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WO2024255822A1 - Liant de dir et utilisation associée - Google Patents

Liant de dir et utilisation associée Download PDF

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Publication number
WO2024255822A1
WO2024255822A1 PCT/CN2024/099135 CN2024099135W WO2024255822A1 WO 2024255822 A1 WO2024255822 A1 WO 2024255822A1 CN 2024099135 W CN2024099135 W CN 2024099135W WO 2024255822 A1 WO2024255822 A1 WO 2024255822A1
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Prior art keywords
dir
binding agent
disease
present application
polypeptide
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English (en)
Chinese (zh)
Inventor
李开诚
李震
史海翔
攸璞
潘晶
马雪飞
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Shanghai Quietd Biotechnology Co Ltd
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Shanghai Quietd Biotechnology Co Ltd
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Publication of WO2024255822A1 publication Critical patent/WO2024255822A1/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans

Definitions

  • the present application relates to the field of biomedicine, and specifically to a binding agent that binds to DIR and/or its functional fragments and its use in improving cognitive impairment.
  • the number of aging population in my country is increasing year by year. Due to the influence of factors such as decreased immunity and vascular sclerosis, the elderly are facing the troubles of various diseases. Among them, the incidence of Alzheimer's disease in the current elderly population cannot be ignored. Dementia patients are not only tortured by the disease themselves, but also put great pressure on their families. Therefore, it is extremely important to develop therapeutic drugs for Alzheimer's disease.
  • the present application provides a novel method for improving cognitive impairment.
  • the present application provides a binding agent that binds to the intron retained splicing product DIR and/or its functional fragment of a DNA damage-inducible transcript 4-like transcript.
  • the DIR and/or its functional fragment are derived from a mammal.
  • the DIR and/or its functional fragment are derived from primates.
  • the DIR and/or its functional fragment are derived from human.
  • the DIR comprises the amino acid sequence shown in SEQ ID NO: 1.
  • the functional fragment of DIR comprises an amino acid sequence encoded by a retained intron in DDIT4L.
  • the functional fragment of DIR comprises an amino acid sequence as shown in any one of SEQ ID NOs: 4-5. Amino acid sequence.
  • the binding agent is a protein and/or a polypeptide.
  • the binding agent comprises the ionotropic glutamate receptor (AMPARs) receptor subunit GluA1 and/or a fragment thereof.
  • AMPARs ionotropic glutamate receptor
  • the GluA1 and/or fragments thereof are derived from mammals.
  • the GluA1 and/or fragments thereof are derived from primates.
  • the GluA1 and/or fragments thereof are of human origin.
  • the GluA1 comprises the amino acid sequence shown in SEQ ID NO: 7 and/or its variants.
  • the binding agent comprises the amino acid sequence shown at positions 198 to 205 of GluA1 and/or a functional variant thereof.
  • the binding agent comprises the amino acid sequence shown in SEQ ID NO: 8 and/or its functional variants.
  • the binding agent is capable of regulating the expression level and/or biological activity of the intron-retained splicing product DIR of the DNA damage-inducible transcript 4-like transcript and/or its functional fragments.
  • the binding agent is capable of reducing the expression level and/or biological activity of the DIR and/or its functional fragment in a subject.
  • the reduction comprises a reduction of at least about 10% in the expression level and/or biological activity of the DIR and/or its functional fragment compared to the original expression level and/or biological activity of the DIR and/or its functional fragment in the subject.
  • the expression level includes the expression level of the gene encoding the DIR/or its functional fragment, the transcription level of the gene encoding the DIR/or its functional fragment and/or the expression level of the DIR/or its functional fragment.
  • the functional fragment of the DIR retains at least a portion of the biological activity of the DIR.
  • the biological activity comprises the ability to reduce the frequency of excitatory postsynaptic current (EPSC) and/or the ability to reduce the amplitude of EPSC.
  • ESC excitatory postsynaptic current
  • the reduction comprises administering the DIR and/or its functional fragment and/or a nucleic acid encoding the DIR and/or its functional fragment, thereby reducing the frequency of excitatory postsynaptic currents (EPSCs) in the subject and/or reducing the amplitude of EPSCs in the subject, compared to the biological activity of the original DIR and/or its functional fragment in the subject.
  • ESCs excitatory postsynaptic currents
  • the biological activity comprises affecting cognitive abilities.
  • the biological activity comprises participation in a signaling pathway associated with A ⁇ deposition, and/or Involved in signaling pathways associated with Tau tangle generation.
  • the biological activity comprises inducing A ⁇ deposition and/or amyloid plaque formation by gelsolin.
  • the DIR and/or its functional fragment induces A ⁇ deposition and/or amyloid plaque formation by binding to gelsolin.
  • the expression level of the DIR and/or its functional fragment is positively correlated with the expression level of A ⁇ .
  • the present application provides an isolated polypeptide comprising the amino acid sequence shown in SEQ ID NO: 8 and/or its variants.
  • the present application provides an immunoconjugate comprising the binding agent described herein and/or the polypeptide described herein.
  • the present application provides a nucleic acid molecule encoding the binding agent described in the present application and/or the polypeptide described in the present application.
  • the present application provides a vector comprising the nucleic acid molecule described in the present application.
  • the present application provides a cell comprising the binding agent described in the present application, the polypeptide described in the present application, the nucleic acid molecule described in the present application and/or the vector described in the present application.
  • the present application provides a pharmaceutical composition
  • a pharmaceutical composition comprising the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein and/or the cell described herein, and optionally a pharmaceutically acceptable carrier.
  • the present application provides a kit comprising the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application and/or the pharmaceutical composition described in the present application.
  • the kit is used to detect DIR in a sample.
  • the present application provides a method for detecting DIR in a sample, the method comprising administering the binding agent described in the present application, the polypeptide molecule described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application, the pharmaceutical composition described in the present application and/or the kit described in the present application.
  • the present application provides the use of the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application and/or the pharmaceutical composition described in the present application in the preparation of a kit.
  • the present application provides the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application and/or the A pharmaceutical composition, use thereof in preparing an agent for preventing and/or treating a disease, wherein the disease includes cognitive impairment.
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein and/or the pharmaceutical composition described herein in the preparation of an agent for preventing and/or treating a disease, wherein the disease includes a neurodegenerative disease.
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein, the pharmaceutical composition described herein and/or the kit described herein in the preparation of a reagent for diagnosing and/or assessing a disease, wherein the disease includes cognitive impairment.
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the pharmaceutical composition described herein and/or the kit described herein in the preparation of a reagent for diagnosing and/or evaluating a disease, wherein the disease includes a neurodegenerative disease.
  • the cognitive impairment comprises cognitive impairment caused by normal aging, Lewis body dementia (LBD), frontotemporal dementia and/or vascular dementia.
  • LBD Lewis body dementia
  • frontotemporal dementia frontotemporal dementia
  • vascular dementia vascular dementia
  • the cognitive impairment inducing diseases include Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and/or Creutzfeldt-Jakob disease (CJD).
  • the cognitive impairment comprises early cognitive impairment (MCI), mid-stage cognitive impairment and late stage cognitive impairment.
  • the cognitive impairment comprises amnestic MCI with impairment of multiple cognitive domains (aMCI-m).
  • the neurodegenerative disease comprises an acute neurodegenerative disease and a chronic neurodegenerative disease.
  • the neurodegenerative disease includes a neurodegenerative disease caused by neuronal death and glial cell homeostasis, a neurodegenerative disease caused by aging, a neurodegenerative disease caused by affected CNS cell function, a neurodegenerative disease caused by abnormal intercellular communication and/or a neurodegenerative disease caused by impaired cell motility.
  • the neurodegenerative disease comprises Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and/or Huntington's disease (HD).
  • MS multiple sclerosis
  • ALS amyotrophic lateral sclerosis
  • HD Huntington's disease
  • the neurodegenerative disease comprises Alzheimer's disease.
  • the neurodegenerative disease comprises early Alzheimer's disease, middle Alzheimer's disease and/or late Alzheimer's disease.
  • the subject comprises a mammal.
  • the subject comprises a human.
  • the subject comprises a patient with a neurodegenerative disease and/or a patient with a cognitive disorder.
  • the subject comprises an Alzheimer's disease patient.
  • the subject is elderly.
  • the agent is formulated for oral administration and/or injection.
  • the present application provides a method for preventing and/or treating cognitive impairment, which comprises administering the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein and/or the pharmaceutical composition described herein to a subject in need thereof.
  • the present application provides a method for preventing and/or treating neurodegenerative diseases, which comprises administering the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein and/or the pharmaceutical composition described herein to a subject in need thereof.
  • the present application provides a method for diagnosing cognitive impairment and/or assessing cognitive impairment, which comprises using the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the pharmaceutical composition described in the present application and/or the kit described in the present application.
  • the present application provides a method for diagnosing a neurodegenerative disease and/or evaluating a neurodegenerative disease, which comprises using the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the pharmaceutical composition described in the present application and/or the kit described in the present application.
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein and/or the pharmaceutical composition described herein in preventing and/or treating cognitive disorders.
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein and/or the pharmaceutical composition described herein in preventing and/or treating neurodegenerative diseases.
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the pharmaceutical composition described herein and/or the kit described herein in the diagnosis and/or assessment of cognitive disorders.
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the pharmaceutical composition described herein and/or the kit described herein in the diagnosis and/or assessment of neurodegenerative diseases.
  • Figures 1A-1D show the identification of DIR described in the present application.
  • Figure 1A shows the formation of the human DDIT4L gene spliceosome described in the present application, wherein NS represents the normal spliceosome and IR represents the DIR formed by abnormal splicing.
  • Figure 1B shows the mRNA identification results of DDIT4L and DIR.
  • Figure 1C shows the results of Western Blot detection of DIR expression using DIR antibody.
  • Figure 1D shows the results of Western Blot detection of DIR expression in blood samples using DIR antibody.
  • FIG. 2 shows the effect of the functional fragment of DIR described in the present application on excitatory postsynaptic current.
  • FIG3 is a schematic diagram showing the structure of the functional fragment of DIR described in the present application.
  • FIG. 4 shows the biological functions of the DIR functional fragments described in the present application.
  • Figures 5A-5I show that DIR induces A ⁇ deposition by binding to gelsolin.
  • Figure 5E Mass spectrometry identified the top 8 molecules in the ⁇ 85 kDa size range. Gelsolin is the most abundant molecule among them.
  • Fig. 5G PLA experiments showed that gelsolin binds to A ⁇ (arrows) in the hippocampus of AD patients.
  • Figures 6A-6D show that DIR does not directly bind to A ⁇ , but induces A ⁇ deposition.
