Blood indicators for Alzheimer's disease

Abstract

Provided herein is a method for identifying a subject suffering from or at risk of developing Alzheimer's disease, comprising determining the presence or amount of 2-aminoethyl dihydrogen phosphate or taurine in blood or blood products obtained from said subject.Also provided herein is a method for preventing, treating, or delaying the onset of Alzheimer's disease.

Description

TECHNICAL FIELD

[0001] Cross-reference of related applications This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 146,031, filed on February 5, 2021, entitled "BLOOD INDICATORS OF ALZHEIMER'S DISEASE", the entire content of which is incorporated herein by reference in its entirety.

[0002] Field of the present invention Certain embodiments of the present invention relate to methods for detecting the presence, absence or amount of 2-aminoethyl dihydrogen phosphate and / or taurine in a sample. Certain embodiments of the present invention also relate to the amount of 2-aminoethyl dihydrogen phosphate and / or taurine, or the ratio of 2-aminoethyl dihydrogen phosphate to taurine in a sample, which is predictive and / or diagnostic of Alzheimer's disease. Certain embodiments of the present invention also relate to methods of monitoring the progression of Alzheimer's disease and / or methods of determining treatment for Alzheimer's disease. BACKGROUND ART

[0003] Ninety-five percent of Alzheimer's disease (AD) cases occur in people aged 60 to 65 years or older. This is generally a period of life when individuals are looking forward to their later years and are ready to make significant contributions to their families and communities. AD not only robs them of their lives, but also imposes the burden of being unable to form or use short-term memory, increasing confusion, disorientation, withdrawal, and increasing dementia that intensifies during their later years before death. Neuropathologically, the onset of AD symptoms is sometimes correlated with high-density tauopathy of neurofibrillary tangles (NFTs) composed of cerebral neuropathology, particularly amyloid-β-42 (Aβ42) plaques and hyperphosphorylated tau deposits in specific brain regions.

[0004] There appears to be a significant latency period between the occurrence of AD-type neuropathology in the brain and the onset of clinical symptoms. A vast amount of 14This originates from the 50-megaton Tsar Bomba explosion by the Soviet Union on October 31, 1961, which released carbon into the atmosphere. 14 Studies using C decay have enabled the precise dating of NFTs and Aβ42 plaques extracted from postmortem brain tissue of AD patients. In two of the six patients studied, NFTs were determined to be 7 years prior to symptom onset, and in one patient, Aβ42 plaques were determined to be 9 years prior to symptom onset. The benefits of discovering and using biomarkers for AD, particularly in the latent period, could be utilized to explore new types of treatments or to try previously failed drugs that may eventually be prescribed much earlier in disease progression.

[0005] Recent meta-analyses have suggested that amyloid deposition, several CSF biomarkers resulting from nerve damage and neuronal loss, and the formation of NFTs, markedly phosphorylated tau (P-tau), amyloid beta (Aβ42), total tau (T-tau), and neurofilamentous light chain proteins (NFLs) are strongly associated with AD. Plasma concentrations of T-tau also showed promise as a potentially excellent indicator of AD, requiring further investigation, while plasma concentrations of Aβ40 and Aβ42 did not appear to be excellent candidates for AD diagnosis. Imaging biomarkers that assess amyloid beta plaques (PiB-PET scan), tau deposition (tau-PET), brain atrophy (structural MRI), memory-related activity patterns (fMRI), and impaired glucose metabolism (FDG-PET) are also currently used as AD biomarkers. [Overview of the Initiative]

[0006] As described herein, plasma concentrations of dihydrogen 2-aminoethyl phosphate (also known as phosphoethanolamine) and / or taurine are not known to be associated with Aβ42, hyperphosphorylated tau, and NFT, and can be distinguished from control blood samples in replication experiments using independent cohorts. Therefore, the amount or ratio of dihydrogen 2-aminoethyl phosphate and / or taurine is often useful in predicting the onset of AD in asymptomatic patients. Furthermore, since the amounts of dihydrogen 2-aminoethyl phosphate and taurine are readily measured using instruments commonly found in most large hospitals for evaluating metabolic diseases, their detection and quantification can be rapidly performed in various clinical settings.

[0007] In certain embodiments, a method for determining the amount of 2-aminoethyl dihydrogen phosphate and / or taurine in a sample is presented herein. In some embodiments, such a method may be used for the diagnosis and early detection of Alzheimer's disease.

[0008] Specific aspects of this technology are further described in the following description, examples, claims, and drawings. [Brief explanation of the drawing]

[0009] [Figure 1] Figure 1 shows the chromatograms from a Hitachi amino acid analyzer for taurine (Tau, 1.607 min) and phosphoethanolamine (2-aminoethyl dihydrogen phosphate, 2.080 min). The pink line indicates the absence of 2-aminoethyl dihydrogen phosphate in the control sample, the blue-green line indicates 2-aminoethyl dihydrogen phosphate in the AD sample, and the dark green line indicates a large taurine peak, which artificially boosts the 2-aminoethyl dihydrogen phosphate and increases the area under the curve profile in Experiment 2. [Figure 2]Figure 2 shows triple quadrupole mass spectrometer chromatography of derivatized and combined early Alzheimer's disease (AD) samples and controls: (a) 3 μl injection of AD, Experiment 1; (b) 6 μl injection of AD, Experiment 1; (c) 6 μl injection of AD, Experiment 2; (d) 6 μl injection of AD with 0.15 ng of 2-aminoethyl dihydrogen phosphate added, Experiment 2; (e) 3 μl injection of control, Experiment 1 (2-aminoethyl dihydrogen phosphate not detected); (f) 6 μl injection of control, Experiment 1; (g) 3 μl injection of 0.004 μg / ml of 2-aminoethyl dihydrogen phosphate standard with a peak at a retention time of 2.05 minutes. [Modes for carrying out the invention]

[0010] Detailed explanation As described herein, plasma doses of 2-aminoethyl dihydrogen phosphate and / or taurine have diagnostic and therapeutic utility, providing insight into whether a subject has or is at risk of developing Alzheimer's disease. Therefore, in some embodiments, methods are provided herein for the treatment of asymptomatic subjects at risk of developing Alzheimer's disease. Such methods can suppress the progression of Alzheimer's disease or delay its onset.

[0011] In some embodiments, the amount of 2-aminoethyl dihydrogen phosphate and / or taurine is determined in the sample. “Sample” is often obtained from a suitable subject using a suitable method. A sample may be isolated from or obtained directly from a subject or a portion thereof. In some embodiments, a sample obtained from a subject is a sample derived from the subject. In certain embodiments, a sample obtained from a subject is a sample obtained from a third party, e.g., a third party who obtained or extracted a sample from the subject. In some embodiments, a sample is obtained directly from an individual or healthcare professional. A sample may be any specimen isolated from or obtained from a subject or a portion thereof. A sample may be any tissue or liquid isolated from or obtained from one or more subjects. Examples of samples, not limited to, include fluids or tissues obtained from or derived from a subject, such as, but not limited to, blood or blood products (e.g., serum, plasma, platelets, buffy coat, lymph, or similar), umbilical cord blood, chorionic villi, amniotic fluid, cerebrospinal fluid (CSF), cerebrospinal fluid, lavage fluids (e.g., lung, stomach, abdominal cavity, glandular duct, ear, arthroscopic), biopsy samples, paracentesis samples, cells (blood cells, lymphocytes, placental cells, stem cells, bone marrow-derived cells, embryonic or fetal cells, neurons) or parts thereof (e.g., mitochondria, nuclei, extracts, lysates, or similar), urine, feces, sputum, saliva, nasal mucosa, prostatic fluid, lavage, semen, lymph, bile, tears, sweat, breast milk, lactation, and similar or combinations thereof. In some embodiments, a sample includes or consists of blood or blood products. In some embodiments, blood products include plasma.

[0012] In certain embodiments described herein, the sample is obtained from a subject, the subject is determined to have or be at risk of having AD, and / or treatment (e.g., administration of drugs) is carried out on the subject. Examples of subjects, not limited to these, include mammals, humans, non-human primates (e.g., apes, gibbons, chimpanzees, orangutans, monkeys, macaques, and similar species), domesticated animals (e.g., dogs and cats), livestock (e.g., horses, cattle, goats, sheep, and pigs), and laboratory animals (e.g., mice, rats, rabbits, and guinea pigs). In some embodiments, the subject is a mammal. In certain embodiments, the subject is a primate. In some embodiments, the subject is a non-human primate. In some embodiments, the subject is a savanna monkey. In certain embodiments, the subject is a human. The subjects may be of any age or developmental stage, such as adults (e.g., 18, 19, 20 or 21 years of age and older), the elderly (e.g., 55 years of age and older, 60 years of age and older, or 65 years of age and older), teenagers (e.g., 12 to 19 years of age), children (e.g., 1 to 12 years of age), infants (e.g., from birth to 1 year of age), or mammals in utero. The subjects may be male or female.

