WO2024122459A1 - 認知機能障害ステージを判定する方法 - Google Patents

認知機能障害ステージを判定する方法 Download PDF

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WO2024122459A1
WO2024122459A1 PCT/JP2023/043089 JP2023043089W WO2024122459A1 WO 2024122459 A1 WO2024122459 A1 WO 2024122459A1 JP 2023043089 W JP2023043089 W JP 2023043089W WO 2024122459 A1 WO2024122459 A1 WO 2024122459A1
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drebrin
level
amount
subject
biological sample
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French (fr)
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WO2024122459A9 (ja
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智明 白尾
祐子 関野
幹夫 東海林
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Alzmed Inc
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Alzmed Inc
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Priority to EP23900574.7A priority Critical patent/EP4632384A1/en
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Priority to CN202380082792.9A priority patent/CN120380345A/zh
Publication of WO2024122459A1 publication Critical patent/WO2024122459A1/ja
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    • 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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G01N2800/2814—Dementia; Cognitive disorders
    • G01N2800/2821—Alzheimer

Definitions

  • the present disclosure relates to a method for determining a stage of cognitive impairment in a subject. More particularly, the present disclosure relates to a method for determining a stage of cognitive impairment based on the level or amount of at least one of drebrin A or a molecule derived from drebrin A.
  • Alzheimer's disease the main cause of dementia, is an irreversible progressive brain disease that is believed to occur when synapses in the brain become dysfunctional.
  • the increase in the number of Alzheimer's disease patients has become a major social concern in recent years, and there is a need to establish early diagnosis methods and therapeutic drugs.
  • Senile plaques have been observed in the postmortem brains of Alzheimer's disease patients, and these are known to be aggregates of "amyloid ⁇ protein" (amyloid plaques).
  • amyloid ⁇ protein amyloid ⁇ protein
  • the deposition of amyloid ⁇ protein is the earliest pathological lesion that can be confirmed pathologically, and it has been reported that amyloid ⁇ protein aggregates and directly exhibits neuronal toxicity.
  • amyloid cascade hypothesis abnormalities in the production and accumulation of amyloid ⁇ protein are widely related to the onset of Alzheimer's disease. This is called the amyloid cascade hypothesis.
  • amyloid ⁇ protein is the main cause of Alzheimer's disease. It has also been found that the accumulation of amyloid ⁇ protein in the brain begins more than 20 years before the onset of the disease.
  • Alzheimer's disease can be caused by reduced cerebral blood flow (depression, etc.), cerebral hemorrhage, cerebral infarction, drug addiction, etc., and in addition to Alzheimer's disease-type dementia, mitochondrial genetic diseases and dementia caused by diabetes are also known.
  • Alzheimer's disease can only be diagnosed definitively by performing a pathological autopsy after death, and diagnosis during life is made by questionnaire tests and imaging tests. In this case, the diagnosis is made by questionnaire tests after the patient feels subjective symptoms such as severe forgetfulness, but an objective diagnosis cannot be made.
  • questionnaire tests and imaging tests can definitively diagnose late-stage dementia, they cannot diagnose early stages such as mild cognitive impairment (MCI), and early treatment cannot be started.
  • MCI mild cognitive impairment
  • the present disclosure provides a technology for easily and accurately determining the stage of cognitive impairment in a subject.
  • the present disclosure provides: (Item 1) A method for determining a stage of cognitive impairment in a subject, comprising: Measuring the level or amount of at least one of drebrin or a molecule derived from drebrin in a biological sample taken from the subject; and determining a stage of cognitive impairment in the subject based on the level or amount. (Item A1) A method for determining a stage of cognitive impairment in a subject, comprising: Measuring the level or amount of at least one of drebrin A or a molecule derived from drebrin A in a biological sample taken from the subject; and determining a stage of cognitive impairment in the subject based on the level or amount.
  • drebrin A or a molecule derived from drebrin A comprises drebrin A-related protein (DARP).
  • DARP drebrin A-related protein
  • the DARP comprises at least the amino acid sequence of positions 221 to 353 of the amino acid sequence of SEQ ID NO:1, and/or at least a portion of the amino acid sequence.
  • (Item A5) The method according to any one of the preceding items, comprising measuring the level or amount of at least one of amyloid beta and phosphorylated tau in the biological sample.
  • the biological sample is selected from the group consisting of cerebrospinal fluid, blood, urine, saliva, and tears.