  • Figures 7A-7C show DIR-mediated A ⁇ plaque formation.
  • the A ⁇ -positive area is larger than the Thioflavin S- and DIR-positive signals.
  • Scale bar 50 ⁇ m.
  • Scale bar 50 ⁇ m.
  • Figures 9A-9L show that plasma DIR is a potential biomarker for AD and aMCI.
  • Figures 10A-10B show ROC analysis of plasma tTau.
  • FIGS 11A-11E show the correlation analysis between plasma DIR and A ⁇ .
  • Figure 11C Three-dimensional visualization of amyloid 18F-AV-45 SUV in I or NC of AD/MC patients. The color bar represents the SUV index obtained from the amyloid 18F-AV-45 image.
  • Plasma DIR values in AD/MCI patients were associated with positive signals in the cortex, especially in the temporal lobe.
  • Figure 11E Model of DIR-induced amyloid plaque deposition and blood secretion. Under pathological conditions, hypoxia leads to abnormal intron retention, which causes the translation of DIR protein. A ⁇ is produced by the proteolytic process of amyloid precursor protein (APP) and has a direct binding to gelsolin. DIR causes A ⁇ deposition by binding to gelsolin, ultimately forming amyloid plaques in the brain. DIR can also be released into the blood system.
  • APP amyloid precursor protein
  • FIGs 12A-12B show the correlation between plasma DIR and A ⁇ 42, and quantitative analysis of the SUV value of A ⁇ -PET.
  • Figure 12B Quantitative analysis of SUV values in AD/MCI patients or NC. *, p ⁇ 0.05.
  • Figures 13A-13B show that GluA1 is a binding target of DIR as described in the present application.
  • Figure 13A shows the results of a co-IP experiment of brain tissues of wild-type mice and DIR knock-in mice.
  • Figure 13B shows that mass spectrometry analysis of the specific binding band of DIR revealed that GluA1 is a binding target of DIR.
  • Figures 14A-14B show that GluA1 described in the present application can bind to DIR.
  • Figure 14A shows that a co-IP experiment was performed using brain tissues of wild-type mice and DIR knock-in mice, and the experimental results showed that DIR can bind to GluA1.
  • Figure 14B shows that DIR and GluA1 were exogenously expressed in HEK293 cells and a co-IP experiment was performed, and the results also showed that DIR can bind to GluA1.
  • Figures 15A-15C show that the fragments derived from GluA1 described in the present application can bind to DIR.
  • Figure 15A shows that through computer simulation, it was found that GluA1 R198-E205 is the binding site of DIR, and GluA1 C204 plays an important role in it.
  • Figure 15B shows that adding a short peptide (R198-E205) of GluA1 at a final concentration of 10uM to the cell lysate co-transfected with GluA1 and DIR can effectively block the binding between GluA1 and DIR.
  • Figure 15C shows that in the case of co-transfected GluA1 Or Co-IP experiments were performed on the cell lysate of GluA1 point mutation (GluA1 C204A ) and DIR, and it was found that the binding of GluA1 C204A to DIR was weakened.
  • FIG. 16 shows that the polypeptide fragment derived from GluA1 described in the present application can reverse the inhibitory effect of sEPSC induced by DIR.
  • FIG. 17 shows that administration of the polypeptide fragment derived from GluA1 described in the present application can improve the learning ability of mice.
  • DDIT4L generally refers to DNA damage-inducible transcript 4-like, which may also be referred to as REDD2/RTP801L. Studies have found that DDIT4L may be associated with cardiac dysfunction. It may also be used to treat gliomas.
  • accession number of the human DDIT4L gene in GenBank is 115265; the accession number of the human DDIT4L protein in GenBank is NP_660287.1.
  • DIR generally refers to the intron retention splicing product of DDIT4L.
  • the splicing reaction of DDIT4L can be seen in Figure 1.
  • amino acid sequence of the DIR can be shown as SEQ ID NO: 1.
  • binding agent generally refers to a natural molecule or non-natural molecule that specifically binds to a target or a portion of a target.
  • Binding agents can include small molecule compounds, polymers and/or biomacromolecules. Binding agents can include proteins, peptides, nucleic acids, sugars, lipids and small molecule compounds.
  • GluA1 may also be referred to as GIRA1 (glutamate ionotropic receptor AMPA type subunit 1), which generally refers to one of the four subtypes of ionotropic glutamate receptors (AMPARs).
  • GIRA1 glutamate ionotropic receptor AMPA type subunit 1
  • AMPARs ionotropic glutamate receptors
  • GluA1 is related to neuronal synaptic transmission and the release of action potentials, and is also closely related to the occurrence and development of AD.
  • the accession number of the human GluA1 gene in GenBank may be 2890; the accession number of the human GluA1 protein in Uniprot may be P42261.
  • the amino acid sequence of the GluA1 may be as shown in SEQ ID NO:7.
  • the "variant" and/or “functional variant” may be, for example, a protein or polypeptide in which one or more amino acids have been substituted, deleted or added in the amino acid sequence of the protein and/or the polypeptide (e.g., a binding agent that specifically binds to DIR or a fragment thereof).
  • the variant may comprise a protein or polypeptide that has been subjected to amino acid changes by at least 1, such as 1-30, 1-20 or 1-10, and for example 1, 2, 3, 4 or 5 amino acid substitutions, deletions and/or insertions.
  • the functional variant may substantially retain the amino acid sequence of the protein or polypeptide prior to the change (e.g., substitution, deletion or addition). The biological properties of the protein or the polypeptide.
  • the functional variant can maintain at least 60%, 70%, 80%, 90%, or 100% of the biological activity (e.g., the ability to specifically bind to DIR) of the protein or the polypeptide before the change.
  • the substitution can be a conservative substitution.
  • the variant can also be a polypeptide covering its functionally active fragments, not limited to the polypeptide containing the functionally active fragment of the protein produced after processing and/or modification occurring in the cell.
  • the "variant" may be a homologue.
  • the homologue may be a protein or polypeptide having at least about 85% (e.g., at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence homology with the amino acid sequence of the protein and/or polypeptide (e.g., a binding agent or fragment thereof that specifically binds to DIR).
  • sequence homology percentage ratio can be calculated in the following manner: two sequences to be compared are compared in a comparison window, determine that there is identical nucleic acid base (for example, A, T, C, G, I) or identical amino acid residue (for example, Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys and Met) number of positions in two sequences to obtain the number of matching positions, the number of matching positions is divided by the total number of positions (that is, window size) in the comparison window, and the result is multiplied by 100, to produce sequence homology percentage ratio.Comparison carried out in order to determine the sequence homology percentage ratio can be realized in a variety of ways known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALI
  • FASTA and BLAST A description of the FASTA algorithm can be found in W.R. Pearson and D.J. Lipman, "Improved tools for biological sequence comparisons," Proc. Natl. Acad. Sci., 85: 2444-2448, 1988; and D.J. Lipman and W.R. Pearson, "Rapid and sensitive protein similarity search,” Science, 227: 1435-1441, 1989.
  • a description of the BLAST algorithm can be found in S. Altschul, W. Gish, W. Miller, E.W. Myers and D. Lipman, "A basic local alignment search tool," Journal of Molecular Biology, 215: 403-410, 1990.
  • polypeptide molecule and “polypeptide” and “peptide” are used interchangeably and generally refer to polymers of amino acid residues.
  • fusion protein generally refers to a polypeptide having at least two parts covalently linked together. Each part can be a polypeptide with different properties.
  • the property can be a biological property, such as in vitro or in vivo activity.
  • the property can also be a simple chemical or physical property, such as binding to a target molecule, catalysis of a reaction, etc.
  • the two parts can be directly connected by a single peptide bond or by a peptide linker.
  • isolated generally refers to an isolated substance obtained from a natural state by artificial means.
  • a certain unisolated polynucleotide or polypeptide naturally exists in a living animal, and a highly purified polynucleotide or polypeptide isolated from this natural state
  • isolated may not exclude the presence of artificial or synthetic substances, nor may it exclude the presence of other impure substances that do not affect the activity of the substance.
  • nucleic acid molecule generally refers to nucleotides of any length in isolated form, either deoxyribonucleotides or ribonucleotides, or analogs isolated from their natural environment or artificially synthesized.
  • vector generally refers to a nucleic acid delivery vehicle into which a polynucleotide encoding a protein can be inserted and the protein can be expressed.
  • a vector can transform, transduce or transfect a host cell so that the genetic material elements it carries are expressed in the host cell.
  • a vector may contain multiple elements that control expression.
  • a vector may also contain a replication initiation site.
  • a vector may also include components that assist it in entering the cell.
  • the term "cell” generally refers to a single cell, cell line or cell culture that may be or has been a recipient of a subject's plasmid or vector, including a nucleic acid molecule described herein or a vector described herein.
  • a cell may include the offspring of a single cell. Due to natural, accidental or intentional mutations, the offspring may not necessarily be identical to the original parent cell (in the form of the total DNA complement or in the genome).
  • a cell may include a cell transfected in vitro with a vector described herein.
  • immunoconjugate generally refers to a substance formed by linking a polypeptide with other active agents, which may be small molecule active agents, such as therapeutic agents, imaging probes or spectroscopy probes.
  • the term "pharmaceutical composition” generally refers to a composition for preventing/treating a disease or condition.
  • the pharmaceutical composition may include an isolated polypeptide as described herein, a nucleic acid molecule as described herein, a vector as described herein, and/or a cell as described herein, and optionally a pharmaceutically acceptable adjuvant.
  • the pharmaceutical composition may also include suitable formulations such as one or more (pharmaceutically effective) carriers.
  • the acceptable ingredients of the composition may be non-toxic to the recipient at the dosage and concentration used.
  • the pharmaceutical composition of the present application includes, but is not limited to, liquid, frozen and lyophilized compositions.
  • pharmaceutically acceptable carrier generally refers to a pharmaceutically acceptable carrier, excipient or stabilizer that is non-toxic to cells or mammals exposed thereto at the doses and concentrations used.
  • Physiologically acceptable carriers may include suitable substances.
  • Pharmaceutically acceptable carriers and vectors used to insert nucleic acids in genetic engineering are generally not the same substance.
  • the term "specific binding” or “specific” generally refers to a measurable and reproducible interaction, such as binding between a target and an antibody, which can determine the presence of a target in the presence of a heterogeneous population of molecules (including biomolecules).
  • an antibody that specifically binds to a target (which may be an epitope) may be an antibody that binds to the target with greater affinity, avidity, more readily, and/or for a greater duration than it binds to other targets.
  • an antibody specifically binds to an epitope on a protein that is conserved among proteins of different species.