[0013] In some embodiments, subjects are suspected of having Alzheimer's disease or are at risk of developing it. In some embodiments, subjects with Alzheimer's disease are subjects diagnosed with Alzheimer's disease by the method described herein. In some embodiments, subjects with Alzheimer's disease are subjects diagnosed with Alzheimer's disease by a healthcare professional (e.g., a physician) based on, for example, the presence of one or more diagnostic symptoms and / or the results of one or more standardized diagnostic tests. Subjects suspected of having Alzheimer's disease are often subjects who have not yet been diagnosed with Alzheimer's disease by a healthcare professional. In some embodiments, subjects suspected of having Alzheimer's disease may exhibit one or more symptoms of Alzheimer's disease, but these symptoms are not conclusive evidence that the subject has Alzheimer's disease. In some embodiments, a subject suspected of having Alzheimer's disease may exhibit one or more symptoms of Alzheimer's disease, but is not diagnosed with Alzheimer's disease because there is insufficient data to definitively demonstrate that the subject has Alzheimer's disease. In some embodiments, a subject suspected of having Alzheimer's disease is a subject suspected of having AD, but has not yet been diagnosed with AD by a healthcare professional.

[0014] In some embodiments, a subject is determined to be at risk of developing Alzheimer's disease by performing the method described herein. In some embodiments, a subject at risk of developing Alzheimer's disease is a subject who is asymptomatic for Alzheimer's disease. In some embodiments, a subject at risk of developing Alzheimer's disease is a subject who has one or more symptoms of Alzheimer's disease, but these symptoms may be mild or transient. In some embodiments, a subject at risk of developing Alzheimer's disease is a subject who has not yet been diagnosed with Alzheimer's disease. In some embodiments, a subject at risk of developing Alzheimer's disease is a subject suspected of having Alzheimer's disease.

[0015] In some embodiments, subjects are healthy subjects. Healthy subjects are often subjects who do not have neurodegenerative diseases or disorders. Healthy subjects are often subjects who do not have and / or show any symptoms of neurodegenerative diseases or disorders. In some embodiments, healthy subjects are subjects who do not have Alzheimer's disease. In some embodiments, healthy subjects are subjects who do not have and / or show any symptoms of Alzheimer's disease. In certain embodiments, healthy subjects are subjects aged 18 to 40 years.

[0016] In some embodiments, the methods described herein identify subjects who have Alzheimer's disease (AD) or are at risk of developing it. In some embodiments, the methods described herein include methods for treating subjects who have been determined to have AD or be at risk of developing it. In some embodiments, subjects are identified by the methods described herein as having Alzheimer's disease or being at risk of developing it, and the methods further include administering appropriate treatment to the subjects (e.g., by administering appropriate drugs).

[0017] In some embodiments, a method for treating Alzheimer's disease includes suppressing or delaying the onset or progression of Alzheimer's disease. In some embodiments, a method for treating Alzheimer's disease includes suppressing or delaying the onset or progression of one or more symptoms of AD. In some embodiments, a method for treating Alzheimer's disease includes reducing or alleviating one or more symptoms of Alzheimer's disease.

[0018] In some embodiments, a method for treating Alzheimer's disease involves administering a therapeutically effective dose of a suitable Alzheimer's drug to the target. Examples of AD drugs, not limited to L-serine, laritrin, phenytoin, lamotrigine, carbamazepine, lidocaine, tetrodotoxin, nitroindazole, sulforaphane or sulforaphane analogues, gabapentin, pregabalin, mirogabalin, gabapentin enacarbil, phenibut, imagabalin, atagabalin, 4-methylpregabalin, PD-217,014, riluzole, edaravone, tetrabenazine, haloperidol, risperidone, quetiapine Apine, amantadine, levetiracetam, clonazepam, citalopram, escitalopram, fluoxetine, sertraline, olanzapine, valoproate, carbamazepine, lamotrigine, vaccines (e.g., amyloid peptides, or fragments or variants thereof, with or without adjuvants, in amounts that elicit an immune response), cholinesterase inhibitors (e.g., donepezil, galantamine, or rivastigmine), memantine, antidepressants, N-methyl D-aspartate (NMDA) antagonists, omega-3 fatty acids, curcumin or curcumin derivatives, vitamin E, sleep aids (e.g., zolpidem, eszopiclone, or zaleplon), anxiolytics (e.g., lorazepam or clonazepam), anticonvulsants (e.g., sodium valproate, carbamazepine, or oxcarbazepine), antipsychotics (e.g., risperidone, quetiapine, or olanzapine), carbidopa / levodopa, amantadine These include dopamine agonists (e.g., pramipexole, ropinirole, rotigotine, or apomorphine), monoamine oxidase B (MAO-B) inhibitors (e.g., selegiline, rasagiline, or safinamide), catechol-O-methyltransferase (COMT) inhibitors (e.g., entacapone or tolcapone), anticholinergics (e.g., benztropine or trihexyphenidyl), and similar drugs, as well as combinations thereof.In some embodiments, the Alzheimer's drug is selected from one or more of the following: laritolin, phenytoin, lamotrigine, carbamazepine, lidocaine, tetrodotoxin, riluzole (RILUTEK®; TIGLUTIK®), edaravone (RADICAVA®), gabapentin, pregabalin, mirogabalin, gabapentin enacarbil, phenibut, imagabalin, atagabalin, 4-methylpregabalin, PD-217,014, trihexyphenidyl, amitriptyline, baclofen, diazepam, L-serine, CK-2127107 (reldesemtiv), nusinersen, onasemnogene abeparvovec (ZOLGENSMA®), NUEDEXTA®, and similar drugs or combinations thereof. In some embodiments, the AD drug comprises L-serine, or a salt thereof, a metabolic precursor, a derivative, a polymer, or a conjugate.

[0019] L-serine In some embodiments, a subject is administered a therapeutically effective amount of L-serine, or a salt thereof, a metabolite, a derivative, or a conjugate thereof. In some embodiments, a subject is administered a therapeutically effective amount of free L-serine, or a salt thereof. A therapeutically effective amount of L-serine or free L-serine may be administered as a pharmaceutical composition comprising one or more pharmaceutical excipients, additives, carriers, and / or diluents. In some embodiments, the methods herein involve administering a therapeutically effective amount of a composition comprising, or essentially comprising, L-serine, or a salt thereof, a metabolite, a derivative, or a conjugate thereof, to a subject. In some embodiments, the methods herein involve administering a therapeutically effective amount of a composition comprising, or essentially comprising, free L-serine, or a salt thereof, a derivative, or a conjugate thereof, to a subject. In some embodiments, the methods herein involve administering a therapeutically effective amount of a polymer of L-serine, or a salt thereof, a derivative, or a conjugate thereof, to a subject. In some embodiments, compositions comprising essentially L-serine, free L-serine, or their salts, precursors, derivatives, or conjugates exclude proteins or protein fractions containing 100%, 99%, less than 98%, less than 95%, less than 90%, less than 80%, less than 70%, less than 60%, or less than 50% L-serine (wt / wt). In some embodiments, compositions comprising essentially L-serine, free L-serine, or their salts, precursors, derivatives, or conjugates exclude proteins or protein fractions containing protein (wt / wt) in proportions greater than 5%, greater than 10%, greater than 20%, greater than 30%, greater than 40%, greater than 50%, or greater than 60%. In some embodiments, compositions comprising essentially L-serine include free L-serine or L-serine polymers having an amino acid content of at least 99%, 98%, 95%, 90%, 85%, or at least 80% L-serine.In some embodiments, the composition consisting essentially of L-serine excludes creatine, creatine pyruvate, guanidinoacetic acid (GA), glycocyamine, N-amidinoglycine, and salts or esters thereof. In some embodiments, the composition consisting essentially of L-serine is a composition comprising free L-serine with a purity of at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%. In certain embodiments, the composition consisting essentially of L-serine, free L-serine, or a salt, precursor, derivative or conjugate of L-serine is a composition that further comprises zinc.

[0020] Free L-serine refers to L-serine or a salt thereof in the form of a single amino acid. In some embodiments, the composition comprises free L-serine with a purity of at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%. In certain embodiments, free L-serine is not covalently bonded to any other amino acid.

[0021] In some embodiments, the composition comprising L-serine may exclude other active ingredients. In some embodiments, the composition may exclude proteins containing L-serine. In some embodiments, the composition may exclude proteins having a molecular weight greater than 10 kDa, greater than 20 kDa, greater than 30 kDa, or greater than 50 kDa. In some embodiments, the composition may exclude proteins containing less than 99%, 98%, 95%, 92%, 90%, 80%, 70%, 60%, or less than 50% L-serine. In some embodiments, the composition may exclude creatine, or any energy metabolism precursor of creatine, such as guanidinoacetic acid (GA), equivalents thereof, and mixtures thereof.

