CN119998661A

CN119998661A - Identifying subjects with or at risk of developing Alzheimer's dementia

Info

Publication number: CN119998661A
Application number: CN202380074014.5A
Authority: CN
Inventors: 马库斯·克罗恩
Original assignee: Universitaet Rostock
Current assignee: Universitaet Rostock
Priority date: 2022-10-18
Filing date: 2023-10-17
Publication date: 2025-05-13
Also published as: EP4605754A1; JP2025537481A; WO2024083822A1; CA3268498A1

Abstract

在第一方面，本发明涉及一种用于鉴别患有阿尔茨海默氏痴呆(AD)或处于发展阿尔茨海默氏痴呆的风险的受试者的方法，所述方法包括：(a)在受试者的第一体液样品中至少确定以克/摩尔为单位的质量为110.0367的化合物的量；(b)在受试者的第二体液样品中确定与(a)中相同的化合物的量；(c)确定根据(a)确定的化合物的量与根据(b)确定的化合物的相应量相比的变化；(d)使用能够基于所述化合物预测受试者的AD的计算机实现的预测算法来分析在(c)中所确定的变化，从而如果预测到AD，则鉴别出患有AD或处于发展阿尔茨海默氏痴呆的风险的受试者；其中所述第一体液样品是在施用硫乙拉嗪或其药用盐之前获得的，并且所述第二体液样品是在对受试者施用硫乙拉嗪或其药用盐之后获得的。In a first aspect, the present invention relates to a method for identifying a subject suffering from Alzheimer's dementia (AD) or at risk of developing Alzheimer's dementia, the method comprising: (a) determining at least the amount of a compound having a mass of 110.0367 in grams per mole in a first body fluid sample of the subject; (b) determining the amount of the same compound as in (a) in a second body fluid sample of the subject; (c) determining the change in the amount of the compound determined according to (a) compared to the corresponding amount of the compound determined according to (b); (d) analyzing the change determined in (c) using a computer-implemented prediction algorithm capable of predicting AD in the subject based on the compound, thereby identifying a subject suffering from AD or at risk of developing Alzheimer's dementia if AD is predicted; wherein the first body fluid sample is obtained before administering thiethylperazine or a pharmaceutically acceptable salt thereof, and the second body fluid sample is obtained after administering thiethylperazine or a pharmaceutically acceptable salt thereof to the subject.

Description

Identifying a subject suffering from or at risk of developing Alzheimer's dementia

In a first aspect, the invention relates to a method for identifying (identifying or recognizing) a subject suffering from or at risk of developing Alzheimer's dementia (Alzheimer ' S DEMENTIA, AD), comprising (a) determining in a first body fluid sample of the subject the amount of at least one compound having masses ：110.0367、232.004、224.084、254.1153、189.9936、189.0096、155.007、355.345、358.159、109.0706、110.0004、145.0226、166.0633、178.0995、190.0631、191.0253、200.0452、212.0536、234.1832、236.1049、259.9982、285.1362、291.0832、301.1313、314.1266、318.1215、319.2486、331.2545、332.1372、334.2146、339.3135、346.1528、350.0956、366.0912、371.8758、440.2751、538.1752、540.2563 and 782.5638 (group 1) in grams/mole selected from the group consisting of (b) analyzing the determined amount of at least one compound using a computer-implemented prediction algorithm capable of predicting AD in the subject based on the at least one compound, such that if AD is predicted, a subject suffering from AD or at risk of developing Alzheimer's dementia is identified. In a second aspect, the invention relates to a method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD). Both the third aspect of the invention and the fourth aspect of the invention relate to a method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD). In a fifth aspect, the invention relates to a second diagnostic use and the sixth aspect of the invention relates to a device suitable for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), while the seventh aspect of the invention relates to a kit for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD).

Background

Accumulation of proteins or protein fragments (peptides) in the brain is a significant feature of age-related neurodegenerative diseases. In Alzheimer's Disease (AD) and cerebral beta-amyloid angiopathy (CAA), the aggregation of beta-amyloid peptide (Abeta) is thought to cause the disease. Regarding the treatment of AD, several active agents are known, such as for example thioethyllazine (Thiethylperazine) [1]. Regarding diagnosis, higher amounts of 40 and 42 amino acid aβ peptides in the brain reflect in part the increase of these peptides in the peripheral blood stream and are used as diagnostic indicators of alzheimer's disease. However, quantification of aβ in blood samples is unreliable (even if extremely sensitive methods such as liquid chromatography-mass spectrometry (LC-MS/MS) are used), and requires aβ immunoprecipitation as an additional and expensive step for sample preparation. Such assays have recently become publicly available and achieve an accuracy of about 77% in predicting the AD indicative brain amyloid status in patients in the age group 65-75 years old, with decreasing accuracy with decreasing age [2]. E. Trushina et al use of UPLCToF-MS based metabolomics Another study analyzed and compared the distribution of metabolites in plasma and cerebrospinal fluid samples from cognitive normal pre-patients (proband), pre-patients with mild cognitive impairment and AD patients [3]. However, while early diagnosis is mentioned as a number (epidemic, catchword), it is also pointed out that the current study is not done much more than to distinguish healthy from sick forerunner. In addition, since different metabolic markers are detected in the plasma and in the cerebrospinal fluid, there are also other differences, such as examining which body fluids.

However, since it is assumed that AD pathology starts up to 20 years before the onset of clinical symptoms, diagnostic methods for detecting signs of AD pathology in younger populations are needed.

Accordingly, it is a potential technical problem to provide means and methods for meeting the above-mentioned needs, in particular to provide methods for diagnosing or pre-diagnosing Alzheimer's dementia in a subject.

Aspect 1-methods for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD)

Accordingly, in a first aspect, the present invention relates to a method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(a) Determining in a first body fluid sample of a subject an amount of at least one compound having masses ：110.0367、232.004、224.084、254.1153、189.9936、189.0096、155.007、355.345、358.159、109.0706、110.0004、145.0226、166.0633、178.0995、190.0631、191.0253、200.0452、212.0536、234.1832、236.1049、259.9982、285.1362、291.0832、301.1313、314.1266、318.1215、319.2486、331.2545、332.1372、334.2146、339.3135、346.1528、350.0956、366.0912、371.8758、440.2751、538.1752、540.2563 and 782.5638 in grams/mole selected from the group consisting of (group 1);

(b) The determined amount of the at least one compound is analyzed using a computer-implemented prediction algorithm capable of predicting AD of the subject based on the at least one compound, thereby identifying the subject as suffering from AD or at risk of developing alzheimer's dementia if AD is predicted.

As used hereinafter, the terms "having," "including," or "comprising," or any arbitrary grammatical variant thereof, are used in a non-exclusive manner. Thus, these terms may refer to either a situation in which no other feature is present in the entity described in this context other than the feature introduced by the terms, or a situation in which one or more other features are present. As an example, the expressions "a has B", "a contains B" and "a includes B" may refer to both a case in which no other element is present in a except B (i.e., a case in which a consists of B only and exclusively) and a case in which one or more other elements are present in entity a except B (such as element C, elements C and D, or even other elements).

Furthermore, as used hereinafter, the terms "preferably," "more preferably," "most preferably," "particularly," "more particularly," "specifically," "more specifically," or similar terms are used in conjunction with optional features without limiting further possibilities. Thus, the features introduced by these terms are optional features and are not intended to limit the scope of the claims in any way. As the skilled person will appreciate, the invention may be implemented using alternative features. Similarly, features introduced by "in one embodiment of the invention" or similar expressions are intended to be optional features, without any limitation concerning other embodiments of the invention, without any limitation concerning the scope of the invention and without any limitation concerning the possibility of combining features introduced in such a way with other optional or non-optional features of the invention.

The term "identifying a subject having Alzheimer's Disease (AD)" is synonymous with "diagnosing Alzheimer's Disease (AD)" in a subject, wherein "identifying" and "diagnosing" as used herein both refer to assessing whether a Ka disease is present. Thus, the presence or absence of a disease in a subject can be diagnosed. Preferably, the term refers to determining or excluding the disease. As will be appreciated by those skilled in the art, although it is preferred that such an assessment is correct for 100% of the subjects under investigation, such an assessment is generally not possible to be correct for 100% of the subjects under investigation. However, the term requires that subjects of statistically significant portion can be correctly evaluated and thereby diagnosed. One skilled in the art can determine whether a portion is statistically significant without further expense using a variety of well-known statistical evaluation tools, such as confidence interval determination, p-value determination, student t-test, mann-Wheatstone test (Mann-WHITNEY TEST), and the like. Details can be found, for example, in Dowdy and Wearden, STATISTICS FOR RESEARCH (research statistics), john Wiley & Sons, new York 1983. Preferred confidence intervals are at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95%. The p value is preferably 0.2 or less, 0.1 or less or 0.05 or less.

The terms "identify" and "diagnose" include individual diagnosis of a disease or symptom thereof as well as continuous monitoring of a patient. Monitoring (i.e., diagnosing the presence or absence of a disease or its concomitant symptoms at different time points) includes monitoring of patients known to have a disease as well as monitoring of subjects known to be at risk of developing a disease, e.g., monitoring of subjects having a genetic susceptibility to a disease or subjects determined to be at risk based on the above identified methods. In addition, monitoring can also be used to determine whether a patient is successfully treated or at least whether symptoms of a disease can be ameliorated over time by a particular therapy. In addition, monitoring may be used for active patient management, including decision-making for hospitalization, intensive care and/or additional qualitative monitoring measures, as well as quantitative monitoring measures.

The term "identifying a subject at risk of developing Alzheimer's disease" refers to assessing the probability that a subject will suffer from Alzheimer's disease within a particular time window (i.e., predictive window). The term preferably relates to predicting whether there is an increased risk of developing Alzheimer's dementia as compared to the average risk of developing Alzheimer's dementia in a population of subjects, rather than giving an accurate probability of the risk.

Alzheimer's Disease (AD) (Alzheimer's disease (Morbus Alzheimer/Alzheimer)'s disease) is a neurodegenerative disease characterized by increased dementia. Methods known to date for determining whether a subject has AD are current diagnostic criteria, preferably based on amyloid positron emission tomography and/or cerebrospinal fluid diagnosis and/or through cognitive testing. Biomarkers associated with AD are biomarkers of neurodegeneration known to the skilled person, such as for example aβ -40, aβ -42 or aβ 42/40 ratio in body fluids (blood, CSF), phosphorylated variants of Tau-proteins (e.g. p-Tau181, p-Tau217 etc.), glial Fibrillary Acidic Protein (GFAP), neurofilament light chain (NfL) etc. Methods for determining whether a subject has AD involve determining cognitive ability in the subject by one or more tests known to the skilled artisan (such as, for example, ADAS-Cog, FCSRT, ADCS-iADL, qoL-AD, CDR, MMSE, etc.).