  • specific binding may include but does not require exclusive binding.
  • expression level generally refers to the protein, RNA or mRNA level of a specific related gene. Any method known in the art can be used to determine the expression level of a specific related gene (e.g., human DDIT4L gene). In the present application, the "expression” generally refers to the process of converting the information encoded by a gene into a structure that exists in a cell and operates in the cell.
  • reverse transcription and amplification analysis e.g., PCR, RT-PCR or quantitative RT-PCR
  • hybridization analysis e.g., Northern blotting, dot blotting, in situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, protein blotting, immunohistochemistry, immunostaining or mass spectrometry
  • Analysis can be performed directly on a biological sample or on a protein/nucleic acid separated from a sample.
  • the term "activity" generally refers to any activity associated with a particular protein.
  • the activity may include any activity associated with, for example, a DIR protein.
  • the activity may include an enzymatic activity associated with a protease.
  • the activity may include a biological activity.
  • the activity may include the binding of a protein to a receptor, for example, the binding may produce a measurable downstream effect.
  • the activity may include any activity that would be attributed to the protein by a person skilled in the art.
  • cognitive impairment generally refers to a progressive loss (including neuronal death) or related diseases and conditions that are believed to be or are involved in neuronal structure and/or function.
  • the characteristics of the cognitive impairment may include damage to cognition (e.g., memory, attention, perception and/or thinking). These disorders may include pathogen-induced cognitive dysfunction, such as HIV-related cognitive dysfunction and Lyme disease-related cognitive dysfunction.
  • Examples of cognitive impairment may include Alzheimer's disease, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), autism, early cognitive impairment (MCI), stroke, traumatic brain injury (TBI) and/or age-related memory impairment (AAMI).
  • ALS amyotrophic lateral sclerosis
  • MCI early cognitive impairment
  • TBI traumatic brain injury
  • AAMI age-related memory impairment
  • neurodegenerative disease generally refers to cognitive disorders such as dementia caused by the gradual loss of neuronal structure and function, including neuronal death and glial cell balance.
  • age e.g., Alzheimer's disease (AD), Parkinson's disease (PD)
  • gene mutations that affect CNS cell function e.g., Huntington's disease, early-onset AD or PD, amyotrophic lateral sclerosis (ALS)
  • AD Alzheimer's disease
  • PD Parkinson's disease
  • ALS amyotrophic lateral sclerosis
  • the neurodegenerative disease may have changes and/or conditions selected from the following: protein misfolding and aggregation; neuroinflammation (e.g., CNS inflammation that occurs under toxic stimulation (e.g., protein aggregation), infection, traumatic injury, or autoimmune signal stimulation); changes in cell signal transduction; acquired aging/cell death (e.g., interrupted apoptotic signal transduction, mitochondrial dysfunction, impaired autophagy, and necrotic bodies activated by stress/inflammation); motor cell damage and epigenetic changes.
  • neuroinflammation e.g., CNS inflammation that occurs under toxic stimulation (e.g., protein aggregation), infection, traumatic injury, or autoimmune signal stimulation)
  • changes in cell signal transduction e.g., acquired aging/cell death (e.g., interrupted apoptotic signal transduction, mitochondrial dysfunction, impaired autophagy, and necrotic bodies activated by stress/inflammation); motor cell damage and epigenetic changes.
  • Alzheimer's disease usually refers to early-onset dementia or senile dementia, a neurodegenerative disease that progresses slowly and worsens over time.
  • the most common early symptom is loss of short-term memory (difficulty remembering recent events).
  • As the disease progresses at least one of the following symptoms may gradually appear: language disorders, disorientation (e.g., getting lost easily), emotional instability, loss of motivation, inability to take care of oneself, and behavioral problems.
  • language disorders e.g., getting lost easily
  • emotional instability e.g., getting lost easily
  • loss of motivation e.g., inability to take care of oneself
  • behavioral problems e.g., a cognitive problems.
  • the true cause of Alzheimer's disease remains unknown. Its progression may be related to the deposition of fibrillar amyloid plaques and Tau proteins in the brain.
  • There is currently no treatment that can stop or reverse the course of the disease but only a few methods may temporarily relieve or improve symptoms.
  • the term "Alzheimer's disease” can be used interchangeably with the term "Alzheimer's disease” in this application.
  • the Alzheimer's disease can include early Alzheimer's disease, mid-Alzheimer's disease and/or late Alzheimer's disease.
  • the learning and memory disorders of the early Alzheimer's disease patients will become more and more obvious, and in some cases, there will be language disorders, execution disorders, cognitive disorders (anognosia) and/or skill execution disorders (apraxia).
  • the mid-Alzheimer's disease patients will lose the ability to live independently and may not be able to carry out most of the daily activities (in some cases, they may suffer from anomia, aphasia, and/or agnosia).
  • the late Alzheimer's disease patients may rely on caregivers in the late stage.
  • the language ability may be completely lost.
  • they may not be able to eat on their own.
  • MCI middle cognitive impairment
  • the MCI may include cognitive impairment that meets the criteria for dementia but exceeds normal aging.
  • MCI is diverse in clinical manifestations, causes, prognosis, and prevalence.
  • MCI may be a pathological stage of Alzheimer's disease. Certain forms of cognitive impairment can be considered early manifestations of neurodegenerative diseases, which will eventually lead to dementia.
  • the MCI may include a subtype selected from the following group: aMCI-s: amnestic MCI with a single cognitive domain impairment; aMCI-m: amnestic MCI with multiple cognitive domain impairment; naMCI-s: non-amnestic MCI with a single cognitive domain impairment; and naMCI-m: non-amnestic MCI with multiple cognitive domain impairment.
  • cognitive impairment due to normal aging generally refers to cognitive impairment due to normal aging.
  • cognitive impairment due to normal aging can manifest as: memory loss, confusion about the location of familiar places, taking longer than usual to complete daily tasks, or changes in mood and personality.
  • Lewy body dementia usually refers to Lewy Body Detmentia.
  • Lewy body dementia is characterized by abnormal buildup of proteins into lumps called Lewy bodies.
  • Lewy body dementia causes a gradual decline in mental abilities. People with Lewy body dementia may experience visual hallucinations and changes in alertness and attention. Other effects include muscle stiffness, slowed movements, difficulty walking, and tremors. People with Lewy bodies in their brains can also have plaques and tangles associated with Alzheimer's disease.
  • 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-level reasoning, expressive language, speech perception, and memory formation.
  • the frontal and temporal lobes of the brain can shrink over time in people with frontotemporal dementia.
  • vascular dementia generally refers to problems with reasoning, judgment, and memory caused by impaired blood flow to the brain.
  • vascular dementia can include dementia caused by factors that increase risk of heart disease and stroke, such as high blood pressure and high cholesterol.
  • multi-infarct type generally refers to small noncortical infarcts caused by occlusion of a single perforator of a large cerebral artery.
  • the multiple infarction type may be a special type of cerebral infarction, also known as ischemic stroke.
  • the multiple infarction type may be manifested as hemisensory disturbance, aphasia, dysarthria, slow movements, and clumsiness (especially fine movements such as writing are more difficult).
  • Parkinson's disease generally refers to a progressive neurodegenerative disease.
  • the clinical features of Parkinson's disease (PD) may include motor symptoms (e.g., tremor, bradykinesia, rigidity, and postural instability), as well as neuropsychiatric and other non-motor manifestations.
  • the non-motor manifestations may include cognitive dysfunction and dementia, mood disorders (e.g., depression, anxiety, apathy), and sleep disorders.
  • CJD usually refers to a transmissible spongiform encephalopathy that occurs in humans.
  • CJD is a disease caused by prion infection.
  • CJD patients may show paranoid behavior, confusion, loss of appetite and weight, depression, and a few patients have visual or auditory abnormalities; in the advanced stage, it manifests as progressive neurological deterioration (such as sensory abnormalities, language disorders and aphasia).
  • MS multiple sclerosis
  • the term “multiple sclerosis (MS)” generally refers to a demyelinating neuropathy.
  • the insulating material i.e., myelin sheath
  • myelin sheath on the surface of nerve cells in the brain or spinal cord of the MS patient is damaged, and the signal transduction of the nervous system is impaired, which can lead to a series of possible symptoms that affect the patient's activities, mind, and even mental state. These symptoms can include double vision, unilateral visual impairment, muscle weakness, dysesthesia, or coordination disorders.
  • ALS myotrophic lateral sclerosis
  • Lou Gehrig's disease and motor neuron disease which is a progressive and fatal neurodegenerative disease.
  • a small number of ALS patients may develop frontotemporal dementia.
  • Some ALS patients will experience degeneration of their sense of sight, vision, touch, smell and taste, and a very small number of ALS patients will also develop dementia.
  • Huntington's disease generally refers to a genetic disorder that causes brain cells to die. As the disease progresses, the incoordination of body movements becomes more pronounced, and abilities gradually deteriorate until movement becomes difficult and speech is impossible. Mental abilities often decline into dementia.
  • senile stage generally refers to the aging stage of a subject.
  • the senile stage may be over 60 years old, over 70 years old, or over 75 years old; for mice, the senile stage may be over 10 months old, for example, over 13 months old or over 18 months old.
  • the subject at the senile stage may have one or more symptoms of learning deficits, memory impairment, memory deficits, and/or brain dysfunction.
  • neuron generally refers to a nerve cell, which is the main functional unit of the nervous system.
  • a neuron can be composed of a cell body and its protrusions, an axon, and one or more dendrites. Neurons can transmit information to other neurons or cells by releasing neurotransmitters at synapses.
  • excitatory postsynaptic current generally refers to the ion flow that causes an excitatory postsynaptic potential (EPSP).
  • the EPSP is a postsynaptic potential that makes the postsynaptic neuron more likely to fire an action potential. This temporary depolarization of the postsynaptic membrane potential caused by the influx of positively charged ions into the postsynaptic cell is the key to opening the ligand Results of gating ion channels.
  • the frequency and/or amplitude of the EPSC can be recorded using voltage clamp.
  • cognition generally refers to the ability of mental behavior to gain knowledge and understanding through thoughts, experiences and feelings.
  • the concept of cognition may not be limited to psychological concepts/domains, and may include, for example, executive function, memory, perception, attention, emotion, motor control and/or interference processing.
  • a ⁇ generally refers to any peptide resulting from the cleavage of ⁇ -amyloid precursor protein (APP) mediated by ⁇ -secretase.
  • the A ⁇ may include peptides of 37, 38, 39, 40, 41, 42, and 43 amino acids, and extend from the ⁇ -secretase cleavage site to amino acids 37, 38, 39, 40, 41, 42, or 43.
