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WO2023068173A1 - BIOMARQUEUR POUR ÉVALUER L'ACCUMULATION D'AMYLOÏDE-β INTRACÉRÉBRALE - Google Patents

BIOMARQUEUR POUR ÉVALUER L'ACCUMULATION D'AMYLOÏDE-β INTRACÉRÉBRALE Download PDF

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Publication number
WO2023068173A1
WO2023068173A1 PCT/JP2022/038295 JP2022038295W WO2023068173A1 WO 2023068173 A1 WO2023068173 A1 WO 2023068173A1 JP 2022038295 W JP2022038295 W JP 2022038295W WO 2023068173 A1 WO2023068173 A1 WO 2023068173A1
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Prior art keywords
tau
nfl
accumulation
amyloid
tau181
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English (en)
Japanese (ja)
Inventor
義明 佐藤
大介 伊東
將 三村
鍾玉 文
肇 田渕
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Keio University
Eisai R&D Management Co Ltd
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Keio University
Eisai R&D Management Co Ltd
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    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • 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

Definitions

  • the present disclosure relates to biomarkers for evaluating the accumulation of brain amyloid ⁇ .
  • Amyloid ⁇ is a type of protein produced in the brain, and the accumulation of amyloid ⁇ in the brain is believed to be closely related to the development of Alzheimer's disease (AD).
  • Tau protein also simply referred to as "tau” is also a type of protein produced in nerve cells such as the brain. Aberrant phosphorylation is believed to be involved in AD.
  • Accumulation of amyloid- ⁇ and abnormally phosphorylated tau (p-Tau) in the brain is clinically assessed by radiological studies such as positron emission tomography (PET) or by biomarkers of amyloid- ⁇ and tau in cerebrospinal fluid. be done.
  • PET positron emission tomography
  • biomarkers of amyloid- ⁇ and tau in cerebrospinal fluid be done.
  • these detection methods are of limited utility as diagnostic tools due to their high invasiveness and high cost.
  • Non-Patent Document 1 discloses that the plasma p-tau-217/T217 ratio and p-tau-217 value distinguish between the amyloid ⁇ -negative group and the amyloid ⁇ -positive group, regardless of cognitive function. It is Non-Patent Document 2 discloses that plasma P-Tau181 is a non-invasive diagnostic and prognostic biomarker for AD. Non-Patent Document 3 discloses that plasma p-Tau181 is useful for AD screening, and that plasma p-Tau181 can be used to select individuals determined to be amyloid ⁇ -positive by PET.
  • Non-Patent Document 4 discloses that plasma p-Tau181 can distinguish AD from young adults and elderly without cognitive impairment.
  • Non-Patent Document 5 discloses that plasma p-Tau181 sorts AD from non-AD lesions with high accuracy.
  • Non-Patent Document 6 discloses that plasma p-Tau217 can distinguish AD from other neurodegenerative diseases.
  • Non-Patent Document 7 there is also a report on prediction of progression to AD in patients with mild cognitive impairment (MCI) using a combined biomarker of plasma p-Tau181 and neurofilament light chain (NFL) (Non-Patent Document 7).
  • the present disclosure provides a low-cost, high-accuracy biomarker for evaluating brain amyloid ⁇ accumulation and a method for evaluating brain amyloid ⁇ accumulation using the biomarker. intended to provide
  • the present inventors have made intensive studies to solve the above problems, and as a result, the combination of phosphorylated tau 181 (p-Tau181) and neurofilament light chain (NFL) in plasma is useful for evaluating the accumulation of amyloid ⁇ in the brain. They are useful biomarkers, and it was found that the accumulation of brain amyloid ⁇ can be evaluated with high diagnostic accuracy by using these two biomarkers.
  • p-Tau181 phosphorylated tau 181
  • NNL neurofilament light chain
  • the disclosure provides a method for assessing brain amyloid- ⁇ accumulation, comprising phosphorylated Tau (p-Tau) 181, p-Tau217, p-Tau231 in a blood sample obtained from a subject. and a combination thereof, measuring the amount of p-Tau and the amount of neurofilament light chain (NFL), and comparing the measured value obtained in the above step with a control value.
  • a method that includes This method is a low-cost and minimally invasive method, and enables a method of evaluating brain amyloid- ⁇ accumulation with high diagnostic accuracy.
  • a combination of predetermined p-Tau (p-Tau181, p-Tau217, p-Tau231) and NFL enables examination without failure while maintaining high specificity.