As used herein, the term "subject" relates to a multicellular animal, preferably to a vertebrate, more preferably to a mammal, more preferably to a human.

A "body fluid sample" is a sample from the group consisting of a blood sample, a urine sample, a liquid sample (CSF) and a mixture of two or more thereof, preferably the body fluid sample is a blood sample.

The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) is an in vitro method.

The term "determined amount" means a determination based on Mass Spectrometry (MS). Mass spectrometry as used herein encompasses all techniques that allow determination of molecular weight (i.e., mass) or mass variable corresponding to a compound (i.e., metabolic analyte). More preferably, mass spectrometry as used herein relates to GC-MS, LC-MS, direct injection mass spectrometry, FT-ICR-MS, CE-MS, HPLC-MS, quadrupole mass spectrometry, any sequential coupled mass spectrometry such as MS-MS or MS-MS, ICP-MS, py-MS, TOF or any combination of the above techniques. How to apply these techniques is well known to those skilled in the art. Further, suitable devices are commercially available. More preferably, mass spectrometry as used herein relates to LC-MS and/or GC-MS, i.e. to mass spectrometry operably connected to a preceding chromatographic separation step. More preferably, mass spectrometry as used herein encompasses LC-MS/MS (liquid chromatography-mass spectrometry/mass spectrometry). Liquid chromatography as used herein refers to all techniques that allow separation of compounds (e.g., separation of at least two metabolic analytes to be determined according to the present invention) in a liquid or supercritical phase. Liquid chromatography is characterized by passing a compound in the mobile phase through the stationary phase. As the compounds pass through the stationary phase at different rates, they are separated over time because each individual compound has its specific retention time (i.e., the time it takes for the compound to pass through the system). Liquid chromatography as used herein also includes HPLC. Devices for liquid chromatography are commercially available, for example from Agilent Technologies, USA. The specific compound and its specific mass and its amount are determined based on the correlation peaks in the mass spectrum, respectively. Such peaks contain characteristic information (i.e., m/z information) of the compound and intensity values related to the abundance of the compound (i.e., its amount) in the sample.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least one compound having a mass in grams/mole is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936 and 189.0096 (group 2).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least a compound having a mass in grams/mole of 110.0367 is determined in (a) and the amount determined is analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the amount of at least two compounds, preferably at least three compounds, more preferably at least four compounds, more preferably at least five compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 is determined in a first body fluid sample of the subject in step (a), and the analysis is performed in (b), wherein more preferably one of the at least two compounds, preferably one of the at least three compounds, more preferably one of the at least four compounds, more preferably one of the at least five compounds is a compound having a mass in grams/mole of 110.0367.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the amount of at least two compounds, preferably at least six compounds, more preferably at least seven compounds, having a mass in grams/mole selected from the group consisting of group 1 or group 2 is determined in step (a) in a first body fluid sample of the subject and analyzed in (b), wherein more preferably one of the at least two compounds, more preferably one of the at least six compounds or one of the at least seven compounds is a compound having a mass in grams/mole of 110.0367.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the amount of at least one further compound having a mass in grams/mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), preferably from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4) is additionally determined in step (a) in a first body fluid sample of the subject and is additionally analyzed in (b).

In a first aspect, the invention also relates to a method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(a) Determining in a first body fluid sample of the subject the amount of at least two compounds having a mass in grams/mole selected from the group consisting of group 3, preferably selected from the group consisting of group 4;

(b) The determined amounts of the at least two compounds are analyzed using a computer-implemented prediction algorithm capable of predicting AD of the subject based on the at least two compounds, thereby identifying the subject as suffering from AD or at risk of developing alzheimer's dementia if AD is predicted.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the first body fluid sample is derived from a subject after administration of thioethylazine or a pharmaceutically acceptable salt thereof to the subject, wherein the sample is preferably taken within 0.5 to 8 hours after TEP administration.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the computer-implemented predictive algorithm uses a supervised machine learning algorithm that has been trained by a training dataset comprising quantitative or qualitative data from the following compounds in body fluid samples from subjects known to suffer from AD and subjects known not to suffer from AD:

At least one, preferably at least two, more preferably at least three, more preferably at least four, more preferably at least five compounds from group 1 or group 2, optionally in combination with at least one further compound from group 3, or optionally in combination with at least one further compound from group 4, or

At least two, more preferably at least six, more preferably at least seven compounds from group 1 or group 2, optionally in combination with at least one further compound from group 3, or optionally in combination with at least one further compound from group 4, or

-At least two compounds from group 3, preferably from group 4.

As noted above, according to current diagnostic criteria, it is preferably determined whether a subject has AD based on amyloid positron emission tomography and/or cerebrospinal fluid diagnosis and/or by cognitive testing.

The term "computer-implemented prediction algorithm" as used herein refers to a prediction algorithm that is automatically executed by a data processing device (such as a computer). The data processing means preferably comprise a tangibly embedded rule for automatically executing a predictive algorithm based on the measurement results obtained in the method of the invention. The measurement results may preferably be stored in a database which may be linked permanently or temporarily to a data processor. Further, the link may preferably be a direct link through wires or a wireless link. A predictive algorithm as referred to herein is an algorithm that determines parameters related to the likelihood that a subject has AD. The predictive algorithm as referred to herein should generate the above parameters and may be further processed to establish a diagnostic result according to the method of the present invention. Preferably, the (computer-implemented prediction) parameters may be determined as set forth elsewhere herein. In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the supervised machine learning algorithm establishes a classification algorithm selected from the group consisting of logistic regression (Logistic Regression), random gradient descent classifier (Stochastic GRADIENT DESCENT CLASSFIER, SGD classifier), decision tree classifier (Decision TREE CLASSFIER), random tree classifier (Random TREE CLASSFIER), limit tree classifier (Extra TREE CLASSFIER), adaBoost classifier (AdaBoost Classfier), multi-layer perceptron classifier (Multi-layer Perceptron Classfier), gradient boost classifier (Gradient Boosting Classfier), gaussian naive bayes (Gaussian Naive Bayes), K nearest neighbor classifier (K Nearest Neighbor Classfier), C-support vector classification (C-Support Vector Classification), quadratic discriminant analysis (Quadratic DISCRIMINANT ANALYSIS), and linear regression (Linear Regression). In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the supervised machine learning algorithm builds and/or applies a logistic regression model based on equation (I)

y = I + C₁*F₁ + C₂*F₂ +…+ C_n*F_n, (I)

Where I is the intercept and C _n is the coefficient of mass F _n.

The computer-implemented prediction parameter here is y. A positive y value indicates the likelihood that the subject has or does not have AD, i.e., is predictive of a corresponding condition. It will be appreciated that this is the case when training based on the definition that the diagnosis of AD is a positive result, so that a larger absolute value means a greater certainty of the prediction, i.e. a larger positive y value indicates a greater likelihood that the subject is suffering from AD, while a negative value of y with a higher absolute value indicates a greater likelihood that the subject is not suffering from AD. On the other hand, in the case of training based on the definition that the AD diagnosis is a negative result, then a larger negative y value indicates a greater likelihood that the subject has AD, while a positive value of y with a higher (absolute) value indicates a greater likelihood that the subject does not have AD.

Each coefficient preferably has a value within a range as shown in table 2 of the experimental part, wherein each range does not include zero-for each specific quality combination, see the relevant row called "range" in table 2. For example, for a combination of masses (each in grams/mole) 110.0367, 254.1153, 189.0096, and 224.084, the intercept I has a value in the range of-50 to +50 (excluding zero), and the coefficient C1 of the first characteristic (i.e., mass = 110.0367/gram/mole) is in the range of-150 to +150 (excluding zero), and so on. Furthermore, it is even more preferred if the coefficients of the respective masses have ratios to each other within the ranges shown in the following table a, more preferably in the following table B, more preferably in the following table C, wherein these ratios more preferably apply to the combination of masses from group 2, optionally with the mass of group 4:

Table A

Table B

Table C

For each of tables A, B and C, the "lower half" refers to the portion below the diagonal indicated by the "//" symbol, and the "upper half" refers to the portion above the line.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 189.0096 g/mol, a compound having a mass of 224.084 g/mol, and a compound having a mass of 254.1153 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 189.993 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.115 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, and a compound having a mass of 254.1153 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 254.1153 g/mol, and a compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, and a compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in (a) and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, and a compound having a mass of 208.0737 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 208.0737 g/mol and a compound having a mass of 702.5682 g/mol are determined in (a) and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, and a compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the amounts of at least a compound having a mass of 110.0367 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 254.1153 g/mol, a compound having a mass of 189.9936 g/mol, a compound having a mass of 189.0096 g/mol and a compound having a mass of 224.084 g/mol are determined in (a) and analyzed in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the amounts of at least a compound having a mass of 110.0367 g/mol, a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 702.5682 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.1153 g/mol are determined in (a), and analyzed in (b).

Aspect 2-method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD) (change-based modeling)

In a second aspect, the invention relates to a method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(b) Determining the amount of at least one compound identical to that in (a) in a second body fluid sample of the subject;

(c) Determining a change in the amount of the at least one compound determined according to (a) as compared to a corresponding amount of the at least one compound determined according to (b);

(d) Analyzing the change determined in (c) using a computer-implemented prediction algorithm capable of predicting AD in a subject based on the at least one compound, thereby identifying a subject suffering from AD or at risk of developing alzheimer's dementia if AD is predicted;

Wherein the first bulk liquid sample is obtained prior to administration of thioethlazine or a pharmaceutically acceptable salt thereof and the second bulk liquid sample is obtained after administration of thioethlazine or a pharmaceutically acceptable salt thereof to a subject.

The definitions indicated above in the section relating to the first aspect apply here also to the section relating to the second aspect of the invention.