  • the A ⁇ may also be an N-terminal truncated type of the above-mentioned peptides, such as pyroglutamic acid forms pE3-40, pE3-42, pE3-43, pE11-42, pE11-43, and the like.
  • Tau generally refers to Tau proteins and components of a wide range of Tau aggregates (e.g., neurofibrillary tangles) associated with the stabilization of microtubules in neural cells.
  • the Tau tangles may include oligomeric and/or fibrous forms of Tau, which are toxic.
  • the Tau may also include all types and forms of Tau (e.g., different alternative splicing forms).
  • the term "subject” generally refers to a human or non-human animal, including but not limited to a cat, dog, horse, pig, cow, sheep, rabbit, mouse, rat or monkey.
  • the term “comprising” generally means including, encompassing, containing or encompassing. In some cases, it also means “for”, “consisting of".
  • the term "about” generally refers to a numerical range of 20% more or less than a particular value.
  • “about X” includes a numerical range of ⁇ 20%, ⁇ 10%, ⁇ 5%, ⁇ 2%, ⁇ 1%, ⁇ 0.5%, ⁇ 0.2% or ⁇ 0.1% of X, where X is a numerical value.
  • DIR intron retention
  • the functional fragment 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., DIR) by at least one amino acid residue.
  • the functional fragment may have a higher (e.g., at least 80%) homology with the parent or reference polypeptide.
  • the homology may include sequence similarity or identity.
  • the homology may be determined using standard techniques known in the art (see, e.g., Smith and Waterman, Adv. Appl. Math.
  • the percentage of identity shared by polynucleotide or polypeptide sequences is determined by direct comparison of sequence information between molecules, the comparison being performed by sequence alignment and using methods known in the art to determine identity.
  • An example of an algorithm suitable for determining sequence similarity is the BLAST algorithm (see Altschul et al., J. Mol. Biol. Journal of Molecular Biology, 215: 403-410 [1990]).
  • Software for performing BLAST analysis may be provided by Publicly available from the National Center for Biotechnology Information (NCBI).
  • the expression level of DIR may include the expression level of the DIR gene, the transcription level of the DIR gene and/or the expression level of the DIR protein.
  • the expression level may include the amount of a polynucleotide, mRNA or amino acid product or protein of a specific gene (e.g., human DDIT4L gene; and/or, a gene (e.g., human DIR gene) encoding human DIR and/or its functional fragments (e.g., DIR-I, and/or, DIR-II)).
  • the expression level may include the amount of a polynucleotide transcribed from a specific gene, a translated protein, or a fragment of a post-translationally modified protein.
  • QDLIR may be used in place of DIR, which may be a form of intron retention produced by abnormal shearing during expression of the human DDIT4L gene.
  • the gene encoding the DIR may be referred to as the DIR gene.
  • the expression level of the functional fragment of DIR may include the expression level of the functional fragment gene encoding DIR, the transcription level of the functional fragment gene encoding DIR, and/or the expression level of the functional fragment protein of DIR.
  • the expression level may include the amount of polynucleotides, mRNA or amino acid products or proteins of a specific gene (e.g., a gene encoding a functional fragment of human DIR (e.g., DIR-I, and/or, DIR-II)).
  • the expression level may include the amount of polynucleotides transcribed from a specific gene (e.g., a gene encoding a functional fragment of human DIR (e.g., DIR-I, and/or, DIR-II)), a translated protein, or a fragment of a post-translationally modified protein.
  • the DIR-I may be an amino acid sequence of IR (i.e., an amino acid sequence encoded by a retained intron, whose amino acid sequence is shown in SEQ ID NO: 3) consisting of the first 27 amino acids from the N-terminus.
  • the amino acid sequence of DIR-I is shown in SEQ ID NO: 4.
  • the DIR-II may be an amino acid sequence of IR consisting of the last 27 amino acids from the C-terminus.
  • the amino acid sequence of DIR-II is shown in SEQ ID NO: 5.
  • the reduction can include that the expression level of the DIR is reduced by at least about 10% compared to the expression level of the original DIR in the subject. For example, it can be reduced by 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.
  • the expression level of DIR can be measured by utilizing substances selected from the following group: primers that specifically amplify the DIR gene, nucleic acid molecules that specifically bind to the DIR gene, nucleic acid molecules that specifically bind to the DIR protein, small molecules that specifically bind to the DIR protein, probes that specifically bind to the DIR protein, and polypeptides that specifically bind to the DIR protein.
  • the expression level of the functional fragment of DIR can be measured by using a substance selected from the following group: a primer that specifically amplifies the functional fragment gene of DIR, a nucleic acid molecule that specifically binds to the functional fragment gene of DIR, a nucleic acid molecule that specifically binds to the functional fragment of DIR (e.g., DIR-I and/or DIR-II), Acid molecules, small molecules that specifically bind to functional fragments of DIR (e.g., DIR-I and/or, DIR-II), probes that specifically bind to functional fragments of DIR (e.g., DIR-I and/or, DIR-II), and polypeptides that specifically bind to functional fragments of DIR (e.g., DIR-I and/or, DIR-II).
  • a primer that specifically amplifies the functional fragment gene of DIR e.g., a nucleic acid molecule that specifically binds to the functional fragment gene of DIR, a nu
  • the expression level of the DIR and/or its functional fragment can be measured by implementing an experiment selected from the following group: reverse transcription and amplification analysis (e.g., PCR, linked RT-PCR or quantitative RT-PCR), hybridization analysis, Northern blotting, dot blotting, in situ hybridization, gel electrophoresis, capillary electrophoresis, column chromatography, protein blotting, immunohistochemistry, immunostaining or mass spectrometry.
  • the expression level of the DIR 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 DIR can also be measured by directly analyzing the biological sample or the protein/nucleic acid isolated from the sample.
  • the activity of the DIR and/or its functional fragment may include the biological activity of the DIR protein.
  • the biological activity may include affecting the excitability of neurons and/or inhibiting the activity of neurons.
  • the biological activity may include inhibiting cognitive ability by inhibiting the excitability of neurons and/or inhibiting the activity of neurons.
  • the biological activity may include being able to reduce the frequency of excitatory postsynaptic current (EPSC), and/or being able to reduce the amplitude of EPSC.
  • the reduction may include being able to reduce the frequency of excitatory postsynaptic current (EPSC) in the subject, and/or reducing the amplitude of EPSC, compared with the biological activity of the original DIR and/or its functional fragment in the subject, applying the DIR and/or its functional fragment and/or encoding the DIR and/or its functional fragment nucleic acid, and/or reducing the amplitude of EPSC in the subject.
  • EPC excitatory postsynaptic current
  • the functional fragment DIR-I of the DIR can reduce the frequency of EPSC.
  • the reduction can include that the frequency of EPSC is reduced by at least about 10% after the DIR-I is applied compared with the frequency of the original EPSC in the subject.
  • it can be reduced by 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.
  • the functional fragment DIR-II of the DIR can reduce the frequency of EPSC.
  • the reduction can include that the amplitude of EPSC is reduced by at least about 10% after the DIR-II is applied compared with the amplitude of the original EPSC in the subject.
  • it can be reduced by 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.
  • the biological activity may include affecting cognitive ability.
  • the biological activity may include participating in a signaling pathway associated with A ⁇ deposition, and/or participating in a signaling pathway associated with Tau tangle generation.
  • the DIR and/or its functional fragment can inhibit cognitive ability.
  • the DIR and/or its functional fragment can inhibit cognitive ability by inhibiting the signaling pathway associated with A ⁇ deposition and/or inhibiting the signaling pathway associated with Tau tangle generation.
  • the reduction in the activity of the DIR and/or its functional fragment may include administering the DIR and/or its functional fragment and/or the nucleic acid encoding the DIR and/or its functional fragment, thereby reducing the cognitive ability of the subject, compared with the biological activity of the original DIR and/or its functional fragment in the subject.
  • the reduction can include that the biological activity of the DIR is reduced by at least about 10% compared to the biological activity of the original DIR in the subject. For example, it can be reduced by 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.
  • the expression level of the DIR and/or its functional fragment may be positively correlated with the A ⁇ level (e.g., the expression level in plasma).
  • the expression level of the DIR and/or its functional fragment may be positively correlated with the expression level of A ⁇ 40.
  • the expression level of the DIR and/or its functional fragment may be positively correlated with the expression level of A ⁇ 42.
  • the expression level of the DIR and/or its functional fragment (e.g., the expression level in plasma) can be positively correlated with the degree of reduction in the cognitive ability of the subject.
  • the expression level of the DIR and/or its functional fragment can increase with the progression of cognitive impairment (e.g., disease progression of MCI and/or AD).
  • the expression level of the DIR and/or its functional fragment (e.g., the expression level in plasma) can be positively correlated with the A ⁇ level (e.g., the A ⁇ -PET expression level in the cortex).
  • the expression level of the DIR and/or its functional fragment can be correlated with the formation of amyloid plaques in AD patients.
  • the expression level of the DIR and/or its functional fragment can be related to the storage of declarative memory. For example, the higher the expression level of the DIR and/or its functional fragment is, the lower the ability of the storage of declarative memory is.
  • the expression level of the DIR and/or its functional fragment can be related to the storage of associative learning. For example, the higher the expression level of the DIR and/or its functional fragment is, the lower the ability of the storage of associative learning is.
  • the reduction of the expression level of the DIR and/or its functional fragment can improve cognitive ability.
  • the reduction of the expression level can include that the expression level of the DIR is reduced by at least about 10% compared with the expression level of the original DIR in the subject. For example, it is possible to reduce 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.
  • the lower the expression level of the DIR and/or its functional fragment the higher the cognitive ability (e.g., cognitive ability measured by novel object recognition behavior experiment).
  • the cognitive ability e.g., cognitive ability measured by novel object recognition behavior experiment.
  • the expression level of the DIR is reduced by at least about 10%, and the cognitive ability can be improved by at least about 10%.
  • it can be improved by 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.
  • the DIR or its functional fragment may be derived from mammals, for example, primates, or humans.
  • the DIR may comprise the amino acid sequence shown in SEQ ID NO: 1.
  • the functional fragment of DIR may include the amino acid sequence encoded by the retained intron in DDIT4L. In the present application, the functional fragment of DIR may include the amino acid sequence shown in SEQ ID NO: 3.
  • the functional fragment of the DIR may comprise the amino acid sequence shown in any one of SEQ ID NO: 4-5.
  • the DIR or its functional fragment can be involved in A ⁇ deposition.
  • the DIR or its functional fragment can induce A ⁇ deposition through gelsolin.