  • (Item A6a) The method according to any one of the preceding claims, wherein the biological sample is cerebrospinal fluid.
  • (Item A10) The method according to any one of the above items, wherein the biological sample is cerebrospinal fluid, and when the level or amount of at least one of the drebrin A or molecules derived from drebrin A is an abnormal value and the level or amount of at least one of the amyloid ⁇ is a standard value, the subject is determined to be cognitively unimpaired (CU) in Alzheimer's disease.
  • (Item A10a) The method according to any one of the preceding items, wherein the drebrin A or a molecule derived from drebrin A is A-DARP.
  • (Item A11) The method according to any one of the above items, wherein the biological sample is cerebrospinal fluid, and the level or amount of at least one of the drebrin A or molecules derived from drebrin A is a standard value, and 1) the level or amount of at least one of the phosphorylated tau is an abnormal value, or 2) the level or amount of at least one of the amyloid ⁇ is an abnormal value, the subject is determined to be cognitively unimpaired (CU) in Alzheimer's disease.
  • the drebrin A or a molecule derived from drebrin A is A-DARP.
  • (Item A12) The method according to any one of the above items, wherein the biological sample is cerebrospinal fluid, and when the level or amount of at least one of the drebrin A or molecules derived from drebrin A is a standard value, the level or amount of at least one of the phosphorylated tau is an abnormal value, and the level or amount of at least one of the amyloid ⁇ is a standard value, the subject is determined to be cognitively unimpaired (CU) in Alzheimer's disease.
  • the drebrin A or a molecule derived from drebrin A is A-DARP.
  • (Item A14) The method according to any one of the above items, wherein the subject is judged to have dementia in Alzheimer's disease when the level or amount of at least one of the drebrin A or molecules derived from drebrin A is a standard value, the level or amount of at least one of the phosphorylated tau is an abnormal value, and the level or amount of at least one of the amyloid beta concentrations is an abnormal value.
  • (Item A15) The method according to any one of the preceding items, wherein when the biological sample is cerebrospinal fluid, the abnormal value of the level or amount of at least one of the drebrin A or molecules derived from drebrin A is about 100 pg/ml or more.
  • a method for determining a stage of cognitive impairment in a subject comprising: determining a stage of cognitive impairment based on the level or amount of at least one component in a biological sample taken from the subject; (a) determining that the subject has mild cognitive impairment (MCI) in Alzheimer's disease when the level or amount of at least one of drebrin A or a molecule derived from drebrin A in a biological sample collected from the subject is an abnormal value; (b) when the biological sample is cerebrospinal fluid, and the level or amount of at least one of drebrin A or a molecule derived from drebrin A in the biological sample collected from the subject is an abnormal value, and the level or amount of at least one of amyloid ⁇ is a standard value, the subject is judged to be cognitively unimpaired (CU) in Alzheimer's disease; (c) when the biological sample is cerebrospinal fluid, and the level or amount of at least one of drebrin A or a
  • (Item C1) A method for using the level or amount of at least one of drebrin A or a molecule derived from drebrin A as an index for determining a stage of cognitive impairment in a subject, comprising: Measuring the level or amount of at least one of drebrin A or a molecule derived from drebrin A in a biological sample taken from the subject; and using the level or amount as an index for determining a stage of cognitive impairment in the subject.
  • the disclosed method of assessment allows for early and simple assessment of the stage of cognitive impairment, which previously relied on imaging tests, etc., leading to the prevention of the onset of these diseases and early treatment.
  • FIG. 1 is a graph showing the results of an analysis of the association between A-DARP and the stage of brain cognitive dysfunction when all cases are used in one embodiment of the present disclosure.
  • FIG. 2 is a graph showing the results of an analysis of the relationship between A-DARP and the stage of brain cognitive dysfunction, excluding patients with diseases other than Alzheimer's disease and patients with complications, in one embodiment of the present disclosure.
  • FIG. 3 is a graph showing the results of an analysis of the relationship between A ⁇ 42 in cerebrospinal fluid and the stage of brain cognitive dysfunction in one embodiment of the present disclosure.
  • FIG. 4 is a diagram showing the results of stage determination of brain cognitive dysfunction in a total of 22 cases including uncomplicated Alzheimer's disease patients and healthy subjects in one embodiment of the present disclosure.
