any one of claims 160 to 170.

172. The non-human animal is a non-human mammal. A non-human animal or a part thereof according to any one of claims 160 to 171.

173. The non-human animal is selected from the group consisting of mouse, rat, rabbit, ape, monkey, pig, cow, sheep, horse, chicken, dog, alpaca and cat; A non-human animal or a part thereof according to any one of claims 160 to 172.

174. The non-human animal is a rodent or a primate. A non-human animal or a part thereof according to any one of claims 160 to 173.

175. The portion comprises an organ, tissue and / or cell of the non-human animal. A non-human animal or a part thereof according to any one of claims 160 to 174.

176. the portion includes a bodily fluid; A non-human animal or a part thereof according to any one of claims 160 to 175.

177. The body fluid is selected from the group consisting of blood, plasma, serum, urine, sweat, tears, saliva, semen and cerebrospinal fluid; 177. A non-human animal or part thereof according to claim 176.

178. The part includes a brain or a part thereof. A non-human animal or a part thereof according to any one of claims 160 to 177.

179. the portion is incapable of developing into a complete non-human animal; A non-human animal or a part thereof according to any one of claims 160 to 178.

180. ATP6V1B2, functionally active fragments thereof and / or nucleic acid molecules encoding them, Isolated cells.

181. The ATP6V1B2 is derived from a human. The cell described in claim 180.

182. The ATP6V1B2 and / or its functionally active fragments comprise an amino acid sequence as set forth in any one of SEQ ID NOs: 8, 10, 11, and 16; A cell according to any one of claims 180 to 181.

183. The gene encoding ATP6V1B2 and / or its functionally active fragment is homozygous or heterozygous; A cell according to any one of claims 180 to 182.

184. The nucleic acid molecule encoding ATP6V1B2 and / or its functionally active fragments comprises the nucleic acid sequence shown in SEQ ID NO: 9 or 17; A cell according to any one of claims 180 to 183.

185. The expression of the ATP6V1B2 and / or its functionally active fragment is regulated by a corresponding endogenous regulatory element in the cell genome; A cell according to any one of claims 180 to 184.

186. Non-human animal cells, A cell according to any one of claims 180 to 185.

187. The non-human animal is a non-human mammal. The cell described in claim 186.

188. The non-human animal is selected from the group consisting of mouse, rat, rabbit, ape, monkey, pig, cow, sheep, horse, chicken, dog, alpaca and cat; A cell according to any one of claims 180 to 187.

189. The non-human animal is a rodent or a primate. A cell according to any one of claims 180 to 188.

190. incapable of developing into a complete non-human animal, A cell according to any one of claims 180 to 189.

191. ATP6V1B2, functionally active fragments thereof and / or nucleic acid molecules encoding them, organization.

192. The ATP6V1B2 is derived from a human. The tissue of claim 191.

193. The ATP6V1B2 and / or its functionally active fragments comprise an amino acid sequence as set forth in any one of SEQ ID NOs: 8, 10, 11, and 16; 193. The tissue according to any one of claims 191 to 192.

194. The gene encoding ATP6V1B2 and / or its functionally active fragment is homozygous or heterozygous; 194. The tissue according to any one of claims 191 to 193.

195. The nucleic acid molecule encoding ATP6V1B2 and / or its functionally active fragments comprises the nucleic acid sequence shown in SEQ ID NO: 9 or 17; 195. The tissue according to any one of claims 191 to 194.

196. is or is derived from gene-edited tissue; 196. The tissue according to any one of claims 191 to 195.

197. A gene knock-in tissue or derived from a gene knock-in tissue.

197. A tissue according to any one of claims 191 to 196.

198. The expression of the ATP6V1B2 and / or its functionally active fragments is regulated by corresponding endogenous regulatory elements in the tissue genome; 202. The tissue according to any one of claims 191 to 197.

199. Non-human animal tissue, 200. The tissue of any one of claims 191 to 198.

200. The non-human animal is a non-human mammal. The tissue described in claim 199.

201. The non-human animal is selected from the group consisting of mouse, rat, rabbit, ape, monkey, pig, cow, sheep, horse, chicken, dog, alpaca and cat; The tissue according to any one of claims 191 to 200.

202. The non-human animal is a rodent or a primate. The tissue according to any one of claims 191 to 201.

203. Including body fluids, 203. The tissue according to any one of claims 191 to 202.

204. The body fluid is selected from the group consisting of blood, plasma, serum, urine, sweat, tears, saliva, semen and cerebrospinal fluid; The tissue of claim 203.

205. Including the brain or parts thereof, A tissue according to any one of claims 191 to 204.

206. incapable of developing into a complete non-human animal, A tissue according to any one of claims 191 to 205.

207. 1. A cell line or cell culture comprising: Derived from a non-human animal or part thereof according to any one of claims 160 to 179, derived from a cell according to any one of claims 180 to 190, and / or derived from a tissue according to any one of claims 191 to 206. Cell lines or cell cultures.

208. The cell culture is a primary cell culture.

208. The cell line or cell culture of claim 207.

209. The cell culture comprises an organoid.

209. A cell line or cell culture according to any one of claims 207 to 208.

210. A non-human animal or part thereof according to any one of claims 160 to 179, a cell according to any one of claims 180 to 190, a tissue according to any one of claims 191 to 206, and / or a cell line or primary cell culture according to any one of claims 207 to 209, composition.