  • the retained intron in DDIT4L i.e., DIR-intron (IR), whose amino acid sequence is shown in SEQ ID NO: 3
  • DIR-intron whose amino acid sequence is shown in SEQ ID NO: 3
  • a ⁇ can contribute to the interaction between DIR and gelsolin.
  • the DIR-intron can participate in A ⁇ deposition.
  • the DIR and/or its functional fragment can bind to gelsolin under pathological conditions, thereby causing A ⁇ deposition and amyloid plaque formation.
  • the present application provides a binding agent that binds to the intron retained splicing product DIR and/or its functional fragment of a DNA damage-inducible transcript 4-like transcript.
  • the binding of the binding agent to DIR may be specific.
  • the binding agent can have a Ka (i.e., the equilibrium association constant for the binding interaction, which is 1/M) of greater than or equal to about 10 5 M -1 ( e.g., greater than or equal to about 10 5 M -1 , greater than or equal to about 10 6 M -1 , greater than or equal to about 10 7 M -1 , greater than or equal to about 10 8 M -1 , greater than or equal to about 10 9 M -1 , greater than or equal to about 10 10 M -1, greater than or equal to about 10 11 M -1 , greater than or equal to about 10 12 M -1, greater than or equal to about 10 13 M -1 , or more); or, have a Ka of less than or equal to about 10 -5 M (e.g., less than or equal to about 10 -5 M, less than or equal to about 10 -6 M, less than or equal to about 10 -7 M, less than or equal to about 10 -8 M, less than or equal to about
  • the binding agent may bind or associate with DIR with an equilibrium dissociation constant Kd of about 10-10 M, less than or equal to about 10-11 M, less than or equal to about 10-12 M, less than or equal to about 10-13 M, or less.
  • Kd equilibrium dissociation constant
  • the binding of the binding agent to DIR and/or its functional fragment may be in vivo or in vitro.
  • the binding agent may be isolated.
  • the binding agent may include a small molecule compound, a polymer and/or a biomacromolecule.
  • the binding agent may include a protein and/or a polypeptide.
  • the binding agent may include an ionotropic glutamate receptor (AMPARs) receptor subunit GluA1 and/or a fragment thereof.
  • AMPARs ionotropic glutamate receptor
  • the binding agent may include GluA1 and/or its fragments derived from various organisms.
  • the organism may include a mammal.
  • the organism may include a primate.
  • the organism may include a human.
  • the GluA1 and/or its functional fragments may include GluA1 and/or its functional fragments derived from humans.
  • the amino acid sequence of the GluA1 derived from humans may be a sequence as shown in SEQ ID NO: 7.
  • the binding agent may include an amino terminal deletion, a carboxyl terminal deletion, and/or an internal deletion or substitution compared to the full length of GluA1, while the remaining amino acid sequence is generally the same as the corresponding position of the full length of GluA1.
  • the fragment of GluA1 may include a truncate of GluA1.
  • the fragment of GluA1 is at least 3, 4, 5, 6, 7, 8, 9, 10, 14, 20, 50, 70, 100, 110, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 905 amino acids in length.
  • the fragment of GluA1 may include all fragments and functional variants thereof that can bind to DIR.
  • the fragment of GluA1 may include at least a portion of the amino acid sequence shown in positions 198 to 205 in human GluA1.
  • the binding agent may include the amino acid sequence shown at positions 198 to 205 of human GluA1.
  • the fragment of GluA1 may include a variant of the amino acid sequence shown at positions 198 to 205 of human GluA1.
  • the fragment of GluA1 may include a functional variant of the amino acid sequence shown at positions 198 to 205 of human GluA1.
  • the fragment of GluA1 may include a polypeptide having at least 60%, 70%, 80%, 90%, or 100% of the biological activity of the polypeptide having the amino acid sequence shown at positions 198 to 205 of human GluA1.
  • the fragment of GluA1 may include a polypeptide having at least 60%, 70%, 80%, 90%, or 100% of the DIR binding activity of the polypeptide having the amino acid sequence shown at positions 198 to 205 of human GluA1.
  • the fragment of GluA1 includes a protein or polypeptide in which one or more amino acids are substituted, deleted or added in the amino acid sequence shown in positions 198 to 205 of human GluA1.
  • the fragment of GluA1 may include a polypeptide having amino acid changes by substitution, deletion and/or insertion of at least 1, such as 1-30, 1-20 or 1-10, and for example 1, 2, 3, 4 or 5 amino acids in the amino acid sequence shown in positions 198 to 205 of human GluA1.
  • the binding agent may include a homolog of the amino acid sequence shown at positions 198 to 205 in human GluA1.
  • the binding agent may include a polypeptide having at least about 85% (e.g., at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence homology to the amino acid sequence shown at positions 198 to 205 in human GluA1.
  • the binding agent can regulate the expression level and/or biological activity of DIR and/or its functional fragment.
  • the binding agent can reduce the expression level and/or biological activity of DIR and/or its functional fragment.
  • the binding agent can reduce the expression level and/or biological activity of the DIR and/or its functional fragment in the subject.
  • the binding agent can reduce the expression level of the DIR gene, the transcription level of the DIR gene and/or the expression level of the DIR protein.
  • the binding agent can reduce, for example, the human DDIT4L gene; and/or, the amount of polynucleotides, mRNA or amino acid products or proteins of genes (e.g., human DIR genes) encoding human DIR and/or its functional fragments (e.g., DIR-I, and/or, DIR-II).
  • the binding agent can reduce, for example, the human DDIT4L gene; and/or, the amount of polynucleotides, translated proteins or fragments of post-translationally modified proteins of genes (e.g., human DIR genes) encoding human DIR and/or its functional fragments (e.g., DIR-I, and/or, DIR-II).
  • genes e.g., human DIR genes
  • encoding human DIR and/or its functional fragments e.g., DIR-I, and/or, DIR-II
  • the reduction can include that the expression level of the DIR is reduced by at least about 10% compared to the expression level of the original DIR in the subject. For example, it can be reduced by 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.
  • the reduction can include that the expression level of the functional fragment of the DIR is reduced by at least about 10% compared with the expression level of the original DIR in the subject. For example, it can be reduced by 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.
  • the binding agent can reduce the biological activity of the DIR and/or its functional fragment.
  • the reduction can include that the biology of the DIR or its functional fragment is reduced by at least about 10% compared with the biological activity of the original DIR in the subject. For example, it can be reduced by 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.
  • the binding agent can increase the frequency of excitatory postsynaptic current (EPSC), and/or increase the amplitude of EPSC.
  • the increase can include the frequency and/or amplitude of EPSC after administering DIR to the subject. Compared to the frequency and/or amplitude of EPSCs after administration of DIR and the binding agent, the frequency and/or amplitude of EPSCs is increased by at least about 1%.
  • it can be increased by at least about 2%, at least about 5%, at least about 10%, 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.
  • the binding agent can improve cognitive ability.
  • the cognitive ability can include cognitive ability that can be measured by a novel object recognition behavior experiment.
  • the cognitive ability can include cognitive ability that can be measured by a water maze behavior experiment.
  • the improvement can include that after the binding agent is administered to the subject, the cognitive ability of the subject is improved.
  • the improvement of the subject's cognitive ability can include that in a water maze behavior experiment, the subject's time in the quadrant where the platform is located is increased, the interval time to enter the quadrant where the platform is located is reduced, and/or the number of times the quadrant where the platform is crossed is increased.
  • the binding agent can inhibit the signaling pathway associated with A ⁇ deposition.
  • the binding agent can inhibit the signaling pathway associated with Tau tangle production.
  • the inhibition can include the expression level and/or biological activity of the molecules originally involved in the signaling pathway associated with A ⁇ deposition and/or the signaling pathway associated with Tau tangle production in the subject, and after the administration of the binding agent, the expression level and/or biological activity of these molecules is reduced by at least about 10%.
  • the binding agent can inhibit A ⁇ deposition and/or amyloid plaque formation through gelsolin.
  • the binding agent can inhibit A ⁇ deposition and/or amyloid plaque formation by inhibiting the binding of DIR/or its functional fragment to gelsolin.
  • the present application provides an isolated polypeptide comprising the amino acid sequence shown in SEQ ID NO: 8 (RLVVVDCE) and/or its functional variants.
  • the isolated polypeptide can bind to DIR.
  • the binding of the isolated polypeptide to DIR can be specific.
  • the isolated polypeptide can be greater than or equal to about 10 5 M -1 (e.g., greater than or equal to about 10 5 M -1 , greater than or equal to about 10 6 M -1 , greater than or equal to about 10 7 M -1 , greater than or equal to about 10 8 M -1 , greater than or equal to about 10 9 M -1 , greater than or equal to about 10 10 M -1 , greater than or equal to about 10 11 M -1, greater than or equal to about 10 12 M -1 , greater than or equal to about 10 13 M -1 or higher) Ka (i.e., the equilibrium association constant for the binding interaction, which is 1/M); or, with a value of less than or equal to about 10 - 5 M (e.g., less than or equal to about 10 -5 M, less than or equal to about 10 -6 M, less than or equal to about 10
  • the isolated polypeptide can be used as a binding agent of DIR.
  • the isolated polypeptide can have one or more functions of a DIR binding agent.
  • the isolated polypeptide can regulate the expression level and/or biological activity of DIR and/or its functional variant.
  • the isolated polypeptide can reduce the expression level and/or biological activity of DIR and/or its functional variant.
  • the isolated polypeptide may include variants and/or homologs of the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a variant of the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a functional variant of the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a polypeptide having at least 60%, 70%, 80%, 90%, or 100% of the biological activity of the polypeptide having the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a polypeptide having at least 60%, 70%, 80%, 90%, or 100% of the activity of binding to DIR of the polypeptide having the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a protein or polypeptide having one or more amino acids substituted, deleted or added in the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a polypeptide having at least 1, such as 1-30, 1-20 or 1-10, such as 1, 2, 3, 4 or 5 amino acids substituted, deleted and/or inserted in the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a homolog of the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide may include a polypeptide having at least about 85% (e.g., at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence homology to the amino acid sequence shown in SEQ ID NO: 8.
  • the isolated polypeptide can be a modified polypeptide.
  • the modification can include glycosylation, acetylation, phosphorylation, acylation, derivatization by known protective/blocking groups, proteolytic cleavage, and/or modification by non-naturally occurring amino acids.
  • the isolated polypeptide can form a multimer.
  • a homodimer can be formed.
  • a heterodimer can be formed with another protein or polypeptide.
  • the multimer can still have the ability to bind to DIR or its functional fragment.
  • the isolated polypeptide may form a fusion protein.