  • a method for evaluating the accumulation of brain amyloid ⁇ comprising: The amount of p-Tau selected from the group consisting of phosphorylated Tau (p-Tau) 181, p-Tau217, p-Tau231 and combinations thereof, and neurofilament light chain (NFL ) and and comparing the measured value obtained in the above step with a control value.
  • the cut-off values are the measured values of the p-Tau and the NFL in a blood sample obtained from a control subject, and the p-Tau and the p-Tau in a blood sample obtained from a subject with amyloid ⁇ accumulation in the brain.
  • the method of [2], wherein the method is determined by comparing the measured values of NFL above.
  • the method according to [2] or [3], wherein the cutoff value of the p-Tau and/or the cutoff value of the NFL is one or more.
  • a method of aiding the diagnosis of Alzheimer's disease (AD), mild cognitive impairment in the setting of AD or preclinical AD comprising: The amount of p-Tau selected from the group consisting of phosphorylated Tau (p-Tau) 181, p-Tau217, p-Tau231 and combinations thereof, and neurofilament light chain (NFL ) and and comparing the measured value obtained in the above step with a control value to evaluate the accumulation of brain amyloid ⁇ .
  • the method of [10] wherein the control value is a predetermined cutoff value.
  • the cut-off values are the measured values of the p-Tau and the NFL in a blood sample obtained from a control subject, and the p-Tau and the p-Tau in a blood sample obtained from a subject with amyloid ⁇ accumulation in the brain.
  • the method according to [11] or [12], wherein the cutoff value of the p-Tau and/or the cutoff value of the NFL is one or more.
  • a method for obtaining an index for assisting diagnosis of Alzheimer's disease (AD), mild cognitive impairment in the background of AD, or preclinical AD comprising: measuring the amount of p-Tau selected from the group consisting of p-Tau181, p-Tau217, p-Tau231 and combinations thereof and the amount of NFL in a blood sample obtained from the subject; and comparing the measured value obtained in the above step with a control value to obtain information on the accumulation of brain amyloid ⁇ .
  • a kit for evaluating the accumulation of brain amyloid ⁇ comprising: An antibody that specifically binds to p-Tau selected from the group consisting of phosphorylated Tau (p-Tau) 181, p-Tau217, p-Tau231 and combinations thereof, and specific to neurofilament light chain (NFL) and an antibody that binds to.
  • p-Tau phosphorylated Tau
  • NNL neurofilament light chain
  • a kit for evaluating the accumulation of brain amyloid ⁇ comprising: phosphorylated tau (p-Tau) 181, p-Tau217, p-Tau231, and a neurofilament light chain (NFL) comprising an antibody that specifically binds to p-Tau selected from the group consisting of combinations thereof;
  • a kit for evaluating the accumulation of brain amyloid ⁇ comprising: p-Tau comprising an antibody that specifically binds to a neurofilament light chain (NFL) and is selected from the group consisting of phosphorylated tau (p-Tau) 181, p-Tau217, p-Tau231 and combinations thereof
  • p-Tau phosphorylated tau
  • a method of treating Alzheimer's disease (AD), mild cognitive impairment in the setting of AD or preclinical AD comprising: The amount of p-Tau selected from the group consisting of phosphorylated Tau (p-Tau) 181, p-Tau217, p-Tau231 and combinations thereof, and neurofilament light chain (NFL ) and A step of comparing the measured value obtained in the above step with a control value to evaluate the accumulation of amyloid ⁇ in the brain; administering a therapeutically effective amount of a therapeutic agent for Alzheimer's disease, mild cognitive impairment in the setting of AD, or preclinical AD to a subject assessed for accumulation of brain amyloid beta.
  • AD Alzheimer's disease
  • p-Tau phosphorylated Tau
  • NNL neurofilament light chain
  • a method of diagnosing Alzheimer's disease (AD), mild cognitive impairment in the setting of AD or preclinical AD comprising: The amount of p-Tau selected from the group consisting of phosphorylated Tau (p-Tau) 181, p-Tau217, p-Tau231 and combinations thereof, and neurofilament light chain (NFL ) and and comparing the measured value obtained in the above step with a control value to evaluate the accumulation of brain amyloid ⁇ .
  • AD Alzheimer's disease
  • p-Tau phosphorylated Tau
  • NNL neurofilament light chain
  • the method of evaluating the accumulation of amyloid ⁇ in the brain of the present disclosure not only has high diagnostic accuracy, but also is low cost, so the range of use can be greatly expanded, and large-scale utilization such as initial diagnosis and health checkup is possible. Be expected. Therefore, in particular, as a primary screening for amyloid ⁇ test, a high true positive rate and diagnostic accuracy can be achieved, and it can be a promising evaluation method for drug discovery based on amyloid ⁇ accumulation in the brain.