  • FIG. 5 is a diagram showing the results of stage determination of brain cognitive dysfunction in a total of 31 cases including Alzheimer's disease patients and healthy subjects in one embodiment of the present disclosure.
  • cognition dysfunction refers to a state in which some kind of substantial disorder has occurred in brain functions related to cognition, such as perception, memory, attention, and executive functions, and is not limited to the presence or absence of symptoms or their severity.
  • drebrin A refers to the adult isoform in neurons of drebrin, an actin-binding protein that is expressed in large amounts in neurons during development. Drebrin is involved in the morphogenesis of neurons, particularly process formation, by changing the properties of actin fibers. In developing and migrating neurons, it is present in the cell body and the entire process, but in mature neurons, it is specifically present in the spine structure (dendritic spine). There are two isoforms of drebrin: embryonic type drebrin E and adult type drebrin A. Drebrin A, which is specifically found in the dendritic spine of mature neurons, has the characteristic of being expressed only in neurons. It is also known that drebrin in dendritic spines is widely lost in dementia diseases such as Alzheimer's disease.
  • a molecule derived from drebrin A refers to a part or fragment of drebrin A, or a molecule obtained by any modification of these.
  • DARP drebrin A-related protein
  • A-DARP refers to a peptide that contains at least the peptide sequence from positions 221 to 353 of drebrin A (SEQ ID NO: 1), or a portion thereof.
  • CDA cerebrospinal fluid
  • PDA refers to A-DARP in the blood.
  • abnormal value when referring to a biomarker refers to a value that is not a standard value, and is also called a "non-standard value.”
  • standard value when referring to a biomarker refers to a value that is normal, and since it is a characteristic of the value alone, even if the value itself is a standard value, it may be normal or may not be normal (disease state).
  • a method for determining the stage of cognitive impairment in a subject comprising the steps of measuring the level or amount of at least one of drebrin A or a molecule derived from drebrin A in a biological sample collected from the subject, and determining the stage of cognitive impairment in the subject based on the level or amount.
  • the stage of cognitive impairment can be determined simply by measuring the level or amount of at least one of drebrin A or a molecule derived from drebrin A in a biological sample, and is therefore useful for preventing the onset of cognitive impairment and for early treatment.
  • the present disclosure relates to a method for the detection of drebrin A (SEQ ID NO: 1) localized in the brain, which is a human drebrin A protein (SEQ ID NO: 1) localized in the brain.
  • DARPs drebrin A related proteins
  • DARPs drebrin A related proteins
  • DARPs drebrin A related proteins
  • drebrin A or a molecule derived from drebrin A may include drebrin A-related protein (DARP).
  • DARP may be a drebrin A or splice variant having at least a part of the Ins2 translation sequence RPYCPFIKASDSGPSSSSSSSSSPPRTPFPYITCHRTPNLSSSLPC (SEQ ID NO: 2) characteristic of drebrin A, or a dimer, oligomer, or polymer composed of them.
  • drebrin A or a molecule derived from drebrin A such as DARP may be used alone in the method of the present disclosure, or one or more molecules may be used in combination in the method of the present disclosure.
  • the total concentration of drebrin A or a molecule derived from drebrin A such as DARP may be measured and used in the method of the present disclosure.
  • the DARP may contain at least the amino acid sequence from positions 221 to 353 of the amino acid sequence of SEQ ID NO: 1, and in one embodiment, the DARP may also contain intact drebrin A from positions 1 to 795 of the amino acid sequence of SEQ ID NO: 1. Methods for measuring intact drebrin A are known, for example, the method described in JP 2016-40540A.
  • the concentration of drebrin A or a molecule derived from drebrin A, such as DARP can be measured by any known method.
  • the concentration of drebrin A or a molecule derived from drebrin A, such as DARP can be measured by, for example, Western blotting, ELISA, immunoprecipitation, etc., using an antibody that recognizes DARP.
  • the anti-DARP antibody may be either a monoclonal antibody or a polyclonal antibody, as long as it has a part of DARP as an epitope, and may be used alone or in combination of two or more types.
  • the present disclosure relates to an antibody that recognizes a drebrin A-specific epitope for use in the method of the present disclosure.
  • Drebrin A-specific epitope means an epitope that is not present in drebrin E but is present in drebrin A, and is preferably a region including at least a part of the Ins2 translation sequence RPYCPFIKASDSGPSSSSSSSSSPPRTPFPYITCHRTPNLSSSLPC (SEQ ID NO: 2), and/or a specific three-dimensional structure that is created because drebrin A contains the Ins2 translation sequence.