[0022] In certain embodiments, the composition comprises L-serine, and examples thereof include, but are not limited to, free L-serine and polymers or polypeptides containing L-serine in weight or amino acid content of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the L-serine polymer or L-serine-containing polypeptide contains 2 to 50,000, 2 to 500, 2 to 100, 2 to 50, 2 to 20, 2 to 15, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, or 2 to 4 L-serine amino acids linked by covalent bonds. In certain embodiments, the composition comprises L-serine, and examples thereof include polymers or polypeptides containing L-serine in content (wt / wt) of 20%-100%, 30%-100%, 35%-100%, 40%-100%, 45%-100%, 50%-100%, 55%-100%, 60%-100%, 65%-100%, 70%-100%, 75%-100%, 80%-100%, 85%-100%, 90%-100%, 95%-100%, 96%-100%, 97%-100%, 98%-100%, or 99%-100% in terms of amino acid content (i.e., L-serine monomer / total amino acid monomer).

[0023] Non-limiting examples of salts of L-serine include sodium salt, potassium salt, calcium salt, magnesium salt, zinc salt, ammonium salt; inorganic salts (such as hydrogen chloride, sodium chloride, potassium chloride, calcium chloride, sodium phosphate, potassium phosphate, sodium bicarbonate and the like); organic salts (such as sodium citrate, citrate, acetate and the like). In certain embodiments, the composition comprises L-serine as alkylated L-serine (e.g., L-serine having an alkyl group, or an alkyl containing, for example, 1 to 20 carbon atoms). In certain embodiments, derivatives of L-serine include L-serine ester, L-serine diester, phosphate ester of L-serine, or sulfuric acid or sulfonic acid ester of L-serine. Non-limiting examples of conjugates of L-serine include pegylated L-serine (e.g., L-serine comprising one or more polyethylene glycol (PEG) moieties), and lipidated L-serine. Non-limiting examples of precursors of L-serine include L-phosphoserine.

[0024] Non-limiting examples of precursors of L-serine include pro-forms of L-serine that are degraded into L-serine monomers by the digestive system of a subject. In some embodiments, L-serine or a conjugate thereof consists of a sustained release version. In some embodiments, the derivative of L-serine is conjugated to a different molecule that forms a prodrug from which L-serine is released after crossing the blood-brain barrier.

[0025] In some embodiments, a composition consisting essentially of L-serine may contain a certain amount of D-serine. For example, a composition consisting essentially of L-serine may contain small amounts of D-serine, e.g., less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% by weight (e.g., wt / wt) or amino acid content (e.g., D-serine / total amino acid content). For example, a composition may contain 0.001% to 30%, 0.005% to 30%, 0.1% to 30%, 1% to 30%, 2% to 30%, 3% to 30%, 4% to 30%, 5% to 30%, 6% to 30%, 7% to 30%, 8% to 30%, 9% to 30%, 10% to 30%, 0.001% to 20%, 0.005% to 20%, 0.1% to 20%, 1% to 20%, 2% to 20%, 3% to 20%, 4% to 20%, 5% to 20%, 6% to 20%, 7% to 20%, 8% to 20%, 9% to 20%, or 10% to 20% of D-serine. In some embodiments, a composition containing or essentially consisting of L-serine does not contain a substantial amount of D-serine. In some embodiments, compositions containing or essentially consisting of L-serine do not contain D-serine.

[0026] Any suitable method may be used to detect and / or quantify the presence and / or amount of 2-aminoethyl dihydrogen phosphate and / or taurine in a sample. In certain embodiments, the presence and / or amount of 2-aminoethyl dihydrogen phosphate and / or taurine in a sample is determined by a method including the use of mass spectrometry (MS).

[0027] As used herein, “amount” refers to the mass, weight, volume, and / or concentration of a substance in a sample. Amount may be an average, mean, maximum, or absolute value. Maximum value may be an average, mean, or absolute maximum value. In certain embodiments, amount is the ratio of the amount of a first substance (e.g., 2-aminoethyl dihydrogen phosphate) in a sample to the amount of a second substance (e.g., taurine) in a sample (e.g., the same sample).

[0028] In some embodiments, the amount is compared to a predetermined threshold. In some embodiments, it is determined whether the amount is below, substantially the same as, or above the predetermined threshold. In some embodiments, the predetermined threshold is the amount of substance in a sample obtained from a healthy subject. In some embodiments, the predetermined threshold is the amount of substance determined in one or more samples obtained from one or more healthy subjects. In some embodiments, the predetermined threshold is the amount of 2-aminoethyl dihydrogen phosphate determined in one or more samples obtained from one or more healthy subjects. In some embodiments, the predetermined threshold is the amount of taurine determined in one or more samples obtained from one or more healthy subjects. In some embodiments, the predetermined threshold is the ratio of 2-aminoethyl dihydrogen phosphate to taurine determined in one or more samples obtained from one or more healthy subjects.

[0029] In certain embodiments, predetermined thresholds for taurine in plasma are amounts of 200 μmol / L (taurine / plasma volume) or less, 175 μmol / L or less, 150 μmol / L or less, 140 μmol / L or less, 130 μmol / L or less, or 125 μmol / L or less. In certain embodiments, predetermined thresholds for 2-aminoethyl dihydrogen phosphate in plasma are amounts of 100 μmol / L (2-aminoethyl dihydrogen phosphate / plasma volume) or less, 90 μmol / L or less, 80 μmol / L or less, 75 μmol / L or less, 70 μmol / L or less, or 60 μmol / L or less.

[0030] In some embodiments, the predetermined threshold, which is the ratio of 2-aminoethyl dihydrogen phosphate to taurine, is an average, mean, or absolute ratio of 0.30 or less, 0.25 or less, 0.20 or less, 0.19 or less, 0.18 or less, 0.17 or less, 0.16 or less, 0.15 or less, or 0.14 or less.

[0031] In certain embodiments, the amount of 2-aminoethyl dihydrogen phosphate and / or taurine in a sample obtained from a subject indicating that the subject has or is at risk of developing AD is approximately 20 times, 19 times, 18 times, 17 times, 16 times, 15 times, 14 times, 13 times, 12 times, 11 times, 10.5 times, 10 times, 9.5 times, and approximately above a predetermined threshold. It is 9 times, approximately 8.5 times, approximately 8 times, approximately 7.5 times, approximately 7 times, approximately 6.5 times, approximately 6 times, approximately 5.5 times, approximately 5 times, approximately 4.5 times, approximately 4 times, approximately 3.5 times, approximately 3 times, approximately 2.9 times, approximately 2.8 times, approximately 2.7 times, approximately 2.6 times, approximately 2.5 times, approximately 2.4 times, approximately 2.3 times, approximately 2.2 times, approximately 2 times, approximately 1.9 times, approximately 1.8 times, approximately 1.7 times, approximately 1.6 times, approximately 1.5 times, approximately 1.4 times, approximately 1.3 times, approximately 1.2, or approximately 1.1 times higher.

[0032] In certain embodiments, the ratio of 2-aminoethyl dihydrogen phosphate to taurine in a sample obtained from a subject indicating that the subject has or is at risk of developing AD is approximately 20 times, 19 times, 18 times, 17 times, 16 times, 15 times, 14 times, 13 times, 12 times, 11 times, 10.5 times, 10 times, 9.5 times, and 9 times higher than a predetermined threshold (ratio) of 2-aminoethyl dihydrogen phosphate to taurine. The ratio of 2-aminoethyl dihydrogen phosphate to taurine is approximately 8.5 times, 8 times, 7.5 times, 7 times, 6.5 times, 6 times, 5.5 times, 5 times, 4.5 times, 4 times, 3.5 times, 3 times, 2.9 times, 2.8 times, 2.7 times, 2.6 times, 2.5 times, 2.4 times, 2.3 times, 2.2 times, 2 times, 1.9 times, 1.8 times, 1.7 times, 1.6 times, 1.5 times, 1.4 times, 1.3 times, 1.2, or 1.1 times higher.

[0033] In some embodiments, the method provided herein may be performed two or more times to monitor the progression of Alzheimer's disease in a subject and / or to monitor the subject's response to treatment for Alzheimer's disease.

[0034] Administration Any suitable method may be used to administer treatment or drugs to the subject. Any suitable formulation and / or route of administration may be used for administering or administering the treatment or drugs disclosed herein (see, for example, Fingl et al. 1975, in “The Pharmacological Basis of Therapeutics,” which is incorporated herein by reference in its entirety). A suitable formulation and / or route of administration may be selected by a healthcare professional (e.g., a physician) taking into consideration, for example, the subject's disease, condition, symptoms, weight, age, and / or overall health. Examples of routes of administration that are not limited to topical or local (e.g., transdermally or cutaneously (e.g., on the skin or epidermis), intraocular or supraocular, intranasal, transmucosal, intraocular, inside the ear (e.g., behind the eardrum)), enteral (e.g., delivered through the gastrointestinal tract, e.g., orally (e.g., as tablets, capsules, granules, liquids, emulsions, lozenges, or combinations thereof), sublingual, by gastrointestinal tube, rectally, and similar), parenteral administration (e.g., parenteral, e.g., intravenous, intra-arterial, intramuscular, intraperitoneal, intradermal, subcutaneous, intracavitary, intracranial, intra-articular, intra-articular cavity, intracardiac (into the heart), intracavernosal injection, intrafocal (into a skin lesion), intramedullary infusion (into the bone marrow), intrathecal (into the spinal canal), intrauterine, intravaginal, intravesical instillation, intravitreal), and similar or combinations thereof.