Thioethylprazine (IUPAC: 2- (ethylsulfanyl) -10- [3- (4-methylpiperazin-1-yl) propyl ] -10H-phenothiazine, CAS:1420-55-9, tep) has been shown to be effective in exporting amyloid beta (aβ) from and through endothelial cells of the blood brain barrier to the periphery in a mouse model of Alzheimer's Disease (AD). Drug activation of ABCC1 has also been shown to reduce amyloid plaque deposition in the brain. As used herein, the term "pharmaceutically acceptable" refers to salts that are pharmaceutically acceptable in the sense of being harmless to their recipients and preferably compatible with the other optional ingredients of their formulations. preferably, the pharmaceutically acceptable salt is a salt of thioethyllazine that causes up to a moderately adverse drug reaction, preferably up to a slightly adverse drug reaction. As used herein, the term "mild" adverse reaction relates to adverse reactions that do not require medical intervention, such as rashes, headaches, digestive disturbances, fatigue, etc., while "moderate" adverse reactions are adverse reactions that require medical intervention but are potentially life threatening. Preferably, the pharmaceutically acceptable salt comprises one or more anions selected from the group consisting of sulfate, pyrosulfate, bisulfate, sulfite, bisulfate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate (decanoate), octanoate, acrylate, formate, isobutyrate, decanoate (caprate), heptanoate, propiolate (propiolate), oxalate, malonate, succinate, suberate, sebacate, fumarate, malate, Maleate, butyne-1, 4-dioate, hexyne-1, 6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylensulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β -hydroxybutyrate, glycolate, tartrate, methanesulfonate, propylsulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate (almondate), adipate, alginate, mucinate (mucate), aspartate, aspartic acid, Benzene sulfonate, butyrate, camphorate, camphorsulfonate, cyclopentane propionate, digluconate, dodecyl sulfate, ethane sulfonate, glucoheptonate (glucoheptanoate), glycerophosphate, hemisulfate, hexanoate, 2-hydroxyethane sulfonate, 2-naphthalene sulfonate, nicotinate, nitrate, palmitate, pectate (pectin), persulfate, picrate, pivalate, salicylate, thiocyanate, toluene sulfonate, arginine (arginate), and undecanoate. More preferably, the pharmaceutically acceptable salt comprises one or more anions selected from the group consisting of chloride, sulfate, methanesulfonate, benzenesulfonate, toluenesulfonate, fumarate, malonate, malate, maleate, succinate, lactate, glycolate, citrate, aspartate and mandelate, preferably malate and/or maleate, more preferably maleate.

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least one compound having a mass in grams/mole, the amount of which is determined in (a) and (b), the variation of which is determined in (c) and analyzed in (d), is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936 and 189.0096 (group 2).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least one compound having a mass in grams/mole, the amount of which is determined in (a) and (b), the variation of which is determined in (c) and analyzed in (d), is at least a compound having a mass in grams/mole of 110.0367.

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, the amounts of at least two compounds, preferably at least three compounds, more preferably at least four compounds, more preferably at least five compounds, having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in (a) and (b), the change in phase is determined in (c), and an analysis is performed in (d), wherein more preferably one of the at least two compounds, preferably one of the at least three compounds, more preferably one of the at least five compounds is a compound having a mass in grams/mole of 110.0367.

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, the amounts of at least two compounds, preferably at least six compounds or at least seven compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in (a) and (b), the change in the phase ratio is determined in (c), and an analysis is performed in (d), wherein more preferably one of the at least two compounds, more preferably one of the at least six compounds or one of the at least seven compounds is a compound having a mass in grams/mole of 110.0367.

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the amount of at least one further compound having a mass in grams/mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), preferably from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4) is additionally determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the method comprises:

(b) Determining the amount of at least two compounds identical to those in (a) in a second body fluid sample of the subject;

(c) Determining a change in the amount of the at least two compounds determined according to (a) as compared to the corresponding amount of the at least two compounds determined according to (b);

(d) Analyzing the change determined in (c) using a computer-implemented prediction algorithm capable of predicting AD in a subject based on the at least one compound, thereby identifying a subject suffering from AD or at risk of developing alzheimer's dementia if AD is predicted.

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 189.0096 g/mol, a compound having a mass of 224.084 g/mol and a compound having a mass of 254.1153 g/mol are determined in (a) and (b), the change in the phase is determined in (c) and analyzed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 189.993 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 254.115 g/mol are determined in (a) and (b), the change in the phase is determined in (c) and analyzed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 315.2409 g/mol and a compound having a mass of 702.5682 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and an analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol and a compound having a mass of 254.1153 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 254.1153 g/mol and a compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and an analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol and a compound having a mass of 702.5682 g/mol are determined in (a) and (b), the change in the phase is determined in (c) and analyzed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol and a compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in the phase is determined in (c) and analyzed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in (a) and (b), the change in the comparison is determined in (c), and the analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 208.0737 g/mol and a compound having a mass of 702.5682 g/mol are determined in (a) and (b), the change in the comparison is determined in (c), and the analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol and a compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amount of a compound having a mass of 110.0367 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 254.1153 g/mol, a compound having a mass of 189.9936 g/mol, a compound having a mass of 189.0096 g/mol and a compound having a mass of 224.084 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and an analysis is performed in (d).

In some preferred embodiments of the method of the second aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 702.5682 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 254.1153 g/mol are determined in (a) and (b), the change in comparison is determined in (c) and analyzed in (d).

The determination of the amount in any of steps (a) and/or (b) is performed by Mass Spectrometry (MS), preferably by tandem mass spectrometry (MS/MS), more preferably by tandem mass spectrometry (LC-MS/MS) in combination with previous liquid chromatography, further details being found in the section relating to the first aspect of the invention. The method is an in vitro method. As used herein, the term "subject" relates to a multicellular animal, preferably to a vertebrate, more preferably to a mammal, more preferably to a human. The body fluid sample is selected from the group consisting of a blood sample, a urine sample, a liquid sample (CSF) and a mixture of two or more thereof, preferably the body fluid sample is a blood sample, and the first body fluid sample and the second body fluid sample are selected independently of each other from the group consisting of a blood sample, a urine sample, a liquid sample (CSF) and a mixture of two or more thereof, respectively, wherein preferably the first body fluid sample and the second body fluid sample are the same kind of sample and are each selected from the group consisting of a blood sample, a urine sample, a liquid sample (CSF) and a mixture of two or more thereof, wherein more preferably the first body fluid sample and the second body fluid sample are each a blood sample (first blood sample and second blood sample).

Aspect 3-method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD) (comparison of amounts before and after TEP administration)

In a third aspect, the invention relates to a method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(c) Comparing the amount of the at least one compound determined according to (a) with a corresponding amount of the at least one compound determined according to (b), wherein a change in the amount of the at least one compound according to (b) compared to the amount determined in (a) is indicative of the subject's Alzheimer's dementia or the subject's risk of developing Alzheimer's dementia;

The definitions indicated above in the section relating to the first aspect apply here also to the section relating to the third aspect of the invention. Thioethyllazine (IUPAC: 2- (ethylsulfanyl) -10- [3- (4-methylpiperazin-1-yl) propyl ] -10H-phenothiazine, CAS:1420-55-9, TEP) and pharmaceutically acceptable salts thereof are also as defined in the section above relating to the second aspect.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the at least one compound having a mass in grams/mole is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936 and 189.0096 (group 2).

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amount of a compound having a mass of 110.0367 g/mole is determined in (a) and (b), and in (c) the comparison is made.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, the amounts of at least two compounds, preferably at least three compounds, more preferably at least four compounds, more preferably at least five compounds, having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in step (a) and (b) in a first body fluid sample of the subject, and the comparison is made in (c), wherein a change in the amounts of each of the at least two compounds, preferably each of the at least three compounds, more preferably each of the at least four compounds, more preferably each of the at least five compounds, compared to the amounts determined in (a) is indicative of the risk of developing alzheimer's disease in the subject or the subject, wherein the change in the amounts of the at least two compounds is preferably at least one of the at least two compounds, preferably at least one of the at least four compounds, preferably at least one of the at least five compounds, more preferably at least one of the compounds, preferably at least one of the compounds, 110.0367 grams.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, a change in the amount of each of the at least two compounds, preferably at least six compounds, preferably each of the at least six compounds, more preferably each of the at least seven compounds, in relation to the amount determined in (a) is determined in steps (a) and (b) in a first body fluid sample of the subject, with a mass in grams/mole selected from the group consisting of group 1 or group 2, and a comparison is made in (c), wherein a change in the amount of each of the at least two compounds, preferably each of the at least six compounds, more preferably each of the at least seven compounds, in relation to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease, wherein the mass of one of the at least two compounds, more preferably one of the at least six compounds, or at least seven compounds is a compound of 110.0367 grams/mole.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the amount of at least one further compound is additionally determined in steps (a) and (b) in the first body fluid sample of the subject, said at least one further compound having a mass in grams/mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), more preferably from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4), and is compared in (c), wherein a change in the amount of at least one compound according to (b) compared to the amount determined in (a) and a change in the amount of at least one further compound according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the method comprises:

(a) Determining in a first body fluid sample of the subject the amount of at least two compounds having a mass in grams/mole selected from the group consisting of group 3, preferably group 4;

(c) Comparing the amount of the at least two compounds determined according to (a) with the corresponding amount of the at least two compounds determined according to (b), wherein a change in the amount of the at least two compounds according to (b) compared to the amount determined in (a) is indicative of the subject's Alzheimer's disease or the subject's risk of developing Alzheimer's disease;

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 189.0096 g/mol, a compound having a mass of 224.084 g/mol and a compound having a mass of 254.1153 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and the comparison is made in (c), wherein a change of at least 1% of the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least amounts of a compound having a mass of 189.993 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 254.115 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and the comparison is made in (c), wherein a change of at least 1% of the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 315.2409 g/mol and a compound having a mass of 702.5682 g/mol are determined in step (a) and (b) in a first body fluid sample of the subject, and a comparison is made in (c), wherein a change in the amount of each of these compounds according to (b) of at least 1% compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol and a compound having a mass of 254.1153 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) of at least 12.5% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 254.1153 g/mol and a compound having a mass of 315.2409 g/mol are determined in step (a) and (b) in a first body fluid sample of the subject, and a comparison is made in (c), wherein a change in the amount of each of these compounds according to (b) of at least 1% compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol and a compound having a mass of 702.5682 g/mol are determined in step (a) and (b) in a first body fluid sample of the subject, and a comparison is made in (c), wherein a change of at least 12.5% in the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to at least 2% of the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and the comparison is made in (c), wherein a change of at least 1% of the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) of at least 12.5% is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to at least 1% of the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 208.0737 g/mol and a compound having a mass of 702.5682 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to at least 1% of the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to at least 40% of the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 254.1153 g/mol, a compound having a mass of 189.9936 g/mol, a compound having a mass of 189.0096 g/mol and a compound having a mass of 224.084 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and a comparison is made in (c), wherein a change in the amount of each of these compounds according to (b) of at least 1% compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments of the method of the third aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amount of a compound having a mass of 110.0367 g/mol, a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 702.5682 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.1153 g/mol are determined in step (a) and (b) in a first body fluid sample of the subject, and a comparison is made in (c), wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) of at least 2% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

Aspect 4-method for identifying a subject suffering from Alzheimer's Dementia (AD) (comparison of amount after TEP administration to reference value)

A fourth aspect of the invention relates to a method for identifying a subject suffering from Alzheimer's Dementia (AD), the method comprising:

(a) Determining in a body fluid sample of a subject an amount of at least one compound having masses ：110.0367、232.004、224.084、254.1153、189.9936、189.0096、155.007、355.345、358.159、109.0706、110.0004、145.0226、166.0633、178.0995、190.0631、191.0253、200.0452、212.0536、234.1832、236.1049、259.9982、285.1362、291.0832、301.1313、314.1266、318.1215、319.2486、331.2545、332.1372、334.2146、339.3135、346.1528、350.0956、366.0912、371.8758、440.2751、538.1752、540.2563 and 782.5638 in grams/mole selected from the group consisting of (group 1);

(b) Comparing the amount of the at least one compound determined according to (a) with the average amount of healthy subjects, wherein a change in the amount of the at least one compound according to (a) compared to the average amount of healthy subjects, preferably based on the average amount of healthy subjects being 100% being at least a 3% change, indicates that the subject is suffering from or is at risk of developing alzheimer's disease;

Wherein the body fluid sample is obtained after administration of thioethyllazine or a pharmaceutically acceptable salt thereof to a subject.