  • the isolated polypeptide may form an immunoconjugate.
  • the fusion protein and/or immunoconjugate may still have the ability to bind to DIR or its functional fragment.
  • the isolated polypeptide can be prepared by any method, including but not limited to, recombinantly produced polypeptides, synthetically produced polypeptides, extracted from cells or tissues and other sources.
  • the present application also provides the use of the isolated polypeptide in the preparation of a medicament for preventing and/or treating a disease or condition, wherein the disease or condition includes cognitive impairment and/or neurodegenerative disease.
  • the present application also provides a method for preventing and/or treating cognitive disorders and/or neurodegenerative diseases, which comprises the following steps: administering the isolated polypeptide to a subject in need thereof.
  • the present application also provides the use of the isolated polypeptide in preventing and/or treating cognitive disorders.
  • the isolated polypeptide may be formulated to be suitable for oral administration and/or injection administration.
  • the present application also provides the use of the isolated polypeptide in preparing a reagent for diagnosing and/or evaluating a disease, wherein the disease includes cognitive disorders and/or neurodegenerative diseases.
  • the present application also provides a method for diagnosing and/or evaluating cognitive disorders and/or neurodegenerative diseases, which comprises using the isolated polypeptide.
  • the present application also provides the use of the isolated polypeptide in diagnosing and/or evaluating neurodegenerative diseases.
  • the present application provides an immunoconjugate, wherein the immunoconjugate comprises the binding agent described in the present application and/or the polypeptide described in the present application.
  • the immunoconjugate may include 1, 2 or more binding agents described herein and/or polypeptides described herein.
  • the immunoconjugates described herein have biological activity.
  • biological activity may include the ability to bind DIR and/or its functional fragments in vivo or in vitro and cause a reaction.
  • the reaction includes but is not limited to reducing the expression level and/or biological activity of DIR and/or its functional fragments, increasing the frequency and/or amplitude of excitatory postsynaptic currents (EPSCs), improving cognitive ability, and inhibiting signaling pathways associated with A ⁇ deposition.
  • EPCs excitatory postsynaptic currents
  • the present application provides one or more nucleic acid molecules, which can encode the conjugate described in the present application and/or the isolated polypeptide described in the present application.
  • each of the one or more nucleic acid molecules can encode a complete polypeptide variant or the fusion protein or immunoconjugate, or a portion thereof.
  • the nucleic acid molecules described herein can be isolated. For example, they can be produced or synthesized by the following methods: (i) amplified in vitro, such as produced by polymerase chain reaction (PCR) amplification, (ii) produced by cloning and recombination, (iii) purified, such as by enzyme digestion and gel electrophoresis fractionation, or (iv) synthesized, such as by chemical synthesis.
  • the isolated nucleic acid is a nucleic acid molecule prepared by recombinant DNA technology. Recombinant DNA and separation Subcloning techniques include those described by Sambrook, J., Fritsch, EF and Maniatis, T.
  • nucleic acids may be prepared from genomic DNA fragments, cDNA and RNA, all of which may be extracted directly from cells or recombinantly produced by various amplification methods (including, but not limited to, PCR and RT-PCR).
  • the application provides one or more vectors comprising the nucleic acid molecules.
  • the vector may contain one or more nucleic acid molecules.
  • the vector may also contain other genes, such as marker genes that allow the vector to be selected in a suitable host cell and under suitable conditions.
  • the vector may also contain expression control elements that allow the coding region to be correctly expressed in a suitable host.
  • control elements are well known to those skilled in the art, for example, may include promoters, ribosome binding sites, enhancers and other control elements that regulate gene transcription or mRNA translation, etc.
  • the one or more nucleic acid molecules described in the application may be operably connected to the expression control elements.
  • the vector may include, for example, a plasmid, a cosmid, a virus, a phage or other vectors commonly used in, for example, genetic engineering.
  • the vector is an expression vector.
  • the present application provides a cell, which may contain one or more binding agents described in the present application, one or more polypeptides described in the present application, a nucleic acid molecule described in the present application, and/or one or more vectors described in the present application.
  • each or each host cell may contain one or more nucleic acid molecules or vectors described in the present application.
  • each or each host cell may contain multiple (e.g., 2 or more) or multiple (e.g., 2 or more) nucleic acid molecules or vectors described in the present application.
  • the vector described in the present application may be introduced into the cell by methods known in the art, such as electroporation, liposome transfection, etc.
  • the present application provides a pharmaceutical composition, which may include the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein and/or the cell described herein, and optionally a pharmaceutically acceptable carrier.
  • the pharmaceutical composition may include a drug product suitable for pharmaceutical uses such as improving learning ability, treating cognitive disorders and/or treating neurodegenerative diseases such as Alzheimer's disease.
  • the pharmaceutical composition may be a composition comprising one or more active ingredients (such as the binding agent described herein) and one or more inert ingredients; and any combination, complex or aggregation of any two or more ingredients, or dissociation of one or more ingredients, or other types of reactions or interactions of one or more ingredients directly or indirectly obtained. Any product.
  • the pharmaceutically acceptable carrier may include sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions immediately before use.
  • suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles may 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 esters such as ethyl oleate.
  • the present application provides a kit comprising the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein and/or the cell described herein; and one or more additional components selected from the following group: analytical buffer, controls, substrates, standards, detection materials, laboratory supplies, equipment, instruments, cells, organs, tissues and user manuals or instructions.
  • the kit can detect DIR in a sample.
  • the kit can detect the expression level and/or biological activity of DIR and/or its functional fragments.
  • the detection kit can include instructions, which record the specific steps of how to use the kit to detect the expression level and/or biological activity of DIR and/or its functional fragments, and/or how to use the test results to determine whether the candidate drug can prevent and/or treat cognitive impairment and/or treat subjects with neurodegenerative diseases.
  • the present application provides the use of the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application and/or the pharmaceutical composition described in the present application in the preparation of an agent for preventing and/or treating a disease.
  • the present application provides a method for preventing and/or treating a disease, which comprises administering to a subject in need thereof the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein, the pharmaceutical composition described herein and/or the reagent for preventing and/or treating a disease described herein.
  • the present application provides the use of the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application and/or the pharmaceutical composition described in the present application in preventing and/or treating diseases.
  • the disease may include cognitive disorders.
  • the disease may include neurodegenerative diseases.
  • the reagents described herein can be administered in any manner.
  • the reagents described herein can be administered orally and/or by injection.
  • the reagents described herein can be formulated into a form suitable for its mode of use.
  • the reagents described herein can be formulated into a form suitable for oral administration and/or by injection.
  • a therapeutically effective dose may include an amount that effectively prevents or improves the symptoms of one or more diseases or conditions or the development of the disease or condition when administered to a subject.
  • a therapeutically effective dose may include an amount of a binding compound sufficient to cause symptom improvement, such as an amount that treats, cures, prevents or improves a related medical condition or increases the speed of treatment, cure, prevention or improvement of such conditions.
  • the therapeutically effective dose refers only to the ingredient.
  • a therapeutically effective dose refers to the combined amount of active ingredients that cause a therapeutic effect, whether in combination, administered sequentially or administered simultaneously.
  • the prevention and/or treatment not only includes the prevention and/or treatment of the disease, but also generally includes preventing the onset of the disease, slowing down or reversing the progression of the disease, preventing or slowing down the onset of one or more symptoms associated with the disease, reducing and/or alleviating one or more symptoms associated with the disease, reducing the severity and/or duration of the disease and/or any symptom associated therewith and/or preventing the disease and/or any symptom associated therewith from further increasing in severity, preventing, reducing or reversing any physiological damage caused by the disease, and any pharmacological effect that is generally beneficial to the patient being treated.
  • the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application, the pharmaceutical composition described in the present application and/or the reagent for preventing and/or treating the disease described in the present application do not need to achieve complete cure or eradication of any symptom or manifestation of the disease.
  • drugs used as therapeutic agents can reduce the severity of a given disease state, but do not need to eliminate every manifestation of the disease to be considered a useful therapeutic agent.
  • the preventive administration of treatment constitutes a feasible preventive agent that does not need to completely and effectively prevent the onset of the disease. Simply reducing the effects of the disease in a subject (e.g., by reducing the number or severity of its symptoms, or by increasing the effectiveness of another treatment, or by producing another beneficial effect), or reducing the likelihood that the disease will occur or worsen, is sufficient.
  • the present application provides a method for detecting DIR in a sample, which method comprises administering the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application, the pharmaceutical composition described in the present application and/or the kit described in the present application.
  • the present application provides the use of the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application, the pharmaceutical composition described in the present application and/or the kit described in the present application in the preparation of a kit.
  • the kit can be used to detect DIR in a sample.
  • the DIR in the sample can be detected by combining the material composition and/or the kit described in the present application with the DIR in the preparation of the kit.
  • the detection can include direct detection.
  • the detection can include indirect detection.
  • the detection of DIR in a sample as described herein may include detecting whether DIR exists in the sample.
  • the detection of DIR in a sample as described herein may include detecting the amount of DIR in the sample.
  • the DIR may include DIR and/or a nucleic acid encoding DIR.
  • the detection of DIR in a sample described in the present application may include contacting the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, the nucleic acid molecule described in the present application, the vector described in the present application, the cell described in the present application, the pharmaceutical composition described in the present application and/or the kit described in the present application with the sample.
  • the detection process may include forming a binding agent-DIR complex.
  • the binding agent-DIR complex can be used for detection and analysis after releasing DIR, or the binding agent-DIR complex can be directly used for detection.
  • the method provided herein may directly detect the sample.
  • the method provided herein may only include contacting the sample with a binding agent.
  • the method provided herein may only include contacting the sample with a binding agent and detecting its binding.
  • the method provided herein may include contacting the sample with a binding agent, removing non-specifically bound substances, and detecting its binding.
  • the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein, the pharmaceutical composition described herein and/or the kit described herein can be connected to a carrier.
  • the binding agent is directly or indirectly connected to a carrier such as a magnetic bead or a resin or a mixture thereof.
  • the binding agent can also be directly or indirectly connected to a solid surface or substrate.
  • the binding agent can also be bound to particles, such as beads or microspheres.
  • the binding agent can also be labeled with the following substances, including but not limited to magnetic markers, fluorescent moieties, enzymes, chemiluminescent probes, metal particles, non-metallic colloidal particles, polymer dye particles, pigment molecules, pigment particles, electrochemically active substances, semiconductor nanocrystals or other nanoparticles (including quantum dots or gold particles).