  • a method of using anti-DARP antibody as a primary antibody and detecting with a labeled secondary antibody that recognizes the primary antibody can be preferably used.
  • the primary antibody is a rabbit antibody
  • a labeled anti-rabbit IgG antibody can be used as a secondary antibody
  • a labeled anti-mouse IgG antibody can be used as a secondary antibody.
  • Anti-DARP antibodies that can be used in the measurement of the present invention are known and already commercially available. Specific examples include the DAS2 antibody described in THE JOURNAL OF COMPARATIVE NEUROLOGY 505:352-362 (2007).
  • the labeling substance in the above-mentioned labeled secondary antibody may include enzymes, radioisotopes, fluorescent substances, luminescent substances, gold colloids, etc.
  • enzymes are preferred from the viewpoint of sensitivity and ease of operation, and horseradish peroxidase (HRP), alkaline phosphatase (AP), glucose oxidase (GOD), etc. are more preferred.
  • TMB 3,3',5,5'-tetramethylbenzidine
  • AP AMPPD (3-(2'-spiroadamantane)-4-methoxy-4-(3''-phosphoryloxy)phenyl-1,2-dioxetane disodium salt), 9-(4-chlorophenylthiophosphoryloxymethylidene)-10-methylacridan disodium salt, etc.
  • substrate for chemiluminescence may be used as the substrate.
  • Other fluorescent dyes such as FITC (fluoresceinisothiocyanate) and rhodamine can also be used as labeling substances.
  • Methods for detecting and quantifying drebrin A or molecules derived from drebrin A vary depending on the labeling method, and can be performed by methods well known and conventional to those skilled in the art.
  • a chromogenic or luminescent substrate can be added, and the absorbance or luminescence intensity can be measured to quantify the substance to be measured.
  • the substance to be measured can be quantified by measuring its fluorescence intensity.
  • a radioisotope is used as the labeling substance, the substance to be measured can be quantified by measuring its radioactivity.
  • the substance to be measured can be quantified by measuring its absorbance.
  • a calibration curve standard curve
  • the measured values may be collated with the calibration curve to calculate the concentrations of drebrin A or molecules derived from drebrin A, such as DARP, in the samples, which may be used for comparison in the method of the present disclosure; however, the absorbance, radioactivity, and luminescence intensity may be directly used for comparison in the method of the present disclosure without calculating the concentrations of drebrin A or molecules derived from drebrin A, such as DARP.
  • the biological sample means a body fluid collected from a subject or a control.
  • the body fluid may be cerebrospinal fluid, blood, urine, saliva, and tears, among which cerebrospinal fluid and blood are preferred.
  • Blood includes serum, plasma, and the like.
  • the biological sample may be directly subjected to measurement of the level or concentration of molecules derived from drebrin A, such as drebrin A or DARP, but preferably is pretreated before measurement to increase the detection sensitivity of molecules derived from drebrin A, such as drebrin A or DARP.
  • the method of the present disclosure may further include a step of measuring the level or amount of at least one of amyloid ⁇ and/or phosphorylated tau in a biological sample.
  • the level or amount of at least one of drebrin A or a molecule derived from drebrin A in a biological sample collected from the subject can be measured to determine the stage of cognitive impairment in the subject.
  • the method of the present disclosure can determine the stage of cognitive impairment into categories such as, for example, dementia due to Alzheimer's disease, mild cognitive impairment (MCI) due to Alzheimer's disease, cognitively unimpaired (CU) due to Alzheimer's disease, and diseases other than Alzheimer's disease, but the stage categories are not limited to these.
  • the method of the present disclosure can also determine the stage of cognitive impairment such as Alzheimer's disease, vascular dementia, dementia with Lewy bodies, Parkinson's disease, corticobasal degeneration, amyotrophic lateral sclerosis, spinocerebellar degeneration, cerebral hemorrhage, cerebral infarction, brain tumor, etc.
  • the subject in the method of the present disclosure may be a subject who has developed cognitive impairment, a subject who has not developed cognitive impairment, and/or a subject whose future cognitive impairment is unknown.
  • a control for comparing the level or amount of at least one of the drebrin A or molecules derived from drebrin A in a subject can be a healthy individual who has a low or almost no risk of developing cognitive dysfunction, or who has not developed cognitive dysfunction.