The definitions given in the section relating to the first aspect and the explanations given in the second and third sections relating to Thioethyllazine (TEP) and pharmaceutically acceptable salts thereof are also applicable here in the section relating to the fourth aspect of the invention.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the at least one compound having a mass in grams/mole is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936 and 189.0096 (group 2).

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), at least the amount of compound of mass 110.0367 in grams/mole is determined in (a) and compared in (b).

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing alzheimer's disease involving the fourth aspect, the amounts of at least two compounds, preferably at least three compounds, more preferably at least four compounds, and more preferably at least five compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in a first bulk liquid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of the at least two compounds, preferably each of the at least three compounds, more preferably each of the at least four compounds, more preferably each of the at least five compounds is indicative of the risk of developing alzheimer's disease in the subject or the subject developing alzheimer's disease, wherein one of the at least two compounds, preferably one of the at least three compounds, more preferably at least four compounds, more preferably at least one of the at least five compounds, more preferably at least one of the compounds is at least 25 grams of the more preferably at least one of the compounds.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, the amounts of at least two compounds, preferably at least six compounds, more preferably at least seven compounds, having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in a first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of the at least two compounds, preferably each of the at least six compounds, more preferably each of the at least seven compounds, is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease, wherein more preferably one of the at least two compounds, more preferably one of the at least six compounds, or one of the at least seven compounds is a compound having a mass in grams/mole of 110.0367.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing alzheimer's disease relating to the fourth aspect, the amount of at least one further compound is determined in step (a) in the first body fluid sample of the subject, said at least one further compound having a mass in grams/mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), more preferably from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4), and is compared in (b), wherein the amount of each of the at least one compound is at least 3% change compared to the average amount of healthy subjects, preferably based on the average amount of healthy subjects being 100%, indicative of alzheimer's disease or the risk of developing alzheimer's disease in the subject.

The fourth aspect of the invention also relates to a method for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(a) Determining in a body fluid sample of a subject the amount of at least two compounds having a mass in grams/mole selected from the group consisting of group 3, preferably selected from the group consisting of group 4;

(b) Comparing the amount of the at least two compounds determined according to (a) with the average amount of healthy subjects, wherein a change in the amount of the at least two compounds according to (a) compared to the average amount of healthy subjects, preferably based on a change of at least 3% as 100% of the average amount of healthy subjects, indicates that the subject is suffering from or is at risk of developing alzheimer's disease;

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, in step (a) at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 189.0096 g/mol, a compound having a mass of 224.084 g/mol and a compound having a mass of 254.1153 g/mol are determined in a first body fluid sample of the subject, and in (b) a comparison is made, wherein a change of at least 3% in the amount of each of these compounds is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, in step (a) at least the amounts of a compound having a mass of 189.993 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 254.115 g/mol are determined in a first body fluid sample of the subject, and in (b) a comparison is made, wherein a change of at least 3% in the amount of each of these compounds is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing alzheimer's disease involving the fourth aspect, at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 315.2409 g/mol and a compound having a mass of 702.5682 g/mol are determined in the first body fluid sample of the subject in step (a), and a comparison is made in (b), wherein a change of at least 3% in the amount of each of these compounds is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol and a compound having a mass of 254.1153 g/mol are determined in the first body fluid sample of the subject in step (a), and in (b) a comparison is made, wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments of the method for identifying a subject suffering from or at risk of developing alzheimer's disease involving the fourth aspect, at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 254.1153 g/mol and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in step (a), and a comparison is made in (b), wherein a change of at least 3% in the amount of each of these compounds is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, in step (a) at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol and a compound having a mass of 702.5682 g/mol are determined in a first body fluid sample of the subject, and in (b) a comparison is made, wherein a change of at least 3% in the amount of each of these compounds is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in step (a), and in (b) a comparison is made, wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, in step (a) at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 315.2409 g/mol are determined in a first body fluid sample of the subject, and in (b) a comparison is made, wherein a change of at least 3% in the amount of each of these compounds is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), in step (a) at least the amount of a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol is determined in a first body fluid sample of the subject, and in (b) a comparison is made, wherein a change of at least 3% in the amount of each of these compounds is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in the first body fluid sample of the subject in step (a), and in (b) a comparison is made, wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), in step (a) at least the amount of a compound having a mass of 208.0737 g/mol and a compound having a mass of 702.5682 g/mol is determined in a first body fluid sample of the subject, and in (b) a comparison is made, wherein a change of at least 12% in the amount of each of these compounds is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in step (a), and in (b) a comparison is made, wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amount of a compound having a mass of 110.0367 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 254.1153 g/mol, a compound having a mass of 189.9936 g/mol, a compound having a mass of 189.0096 g/mol and a compound having a mass of 224.084 g/mol are determined in the first body fluid sample of the subject in step (a), and a comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the subject's risk of developing alzheimer's disease.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing alzheimer's disease, at least the amount of a compound having a mass of 110.0367 g/mol, a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 702.5682 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, a compound having a mass of 232.004 g/mol and a compound having a mass of 254.1153 g/mol are determined in step (a) in a first body fluid sample of the subject, and are compared in step (b), wherein a change of at least 3% in the amount of each of these compounds is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

In some preferred embodiments relating to the method of the fourth aspect for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the average amount of healthy subjects is a reference value from healthy subjects of the literature, or a reference value previously determined from healthy subjects, or a reference value from a body fluid sample of the subject prior to TEP administration.

Diagnostic use of 5 th aspect 2

A fifth aspect of the invention relates to a combination of at least one compound having a mass in grams/mole selected from the group consisting of group 1 or group 2 or as defined in any of the preferred embodiments described in the previous section relating to the first, second, third or fourth aspect of the invention, and optionally at least one compound having a mass in grams/mole selected from the group consisting of group 3, preferably selected from the group consisting of group 4, for use in diagnosing AD in a subject or for determining the risk of a subject developing AD.

Group 1, group 2, group 3 and group 4 are as defined above in the section relating to the first, second, third or fourth aspect of the invention.

In some preferred embodiments of at least one compound used according to the fifth aspect, at least two compounds having a mass in grams per mole selected from the group consisting of group 1 or group 2 as described above in the section relating to the first, second, third or fourth aspect of the invention are used, preferably at least four compounds having a mass in grams per mole selected from the group consisting of group 1 or group 2 as defined in any of the preferred embodiments as described above in the section relating to the first, second, third or fourth aspect of the invention, more preferably at least three compounds having a mass in grams per mole selected from the group consisting of group 1 or group 2 as defined in any of the preferred embodiments as described above in the section relating to the first, second, third or fourth aspect of the invention, more preferably at least four compounds having a mass in grams per mole selected from the group consisting of group 1 or group 2 as defined above in any of the preferred embodiments relating to the first, second, third or fourth aspect of the invention, and at least one compound having a mass in grams per mole selected from the group 1 or group 4 as defined in any of the preferred embodiments as described above in any of the group 1 or group 2 of the fourth aspect of the invention.

In some preferred embodiments of at least one compound used according to the fifth aspect, at least two compounds having a mass in grams/mole selected from the group consisting of any of the preferred embodiments defined in the section relating to the first, second, third or fourth aspects of the invention, preferably at least seven compounds having a mass in grams/mole selected from the group consisting of any of the preferred embodiments defined in the section relating to the first, second, third or fourth aspects of the invention, each optionally in combination with at least one compound having a mass in grams/mole selected from the group consisting of the group 1 or the group 2, more preferably selected from the group consisting of any of the preferred embodiments defined in the section relating to the first, second, third or fourth aspects of the invention, group 1 or group 2.

In some preferred embodiments of the at least one compound for use according to the fifth aspect, at least two compounds having a mass in grams/mole selected from the group consisting of group 3 or group 4 are used for diagnosing AD in a subject or for determining the risk of a subject developing AD.

Diagnostic use (alternative wording)

The fifth aspect of the invention also relates to the use of at least one compound having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of the preferred embodiments described above in the section relating to the first, second, third or fourth aspects of the invention, and optionally in combination with at least one compound having a mass in grams/mole selected from the group consisting of group 3 or group 4, in a sample of a subject for diagnosing AD in the subject or for determining the risk of the subject developing AD.

In some preferred embodiments of the use according to the fifth aspect, a combination of at least two, preferably at least three, more preferably at least four, more preferably at least five compounds having a mass in grams/mole selected from the group consisting of any of the preferred embodiments set 1 or set 2 as defined above in the section relating to the first, second, third or fourth aspects of the invention in a sample of a subject is used for diagnosing AD in the subject or for determining the risk of the subject developing AD.

In some preferred embodiments of the use according to the fifth aspect, the sample of the subject has at least two, preferably at least six, more preferably at least seven compounds of mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of the preferred embodiments described above in the section relating to the first, second, third or fourth aspects of the invention and optionally in combination with at least one compound of mass in grams/mole selected from the group consisting of group 3 or group 4 for diagnosing AD in the subject or for determining the risk of the subject developing AD.

In some preferred embodiments of the use according to the fifth aspect, at least two compounds having a mass in grams/mole selected from the group consisting of group 3 or group 4 in a sample of a subject are used for diagnosing AD in the subject or for determining the risk of the subject developing AD.