[0035] In some embodiments, administering a drug to a subject includes providing the drug to the subject, for example, with respect to self-administration or administration to the subject by another person (e.g., a non-medical professional). In another example, the drug may be provided as instructions written by a physician (e.g., a prescription) authorizing the provision of the drug or treatment described herein to a patient. In yet another example, the drug may be provided to a subject, where the subject self-administers the composition, for example, orally, intravenously, or via an inhaler.

[0036] Alternatively, drugs may be administered by topical methods rather than systemic methods, for example, by direct application to the skin, mucous membranes, or areas of therapeutic interest, including the use of depot formulations or sustained-release formulations.

[0037] In certain embodiments, the drug is administered alone (for example, as a single active ingredient (AI) or, for example, as a single active ingredient (API)). In other embodiments, the drug is administered in combination with one or more further AI / APIs, for example, as two separate compositions, or as a single composition in which one or more further AI / APIs are mixed with the drug in a pharmaceutical composition or formulated together.

[0038] In some embodiments, the amount of Alzheimer's disease drug administered to a subject is a therapeutically effective dose. In some embodiments, the therapeutically effective dose of the drug is the amount necessary to obtain an effective therapeutic outcome. In certain embodiments, the therapeutically effective dose of the drug is sufficient to treat one or more symptoms of Alzheimer's disease, reduce their severity, suppress or delay their onset, or alleviate and / or reduce them. Determining the therapeutically effective dose is well within the capabilities of those skilled in the art, particularly in light of the detailed disclosures provided herein.

[0039] In certain embodiments, the therapeutically effective dose is an amount high enough to provide an effective therapeutic effect (e.g., a beneficial therapeutic effect) and low enough to minimize undesirable adverse reactions. Therefore, in certain embodiments, the therapeutically effective dose of a drug may vary depending on the subject, often depending on the subject's age, weight, overall health or condition, the severity of the condition being treated, and / or the specific combination of drugs administered to the subject. Thus, in some embodiments, the therapeutically effective dose is determined empirically. Therefore, in certain embodiments, the therapeutically effective dose of a drug administered to a subject may be determined by a person skilled in the art based on amounts found effective in animal studies or clinical research, the physician's experience, and / or proposed dose ranges or administration guidelines.

[0040] In certain embodiments, a therapeutically effective amount of L-serine or a composition disclosed herein comprises one or more doses (to be administered to a subject) of L-serine, or a salt, precursor, derivative or conjugate thereof, per kg of body weight of the subject, in amounts of at least 0.1 mg / kg, at least 5 mg / kg, at least 10 mg / kg, at least 15 mg / kg, at least 20 mg / kg, at least 25 mg / kg, at least 50 mg / kg, at least 1000 mg / kg, at least 250 mg / kg, at least 500 mg / kg, at least 1000 mg / kg, at least 5000 mg / kg, or at least 7500 mg / kg.

[0041] In some embodiments, administering a therapeutically effective amount of an Alzheimer's disease drug or composition disclosed herein involves administering an appropriate dose every hour, every two hours, every four hours, every six hours, every eight hours, or every twelve hours. In certain embodiments, the Alzheimer's disease drug may be administered at least once, twice, three times, four times, five times, or six times per day, for example, 1 to 12 times per day, 1 to 8 times per day, or 1 to 4 times per day. In certain embodiments, the Alzheimer's disease drug disclosed herein may be administered once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, eleven times, or twelve times per day. The Alzheimer's disease drug may be administered in a single dosage form or in one or more dosage forms. The daily dose may be achieved in the form of a single dose or in the form of multiple partial doses.

[0042] Alzheimer's disease medications can be administered daily or on a schedule that includes days off. For example, administration may be every other day, or it may be administered for 2, 3, 4, or 5 consecutive days a week, followed by 1 to 5 days off.

[0043] Alzheimer's disease drugs may be administered for at least one day, at least two days, at least three days, at least four days, at least five days, at least one week, at least two weeks, at least three weeks, at least one month, at least two months, at least three months, at least six months, at least one year, at least two years, or longer, or for any extended period, in order to further improve, maintain, or sustain the therapeutic effect. In certain embodiments, Alzheimer's disease drugs are administered for periods ranging from one week to ten years or longer. In some embodiments, administering a therapeutically effective amount of a drug, or a pharmaceutical composition containing a drug, includes administering appropriate doses at the frequency or intervals necessary to obtain effective therapeutic outcomes. In some embodiments, administering a therapeutically effective dose of a drug or pharmaceutical composition disclosed herein involves administering an appropriate dose at intervals of one hour, two hours, four hours, six hours, three times a day, two times a day, once a day, six times a week, five times a week, four times a week, three times a week, two times a week, once a week, in combination thereof, and / or at regular or irregular intervals thereof, and / or simply at a frequency or interval as required or recommended by a healthcare professional. In some embodiments, a therapeutically effective dose of a drug or a pharmaceutical composition containing a therapeutically effective dose of a drug is administered continuously, for example, by intravenous administration. [Examples]

[0044] Example 1 method The plasma used in this experiment was derived from the initial blood draws from two batches of control samples collected by Innovative Research Inc. (Novi, Michigan, USA) and from an FDA-approved Phase IIa human clinical trial (NCT03062449) for early-stage Alzheimer's disease patients. The independent samples were divided into two cohorts, each using different individuals. The first cohort included n=11 early-stage Alzheimer's patients and n=11 controls. The second cohort also used different individuals, including n=14 different Alzheimer's patients and n=14 controls.

[0045] Plasma was collected in a K2-EDTA tube and inverted several times after collection. The sample was immediately centrifuged at 2000xg for 15 minutes at 4°C. The time between blood collection and freezing was less than 1 hour, and the sample was stored at -80°C. The plasma sample was thawed in a refrigerator (4°C) for 2 hours and mixed with an equal volume of cold 10% (w / v) trichloroacetic acid (TCA). The sample was allowed to precipitate at room temperature for 2 hours, followed by centrifugation at 14,000xg for 5 minutes. The supernatant was removed by pipette and filtered through a centrifugal filter (0.2 μm PVDF, Millipore Ultrafree®-MC) at 14,000xg for 5 minutes.

[0046] Underivativeized supernatant (20 μL) was injected at 135°C into a Hitachi Amino Acid Analyzer L8900 equipped with a Hitachi Reaction column (PN 855-3533), a high-speed biofluid analysis column Li-type resin #2622SC 6mm ID x 40L 060928C (PN855-4515), and an Ammonia Filter column (Ion exchange 4.6 x 40 Column #2650L, PN 855-3523). Hitachi pre-made buffers were used as follows: (B1) PF-1 / AN0-5031, (B2) PF-2 / AN0-5032, (B3) PF-3 / AN0-5033, (B4) PF-4 / AN0-5034, (B6) PF-5 / AN0-5035, (R1) ninhydrin solution, and (R2) lithium acetate dihydrate ninhydrin buffer. The washing solution contained (B5) 5% ethanol, (R3) 10% methanol, and (C1) 10% methanol. Separation was performed using pump 1 with a flow rate of 0.54 mL / min, pump 2 with a flow rate of 0.47 mL / min, a reactor column set to 135°C, and a gradient elution of 146 minutes: 0.00 min = 100% B1, column oven temperature 35°C, 50% R1, 50% R2; 16.0 min = 100% B1; 16.1 min = 81% B1, 19% B2; column oven 58°C; 41.0 min = column oven 32°C; 57.0 min = column oven 70°C; 69.5 min = column oven 65°C; 70.0 min = 15% B1, 75% B2, 10% B3; 88.0 min = column oven 60°C; 94.0 min = 20% B2, 80% B4; 94.1 min = 25% B2, 75% B4; 109.0 min = 25% B2, 75% B4, Column oven 70℃; 109.1 min = 100% B4; 123.0 min = 100% B4; 123.1 min = 100% B6; 127.0 min = 50% R1, 50% R2; 127.1 min = 100% R3; 129.0 min = 100% B6, 129.1 min = 100% B1; Column oven 35℃; 132.0 min = 100% R3; 132.1 min = 50% R1, 50% R2; 146.0 min = 100% Achieved by B1.Amino acid standards (Sigma A6407 - acidic and neutral amino acids + Sigma A6282 - basic amino acids) were mixed at equal concentrations, and calibration curves were created at concentrations of 5, 10, 25, 50, 100, 250, 500, and 1000 μmol / L. All amino acid curves were linear within this range (R). 2 =99%. Furthermore, retention times were checked for dihydrogen 2-aminoethyl phosphate (Sigma-Aldrich P0503-1G) and taurine (Sigma-Aldrich T0625-10G). The lower limit curve (1.4, 3.5, 7.1, 14.2, 70.9 μmol / L) is for dihydrogen 2-aminoethyl phosphate at 1.4 μmol / L (R 2 We showed that it is linear up to 99.9%.