  • accumulation of brain amyloid ⁇ can be evaluated with high diagnostic accuracy and at low cost.
  • FIG. 3 is a diagram showing the concentration of p-Tau181 in the blood of 60 cases in the negative group and 34 cases in the positive group in an amyloid ⁇ PET test.
  • FIG. 2 is a diagram showing the concentrations of p-Tau181 and NFL in the blood of 60 cases in the negative group and 34 cases in the positive group in an amyloid ⁇ PET test.
  • the method of assessing the accumulation of brain amyloid- ⁇ comprises phosphorylated Tau181 (p-Tau181), phosphorylated Tau217 (p-Tau217), phosphorylated Tau231 ( measuring the amount of p-Tau selected from the group consisting of p-Tau231) and combinations thereof, and the amount of neurofilament light chain (NFL), and comparing the measured value obtained in the above step with a control value. and the step of
  • a subject means mammals including, but not limited to, humans, chimpanzees, orangutans, gorillas, baboons, monkeys, mice, rats, pigs, horses, dogs, and cows.
  • the subject is human in one embodiment.
  • the blood sample may contain the biomarkers to be measured, p-Tau and NFL selected from the group consisting of p-Tau181, p-Tau217, p-Tau231 and combinations thereof.
  • a blood sample may be whole blood, serum, or plasma, and may be a blood sample obtained from a human subject.
  • Amyloid ⁇ (also referred to as “A ⁇ ”) is a type of protein made in the brain known to those skilled in the art. Assessing the accumulation of cerebral amyloid ⁇ means determining whether or not a greater accumulation of cerebral amyloid ⁇ is observed in comparison with control subjects. Accumulation of cerebral amyloid ⁇ means that amyloid ⁇ is not excreted in the brain and accumulates in the brain. A control subject means a brain amyloid ⁇ -negative subject without accumulation of brain amyloid ⁇ . Accumulation of brain amyloid- ⁇ is conventionally assessed clinically by radiological studies such as positron emission tomography (PET) or by biomarkers of amyloid- ⁇ and phosphorylated tau in cerebrospinal fluid.
  • PET positron emission tomography
  • brain amyloid ⁇ if the accumulation of brain amyloid ⁇ is found to be greater than the diagnostic criteria according to the visual judgment criteria of the 18 F-Florbetaben manufacturer, brain amyloid ⁇ If the test is positive and the accumulation of brain amyloid ⁇ is less than the diagnostic criteria, it is determined to be negative for brain amyloid ⁇ .
  • Accumulation of amyloid ⁇ in the brain is considered to be one of the main pathologies mainly seen in Alzheimer's disease (AD) and mild cognitive impairment (MCI due to AD) in the background of AD.
  • AD Alzheimer's disease
  • MCI mild cognitive impairment
  • CBS corticobasal ganglia syndrome
  • FDD frontotemporal dementia
  • PSP progressive supranuclear palsy
  • PD Parkinson's disease or Parkinson's syndrome
  • the evaluation method of the present embodiment is positive, accumulation of brain amyloid ⁇ is considered to be suspected regardless of the clinically diagnosed disease name. Therefore, the evaluation method of this embodiment is useful as a simple primary screening method for the accumulation of brain amyloid ⁇ .
  • the clinical diagnostic criteria for these neurological diseases are known to those skilled in the art, and include, for example, the diagnostic criteria described in the Examples.
  • p-Tau181 is phosphorylated Tau threonine 181
  • p-Tau217 is Tau threonine 217 phosphorylated
  • p-Tau231 is Tau threonine 231 phosphorylated. is.
  • the amount of any two of p-Tau181, p-Tau217, and p-Tau231 may be measured, and all three p-Tau may be measured.
  • Two p-Tau combinations include p-Tau181 and p-Tau217, p-Tau181 and p-Tau231, and p-Tau217 and p-Tau231.
  • the amount of p-Tau, ie, p-Tau181, p-Tau217, and p-Tau231, in a blood sample can be measured according to known protein quantification methods. For example, absorbance measurement and immunological measurement methods using antigen-antibody reaction such as enzyme-linked immunosorbent assay (ELISA), enzyme immunoassay (EIA), radioimmunoassay (RIA), immunofluorescence method, immunoprecipitation method, and , liquid chromatography/mass spectrometry (LC/MS).
  • ELISA enzyme-linked immunosorbent assay
  • EIA enzyme immunoassay
  • RIA radioimmunoassay
  • immunofluorescence method immunoprecipitation method
  • LC/MS liquid chromatography/mass spectrometry
  • SIMOA registered trademark