  • the threshold can be set based on the level or amount of at least one of the drebrin A or molecules derived from drebrin A in such a healthy individual.
  • the method of the present disclosure can determine the stage of cognitive impairment based on the following criteria:
  • the subject can be determined to have mild cognitive impairment (MCI) in Alzheimer's disease.
  • MCI mild cognitive impairment
  • the biological sample is cerebrospinal fluid
  • the level or amount of at least one of drebrin A or a molecule derived from drebrin A is a standard value, and 1) the level or amount of at least one of phosphorylated tau is an abnormal value, or 2) the level or amount of at least one of amyloid ⁇ is an abnormal value
  • the subject can be determined to be cognitively unimpaired (CU) in Alzheimer's disease.
  • CU cognitively unimpaired
  • the subject when the biological sample is cerebrospinal fluid, and at least one level or amount of drebrin A or a molecule derived from drebrin A is a standard value, at least one level or amount of phosphorylated tau is an abnormal value, and at least one level or amount of amyloid ⁇ is a standard value, the subject can be determined to be cognitively unimpaired (CU) in Alzheimer's disease.
  • CU cognitively unimpaired
  • A-DARP as drebrin A or a molecule derived from drebrin A.
  • drebrin A or a molecule derived from drebrin A, amyloid beta, and phosphorylated tau are all within standard values, it can be determined that there is no brain damage.
  • the subject when the level or amount of at least one of drebrin A or a molecule derived from drebrin A is a standard value, the level or amount of at least one of phosphorylated tau is an abnormal value, and the level or amount of at least one of amyloid ⁇ concentrations is an abnormal value, the subject can be determined to have dementia in Alzheimer's disease.
  • the stage of cognitive impairment in a subject can be determined based on the criteria set forth in the table below.
  • the standard and abnormal values of the level or amount of at least one of drebrin A or a molecule derived from drebrin A, amyloid ⁇ , and/or phosphorylated tau used in determining the stage of cognitive impairment by the method of the present disclosure can be set appropriately depending on the type of biological sample used in the determination, the type of DARP used in the determination, the type of stage divided by the determination, the characteristics of the subject, etc.
  • the abnormal value of the level or amount of at least one of drebrin A or a molecule derived from drebrin A can be about 100 pg/ml or more.
  • the abnormal value of at least one level or amount of amyloid beta can be about 900 pg/ml or less. In one embodiment, when the biological sample is cerebrospinal fluid, the abnormal value of at least one level or amount of phosphorylated tau can be about 50 pg/ml or more.
  • the abnormal value of at least one of the levels or amounts of the drebrin A or the molecule derived from drebrin A can be about 2000 pg/ml or less.
  • the abnormal value of at least one of the levels or amounts of the drebrin A or the molecule derived from drebrin A can be about 1000 pg/ml or more, or about 400 pg/ml or less.
  • abnormal value of at least one of the levels or amounts of the drebrin A or the molecule derived from drebrin A may vary depending on the measurement method or the subject of measurement, and an "abnormal value" refers to a value that is not a standard value when referring to a biomarker, and is also called a "non-standard value.”
  • an agent in another aspect of the present disclosure, includes an antibody that recognizes a drebrin A-specific epitope for determining a stage of cognitive impairment in a subject based on the level or amount of at least one of drebrin A or a molecule derived from drebrin A.
  • a method for using the level or amount of at least one of drebrin A or a molecule derived from drebrin A as an index for determining the stage of cognitive impairment in a subject comprising the steps of measuring the level or amount of at least one of drebrin A or a molecule derived from drebrin A in a biological sample collected from the subject, and using the level or amount as an index for determining the stage of cognitive impairment in the subject.
  • an appropriate selection can be made from the embodiments described elsewhere in this specification.
  • Short Protocols in Molecular Biology A Compendium of Methods from Current Protocols in Molecular Biology, Greene Pub. Associates; Ausubel, F. M. (1995). Short Protocols in Molecular Biology: A Compendium of Methods from Current Protocols in Molecular Biology, Greene Pub. Associates; Innis, M. A. et al. (1995). PCR Strategies, Academic Press; Ausubel, F. M. (1999). Short Protocols in Molecular Biology: A Compendium of Methods from Current Protocols in Molecular Biology, Wiley, and annual updates; Sninsky, J. J. et al. (1999).