[0047] Confirmation of 2-aminoethyl dihydrogen phosphate in the samples was performed by triple quadrupole mass spectrometry. Plasma samples from AD patients and controls were deproteinized as described above and derivatized with 6-aminoquinolyl-N-hydroxysuccinimidylcarbamate (AQC, WAT052880, Waters Corp, Milford, MA) using 10 μl of supernatant, 70 μl of borate buffer, and 20 μl of AQC. AD blood samples were mixed together with the control samples to form a single sample, which was then analyzed using a Thermo TSQ Quantiva® triple quadrupole mass spectrometer after multiple injections per sample. The mass spectrometer was equipped with a Thermo Vanquish® pump, autosampler, H-ESI probe, and a heated column compartment set to 65°C. A Thermo Scientific OPTON-10005 Genius 3022 dual N2 generator (Peak Scientific, Billerica, MA) supplied purified nitrogen to the mass spectrometer. Separation was performed using a Kinetex C-18 column (1.7 μm 100A, 100 x 2.1 mm, 00D-4475-AN); and gradient elution (flow rate 0.5 ml / min, mobile phase A = 20 mM ammonium acetate (Fisher Scientific A11450) ≥ 99% adjusted to pH 5.0 with glacial acetic acid (Fisher Scientific A385-500), B = 100% methanol ≥ 99.9% Chromasolv; 34885-4x4, Honeywell Burdick & Jackson, Muskegon, MI): Initial conditions 99.8% A, 1.0 min 99.8% A, 1.5 min 40% A, 2.0 min 25% A, 2.5 min 25% A, 2.6 min 10% A, 5.5 min 10% A, 6 min 98% A, 8 min 98% This was achieved using A (all curves set to 5). The mass spectrometer was run in positive mode (4400V) with the following settings: spray voltage = electrostatic; sheath gas = 5.6 Arb; auxiliary gas = 23.5 Arb; sweep gas = 1.2 Arb; ion transfer tube = 150°C; evaporation temperature = 450°C; Q1 resolution = 0.7 FWHM; Q3 resolution = 0.7 FWHM; and CID gas = 2 mTorr.Scanning of derivatized dihydrogen dihydrogen 2-aminoethyl phosphate was performed using the following transitions (312–171.2 m / z collision energy (CE) 17.1, 312–142 m / z CE 17.1, 312–312 m / z CE 2, RF lens 90). This experiment was repeated a second time to ensure reproducibility. The detection limit of this method was 0.004 μg / ml, and the lower limit of quantification was 0.01 μg / ml. Calibration curves were created at concentrations of 20 pg / ml, 40 pg / ml, 0.2 ng / ml, 0.4 ng / ml, 2 ng / ml, 4 ng / ml, 10 ng / ml, 20 ng / ml, 0.1 μg / ml, and 0.2 μg / ml, with linearity of 99% R. 2 It had a value.

[0048] Results and Discussion Dihydrogen dihydrogen phosphate 2-aminoethyl consists of a phosphate group bonded to the second carbon of ethanolamine and has a molecular weight of 141.06 g / mol. It can be biosynthesized from ethanolamine (Human Metabolome Database; HMDB0000224), is involved in phosphotidylcholine synthesis, and is a precursor of phosphotidylethanolamine. It is a solid with a melting point of 242°C.

[0049] In the first Hitachi amino acid analyzer experiment, consistently high peaks of 2-aminoethyl dihydrogen phosphate were observed in AD patients (n=11), while 2-aminoethyl dihydrogen phosphate in controls (n=11), if present, was below the detection limit. This experiment was repeated using plasma from a new cohort of AD patients (n=14) and a new set of control plasma (n=14). In the second experiment, excessively large taurine peaks were observed in eight control plasma samples. Since taurine separates on a C-18 column early in chromatography and then elutes near 2-aminoethyl dihydrogen phosphate (at 1.6 and 2.0 mins, respectively; Figure 1), high taurine concentrations in some samples caused separation problems between these two peaks. This artificially enlarged the area under the curve for 2-aminoethyl dihydrogen phosphate in these samples. To correct this problem, the 2-aminoethyl dihydrogen phosphate / taurine ratio was calculated in both experiments using the Mann-Whitney two-tailed U test, and a significant difference (p<0.0001) was identified between the ratio in AD patients and the ratio in controls (Table 1). [Table 1]

[0050] Plasma taurine concentrations in normal individuals are generally considered low, ranging from 25 to 150 μmol / L (Jacobsen and Smith 1968). Measurements of plasma taurine concentrations in the samples in this experiment showed considerable individual variation (3 to 403 μmol / L), with 8 out of 50 samples exceeding 150 μmol / L. We attributed these high values ​​in the eight samples to plasma contamination by taurine-rich platelets or leukocytes (Jacobsen and Smith 1968). While all these samples originated from a single batch of controls received from Innovative Research Inc., it should be noted that none of the samples from the Phase II AD trial showed the same pattern of contamination, and none of the controls from the first batch received from Innovative Research Inc. exhibited the same problem.

[0051] It cannot be ruled out that taurine levels were naturally elevated in the eight control patients for some reason. High taurine concentrations have been shown to result in increased extracellular 2-aminoethyl dihydrogen phosphate. In an in vivo rabbit model, administration of taurine (2 mM) to the dorsal hippocampus resulted in an observed increase in 2-aminoethyl dihydrogen phosphate over time (Lehmann and Hamberger 1984). That study concluded that taurine and 2-aminoethyl dihydrogen phosphate together may stabilize the synaptosome membrane calcium flux as a decrease in phospholipid content in these tissues, which may lead to decreased calcium transport (Lehmann and Hamberger 1984). However, if naturally occurring taurine levels in plasma are high, this can be corrected by adjusting the 2-aminoethyl dihydrogen phosphate / taurine ratio.

[0052] For comparison with other studies, it should be noted that only a small number of studies reporting plasma amino acid concentrations also report 2-aminoethyl dihydrogen phosphate concentrations. In such reports, the reference range for healthy adults is from "not detected" to 69 μmol / L (Perry and Hansen 1969, McGale et al. 1977, Slocum and Cummings 1991, Tan and Gajra 2006). The Mayo Clinic has reported reference ranges for amino acids, indicating that normal 2-aminoethyl dihydrogen phosphate concentrations are as follows: less than 12 for adults 18 years and older, less than 5 for children 2–17 years, and less than 6 for children under 2 years (URL: https: / / neurology.testcatalog.org / show / AAQP). Amino acid quantification is highly dependent on the method used, and variations in recovery, processing, storage methods, and the instruments used for detection introduce variability. The strength of our research lies in the fact that all samples were collected using the same protocol and analyzed with the same instruments, resulting in highly comparable data.

[0053] Dihydrogen dihydrogen 2-aminoethyl phosphate was significantly observed in AD plasma but not recognizable in control samples, making it a promising blood biomarker for AD. Since blood sampling is routine in clinical practice, it could potentially be used as a diagnostic indicator and can be detected using standard amino acid analyzers available in most large hospitals and diagnostic laboratories. Furthermore, accumulation of dihydrogen dihydrogen 2-aminoethyl phosphate may reveal important aspects of cellular pathways that are impaired in AD.

[0054] Confirmation of dihydrogen 2-aminoethyl phosphate was performed by mass spectrometry in two experiments. Dihydrogen 2-aminoethyl phosphate was present, and its concentration in the AD samples was 5 to 10 times higher than that in the control plasma samples. Dihydrogen 2-aminoethyl phosphate in the first Hitachi amino acid analyzer analysis was also 10 times higher in the AD samples than in the controls.

[0055] In the first mass spectrometer experiment analysis, the area under the curve for the combined AD sample was 10 times larger than that of the combined control sample, with slight variation between injections (mean AD = 208 ± 13 (standard deviation), mean control = 27 ± 13, n=5 injections; n=11 AD samples; n=10 control samples; Figures 2a, b, e, f). In the second replication, the AD sample had 5 times the area compared to the control sample (mean 55 ± 9 AD; 10 ± 6 control; n=4 injections; n=11 AD samples; n=11 control; Figures 2c, d). The area exceeded the limit of quantification (LOQ) only in the AD sample in the first experiment (0.1 μg / ml), while all others were below the LOQ but above the limit of detection (LOD).