Aspect 6-apparatus

A sixth aspect of the invention relates to a device suitable for identifying a subject suffering from or at risk of developing Alzheimer's Dementia (AD), the device comprising:

(a) An analysis unit capable of determining an amount of at least one compound having a mass in grams/mole selected from the group consisting of group 1 in a first liquid-in-liquid sample of the subject;

(b) A data processor for analyzing the determined amount of the at least one compound using a prediction algorithm implemented in the data processor, the prediction algorithm being capable of predicting the AD of the subject based on the at least two compounds, thereby identifying the subject suffering from AD if AD is predicted, or identifying the subject at risk of developing alzheimer's dementia if AD is predicted.

Preferred embodiments of the device, in particular of the device relating to a compound having a specific mass in grams per mole and its specific group (such as group 2, group 3 and group 4) are as described above in the section relating to the first, second, third or fourth aspect of the invention. Furthermore, the definitions indicated above in these sections apply here also to the sections relating to the device.

Kit of aspect 7

In a seventh aspect, the present invention relates to a kit for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), said kit comprising a reference sample having at least one compound of a mass in grams/mole selected from the group consisting of group 1 or group 2, optionally a reference sample having at least one compound of a mass in grams/mole selected from the group consisting of group 3, or optionally a reference sample having at least one compound of a mass in grams/mole selected from the group consisting of group 4, and optionally one or more reagents for sample preparation and/or extraction.

The invention is further illustrated by the following embodiments and combinations of embodiments (as indicated by the respective dependencies and reverse references). In particular, it is noted that in each case where the scope of embodiments is mentioned, for example in the context of a term such as "method according to any of embodiments 1 to 4", each embodiment within this scope is meant to be explicitly disclosed to the skilled person, i.e. the wording of the term should be understood by the skilled person as synonymous with "method according to any of embodiments 1,2, 3 and 4".

1. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(b) Analyzing the determined amount of the at least one compound using a computer-implemented prediction algorithm capable of predicting the AD of the subject based on the at least one compound, thereby identifying the subject as suffering from AD or at risk of developing alzheimer's dementia if AD is predicted,

Wherein preferably the first body fluid sample is derived from a subject after administration of Thioethyloxazine (TEP) or a pharmaceutically acceptable salt thereof to the subject, wherein the sample is preferably taken within 0.5 to 8 hours after administration of TEP.

2. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to embodiment 1, wherein the at least one compound having a mass in grams/mole is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936 and 189.0096 (group 2).

3. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to embodiment 1 or 2, wherein at least a compound of mass 110.0367 in grams/mole is determined in (a) and the amount determined is analyzed in (b).

4. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 1 to 3, wherein the amount of at least two compounds, preferably at least three compounds, more preferably at least four compounds, more preferably at least five compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 is determined in the first body fluid sample of the subject in step (a), and is analyzed in (b), wherein more preferably one of the at least two compounds, preferably one of the at least three compounds, more preferably one of the at least five compounds is a compound having a mass in grams/mole of 110.0367.

5. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 1 to 4, wherein the amount of at least two compounds, preferably at least six compounds, more preferably at least seven compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 is determined in a first body fluid sample of the subject in step (a), and analyzed in (b), wherein more preferably one of the at least two compounds, more preferably one of the at least six compounds or one of the at least seven compounds is a compound having a mass in grams/mole of 110.0367.

6. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 1 to 4, wherein the amount of at least one further compound having a mass in grams/mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), preferably selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4) is additionally determined in step (a) in a first body fluid sample of the subject and is additionally analyzed in (b).

7. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

8. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 1-7, wherein the computer-implemented predictive algorithm uses a supervised machine learning algorithm that has been trained by a training dataset comprising quantitative or qualitative data from the following compounds in body fluid samples from subjects known to suffer from AD and subjects known not to suffer from AD:

-At least two compounds from group 3, preferably from group 4.

9. The method of embodiment 8, wherein the supervised machine learning algorithm establishes a classification algorithm selected from the group consisting of logistic regression, random gradient descent classifier (SGD classifier), decision tree classifier, random tree classifier, limit tree classifier, adaBoost classifier, multi-layer perceptron classifier, gradient boost classifier, gaussian naive bayes, K nearest neighbor classifier, C-support vector classification, quadratic discriminant analysis, and linear regression.

10. The method of embodiment 9, wherein the supervised machine learning algorithm builds and/or applies a logistic regression model based on equation (I)

y = I + C₁*F₁ + C₂*F₂ +…+ C_n*F_n,(I)

Where I is the intercept and C _n is the coefficient of mass F _n.

12. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 189.0096 g/mol, the compound having a mass of 224.084 g/mol, and the compound having a mass of 254.1153 g/mol are determined in (a), and analyzed in (b).

13. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 189.993 g/mol, the compound having a mass of 224.084 g/mol, the compound having a mass of 232.004 g/mol, and the compound having a mass of 254.115 g/mol are determined in (a), and analyzed in (b).

14. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 175.0633 g/mol, the compound having a mass of 224.084 g/mol, the compound having a mass of 232.004 g/mol, the compound having a mass of 315.2409 g/mol, and the compound having a mass of 702.5682 g/mol are determined in (a), and analyzed in (b).

15. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 175.0633 g/mol, the compound having a mass of 208.0737 g/mol, and the compound having a mass of 254.1153 g/mol are determined in (a), and analyzed in (b).

16. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 175.0633 g/mol, the compound having a mass of 224.084 g/mol, the compound having a mass of 254.1153 g/mol, and the compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

17. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 175.0633 g/mol, the compound having a mass of 208.0737 g/mol, the compound having a mass of 315.2409 g/mol, and the compound having a mass of 702.5682 g/mol are determined in (a), and analyzed in (b).

18. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 175.0633 g/mol, and the compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

19. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 208.0737 g/mol, the compound having a mass of 232.004 g/mol, and the compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

20. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 175.0633 g/mol and the compound having a mass of 208.0737 g/mol are determined in (a), and the analysis is performed in (b).

21. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 175.0633 g/mol, and the compound having a mass of 208.0737 g/mol are determined in (a), and analyzed in (b).

22. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 208.0737 g/mol and the compound having a mass of 702.5682 g/mol are determined in (a), and the analysis is performed in (b).

23. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 175.0633 g/mol, the compound having a mass of 208.0737 g/mol, and the compound having a mass of 315.2409 g/mol are determined in (a), and analyzed in (b).

24. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 232.004 g/mol, the compound having a mass of 254.1153 g/mol, the compound having a mass of 189.9936 g/mol, the compound having a mass of 189.0096 g/mol, and the compound having a mass of 224.084 g/mol are determined in (a), and analyzed in (b).

25. The method according to any one of embodiments 1 to 10, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 110.0367 g/mol, the compound having a mass of 175.0633 g/mol, the compound having a mass of 702.5682 g/mol, the compound having a mass of 208.0737 g/mol, the compound having a mass of 315.2409 g/mol, the compound having a mass of 232.004 g/mol, and the compound having a mass of 254.1153 g/mol are determined in (a), and analyzed in (b).

25. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

26. The method of embodiment 25, wherein the at least one compound having a mass in grams per mole (the amount of which is determined in (a) and (b), the variation of which is determined in (c) and analyzed in (c)) is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936 and 189.0096 (group 2).

27. The method of embodiment 25 or 26, wherein at least one compound having a mass in grams/mole (the amount of which is determined in (a) and (b), the variation of which is determined in (c), and analyzed in (c)) is at least a compound having a mass in grams/mole of 110.0367.

28. The method of any one of embodiments 25 to 27, wherein the amounts of at least two compounds, preferably at least three compounds, more preferably at least four compounds, more preferably at least five compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in (a) and (b), the change in comparison is determined in (c), and an analysis is performed in (d), wherein more preferably one of the at least two compounds, preferably one of the at least three compounds, more preferably one of the at least four compounds, more preferably one of the at least five compounds is a compound having a mass in grams/mole of 110.0367.

29. The method according to any one of embodiments 25 to 28, wherein the amounts of at least two compounds, preferably at least six compounds or at least seven compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d), wherein more preferably one of the at least two compounds, more preferably one of the at least six compounds or one of the at least seven compounds is a compound having a mass in grams/mole of 110.0367.

30. The method according to any one of embodiments 25 to 29, wherein the amount of at least one further compound having a mass in grams/mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), preferably selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4) is additionally determined in (a) and (b), the change in the phase ratio is determined in (c), and the analysis is performed in (d).

31. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

32. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 189.0096 g/mol, a compound having a mass of 224.084 g/mol, and a compound having a mass of 254.1153 g/mol are determined in (a) and (b), the change in phase is determined in (c), and analyzed in (d).

33. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 189.993 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.115 g/mol are determined in (a) and (b), the change in phase is determined in (c), and analyzed in (d).

34. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d).

35. The method of any one of embodiments 25 to 30, wherein at least the amounts of the compound having a mass of 175.0633 g/mol, the compound having a mass of 208.0737 g/mol, and the compound having a mass of 254.1153 g/mol are determined in (a) and (b), the change in phase is determined in (c), and the analysis is performed in (d).

36. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 254.1153 g/mol, and a compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in the phase is determined in (c), and the analysis is performed in (d).

37. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in (a) and (b), the change in phase is determined in (c), and analyzed in (d).

38. The method of any one of embodiments 25 to 30, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 175.0633 g/mol, and the compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in phase is determined in (c), and the analysis is performed in (d).

39. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in phase is determined in (c), and analyzed in (d).

40. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in (a) and (b), the change in phase is determined in (c), and the analysis is performed in (d).

41. The method of any one of embodiments 25 to 30, wherein at least the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 175.0633 g/mol, and the compound having a mass of 208.0737 g/mol are determined in (a) and (b), the change in phase is determined in (c), and the analysis is performed in (d).

42. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 208.0737 g/mol and a compound having a mass of 702.5682 g/mol are determined in (a) and (b), the change in phase is determined in (c), and the analysis is performed in (d).

43. The method of any one of embodiments 25 to 30, wherein at least the amounts of the compound having a mass of 175.0633 g/mol, the compound having a mass of 208.0737 g/mol, and the compound having a mass of 315.2409 g/mol are determined in (a) and (b), the change in phase is determined in (c), and the analysis is performed in (d).

44. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 254.1153 g/mol, a compound having a mass of 189.9936 g/mol, a compound having a mass of 189.0096 g/mol, and a compound having a mass of 224.084 g/mol are determined in (a) and (b), the change in phase is determined in (c), and an analysis is performed in (d).

45. The method of any one of embodiments 25 to 30, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 702.5682 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.1153 g/mol are determined in (a) and (b), a change in phase is determined in (c), and an analysis is performed in (d).

46. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

47. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to embodiment 46, wherein the at least one compound having a mass in grams/mole is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936, and 189.0096 (group 2).

48. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to embodiment 46 or 47, wherein at least the amount of a compound having a mass of 110.0367 g/mole is determined in (a) and (b) and compared in (c).

49. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46 to 48, wherein in steps (a) and (b) the amounts of at least two compounds, preferably at least three compounds, more preferably at least four compounds, more preferably at least five compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 are determined in a first body fluid sample of the subject, and in (c) the comparison is made, wherein the change in the amount of each of at least two compounds, preferably each of at least three compounds, more preferably each of at least five compounds, compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease, wherein the change in the amount is indicative of the risk of developing alzheimer's disease in terms of at least one of the compounds, preferably at least one of at least three compounds, more preferably at least one of at least five compounds, more preferably at least one of five compounds, more preferably at least one of 5 of compounds.

50. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46 to 49, wherein a change in the amount of each of the at least two compounds, preferably at least six compounds, or at least seven compounds according to (b) as compared to the amount determined in (a) is determined in a first body fluid sample of the subject in steps (a) and (b) having a mass in grams/mole selected from the group consisting of group 1 or group 2, and wherein the comparison in (c) is performed, wherein a change in the amount of each of the at least two compounds, preferably each of the at least six compounds, more preferably each of the at least seven compounds according to (b) as compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the subject's risk of developing alzheimer's disease, wherein more preferably the mass of one of the at least two compounds, more preferably one of the at least six compounds, or at least seven compounds is a compound of 110.0367 grams/mole.

51. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46 to 50, wherein in steps (a) and (b) the amount of at least one further compound is additionally determined in the first body fluid sample of the subject, said at least one further compound having a mass in grams per mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), more preferably from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4), and is compared in (c), wherein a change in the amount of at least one compound according to (b) compared to the amount determined in (a) and a change in the amount of at least one further compound according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

52. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(a) Determining in a first body fluid sample of the subject an amount of at least two compounds having a mass in grams/mole selected from the group consisting of group 3;

53. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 189.0096 g/mol, a compound having a mass of 224.084 g/mol, and a compound having a mass of 254.1153 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change of at least 1% of the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

54. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 189.993 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.115 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change of at least 1% of the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

55. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and wherein a change in the amount of each of these compounds according to (b) as compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

56. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, and a compound having a mass of 254.1153 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change of at least 12.5% in the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the subject's risk of developing alzheimer's disease.

57. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 254.1153 g/mol, and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and wherein a change in the amount of each of these compounds according to (b) as compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

58. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change of at least 12.5% in the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

59. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, and a compound having a mass of 315.2409 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change of at least 2% in the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the subject's risk of developing alzheimer's disease.

60. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 315.2409 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change of at least 1% of the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

61. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein at least the amounts of a compound having a mass of 175.0633 g/mol and a compound having a mass of 208.0737 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and the comparison is made in (c), wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

62. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, and a compound having a mass of 208.0737 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) of at least 1% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

63. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 208.0737 g/mol and a compound having a mass of 702.5682 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) of at least 1% is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

64. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein in steps (a) and (b) at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, and a compound having a mass of 315.2409 g/mol are determined in a first body fluid sample of the subject, and in (c) a comparison is made, wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) of at least 40% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease.

65. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein at least the amount of a compound having a mass of 110.0367 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 254.1153 g/mol, a compound having a mass of 189.9936 g/mol, a compound having a mass of 189.0096 g/mol, and a compound having a mass of 224.084 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and wherein a change in the amount of each of these compounds according to (b) compared to the amount determined in (a) is indicative of alzheimer's disease in the subject or the risk of developing alzheimer's disease in the subject.

66. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 46-51, wherein at least a compound having a mass of 110.0367 g/mol, a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 702.5682 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.1153 g/mol are determined in the first body fluid sample of the subject in steps (a) and (b), and wherein a change in the amount of each of these compounds according to (b) from the amount determined in (a) of at least 2% is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease in the subject.

67. A method for identifying a subject having Alzheimer's Dementia (AD), the method comprising:

68. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to embodiment 67, wherein the at least one compound having a mass in grams/mole is selected from the group consisting of 110.0367, 232.004, 224.084, 254.1153, 189.9936, and 189.0096 (group 2).

69. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to embodiment 67 or 68, wherein at least the amount of compound of mass 110.0367 in grams/mole is determined in (a) and (b) and compared in (c).

70. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 67 to 69, wherein a change in the amount of each of the at least two compounds, preferably at least three compounds, more preferably at least four compounds, more preferably at least five compounds, in the first body fluid sample of the subject is determined in step (a) having a mass in grams/mole selected from the group consisting of group 1 or group 2, wherein a change in the amount of each of the at least two compounds, preferably each of the at least three compounds, more preferably each of the at least four compounds, more preferably each of the at least five compounds is indicative of the risk of developing alzheimer's disease in the subject or the subject is at least 25 grams of each of the at least one compound, more preferably at least four compounds.

71. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 67 to 70, wherein in step (a) the amount of at least two compounds, preferably at least six compounds, more preferably at least seven compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 is determined in a first body fluid sample of the subject, and in (b) the comparison is made, wherein a change in the amount of each of the at least two compounds, preferably each of the at least six compounds, more preferably each of the at least seven compounds, is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease, wherein more preferably one of the at least two compounds, more preferably one of the at least six compounds, or one of the at least seven compounds is a compound having a mass in grams/110.0367.

72. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 67 to 71, wherein in steps (a) and (b) the amount of at least one other compound is determined in the first body fluid sample of the subject, said at least one other compound having a mass in grams/mole selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682, 161.0477 and 97.9769 (group 3), more preferably selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4), and is compared in (c), wherein a change in the amount of at least one compound according to (b) compared to the amount determined in (a) and a change in the amount of at least one other compound according to (a) compared to the average amount of healthy subjects, preferably a change of at least 3% based on the average amount of healthy subjects as 100%, indicates dementia or the risk of developing alzheimer's disease in the subject.

73. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

74. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 189.0096 g/mol, a compound having a mass of 224.084 g/mol, and a compound having a mass of 254.1153 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

75. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 189.993 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.115 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

76. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in the first body fluid sample of the subject in step (a), and wherein a change of at least 3% in the amount of each of these compounds is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease in the subject is compared in (b).

77. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, and a compound having a mass of 254.1153 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

78. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 224.084 g/mol, a compound having a mass of 254.1153 g/mol, and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in step (a), and wherein a change of at least 3% in the amount of each of these compounds is indicative of the subject's alzheimer's disease or the risk of developing alzheimer's disease in the subject is compared in (b).

79. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, and a compound having a mass of 702.5682 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

80. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

81. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

82. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 175.0633 g/mole and a compound having a mass of 208.0737 g/mole are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

83. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, and a compound having a mass of 208.0737 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

84. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 208.0737 g/mole and a compound having a mass of 702.5682 g/mole are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 12% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

85. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amounts of a compound having a mass of 175.0633 g/mol, a compound having a mass of 208.0737 g/mol, and a compound having a mass of 315.2409 g/mol are determined in the first body fluid sample of the subject in step (a), and the comparison is made in (b), wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the risk of the subject developing alzheimer's disease.

86. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least the amount of a compound having a mass of 110.0367 g/mol, a compound having a mass of 232.004 g/mol, a compound having a mass of 254.1153 g/mol, a compound having a mass of 189.9936 g/mol, a compound having a mass of 189.0096 g/mol, and a compound having a mass of 224.084 g/mol is determined in the first body fluid sample of the subject in step (a), and wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the subject's risk of developing alzheimer's disease is compared in step (b).

87. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any one of embodiments 67-72, wherein at least a compound having a mass of 110.0367 g/mol, a compound having a mass of 110.0367 g/mol, a compound having a mass of 175.0633 g/mol, a compound having a mass of 702.5682 g/mol, a compound having a mass of 208.0737 g/mol, a compound having a mass of 315.2409 g/mol, a compound having a mass of 232.004 g/mol, and a compound having a mass of 254.1153 g/mol are determined in the first body fluid sample of the subject in step (a), and wherein a change in the amount of each of these compounds of at least 3% is indicative of the subject's alzheimer's disease or the subject's risk of developing alzheimer's disease is compared in step (b).

88. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to any of embodiments 67-72, wherein the average amount of healthy subject values is a reference value from a healthy subject of the literature, or a reference value previously determined from a healthy subject, or a reference value from a body fluid sample of a subject prior to TEP administration.

89. The method according to any one of embodiments 1 to 88, wherein the determining of the amount in any one of steps (a) and/or (b) is performed by Mass Spectrometry (MS), preferably by tandem mass spectrometry (MS/MS), more preferably by tandem mass spectrometry (LC-MS/MS) combined with the preceding liquid chromatography.

90. The method of any one of embodiments 1 to 89, which is an in vitro method.

91. The method of any one of embodiments 1 to 90, wherein the subject is a human.

92. The method according to any one of embodiments 1 to 91, wherein the body fluid sample is selected from the group consisting of a blood sample, a urine sample, a liquid sample (CSF) and a mixture of two or more thereof, preferably the body fluid sample is a blood sample, and the first body fluid sample and the second body fluid sample are each independently selected from the group consisting of a blood sample, a urine sample, a liquid sample (CSF) and a mixture of two or more thereof, respectively, wherein preferably the first body fluid sample and the second body fluid sample are the same kind of sample and are each selected from the group consisting of a blood sample, a urine sample, a liquid sample (CSF) and a mixture of two or more thereof, wherein more preferably the first body fluid sample and the second body fluid sample are each a blood sample (first blood sample and second blood sample).

93. At least one compound having a mass in grams per mole selected from the group consisting of group 1 or (group 2) or as defined in any of claims 1 to 92, and optionally in combination with at least one compound having a mass in grams per mole selected from the group consisting of group 3, preferably selected from the group consisting of group 4, for use in diagnosing AD in a subject or for determining the risk of a subject developing AD.

94. At least one compound used according to embodiment 93, wherein at least two compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92, preferably at least three compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92, more preferably at least four compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92, more preferably at least five compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92;

Each optionally in combination with at least one compound having a mass in grams per mole selected from the group consisting of group 3 or group 4.

95. At least one compound used according to embodiment 93, wherein at least two compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92, preferably at least six compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92, more preferably at least seven compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92 are used;

96. At least two compounds having a mass in grams/mole selected from the group consisting of group 3 or group 4 for use in diagnosing AD in a subject or for determining the risk of a subject developing AD.

Diagnostic use (alternative wording)

97. Use of at least one compound having a mass in grams per mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92, and optionally in combination with at least one compound having a mass in grams per mole selected from the group consisting of group 3 or group 4, in a sample of a subject for diagnosing AD in the subject or for determining the risk of the subject developing AD.