  • gene synthesis and fragment synthesis services such as GeneArt, GenScript, Integrated DNA Technologies (IDT) can be used, and other references include, for example, Gait, M. J. (1985). Oligonucleotide Synthesis: A Practical Approach, IRL Press; Gait, M. J. (1990). Oligonucleotide Synthesis: A Practical Approach, IRL Press; Eckstein, F. (1991). Oligonucleotides and Analogues: A Practical Approach, IRL Press; Adams, R. L. et al. (1992). The Biochemistry of the Nucleic Acids, Chapman &Hall; Shabarova, Z. et al. (1994).
  • the concentrations of amyloid ⁇ 40 and amyloid ⁇ 42 in the cerebrospinal fluid were measured by sandwich ELISA using the Human ⁇ Amyloid (1-40) ELISA Kit WakoII and Human/Rat ⁇ Amyloid (1-42) ELISA Kit WakoHigh-Sensitive (Wako Pure Chemical Industries, Osaka, Japan) according to the manufacturer's instructions.
  • Total tau and phosphorylated tau in the cerebrospinal fluid were measured using INNOTEST hTAU Ag and PHOSPHO-TAU (181P) (FUJIREBIO Inc, Tokyo, Japan).
  • DARP was measured by sandwich ELISA using a combination of antibodies capable of specifically detecting drebrin A. Specifically, the hDrebrin-A ELISA Kit sold and distributed by the applicant (Almed Co., Ltd.) was used.
  • Brain cognitive dysfunction stage determination The values of phosphorylated tau, amyloid beta 42, and DARP in patients without complications obtained as described above were divided into standard and abnormal values.
  • a threshold was determined for the A-kit measurement value CDA (corresponding to A-DARP) in cerebrospinal fluid, and the value was divided into standard and abnormal values.
  • Example 1 Determination of brain cognitive dysfunction stage by A-DARP using 50 cases
  • the results of an analysis of the relationship between A-DARP and the stage of brain cognitive dysfunction in 50 cases using a nonparametric Kruskal-Wallis test are shown in FIG.
  • Example 2 Assessment of brain cognitive dysfunction stage by A-DARP in patients with uncomplicated Alzheimer's disease
  • the relationship between the measured values using the hDrebrinA ELISA kit and the stage of brain cognitive impairment was analyzed, excluding patients with other than Alzheimer's disease and patients with complications. The results are shown in Figure 2.
  • a ⁇ 42 in cerebrospinal fluid was significantly different among the mild cognitive impairment in Alzheimer's disease (ADMCI) group, the dementia (ADD) group, and the normal group. These results confirmed that A ⁇ 42 in cerebrospinal fluid can be used to determine MCI or dementia in Alzheimer's disease.
  • Example 4 Determination of brain cognitive dysfunction stage by combination of amyloid ⁇ 42, phosphorylated tau, and DARP
  • MCI was assessed for amyloid ⁇ 42, phosphorylated tau, and A-DARP (CDA) alone in the cerebrospinal fluid of Alzheimer's disease patients, as well as for MCI when each was combined.
  • Figure 4 shows the results of MCI assessment for amyloid ⁇ 42, phosphorylated tau, and A-DARP (CDA) alone in a total of 22 cases including Alzheimer's disease patients without complications and healthy individuals, as well as the results of MCI assessment for a total of 22 cases including Alzheimer's disease patients without complications and healthy individuals, when each was combined.
  • Figure 5 shows the MCI assessment results for amyloid ⁇ 42, phosphorylated tau, and A-DARP (CDA) alone in a total of 31 cases, including both Alzheimer's disease patients and healthy individuals, as well as the MCI assessment results when each was combined in a total of 31 cases, including both Alzheimer's disease patients and healthy individuals.
  • CDA A-DARP
  • the disclosed method of assessment allows for early and simple assessment of the stage of cognitive impairment, which previously relied on imaging tests, etc., leading to the prevention of the onset of these diseases and early treatment. As such, it is expected to be applied in the medical and pharmaceutical fields.
  • SEQ ID NO: 1 Amino acid sequence of human drebrin A
  • SEQ ID NO: 2 INS2 sequence

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PCT/JP2023/043089 2022-12-05 2023-12-01 認知機能障害ステージを判定する方法 Ceased WO2024122459A1 (ja)

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