[0056] Although the peak was small in MS / MS analysis, its area doubled in proportion to the injection volume (Figure 2a, b). It was noted that the low-concentration spike increased only the peak at 2.05 minutes, corresponding to the retention time of the 2-aminoethyl dihydrogen phosphate standard (Figure 2c, d, g). The identity of the peak at 2.10 minutes, which is observed only in plasma samples but not with the 2-aminoethyl dihydrogen phosphate standard, remains unknown (Figure 2).

[0057] Dihydrogen diaminoethyl phosphate is important for the structure and function of cell membranes and is a precursor of phosphatidylethanolamine and phosphatidylcholine in mammals. Dihydrogen diaminoethyl phosphate is essential for the biosynthesis of mammalian glycosylphosphatidylinositol-anchored proteins (GPIs), which bind proteins to the plasma membrane, and is highly evolutionarily conserved (Kamitani et al., 1992). The biological activity of GPIs is not fully understood, but they may be involved in intercellular communication, cellular signaling, signal transduction, and lipid raft transport (Paulick et al., 2008). Studies have suggested that phosphatidylethanolamine is the direct raw material for dihydrogen diaminoethyl phosphate in GPI synthesis (Menon et al., 1993).

[0058] In cell culture, diaminoethyl dihydrogen phosphate has been shown to inhibit mitochondrial respiration and induce apoptosis by disrupting mitochondrial membrane potential (Modica-Napolitano and Renshaw 2004, Ferreira et al. 2013). The effects of increased blood diaminoethyl dihydrogen phosphate concentrations are completely unknown. The inverse finding that diaminoethyl dihydrogen phosphate concentrations in the brain are lower in AD patients than in controls is intriguing (Ellison et al. 1987). In that study, diaminoethyl dihydrogen phosphate concentrations were significantly lower in the temporal cortex (64%, Brodmann area 21), frontal cortex (48%, Brodmann area 9), and hippocampus (40%) of postmortem AD brain tissue, but not in the parietal cortex (Brodmann areas 3-12) or occipital cortex (Brodmann area 17). In Huntington's disease, dihydrogen 2-aminoethyl phosphate was lower in both CSF and postmortem brain tissue compared to control tissue. Significant differences were observed in the caudate nucleus, putamen, and nucleus accumbens, but not in the prefrontal cortex (Ellison et al. 1987, Koroshetz et al., 1997). Similarly, a significant decrease in dihydrogen 2-aminoethyl phosphate was also confirmed in the putamen of Parkinson's disease (PD) patients compared to control brain tissue, as confirmed by proton magnetic resonance imaging experiments (Hattingen et al., 2009). Hattingen et al., 2009 suggest this reflects a decrease in membrane turnover due to mitochondrial dysfunction. Mitochondrial dysfunction is considered by some researchers to be a contributing factor to AD (Sheng et al., 2012). The importance of 2-aminoethyl dihydrogen phosphate as a substrate in cell membrane synthesis, and the fact that its concentration increases in the blood rather than the brain of AD patients, suggests that further research into this molecule may provide new insights into the study of neurodegeneration.

[0059] One pathway of interest, intersecting with a Phase IIa clinical trial for early Alzheimer's disease (NCT03580616) testing whether L-serine can slow the progression of AD, is the denovos sphingolipid biosynthesis pathway, in which serine palmitoyltransferase catalyzes the reaction of L-serine with palmitoylcoenzyme A to form a sphingoid base. It is interesting that the degradation of both sphinganine and sphingosine leads to the production of dihydrogen dihydrogen 2-aminoethyl phosphate (Duan and Merrill 2015; Harrison et al., 2018). Furthermore, in vitro studies have shown that increased sphinganine kinases reduce the concentration of free sphinganine, leading to disruption of axonal elongation in cultured hippocampal neurons (van Echten-Deckert 1998). Sphingolipid metabolism is thought to be highly regulated because its metabolites are involved in cellular signaling (van Echten-Deckert 1998). When the sphingolipid biosynthesis pathway is disrupted in AD, an increase in downstream 2-aminoethyl dihydrogen phosphate may occur. Furthermore, it may be necessary to further advance the pathway by requiring more L-serine (Metcalf et al., 2018). In addition, changes in the phospholipid composition of the plasma membrane can cause abnormal signaling, and membrane disruption has also been suggested to be a cause of neurodegeneration in AD (Wells et al., 1995; Gonzalez-Dominguez et al., 2014). Studies predicting an increase in 2-aminoethyl dihydrogen phosphate in this pathway may provide new insights into the pathogenesis of Alzheimer's disease, as empirically demonstrated in our experiments.

[0060] In a rare genetic disorder known as hypophosphatasia, individuals primarily exhibit elevated levels of 2-aminoethyl dihydrogen phosphate in their urine (Goyer 1963, Imbard 2012, Whyte 2016, Salles 2020). The disorder is thought to be caused by mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) gene (Salles 2020). The symptoms of this disorder are highly diverse but always involve problems with bone and dental mineralization, and are frequently accompanied by neurological complications (Linglart and Biosse-Duplan 2016). References BAYRAM, et al., “Current understanding of magnetic resonance imaging biomarkers and memory in Alzheimer's disease”, Alzheimer's & Dementia Translational Research & Clinical Interventions, 2018, doi:10.1016 / j.trci.2018.04.007. CUMMINGS, et al., “Alzheimer's drug-development pipeline: few candidates, frequent failures”, AD Research & Therapy”, August 2014, pp. 1-7, Vol. 6(4). ELLISON, et al., “Phosphoethanolamine and ethanolamine are decreased in Alzheimer's disease and Huntington's disease”, Brain Research, 11 August 1987, pp. 389-392, Vol. 417(2). FERREIRA, et al., “Synthetic 2-aminoethyl dihydrogen phosphate induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells through the mitochondrial pathway”, Biomedicine & Pharmacotherapy, 01 July 2013, pp. 481-487, Vol. 67(6). GONZALEZ-DOMINGUEZ, et al., “Metabolomic study of lipids in serum for biomarker discovery in Alzheimer's disease using direct infusion mass spectrometry”, Journal of Pharmaceutical and Biomedical Analysis, 2014, doi:10.1016 / j.jpba.2014.05.023. GOYER, RA, “Ethanolamine phosphate excretion in a family with hypophosphatasia”, Archives of Disease in Childhood, June 1963, pg. 205, Vol. 38(199). HARRISON, et al., “Sphingolipid biosynthesis in man and microbes”, Nat Prod Rep., 19 September 2018, pp. 921-954, Vol. 35(9). doi:10.1039 / c8np00019k. PMID: 29863195; PMCID: PMC6148460. HATTINGEN, et al., “Phosphorus and proton magnetic resonance spectroscopy demonstrates mitochondrial dysfunction in early and advanced Parkinson's disease”, Brain, December 2009, pp. 3285-3297, Vol 132(12), doi:10.1093 / brain / awp293. IMBARD, et al., “2-aminoethyl dihydrogen phosphate normal range in pediatric urines for hypophosphatasia screening”, Clinical Chemistry and Laboratory Medicine, 01 December 2012, pp. 2231-2233, Vol. 50(12). JACK JR., “Hypothetical model of dynamic biomarkers of the Alzheimer's pathological cascade”, The Lancet Neurology, 01 January 2010, pp. 119-128, Vol. 9(1). JACOBSEN, et al., “Biochemistry and physiology of taurine and taurine derivatives”, Physiological Reviews, April 1968, pp. 424-511, Vol. 48(2). KAMITANI, et al., “Complexity of ethanolamine phosphate addition in the biosynthesis of glycosylphosphatidylinositol anchors in mammalian cells”, Journal of Biological Chemistry, December 1992, pp. 24611-24619, Vol. 267(34). KONDO, et al., “Identification of the phytosphingosine metabolic pathway leading to odd-numbered fatty acids”, Nature Communications, 27 October 2014, pp. 1-11, Vol. 5:5338. KOROSHETZ, et al., “Energy metabolism defects in Huntington's disease and effects of coenzyme Q10”, Annals of Neurology: Official Journal of the American Neurological Association and the Child Neurology Society, February 1997, pp. 160-165, Vol. 41(2). KUMAR, et al., “The sphingolipid degradation product trans-2-hexadecenal induces cytoskeletal reorganization and apoptosis in a JNK-dependent manner”, Cellular Signalling, July 2011, pp. 1144-1152, Vol. 23(7) 2011. LAWRENCE, et al., “A Systematic Review of Longitudinal Studies Which Measure Alzheimer's Disease Biomarkers”, Journal of Alzheimer's Disease, 2017, pp. 1359-1379, Vol. 59(4), doi:10.3233 / jad-170261. LEHMANN, et al., “A possible interrelationship between extracellular taurine and 2-aminoethyl dihydrogen phosphate in the hippocampus”, Journal of Neurochemistry, May 1984, pp. 1286-1290, Vol. 42(5). LINGLART, et al., “Hypophosphatasia”, Current Osteoporosis Reports, 2016, pp. 95-105, Vol. 14(3). LOVELL, et al., “Use of bomb pulse carbon-14 to age senile plaques and neurofibrillary tangles in Alzheimer's disease”, Neurobiology of Aging, 2002, pp. 179-186, Vol. 23(2). MCCONATHY, et al., “Imaging Biomarkers Associated with Cognitive Decline: A Review”, Biological Psychiatry”, 15 April 2015, pp. 658-692, Vol. 77(8), doi:10.1016 / j.biopsych.2014.08.024. MEHTA, et al, “Why do trials for Alzheimer's drugs keep failing? A discontinued drug perspective for 2010-2015:, Expert Opinion on Investigational Drugs, June 2017, pp. 735-739, Vol. 26(6). MENON, et al., “Phosphatidylethanolamine is the donor of the terminal 2-aminoethyl dihydrogen phosphate group in trypanosome glycosylphosphatidylinositols”, The EMBO Journal 1993, doi:10.1002 / j.1460-2075.1993.tb05839.x. METCALF, et al., “L-Serine: a naturally-occurring amino acid with therapeutic potential”, Neurotoxicity Research”, January 2018, pp. 213-221, Vol. 33(1). MODICA-NAPOLITANO, et al., “Ethanolamine and 2-aminoethyl dihydrogen phosphate inhibit mitochondrial function in vitro: implications for mitochondrial dysfunction hypothesis in depression and bipolar disorder”, Biological Psychiatry, February 2004, pp. 273-277, Vol. 55(3). MULLANE, et al, “Alzheimer's Disease (AD) therapeutics-1: Repeated clinical failures continue to question the amyloid hypothesis of AD and the current understanding of AD causality”, Biochemical Pharmacology, December 2018, pp. 359-375, 1;158. PAULICK, et al., “The glycosylphosphatidylinositol anchor: a complex membrane-anchoring structure for proteins”, Biochemistry, July 2008, pp. 6991-7000, Vol. 47(27). OLSSON et al., “CSF and blood biomarkers for the diagnosis of Alzheimer's disease: a systematic review and meta-analysis”, The Lancet Neurology, 01 June 2016, pp. 673-684, Vol. 15(7). SALLES, JP, “Hypophosphatasia: Biological and Clinical Aspects, Avenues for Therapy”, The Clinical Biochemist Reviews, February 2020, pg. 13, Vol. 41(1). SCHINDLER, et al., “High-precision plasma β-amyloid 42 / 40 predicts current and future brain amyloidosis”, Neurology, 2019, pp. e1647-e1659, Vol. 93, available at: http: / / www.ncbi.nlm.nih.gov / pubmed / 31371569. SHENG, et al., “Impaired mitochondrial biogenesis contributes to mitochondrial dysfunction in Alzheimer's disease”, Journal of Neurochemistry, 2012, doi:10.1111 / j.1471-4159.2011.07581.x. VAN ECHTEN-DECKERT, et al., “1-Methylthiodihydroceramide, a Novel Analog of Dihydroceramide, Stimulates Sphinganine Degradation Resulting in Decreased de Novo Sphingolipid Biosynthesis”, Journal of Biological Chemistry, 1998, doi:10.1074 / jbc.273.2.1184. WELLS, et al., “Neural membrane phospholipids in Alzheimer disease”, Neurochemical Research, 01November 1995, pp. 1329-1333, Vol. 20(11). WHYTE, M., “Hypophosphatasia - aetiology, nosology, pathogenesis, diagnosis and treatment”, Nat Rev Endocrinol, 2016, pp. 233-246, https: / / doi.org / 10.1038 / nrendo.2016.14.