  • substances including but not limited to magnetic markers, fluorescent moieties, enzymes, chemiluminescent probes, metal particles, non-metallic colloidal particles, polymer dye particles, pigment molecules, pigment particles, electrochemically active substances, semiconductor nanocrystals or other nanoparticles (including quantum dots or gold particles).
  • the present application provides the use of the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein, the pharmaceutical composition described herein and/or the kit described herein in the preparation of a reagent for diagnosing and/or evaluating a disease.
  • the present application provides a method for diagnosing and/or assessing a disease, which comprises administering to a subject in need thereof the binding agent described herein, the polypeptide described herein, the immunoconjugate described herein, the nucleic acid molecule described herein, the vector described herein, the cell described herein, the pharmaceutical composition described herein and/or the reagent for diagnosing and/or assessing a disease described herein.
  • the present application provides the binding agent described in the present application, the polypeptide described in the present application, the immunoconjugate described in the present application, Use of the nucleic acid molecules, vectors, cells and/or pharmaceutical compositions described in the application in diagnosing and/or evaluating diseases.
  • the diagnosing and/or assessing a disease can include assessing a disease process.
  • the disease can include a cognitive disorder.
  • the disease can include a neurodegenerative disease.
  • the diagnosis and/or assessment of the disease may include detecting the content of DIR and/or nucleic acid encoding the DIR in a sample from a subject in need.
  • the present application diagnoses and/or assesses the disease, and may include comparing the content of the DIR and/or nucleic acid encoding the DIR in the sample of the subject with a normal control value, wherein the normal control value includes the content of the DIR and/or nucleic acid encoding the DIR in a normal subject.
  • the normal subject does not suffer from the cognitive disorder and/or the neurodegenerative disease.
  • the subject when the content of the DIR and/or nucleic acid encoding the DIR in the sample of the subject is significantly higher than the normal control value, the subject is diagnosed as suffering from the cognitive disorder and/or the neurodegenerative disease.
  • the present application diagnoses and/or assesses the disease, and may include: comparing the content of the DIR and/or nucleic acid encoding the DIR in the sample of the subject with an early control value, wherein the early control value includes the content of the DIR and/or nucleic acid encoding the DIR measured by the same subject before.
  • the subject has been diagnosed with the cognitive disorder and/or the neurodegenerative disease.
  • the subject when the content of DIR and/or nucleic acid encoding DIR in the sample of the subject is significantly higher than the early control value, the subject is diagnosed as having the cognitive disorder and/or aggravated disease progression of the neurodegenerative disease.
  • the diagnosis and/or assessment of the disease described in the present application may also include the following steps: detecting the content of markers associated with cognitive impairment and/or neurodegenerative diseases in the sample derived from the subject.
  • the markers associated with cognitive impairment and/or neurodegenerative diseases include AD7C-NTP, pTau-181, pTau-217 and/or A ⁇ 1-42.
  • diagnosis and/or assessment of a disease described herein may also include observing brain imaging of the subject.
  • the sample described in the present application may include a blood sample and/or a tissue or cell sample.
  • the sample described in the present application may include whole blood, serum, plasma and/or cerebrospinal fluid.
  • the subject may include a mammal.
  • the subject may include a rodent and/or a primate, for example, the subject may include a human.
  • the subject may include a patient with cognitive impairment and/or a patient with a neurodegenerative disease.
  • the neurodegenerative disease may include acute neurodegenerative disease and chronic neurodegenerative disease.
  • the neurodegenerative disease may include neurodegenerative disease caused by neuronal death and glial cell homeostasis, neurodegenerative disease caused by aging, neurodegenerative disease caused by CNS cell function being affected, neurodegenerative disease caused by abnormal intercellular communication and/or neurodegenerative disease caused by impaired cell movement.
  • the subject may include a patient with a neurodegenerative disease.
  • the subject may include a patient with Alzheimer's disease.
  • the patient with Alzheimer's disease may be in the early, early, middle or late stages of Alzheimer's disease.
  • the cognitive impairment may include early cognitive impairment (MCI), mid-term cognitive impairment and late cognitive impairment.
  • MCI early cognitive impairment
  • the cognitive impairment may include cognitive impairment caused by normal aging, lewy body dementia (LBD), frontotemporal dementia and/or vascular dementia.
  • LBD lewy body dementia
  • the induced disease of the cognitive impairment may include Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and/or Creutzfeldt-Jakob disease (CJD).
  • the cognitive impairment may include amnestic MCI multi-cognitive domain impairment (aMCI-m).
  • the subject may include a patient with cognitive impairment.
  • the subject may suffer from early cognitive impairment (MCI) (e.g., loss of short-term memory, difficulty in expressing or understanding abstract things, mood or behavior volatility, difficulty in learning new things and following complex instructions, decreased judgment and/or basic self-care requiring reminders from others), mid-term cognitive impairment (e.g., confusion of long-term memory and reality memory, inability to express one's meaning, behavioral personality changes or emotional instability and/or requiring others to assist in self-care) or late cognitive impairment (e.g., memory loss, physical activity and mental state decline, inability to express or communicate effectively, inability to take care of oneself and/or biological clock confusion).
  • MCI early cognitive impairment
  • mid-term cognitive impairment e.g., confusion of long-term memory and reality memory, inability to express one's meaning, behavioral personality changes or emotional instability and/or requiring others to assist in self-care
  • late cognitive impairment e.g., memory loss, physical activity and mental state decline, inability to express or communicate
  • the subject may suffer from a disease that can cause the cognitive impairment to be induced.
  • the subject may suffer from Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and/or Creutzfeldt-Jakob disease (CJD).
  • CJD Creutzfeldt-Jakob disease
  • the subject may be in the elderly stage.
  • the subject has shown cognitive impairment caused by normal aging.
  • the subject has shown symptoms of early cognitive impairment (MCI).
  • MCI early cognitive impairment
  • the subject has shown symptoms of a neurodegenerative disease (e.g., Alzheimer's disease).
  • the subject may suffer from Alzheimer's disease.
  • the subject may be in the early stage of Alzheimer's disease, the middle stage of Alzheimer's disease and/or the late stage of Alzheimer's disease.
  • the disease may include cognitive impairment.
  • the cognitive impairment may include early cognitive impairment (MCI), mid-term cognitive impairment and late cognitive impairment.
  • MCI early cognitive impairment
  • the cognitive impairment may include cognitive impairment caused by normal aging, Lewis body dementia (LBD), frontotemporal lobe dementia and/or vascular dementia.
  • LBD Lewis body dementia
  • the induced disease of the cognitive impairment may include Alzheimer's disease, multi-infarct type, Parkinson's disease, AIDS and/or Creutzfeldt-Jakob disease (CJD).
  • the cognitive impairment may include amnestic MCI multi-cognitive domain impairment (aMCI-m).
  • the disease may include a neurodegenerative disease.
  • the neurodegenerative disease may include acute neurodegenerative disease and chronic neurodegenerative disease.
  • the neurodegenerative disease may include neurodegenerative disease caused by neuronal death and glial cell homeostasis, Neurodegenerative diseases caused by aging, neurodegenerative diseases caused by impaired CNS cell function, neurodegenerative diseases caused by abnormal communication between cells and/or neurodegenerative diseases caused by impaired cell motility.
  • AD Alzheimer's disease
  • the normal splicing body of the human DDIT4L gene will remove the intron part and connect the two adjacent exons during the formation of mRNA.
  • intron retention IR
  • NS represents the normal splicing body
  • IR represents the DIR formed by abnormal splicing
  • DIR open reading frame of DIR was cloned and expressed exogenously, and it was found that DIR can be normally translated into the corresponding DIR protein in vitro (its amino acid sequence is shown in SEQ ID NO: 1), and the protein can be secreted outside the cell.
  • the open reading frame of DIR (SEQ ID NO: 1) was constructed into the pCMV-flag vector, and the Flag-DIR plasmid and the control flag plasmid were transfected into HEK293 cells, respectively. After 48 hours, the cell culture medium and cell lysate were taken for Western Blot, and the expression of DIR was detected with a DIR-specific antibody, and actin was used as a control. The results are shown in Figure 1C.
  • DIR in human blood was detected. Blood from 2 normal subjects and 7 AD patients of different ages (aged 82, 55, 65, 53, 78, 68 and 58 years old) was taken for Western Blot. DIR-specific antibodies were used to detect its expression, and the Ponceau red-stained band was used as a loading control. The results are shown in Figure 1D. It can be seen that DIR can be specifically present in AD patients.
  • the plasmid expressing DIR was transfected into U87 cells. After 48 hours, the cells were lysed and the cell lysate was divided into two parts. Equal amounts of IgG and DIR antibody were added to each part for immunoprecipitation. The experimental products were separated by SDS-PAGE. Separate and soak the separated PAGE gel in Coomassie Brilliant Blue dye for 1 hour, then use decolorizing solution to wash away the dye that non-specifically adheres to the surface of the PAGE gel until the bands can be clearly seen in the PAGE gel. Then cut off the specific bands and perform mass spectrometry analysis to obtain the candidate molecule gelsolin.
  • the open reading frame of gelsolin (SEQ ID NO: 11) was then cloned into the pEGFP-N3 vector to obtain the Gelsolin-GFP plasmid, and the gelsolin-GFP and Flag or Flag-DIR plasmid were co-transfected into HEK293 cells. The cells were lysed after 48 hours, and the lysate was obtained and used for immunoprecipitation experiments with Flag antibodies. Finally, the binding of gelsolin and DIR was detected by the GFP antibody.
  • Brain slices were prepared from brain tissues of adult C57BL/6 mice, and brain slices from the hippocampus were taken for electrophysiological recording. Changes in postsynaptic excitatory currents in brain slices were recorded before and after administration of IR peptides.
  • DIR i.e., QDLIR, whose amino acid sequence is shown in SEQ ID NO: 1 contains the amino acid sequence encoded by the first exon (whose amino acid sequence is shown in SEQ ID NO: 2) and the amino acid sequence encoded by the retained intron (whose amino acid sequence is shown in SEQ ID NO: 3).
  • the amino acid sequence (IR) encoded by the retained intron can be divided into two parts, namely IR-I (i.e., DIR-I, whose amino acid sequence is shown in SEQ ID NO: 4) consisting of the first 27 amino acids from the N-terminus and DIR-II (i.e., DIR-II, whose amino acid sequence is shown in SEQ ID NO: 5) consisting of the last 27 amino acids.
  • IR-I i.e., DIR-I, whose amino acid sequence is shown in SEQ ID NO: 4
  • DIR-III i.e., DIR-II, whose amino acid sequence is shown in SEQ ID NO: 5
  • the IR fragment was synthesized, brain slices of wild-type mice were incubated, and excitatory postsynaptic currents (EPSCs) were recorded (for specific methods, see Pavel et al., Curr Biol. Contemporary Biology, 26:2194-2201 [2016]).