211. A method for the preparation of a non-human animal or part thereof according to any one of claims 160 to 179, a cell according to any one of claims 180 to 190, a tissue according to any one of claims 191 to 206, and / or a cell line or primary cell culture according to any one of claims 207 to 209, and one or more additional components selected from the group consisting of analytical buffers, controls, substrates, standards, detection materials, laboratory supplies, equipment, devices, cells, organs, tissues and user manuals or instructions. Reagent kit.

212. 1. A method for screening for drugs that can improve learning ability, treat cognitive disorders, and / or treat neurodegenerative diseases in a subject, comprising: A method comprising administering a candidate drug to a non-human animal or part thereof described in any one of claims 160 to 179, a cell described in any one of claims 180 to 190, a tissue described in any one of claims 191 to 206, and / or a cell line or primary cell culture described in any one of claims 207 to 209, and measuring the effect of the candidate drug on the expression level and / or activity of ATP6V1B2 in a subject.

213. 1. A method for screening for drugs that can improve learning ability, treat cognitive disorders, and / or treat neurodegenerative diseases in a subject, comprising: A method comprising: (i) administering a candidate drug to a non-human animal or part thereof described in any one of claims 160 to 179, a cell described in any one of claims 180 to 190, a tissue described in any one of claims 191 to 206, and / or a cell line or primary cell culture described in any one of claims 207 to 209; (ii) measuring the effect of the candidate drug on the expression level and / or activity of ATP6V1B2 in a subject; and (iii) if the expression level and / or activity of ATP6V1B2 is improved after administration of the candidate drug, the candidate drug enables improvement of the learning ability of the subject, treatment of a cognitive impairment, and / or treatment of a neurodegenerative disease.

214. The expression level of ATP6V1B2 includes the expression level of the ATP6V1B2 gene, the transcription level of the ATP6V1B2 gene, and / or the expression level of the ATP6V1B2 protein; 214. The method according to any one of claims 212 to 213.

215. The improvement includes an improvement in the expression level and / or activity of ATP6V1B2 by at least about 10% compared to the expression level and / or activity of ATP6V1B2 native to the subject.

215. The method of any one of claims 212 to 214.

216. The learning ability includes cognitive ability, motor ability, memory ability and / or spatial exploration ability; 216. The method of any one of claims 212 to 215.

217. The cognitive impairment includes mild cognitive impairment (MCI), intermediate cognitive impairment, and late cognitive impairment; 217. The method of any one of claims 212 to 216.

218. The cognitive impairment includes cognitive impairment caused by normal aging, Lewis physical dementia (LBD), frontotemporal dementia and / or vascular dementia; 218. The method of any one of claims 212 to 217.

219. The cognitive impairment-inducing disease includes Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and / or Creutzfeldt-Jakob disease (CJD); 219. The method of any one of claims 212 to 218.

220. The neurodegenerative disease includes acute neurodegenerative diseases and chronic neurodegenerative diseases.

220. The method of any one of claims 212 to 219.

221. The neurodegenerative diseases include neurodegenerative diseases caused by neuronal death and glial cell homeostasis, neurodegenerative diseases caused by aging, neurodegenerative diseases caused by effects on CNS cell function, neurodegenerative diseases caused by abnormal communication between cells and / or neurodegenerative diseases caused by impaired cell motility.

221. The method of any one of claims 212 to 220.

222. The neurodegenerative disease includes Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and / or Huntington's disease (HD); 222. The method of any one of claims 212 to 221.

223. Use of a non-human animal or part thereof according to any one of claims 160 to 179, a cell according to any one of claims 180 to 190, a tissue according to any one of claims 191 to 206 and / or a cell line or primary cell culture according to any one of claims 207 to 209 in the manufacture of an identification and / or screening system, The use, wherein the system is used to screen for drugs and / or biomarkers that enable improvement of a subject's learning ability, treatment of cognitive impairment, and / or treatment of a neurodegenerative disease.

224. Used to screen for drugs and / or biomarkers that can improve a subject's learning ability, treat cognitive disorders, and / or treat neurodegenerative diseases; A non-human animal or part thereof according to any one of claims 160 to 179, a cell according to any one of claims 180 to 190, a tissue according to any one of claims 191 to 206, and / or a cell line or primary cell culture according to any one of claims 207 to 209.

225. A method for preparing a non-human animal or a part thereof according to any one of claims 160 to 179, a cell according to any one of claims 180 to 190, or a tissue according to any one of claims 191 to 206, comprising: A method comprising comprising comprising comprising containing and / or expressing said ATP6V1B2 and / or functionally active fragments thereof in a non-human animal or part, cell or tissue thereof.

226. 1. A method for producing a disease model, comprising: A method comprising comprising comprising containing and / or expressing ATP6V1B2 and / or a functionally active fragment thereof in a non-human animal or part, cell or tissue thereof.

227. The ATP6V1B2 and / or its functionally active fragments comprise an amino acid sequence as set forth in any one of SEQ ID NOs: 8, 10, 11, and 16; 227. The method of any one of claims 225 to 226.

228. transfecting a nucleic acid sequence encoding said ATP6V1B2 and / or a functionally active fragment thereof; 228. The method of any one of claims 225 to 227.

229. The nucleic acid sequence encoding ATP6V1B2 and / or its functionally active fragments comprises the nucleic acid sequence shown in SEQ ID NO: 9 or 17; The method of claim 228.

230. said transfecting comprising transfecting an adeno-associated virus comprising said nucleic acid sequence; 230. The method of any one of claims 228 or 229.

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