[0061] Each patent, patent application, publication, or any other reference or document cited herein is incorporated by reference in its entirety. In the event of any conflict, the specification containing the definitions shall prevail.

[0062] No citation of any patent, patent application, publication, or any other document constitutes an endorsement that any of the foregoing is relevant prior art, nor does it constitute any endorsement of the content or date of such publication or document.

[0063] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention pertains. Similar or equivalent methods and materials to those described herein may be used in carrying out or testing the present invention, but suitable methods and materials are described herein.

[0064] All features disclosed herein may be combined in any combination. Each feature disclosed herein may be replaced by an alternative feature that serves the same, equivalent, or similar purpose. Therefore, unless expressly otherwise specified, the disclosed features (e.g., antibodies) are examples of a genus of equivalent or similar features.

[0065] All numbers or ranges of numbers used herein include integers and fractions of numbers or integers within such ranges unless the context expressly indicates otherwise. Furthermore, where a list of numbers is given herein (e.g., about 50%, 60%, 70%, 80%, 85%, or 86%), the list includes all of its intermediate and decimal values ​​(e.g., 54%, 85.4%). Thus, for illustrative purposes only, a reference to 80% or more includes 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, etc., as well as 81.1%, 81.2%, 81.3%, 81.4%, 81.5%, etc., and 82.1%, 82.2%, 82.3%, 82.4%, 82.5%, etc.

[0066] References to integers preceded by "more" or "less than" include all numbers greater than or less than the number being referred to, respectively. Therefore, for example, "less than 100" includes all numbers up to the digit 1, such as 99, 98, 97, etc., and "less than 10" includes all numbers up to the digit 1, such as 9, 8, 7, etc.

[0067] All numbers or ranges used herein include values ​​and integer fractions within such ranges, as well as integer fractions within such ranges, unless the context makes it clear otherwise. For illustrative purposes, a reference to a numerical range such as 1 to 10 includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 1.1, 1.2, 1.3, 1.4, 1.5, etc. Similarly, a reference to a range of 1 to 50 includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc., up to 50 (including 50), and 1.1, 1.2, 1.3, 1.4, 1.5, etc., and 2.1, 2.2, 2.3, 2.4, 2.5, etc.

[0068] A reference to a range includes a range that combines the boundary values ​​of different ranges within that range. Therefore, for illustrative purposes, for example, 1-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-75, 75-100, 100-150, 150-200, 200-250, 250-300, 300-400, 400-500, 500-750, 750-1,000, 1,000-1,500, 1,500-2,000, 2,000-2,500, 2,500 References to ranges such as ~3,000, 3,000~3,500, 3,500~4,000, 4,000~4,500, 4,500~5,000, 5,500~6,000, 6,000~7,000, 7,000~8,000, or 8,000~9,000 include ranges such as 10~50, 50~100, 100~1,000, 1,000~3,000, 2,000~4,000, etc.

[0069] Modifications may be made to the above-described content without departing from the basic aspects of the Art. Although the Art is described in substantial detail in relation to one or more specific embodiments, those skilled in the art will recognize that modifications may be made to the embodiments specifically disclosed in this application, but such modifications and improvements will be within the scope and spirit of the Art.

[0070] The present invention is disclosed herein using affirmative language to describe a number of embodiments and aspects. The present invention also specifically includes embodiments in which the subject matter of a particular patent invention, such as a substance or material, steps and conditions of a method, protocol, or procedure, is completely or partially excluded. For example, in certain embodiments or aspects of the present invention, a material and / or steps of a method are excluded. Therefore, aspects that are not explicitly excluded in the present invention are also disclosed herein, even though it is not expressed herein in general what the present invention does not include.

[0071] The technologies described herein descriptively can be adequately implemented without any elements not specifically disclosed herein. Therefore, for example, in each example herein, any of the terms “including,” “essentially consisting of,” and “consisting of” may be substituted for any of the other two terms. Some embodiments of the technologies described herein can be adequately implemented without any elements not specifically disclosed herein. Therefore, in some embodiments, the term “including” may be substituted for “essentially consisting of,” or “consisting of,” or any grammatical variation thereof. A “essentially consisting of” composition means a composition comprising only the active ingredient described in the claim (e.g., active ingredient (AI) or active ingredient (API); e.g., L-serine, or a salt thereof, metabolic precursor, derivative, or conjugate), which may include other components such as formulation materials, additives, additives, carriers, preservatives, diluents, solvents, fillers, salts, buffers, coatings, binders, and lubricants, excluding other APIs not described in the claim.

[0072] The terms “a” or “an” may refer to one or more elements they modify unless the context makes it clear that one or more of the elements being described are being referred to (for example, “a reagent” may mean one or more reagents). The term “about” as used herein refers to a value within 10 percent of the underlying parameter (i.e., plus or minus 10%) and modifies each of the values ​​when the term “about” is used at the beginning of a set of values ​​(i.e., “about 1, 2 and 3” refers to about 1, about 2 and about 3). For example, a weight of “about 100 grams” may include weights from 90 grams to 110 grams. The term “substantially” as used herein refers to a value modifier meaning “at least 95%”, “at least 96%”, “at least 97%”, “at least 98%”, or “at least 99%”, and may include 100%. For example, a composition that is substantially free of X may contain less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of X, and / or X may not be present in the composition or may not be detectable.