  • ESCs excitatory postsynaptic currents
  • IR-I and DIR-II fragments were synthesized separately to screen the functional regions.
  • the results showed that IR-I could reduce the frequency of EPSC but did not affect the amplitude of EPSC; although DIR-II had no significant effect on the amplitude and frequency of EPSC.
  • DIR-II has a tendency to reduce the amplitude of EPSC and a tendency to increase the frequency of EPSC (see Figure 4).
  • IR-I can reduce the frequency of EPSC, while DIR-II can To reduce EPSC amplitude.
  • DIR was mainly co-localized with A ⁇ in the hippocampus (see Figure 5A). Interaction between DIR and A ⁇ was observed in transfected HEK293T cells, but DDIT4L (including DIR-exon) could not interact with A ⁇ (see Figure 5B, Figure 6A).
  • DIR-intron i.e., the amino acid sequence encoded by the retained intron, whose amino acid sequence is shown in SEQ ID NO: 3
  • DIR-intron may be the main region for DIR to interact with A ⁇ .
  • PKA Proximity ligation analysis
  • DIR was transfected into human brain cell lines, and the cell lysate was immunoprecipitated with rabbit DIR antibody (purchased from Jier Biochemical (Shanghai) Co., Ltd.). The results showed that a co-immunoprecipitated immunoreactive band with a molecular weight of about 85kDa was obtained (see Figure 5D). The bands were extracted and further analyzed by mass spectrometry. The analysis determined 12-15 peptides that matched human gelsolin (85kDa) and covered 35% of the gelsolin sequence (see Figure 5E).
  • Gelsolin has been shown to interact with A ⁇ 42 to form a complex that is transported into the circulation. Studies have shown that DIR interacts with gelsolin in DIR knock-in mice, HEK293T cells, and human tissues (see Figure 5F, G, Figure 6C).
  • Thioflavine S is a specific marker for A ⁇ plaques in the brains of AD patients.
  • thioflavine S failed to label amyloid plaques in the hippocampus of DIR knock-in mice (see Figure 7A) because mouse A ⁇ sequences cannot be detected with thioflavine S.
  • amyloid plaques in hippocampal slices of DIR knock-in mice incubated with human A ⁇ 40 peptide for 6 hours could be stained with Thioflavin S (see FIG7A ), and were consistent with the results of Thioflavin S staining of human A ⁇ plaques.
  • DIR may be an initiator of A ⁇ deposition and amyloid plaque formation (see Figure 11E).
  • Plasma DIR level as a diagnostic indicator
  • the concentration of DIR in the plasma of naMCI, aMCI and AD patients gradually increased (see Figure 9A).
  • the concentration of DIR in plasma was found to be positively correlated with the concentration of pTau181, but negatively correlated with the A ⁇ 42/40 ratio, and negatively correlated with the severity of cognitive decline in the entire clinical range from normal cognition to naMCI, aMCI and AD.
  • the AUC A ⁇ 42/40 ratio in plasma only reached 71% for distinguishing AD patients from normal controls (see Figure 9G).
  • the comprehensive model combining DIR, pTau, and A ⁇ 42/40 showed the potential to distinguish AD patients from cognitively normal controls, with an AUC of 96%, which was higher than the AUC (88%) obtained using pTau and A ⁇ 42/40 (see Figure 9H).
  • Figure 12A shows that the brain tissues of wild-type mice (C57BL/6J) and DIR knock-in mice were used for co-immunoprecipitation experiments (co-IP experiments), followed by Coomassie Brilliant Blue staining, and the specific binding bands of DIR were found.
  • Figure 12B shows that the specific binding bands of DIR were subjected to mass spectrometry analysis, and GluA1 was found to be a binding target of DIR.
  • FIG. 13A shows that the brain tissues of wild-type mice and DIR knock-in mice were used for co-IP experiments, and the experimental results showed that DIR can bind to GluA1.
  • Figure 13B shows that DIR and GluA1 were exogenously expressed in HEK293 cells and co-IP experiments were performed, and the results also showed that DIR can bind to GluA1.
  • Example 8 The polypeptide fragment (R198-E205) derived from GluA1 can bind to DIR
  • FIG14A shows that computer simulation revealed that GluA1 R198-E205 is the binding site of DIR, and GluA1 C204 plays an important role therein.
  • Figure 14B shows that adding a short peptide of GluA1 (R198-E205) at a final concentration of 10 uM to the cell lysate co-transfected with GluA1 and DIR can effectively block the binding between GluA1 and DIR, indicating that GluA1 R198-E205 is the binding site of DIR, suggesting that GluA1 R198-E205 can be developed into a peptide drug.
  • Figure 14C shows that in the Co-IP experiment in the cell lysate co-transfected with GluA1 or GluA1 point mutation (GluA1 C204A ) and DIR, it was found that the binding of GluA1 C204A to DIR was weakened, indicating that the C204 site plays an important role in the binding of GluA1 and DIR.
  • Example 9 Peptide fragments derived from GluA1 can reverse the inhibitory effect of DIR-induced sEPSC
  • mouse brain tissue was placed in pre-cooled artificial cerebrospinal fluid (ACSF), and 95% O 2 and 5% CO 2 were continuously filled, and then cut into tissue slices of about 350 microns thick using a Leica vibrating slicer. After selecting a suitable brain slice, it was placed in an observation chamber filled with artificial cerebrospinal fluid. The observation chamber was fixed under the field of view of a microscope, and fresh artificial cerebrospinal fluid was continuously perfused to maintain the activity of the brain tissue. Finally, electrodes were used to record the discharge of action potentials in the hippocampus. The results showed that after giving 5uM IR, the frequency and amplitude of the action potential discharge were reduced. After giving 5uM IR and 10uM GluA1 R198-E205 at the same time, the frequency and amplitude of the action potential discharge could be partially increased.
  • ASF artificial cerebrospinal fluid
  • Example 10 Administration of a polypeptide fragment derived from GluA1 can improve the learning ability of mice
  • This example shows that administration of a polypeptide fragment derived from GluA1 can improve the learning ability of mice.
  • the water maze experiment was performed on 3-month-old DIR homozygous mice. On the day of the experiment, the mice were acclimated to the experimental room 30 minutes in advance. The experiment was divided into training day and experimental day. The training day was 5-6 days, and the experimental day was 1 day. Before the experiment, the water tank was filled with 30 cm of water, the water temperature was maintained at 19-22 ° C, and talcum powder was added to the water to make it turbid. The water tank was divided into four directions: N, S, E and W. The platform was placed in the NE quadrant. The daily entry point of the mice was selected from a non-repeating combination of S, W, NW and SE.
  • the platform was placed 0.5 cm below the water.
  • the mouse entered the water from the designated entry point, facing the wall of the water tank, and the timer started for 1 minute at the same time. Stop timing when the mouse reaches the platform. If the mouse does not reach the platform within 1 minute, place the mouse on the platform or guide it onto the platform. After the mouse stays on the platform for 30 seconds, take it out, wipe it dry, and return it to the cage. There should be a 30-minute interval before the next entry into the water.
  • mice Place the mouse at a new entry point and repeat the above experiment 3 times.
  • the experimenter records the time it takes the mouse to get on the platform in each experiment. On the second day, repeat the above experiment. Train 4 times a day for a total of 5 to 6 days.
  • Experimental day Conduct the experiment 24 hours after the end of the training day. Remove the platform. The mouse enters the water from the SW quadrant, facing the wall of the water tank. Take the mouse out 1 minute later, wipe it dry, and return it to the cage. After the experiment, use Etho Vision XT 14 software to analyze the mouse behavior video and collect data The data included: escape latency to enter the target quadrant, number of times crossing the platform position, and time spent in the target and non-target quadrants. The experimental operators and data analysts were double-blind.
  • mice The experimental results showed that administration of GluA1 R198-E205 (1 mg/kg, continuous intravenous administration for 7 days) can improve the learning ability of mice. This includes an increase in the time spent in the quadrant where the platform is located, a decrease in the interval time to enter the quadrant where the platform is located, and an increase in the number of times crossing the quadrant where the platform is located.

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Abstract

L'invention concerne un liant qui se lie à un produit d'épissage de rétention d'intron DIR de type transcrit 4 inductible par les dommages à l'ADN et/ou un fragment fonctionnel associé, ainsi que l'utilisation du liant.
PCT/CN2024/099135 2023-06-15 2024-06-14 Liant de dir et utilisation associée Pending WO2024255822A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100008921A1 (en) * 2005-06-16 2010-01-14 Evotec Neurosciences Gmbh Diagnostic and Therapeutic Target Adarb2 Proteins for Neurodegenerative Diseases
CN110520418A (zh) * 2017-02-10 2019-11-29 加的夫大学学院咨询有限公司 Ampa受体增强剂
CN112168969A (zh) * 2020-09-29 2021-01-05 上海联慧脑智工程中心 Ddit4l及其功能小肽对胶质母细胞瘤的抑制作用
WO2023143425A1 (fr) * 2022-01-25 2023-08-03 上海魁特迪生物科技有限公司 Méthode d'amélioration de troubles cognitifs

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100008921A1 (en) * 2005-06-16 2010-01-14 Evotec Neurosciences Gmbh Diagnostic and Therapeutic Target Adarb2 Proteins for Neurodegenerative Diseases
CN110520418A (zh) * 2017-02-10 2019-11-29 加的夫大学学院咨询有限公司 Ampa受体增强剂
CN112168969A (zh) * 2020-09-29 2021-01-05 上海联慧脑智工程中心 Ddit4l及其功能小肽对胶质母细胞瘤的抑制作用
WO2023143425A1 (fr) * 2022-01-25 2023-08-03 上海魁特迪生物科技有限公司 Méthode d'amélioration de troubles cognitifs

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE GenBank 12 March 2023 (2023-03-12), "Homo sapiens glutamate ionotropic receptor AMPA type subunit 1 (GRIA1), transcript variant 1, mRNA", XP093247876, Database accession no. NM_000827.4 *
DATABASE GenPept 12 March 2023 (2023-03-12), "glutamate receptor 1 isoform 1 precursor [Homo sapiens]", XP093247869, Database accession no. NP_000818.2 *
LI, J.B. ET AL.: "p85S6K Sustains Synaptic GluA1 to Ameliorate Cognitive Deficits in Alzheimer’s Disease", TRANSLATIONAL NEURODEGENERATION, vol. 12, 9 January 2023 (2023-01-09), XP021313027, DOI: 10.1186/s40035-022-00334-w *

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