98. The use of embodiment 97, wherein at least two, preferably at least three, more preferably at least four, more preferably at least five compounds having a mass in grams per mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 93, and optionally in combination with at least one compound having a mass in grams per mole selected from the group consisting of group 3 or group 4, in a sample of a subject are used for diagnosing AD in the subject or for determining the risk of the subject developing AD.

99. The use of embodiment 97, wherein at least two, preferably at least six, more preferably at least seven compounds having a mass in grams/mole selected from the group consisting of group 1 or group 2 as defined in any of embodiments 1 to 92, and optionally in combination with at least one compound having a mass in grams/mole selected from the group consisting of group 3 or group 4, in a sample of a subject are used for diagnosing AD in the subject or for determining the risk of the subject developing AD.

100. Use of at least two compounds having a mass in grams/mole selected from the group consisting of group 3 or group 4 in a sample of a subject for diagnosing AD in the subject or for determining the risk of the subject developing AD.

101. A device suitable for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the device comprising:

102. A kit for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the kit comprising a reference sample having at least one compound in grams per mole selected from the group consisting of group 1 or group 2, optionally a reference sample having at least one compound in grams per mole selected from the group consisting of group 3, or optionally a reference sample having at least one compound in grams per mole selected from the group consisting of group 4, and optionally one or more reagents for sample preparation and/or extraction.

The present invention is further illustrated by the following reference examples, comparative examples and examples.

Examples

The following examples should merely illustrate the invention. They should not be construed as limiting the scope of the invention in any way.

I method

I.1 Treatment of

15 Human subjects (7 healthy, 8 suffering from Alzheimer's disease) were treated with 6.5 mg of Thioethyllazine (TEP). Blood samples were collected at 0, 0.5, 1, 1.5, 2,4 and 8 hours after administration. AD subjects were categorized by a simple mental state examination (Mini-MENTAL STATE Examination, MMSE) score of 25-18. Furthermore, the diagnosis of AD was confirmed by recommended examination of "dementia" according to German DGN/DGPPN S guidelines, by:

Psychological and cognitive tests

Lumbar puncture of subjects with uncertain diagnosis of AD after CNS imaging

Clinical dementia rating (overall CDR) of 0.5 or 1, and memory box score of at least 0.5.

Isolated or major episodic memory deficit, manifested by memory in the logical memory sub-scale (delayed segment recall) from West memory scale-III (Wechsler Memory Scale-III) manifested by 1 standard deviation lower than the age-adjusted specification.

I.2 Blood sampling

Samples of 10 ml venous blood were collected into standardized tubes containing EDTA and gently shaken 1-2 times. The samples were centrifuged at 1600 g min at room temperature for 10min as soon as possible (up to 30 min). The plasma was aliquoted into 6 parts (0.5 ml). All aliquots were frozen in a cryocooler (-80 ℃) within 10 minutes after centrifugation and stored until use.

I.3 Non-targeted metabonomics

All procedures performed in this section were completed by BGI Genomics. Non-targeted metabonomics refers to high throughput analysis of metabolic states of biological systems (see Matthaios-Emmanouil P Papadimitropoulos, Catherine G Vasilopoulou, Christoniki Maga-Nteve, Maria I Klapa in Methods Mol Biol. 2018; 1738:133-147; doi: 10.1007/978-1-4939-7643-0_9.).

1. Main instrument and reagent

Ultra high performance liquid chromatography (UHF-LC) Waters 2D UPLC,Waters,USA

High resolution mass spectrometer Q Exactive, thermo FISHER SCIENTIFIC, USA

ACQUITY UPLC BEH C18 chromatographic column 1.7 μm,2.1*100 mm,Waters,USA

Low temperature high speed Centrifuge, centrifuge 5430, eppendorf

Vortex instrument QL-901, kylin-bell Lab Instruments Co., ltd., china

Ultra-pure water system Milli-Q INTEGRAL, millipore Corporation, USA

Vacuum concentrator Maxi Vacbeta GENE COMPANY

The internal standard mixture (IS) contains L-leucine-d 3, L-phenylalanine (13C9,99%), L-tryptophan-d 5, progesterone-2, 3,4-13C3

MS grade methanol (A454-4) and acetonitrile (A996-4) were purchased from Thermo FISHER SCIENTIFIC (USA). Formic acid was purchased from DIMKA (50144-50 ml, USA), and ammonium formate (17843-250G,Honeywell Fluka,USA) was obtained from Fluka. Ultrapure water was filtered through a Milli-Q system.

2. Metabolite extraction

Metabolite extraction was performed mainly following the previously reported methods [ [4] [5]. Briefly, 100. Mu.L of plasma samples were extracted by direct addition of 300. Mu.L of pre-chilled methanol and acetonitrile (2:1, v/v), and internal standard mixture 1 (IS 1) and internal standard mixture 2 (IS 2) were added for quality control of sample preparation. After vortexing for 1 minute and incubation at-20 ℃ for 2 hours, the samples were centrifuged at 4000 rpm for 20 minutes, and the supernatant was then transferred for vacuum freeze-drying. The metabolites were resuspended in 150 μl of 50% methanol and centrifuged at 4000 rpm for 30 minutes and the supernatant transferred to an autosampler vial for LC-MS analysis. Quality Control (QC) samples were prepared by pooling the same volume of each sample to evaluate reproducibility of the entire LC-MS analysis.

3LC-MS/MS analysis

3.1. Chromatographic conditions

Samples were analyzed on a Waters 2D UPLC (Waters, USA) coupled to a Q-Exactive mass spectrometer (Thermo FISHER SCIENTIFIC, USA) with a heated electrospray ionization (HESI) source and controlled by an Xcalibur 2.3 software program (Thermo FISHER SCIENTIFIC, waltham, MA, USA). Chromatographic separation was performed on Waters ACQUITY UPLC BEH C column (1.7 μm,2.1 mm × 100 mm,Waters,USA) and the column temperature was maintained at 45 ℃. In the positive mode, the mobile phase consisted of 0.1% formic acid (a) and acetonitrile (B), and in the negative mode, the mobile phase consisted of 10 mM ammonium formate (a) and acetonitrile (B). The gradient conditions are shown as 0-1 min, 2% B, 1-9 min, 2% -98% B, 9-12 min, 98% B, 12-12.1 min, 98% B to 2% B, and 12.1-15 min, 2% B. The flow rate was 0.35 mL/min and the injection volume was 5. Mu.L.

3.2 Mass spectrometry conditions

The mass spectral settings for the positive/negative ionization mode are shown as spray voltage 3.8/-3.2 kV, sheath gas flow rate 40 arbitrary units (arb), assist gas (auxgas) flow rate 10 arb, assist gas heater temperature 350 ℃, capillary temperature 320 ℃. The full scan range is 70-1050 m/z with a resolution of 70,000 and the Automatic Gain Control (AGC) target for MS acquisition is set to 3e6 with a maximum ion implantation time of 100 MS. The first 3 precursors were selected for subsequent MSMS fragmentation with a maximum ion implantation time of 50 ms and a resolution of 17,500 and agc of 1e5. The step normalized collision energies were set to 20, 40 and 60 eV.

In order to provide more reliable experimental results during instrument testing, samples were randomly ordered to reduce systematic errors. One QC sample was interspersed with every 10 samples.

4. Data preprocessing

Raw mass spectra data (raw files) collected by LC-MS/MS were imported Compound Discoverer 3.1.1 (Thermo FISHER SCIENTIFIC, USA) for data processing including peak extraction, intra-and inter-group retention time correction, additive ion pooling (additive ion pooling), deletion value filling, background peak labeling, and metabolite identification. Finally, information about the molecular weight of the compound, retention time, peak area and discrimination results is derived. The identification of metabolites was a comprehensive result of the BGI self-created standard library, mzCloud and CHEMSPIDER (HMDB, KEGG, lipidMaps) databases.

The result of Compound Discoverer 3.1.1 derivation is imported metaX for data preprocessing, which includes 1. Normalization of the data using probability quotient normalization (Probabilistic Quotient Normalization, PQN [6 ]) to obtain relative peak areas. 2. Batch effects were corrected using QC-RLSC [ [4] (Quality control-based robust LOESS signal correction) based robust LOESS signal correction, 3. CV (coefficient of variation) of the relative peak areas in all QC samples were calculated and compounds with CV greater than 30% were deleted. Probability Quotient Normalization (PQN) is to calculate the average value of all QC samples for each feature to obtain a reference vector, calculate the median value (mean) between the reference vector and each sample, and obtain a coefficient vector associated with each sample, and then normalize by dividing each sample by the median value of the coefficient vector (the median value of each sample is different).

QC-RLSC [ [4] A local polynomial regression fit signal correction based on QC sample information (QC-RLSC).

The data quality was assessed by the reproducibility of QC sample detection. The content includes chromatogram overlap, PCA, peak number and peak response intensity differences of QC samples.

5. Partial least squares-discriminant analysis (PLS-DA)

A partial least squares regression method is used to build a model of the relationship between metabolite expression and sample class to achieve modeling predictions for the sample class. The PLS-DA model between the comparison groups (two groups) was established after log2-log conversion of the data, and the method for scaling (scaling) was Par. 7-fold cross-validation (7-fold cross-validation) was performed during modeling. To evaluate the model, PLS-DA model [7] [8] was subjected to 200 response substitutions.

The data were used for fold change analysis and T-test. Fold Change (FC) was obtained by fold change analysis, and p-value was obtained by student T-test, wherein p-value was corrected for False Discovery Rate (FDR) to obtain q-value.

The comparison was performed between a healthy precursor before administration of thioethlazine and a precursor with alzheimer's disease before administration of thioethlazine, and between a healthy precursor after administration of thioethlazine and a precursor with alzheimer's disease after administration of thioethlazine.

6. Database and authentication

Databases used for compound identification include mzCloud, chemspider, the Human Metabolome Database (HMDB), the Kyoto gene and genome encyclopedia (Kyoto Encyclopedia of Genes and Genomes, KEGG), LIPIDMAPS databases.

II identification of metabolites/quality of diagnostic value

1. Software for providing a plurality of applications

Python 3.9.5 was run as Spyder.3.2 IDE with the library scikit-learn, pandas and numpy. Windows 11,Microsoft Excel 365.

2. Data processing and filtering

After all blood samples were treated using the non-targeted metabolome program, the Area Under Curve (AUC) values for each mass (x) were normalized using the following formula:

z = (x - u) / s

where u is the average of all samples and s is the standard deviation of all samples. The normalized data represents dataset 1.