  • p-Tau181 Advantage V2 Kit manufactured by Quanterix
  • the amount of p-Tau is a measured value (absolute value).
  • the amount of NFL in a blood sample can be measured according to known protein quantification methods, similar to p-Tau. Examples thereof include absorbance measurement, immunological measurement methods using antigen-antibody reactions such as ELISA, EIA, RIA, immunofluorescence, immunoprecipitation, and LC/MS. In addition, it can be easily quantified using a commercially available measurement kit such as SIMOA (registered trademark) NFL Advantage (SR-X) Kit (manufactured by Quanterix).
  • SIMOA registered trademark
  • SR-X NFL Advantage
  • the amount of NFL is a measured value (absolute value).
  • the cutoff value of p-Tau181 and NFL if one cutoff value is 1 and the other cutoff value is 2 or more, for example, the cutoff value is the NFL cutoff value 1 and p-Tau181 cutoff value 1 and p-Tau181 cutoff value 2 (where p-Tau181 cutoff value 1 ⁇ p-Tau181 cutoff value 2), the measured value of p-Tau181 is p-Tau181 If the cutoff value is 2 or more, and the measured value of NFL is less than the NFL cutoff value 1, and the measured value of p-Tau181 is p-Tau181 cutoff value 1 or more p-Tau181 cutoff value If it is less than 2, it may be determined as brain amyloid ⁇ -positive, and otherwise, it may be determined as brain amyloid ⁇ -negative.
  • a method to aid in diagnosing Alzheimer's disease (AD), mild cognitive impairment in the setting of AD, or preclinical AD comprises p-Tau181, p-Tau217, p- A step of measuring the amount of p-Tau selected from the group consisting of Tau231 and combinations thereof and the amount of NFL, and comparing the measured value obtained in the above step with a control value to determine the accumulation of brain amyloid ⁇ and evaluating.
  • the kit for evaluating accumulation of brain amyloid ⁇ comprises p-Tau selected from the group consisting of phosphorylated tau (p-Tau) 181, p-Tau217, p-Tau231, and combinations thereof.
  • p-Tau phosphorylated tau
  • NNL neurofilament light chain
  • the above antibody may be any antibody or antigen-binding fragment thereof that specifically binds to p-Tau or NFL, and may be a polyclonal antibody or a monoclonal antibody. Also, in one embodiment, the antibody is a detectably labeled antibody, and labeling can be accomplished by methods known to those skilled in the art, depending on the method of measuring antigen-antibody binding.
  • the method of diagnosing Alzheimer's disease, mild cognitive impairment in the setting of AD, or preclinical AD comprises: a step of measuring the amount of p-Tau and the amount of NFL selected from the group consisting of; comparing the measured value obtained in the above step with a control value to evaluate the accumulation of brain amyloid ⁇ ; include.
  • the diagnostic accuracy of the combination of p-Tau181 and NFL was compared with that of p-Tau181 alone.
  • the specificity of diagnosis by the combination of p-Tau181 and NFL obtained when using the above cutoff value is 70%, and the diagnostic accuracy is higher than 47% of the specificity of diagnosis with p-Tau181 alone. increased by 23%. Therefore, when the cut-off value for diagnosing without false negatives is set by the combination marker of p-Tau181 and NFL, the accuracy of diagnosis is greatly increased compared to the case of diagnosis with p-Tau181 alone. was done.
  • the breakdown of the clinical diagnosis of 57 subjects is MCI (14), AD (19), CBS (4 people), FTD (10 people), PD (2 people), PSP (4 people) and senile mental illness (4 people). There were 26 cases in the group.

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Abstract

Compte tenu des problèmes de l'état de la technique, la présente invention aborde le problème consistant à fournir un biomarqueur pour évaluer l'accumulation d'amyloïde-β intracérébrale à faible coût et avec une précision de diagnostic élevée, et sur un procédé d'évaluation de l'accumulation d'amyloïde-β intracérébrale à l'aide dudit biomarqueur. Ce procédé pour évaluer l'accumulation d'amyloïde-β intracérébrale comprend : une étape de mesure de la quantité d'une protéine tau phosphorylée (p-Tau) choisie dans le groupe constitué par p-Tau181, p-Tau217, p-Tau231, et une combinaison de ceux-ci, et la quantité de chaîne légère neurofilamentaire dans un échantillon de sang obtenu à partir d'un sujet ; et une étape de comparaison de la valeur de mesure obtenue dans l'étape susmentionnée avec une valeur de commande.
PCT/JP2022/038295 2021-10-18 2022-10-14 BIOMARQUEUR POUR ÉVALUER L'ACCUMULATION D'AMYLOÏDE-β INTRACÉRÉBRALE Ceased WO2023068173A1 (fr)

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