Thereafter, a correlation analysis of all the qualities within the dataset 1 is performed. In case an absolute pair-wise correlation exceeding 0.95 is found, one quality of the pair is excluded from further analysis. The remaining data represents dataset 2.

Then, after running PLS-DA, dataset 2 was continuously filtered according to student's T test for (i) not significantly different mass between healthy subjects and AD subjects at the pre-TEP time point and (ii) significantly different mass at the post-TEP time point to generate dataset 3.

Dataset 3 is then checked against the quality found in dataset 1 to have a highly relevant quality. Four such masses were identified and the relevant masses excluded from data set 2 were reintroduced into data set 3 to yield the final data set containing 45 masses.

3. Analyzing predictive value of individual masses

AUC data at 2 hours after TEP administration for each of the 45 masses selected were normalized to the corresponding mean (X _{Quality of i}= AUC_{Quality of I} X100/mean _{All health qualities i}) for healthy individuals for better visualization. The 32 masses were significantly different in the samples obtained from AD subjects than in the samples from healthy subjects. For 42 of the 45 masses, an absolute change of 3% (relative to the average of healthy subjects) was sufficient to diagnose AD, with an accuracy of at least 67% (table 1).

TABLE 1

TABLE 1-continuous 1

TABLE 1-continuous 2

TABLE 1-3

TABLE 1-Xue 4

4. Screening for quality combinations that can predict disease states

To improve the accuracy and robustness of such diagnostic methods, the final dataset is further analyzed and processed using supervised machine learning algorithms to evaluate the diagnostic potential of all possible combinations of up to 7 qualities (excluding combinations that would contain highly correlated qualities), and to extract combinations with high diagnostic value. Because of the small queue size, rigorous methods of training and testing classifier models have been utilized to estimate the diagnostic robustness of a particular quality combination to unknown samples.

The scikit-LEARN GRIDSEARCH pipeline with nested cross-validation method was used to evaluate the best quality combination and classifier for identifying AD patients. Within the GRIDSEARCH pipeline, supervised learning methods have been screened for logistic regression, random gradient descent classifier (SGD classifier), decision tree classifier, random tree classifier, limit tree classifier, adaBoost classifier, multi-layer perceptron classifier (MLP classifier), gradient lifting classifier, gaussian naive Bayes (GaussianNB), K-nearest neighbor classifier, C-Support Vector Classification (SVC), quadratic discriminant analysis, and linear regression. For each method, a hierarchical tri-fold data segmentation (inner loop) was used to perform hyper-parametric optimization and training on 70% of the complete dataset. The classifier with the highest area under the ROC curve (AUC) was then evaluated on the remaining 30% of the dataset using leave-one-out (outer loop) to assess the prevalence and accuracy of model and mass combinations.

Results

Screening indicated that a large number of combinations of 1 to 7 out of 45 masses found was most valuable for accurate prediction of disease states. Table 2 below shows exemplary choices of quality combinations for predicting disease states with high accuracy. Since the computation of predictions for most classifiers is difficult to delineate or not at all, only the logistic regression algorithm was selected for this table 2. In most cases, however, the decisive factor for accurate prediction is the combination of qualities, not the algorithm used for prediction, which again highlights the effectiveness of the selected quality. This fact is illustrated in table 4, where key diagnostic parameters for different classifiers using the same quality combination are shown.

For disease prognosis, the logistic regression equation should be interpreted as follows:

y = I + C₁*F₁ + C₂*F₂ +…+ C_n*F_n,

Where I is the intercept and C _n is the coefficient of mass F _n. Positive y values are predictive of alzheimer's disease, while negative values predict the non-diseased state. A larger absolute value means a greater certainty of prediction.

For example, in the case of the first mass combination shown in table 2, the formula is as follows:

Where [ FeatX ] is the concentration (standard scaling or absolute) of the corresponding mass in the sample.

TABLE 2

Without being bound by this theory, it is believed that of the 45 compounds represented above by mass in grams/mole, the following 5 compounds are related to the chemical compounds listed in table 3 below:

TABLE 3 Table 3

Structurally related compounds having specific masses (in g/mol)

TABLE 4 Table 4

Reference to the literature

[1] US 2015/0374711 A1。

[2] SCHINDLER S E Bollinger J G, ovod V et al, high-precision plasma beta-amyoid 42/40 predicts current and future brain amyloidosis (High precision plasma beta-amyloid 42/40 predicts current and future brain amyloidoses). Neurology 2019,93 (17): e1647-e1659.

[3] Eugenia Trushina E,Dutta T,Persson X-M T,Mielke M M,Petersen R C,Identification of Altered Metabolic Pathways in Plasma and CSF in Mild Cognitive Impairment and Alzheimer's Disease Using Metabolomics( Metabonomics was used to identify altered metabolic pathways in plasma and cerebrospinal fluid in mild cognitive impairment and alzheimer's disease.) PlosOne, 5 months 2013, 8 th reel, 5 th stage, e63644.

[4] Dunn W B, broadhurst D, begley P et al ,Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry( use a procedure of large scale metabolic analysis of serum and plasma using gas and liquid chromatography in combination with mass spectrometry) Nature Protocols,2011,6 (7): 1060-1083.

[5] SARAFIAN M H, gaudin M, lewis M R et al ,Objective Set of Criteria for Optimization of Sample Preparation Procedures for Ultra-High Throughput Untargeted Body fluid Plasma Lipid Profiling by Ultra Performance Liquid Chromatography-Mass Spectrometry( objective standard set for sample preparation procedure optimization by ultra high throughput non-targeted body fluid plasma lipid analysis by ultra high performance liquid chromatography-mass spectrometry ANALYTICAL CHEMISTRY,2014,86 (12): 5766-5774.

[6] Guida R D Engel J, allwood J W et al ,Non-targeted UHPLC-MS metabolomic data processing methods: a comparative investigation of normalisation,missing value imputation,transformation and scaling[J] ( non-targeted UHPLC-MS metabonomics data processing method comparison study of normalization, deletion value interpolation, conversion and scaling [ J ]) Metabolomics,2016,12 (5): 93.

[7] Barker, m., rayens, w., PARTIAL LEAST squares for discrimination (partial least squares method for discrimination). J Chemometr 2003,17 (3): 166-173.

[8] Westerhuis J A, hoefsloot H C J, smit S et al ASSESSMENT OF PLSDA CROSS VALIDATION [ J ] (evaluation of PLSDA cross-validation [ J ]) Metabolomics,2008,4 (1): 81-89.

Claims

(a) Determining at least an amount of a compound having a mass of 110.0367 in grams per mole in a first body fluid sample of the subject;

(b) Determining the amount of the same compound as in (a) in a second body fluid sample of the subject;

(c) Determining a change in the amount of the compound determined according to (a) as compared to a corresponding amount of the compound determined according to (b);

(d) Analyzing the change determined in (c) using a computer-implemented prediction algorithm capable of predicting AD in a subject based on the compound, thereby identifying a subject suffering from AD or at risk of developing alzheimer's dementia if AD is predicted;

Wherein the first bulk liquid sample is obtained prior to administration of thioethyllazine or a pharmaceutically acceptable salt thereof and the second bulk liquid sample is obtained after administration of thioethyllazine or a pharmaceutically acceptable salt thereof to the subject.

2. The method for identifying a subject having or at risk of developing Alzheimer's Disease (AD) according to claim 1, wherein the first body fluid sample is derived from a subject after administration of thioethyllazine or a pharmaceutically acceptable salt thereof to the subject.

3. The method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to claim 1 or 2, wherein the amount of at least one further compound, the mass in grams/mole of which is selected from the group consisting of 208.0737, 315.2409, 175.0633, 702.5682 (group 4), is additionally determined in step (a) in a first body fluid sample of the subject and additionally analyzed in step (b).

4. A method for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD) according to any one of claims 1 to 3, wherein the computer-implemented predictive algorithm uses a supervised machine learning algorithm that has been trained by a training dataset comprising quantitative or qualitative data from the following compounds in body fluid samples from subjects known to suffer from AD and subjects known not to suffer from AD:

A compound of mass 110.0367 in grams per mole, optionally in combination with at least one other compound from group 4.

5. The method according to any one of claims 1 to 4, wherein the amounts of the compound having a mass of 110.0367 g/mol, the compound having a mass of 175.0633 g/mol and the compound having a mass of 208.0737 g/mol are determined at least in (a) and analyzed in (b).

6. A method for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the method comprising:

(a) Determining in a first body fluid sample of the subject an amount of a compound having a mass in grams/mole of 110.0367;

(c) Comparing the amount of the compound determined according to (a) with a corresponding amount of the compound determined according to (b), wherein a change in the amount of the compound according to (b) compared to the amount determined in (a) is indicative of the subject's Alzheimer's dementia or the subject's risk of developing Alzheimer's dementia;

7. A method for identifying a subject having Alzheimer's Dementia (AD), the method comprising:

(a) Determining in a body fluid sample of the subject an amount of a compound having a mass 110.0367 in grams/mole;

(b) Comparing the amount of the compound determined according to (a) with an average amount of healthy subjects, wherein a change in the amount of the compound according to (a) compared to the average amount of healthy subjects, preferably based on the average amount being at least 3% change as 100% healthy subjects, indicates that the subject is suffering from or at risk of developing alzheimer's disease;

Wherein the body fluid sample is obtained after administration of thioethyloxazine or a pharmaceutically acceptable salt thereof to the subject.

8. A compound having a mass in grams/mole of 110.0367 and optionally in combination with at least one compound selected from group 4 having a mass in grams/mole for use in diagnosing AD in a subject or for determining the risk of a subject developing AD.

9. Use of at least one compound of mass 110.0367 in grams per mole and optionally in combination with at least one compound of mass selected from group 4 in grams per mole in a sample of a subject for diagnosing AD in the subject or for determining the risk of the subject developing AD.

10. A device suitable for identifying a subject having or at risk of developing Alzheimer's Dementia (AD), the device comprising:

(a) An analysis unit capable of determining an amount of at least one compound of mass 110.0367 in grams/mole in a first body fluid sample of the subject;

(b) A data processor for analyzing the determined amounts of the compounds using a prediction algorithm implemented in the data processor, the prediction algorithm being capable of predicting the AD of a subject based on the at least two compounds, thereby identifying a subject suffering from AD if AD is predicted, or identifying a subject at risk of developing alzheimer's dementia if AD is predicted.

11. A kit for identifying a subject suffering from or at risk of developing Alzheimer's Disease (AD), the kit comprising a reference sample of a compound having a mass of 110.0367 in grams per mole, optionally a reference sample of at least one compound selected from the group consisting of group 4 in grams per mole, and optionally one or more reagents for sample preparation and/or extraction.