[0073] Therefore, although this technology is specifically disclosed by representative embodiments and optional features, it should be understood that those skilled in the art can use modifications and variations of the concepts disclosed herein, and such modifications and variations will be considered to be within the scope of this technology. Furthermore, the present invention encompasses the following aspects. 1. A method for identifying a person who has Alzheimer's disease or is at risk of developing it, wherein the method is (a) Determine the amount of 2-aminoethyl dihydrogen phosphate or taurine in the sample obtained from the subject, and (b) Determining whether the subject has Alzheimer's disease or is at risk of developing it, according to the amount of 2-aminoethyl dihydrogen phosphate or taurine in the sample. Methods that include... 2. The method according to item 1, wherein the amount of 2-aminoethyl dihydrogen phosphate is determined. 3. The method according to item 1, wherein the amount of taurine is determined. 4. The method according to item 1, wherein the amounts of dihydrogen diaminoethyl and taurine are determined. 5. The method according to item 1, wherein the subject is determined to have or be at risk of developing Alzheimer's disease in (b) if the amount of 2-aminoethyl dihydrogen phosphate or taurine exceeds a predetermined threshold. 6. The amount of 2-aminoethyl dihydrogen phosphate or taurine exceeding the predetermined threshold is approximately 20 times, 19 times, 18 times, 17 times, 16 times, 15 times, 14 times, 13 times, 12 times, 11 times, 10.5 times, 10 times, 9.5 times, 9 times, 8.5 times, 8 times, 7.5 times, 7 times, and 6.5 times greater than the predetermined threshold. The method according to item 5, which is approximately 6 times, approximately 5.5 times, approximately 5 times, approximately 4.5 times, approximately 4 times, approximately 3.5 times, approximately 3 times, approximately 2.9 times, approximately 2.8 times, approximately 2.7 times, approximately 2.6 times, approximately 2.5 times, approximately 2.4 times, approximately 2.3 times, approximately 2.2 times, approximately 2 times, approximately 1.9 times, approximately 1.8 times, approximately 1.7 times, approximately 1.6 times, approximately 1.5 times, approximately 1.4 times, approximately 1.3 times, approximately 1.2 times, or approximately 1.1 times higher. 7. The method according to item 5, wherein the predetermined threshold is the average, mean, absolute, or maximum amount of 2-aminoethyl dihydrogen phosphate or taurine present in one or more healthy control subjects. 8. The method according to claim 1, wherein the determination of the presence or amount in (a) comprises determining the ratio of 2-aminoethyl dihydrogen phosphate to taurine, and the determination in (b) comprises determining whether the subject has or is at risk of developing Alzheimer's disease according to the ratio of 2-aminoethyl dihydrogen phosphate to taurine in the sample. 9. The method of paragraph 8, wherein the subject is determined to have or be at risk of developing Alzheimer's disease in (b) if the ratio of 2-aminoethyl dihydrogen phosphate to taurine exceeds a predetermined threshold. 10. The ratio of 2-aminoethyl dihydrogen phosphate to taurine that exceeds the predetermined threshold is approximately 20 times, 19 times, 18 times, 17 times, 16 times, 15 times, 14 times, 13 times, 12 times, 11 times, 10.5 times, 10 times, 9.5 times, 9 times, 8.5 times, 8 times, 7.5 times, 7 times, 6.5 times, The method according to item 9, which is a ratio that is approximately 6 times, approximately 5.5 times, approximately 5 times, approximately 4.5 times, approximately 4 times, approximately 3.5 times, approximately 3 times, approximately 2.9 times, approximately 2.8 times, approximately 2.7 times, approximately 2.6 times, approximately 2.5 times, approximately 2.4 times, approximately 2.3 times, approximately 2.2 times, approximately 2 times, approximately 1.9 times, approximately 1.8 times, approximately 1.7 times, approximately 1.6 times, approximately 1.5 times, approximately 1.4 times, approximately 1.3 times, approximately 1.2 times, or approximately 1.1 times higher. 11. The method according to item 9, wherein the predetermined threshold is the average, mean, absolute, or maximum amount of 2-aminoethyl dihydrogen phosphate or taurine present in one or more healthy control subjects. 12. In response to the determination that the subject has Alzheimer's disease or is at risk of developing it, (c) Administering a therapeutically effective dose of Alzheimer's disease drug to the subject. The method described in item 1, further comprising: 13. The Alzheimer's disease drug mentioned above is L-serine, laritrin, phenytoin, lamotrigine, carbamazepine, lidocaine, tetrodotoxin, nitroindazole, sulforaphane or sulforaphane analogues, gabapentin, pregabalin, mirogabalin, gabapentin enacarbil, phenibut, imagabalin, atagabalin, 4-methylpregabalin, PD-217,014, riluzole, edaravone, tetrabenazine, haloperidol, risperidone, quetiapine, amantadine, levetiracetam, clonazepam, citalopram, escitalopram, fluoxetine, sertraline, olanzapine, valoproate, carbamazepine, lamotrigine, vaccines, cholinesterase inhibitors, memantine, antidepressants, N-methyl The method according to item 12, selected from D-aspartic acid (NMDA) antagonists, omega-3 fatty acids, curcumin or curcumin derivatives, vitamin E, hypnotics, anxiolytics, anticonvulsants, antipsychotics, carbidopa / levodopa, amantadine, dopamine agonists, monoamine oxidase B (MAO-B) inhibitors, catechol O-methyltransferase (COMT) inhibitors, and anticholinergics. 14. The method according to item 12, wherein the Alzheimer's disease drug is selected from L-serine, laritrin, phenytoin, lamotrigine, carbamazepine, lidocaine, tetrodotoxin, riluzole, edaravone, gabapentin, pregabalin, mirogabalin, gabapentin enacarbil, phenibut, imagabalin, atagabalin, 4-methylpregabalin, PD-217,014, trihexyphenidyl, amitriptyline, baclofen, diazepam, and CK-2127107. 15. The method according to item 12, wherein the Alzheimer's disease drug comprises L-serine. 16. The method according to item 1, wherein the sample comprises blood or a blood product. 17. The method according to item 16, wherein the blood product contains or consists of plasma. 18. The method according to item 1, wherein the subject is a mammal. 19. The method according to item 18, wherein the mammal is a non-human primate. 20. The method according to item 19, wherein the non-human primate is a savanna monkey. 21. The method according to item 1, wherein the subject is a human. 22. The method according to item 1, wherein the subject is asymptomatic for Alzheimer's disease. 23. The method according to item 1, further comprising monitoring the progression of Alzheimer's disease in the subject, wherein the method is performed two or more times. 24. The method of paragraph 1, wherein the subject has not been diagnosed with Alzheimer's disease prior to the determination in (b). 25. The method described in item 1, which is an ex vivo method. 26. The method described in item 1, which is an in vitro method.

Claims

1. A biomarker for identifying individuals who have Alzheimer's disease or are at risk of developing it, wherein the biomarker comprises 2-aminoethyl dihydrogen phosphate, where the biomarker is (a) Determine the amount of 2-aminoethyl dihydrogen phosphate in the sample obtained from the subject, and (b) If the amount of 2-aminoethyl dihydrogen phosphate in the sample exceeds the average or mean of 2-aminoethyl dihydrogen phosphate in a healthy control sample, it is determined that the sample is from a person with Alzheimer's disease or at risk of developing it. A biomarker used in a method that includes the following.

2. The amount of dihydrogen diaminoethyl phosphate was multiplied by 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10.5, 10, 9.5, 9, 8.5, 8, 7.5, 7, 6.5, 6, 5.5, 5, 4.5, 4, and 3.5 times the average or mean of dihydrogen diaminoethyl phosphate in the healthy control sample. The biomarker according to claim 1, wherein the sample is determined to be derived from a person having or at risk of developing Alzheimer's disease in (b) if it exceeds by an amount 3 times, 2.9 times, 2.8 times, 2.7 times, 2.6 times, 2.5 times, 2.4 times, 2.3 times, 2.2 times, 2 times, 1.9 times, 1.8 times, 1.7 times, 1.6 times, 1.5 times, 1.4 times, 1.3 times, 1.2 times, or 1.1 times higher.

3. The biomarker according to claim 1, wherein in (a) taurine in the sample is further used as a biomarker, and the determination in (b) is further based on the ratio of 2-aminoethyl dihydrogen phosphate to taurine.

4. The biomarker according to claim 3, wherein the ratio of 2-aminoethyl dihydrogen phosphate to taurine exceeds a predetermined threshold, and the sample is determined to be derived from a subject who has or is at risk of developing Alzheimer's disease, in (b).

5. The biomarker according to claim 4, wherein the ratio of 2-aminoethyl dihydrogen phosphate to taurine that exceeds the predetermined threshold is 20 times, 19 times, 18 times, 17 times, 16 times, 15 times, 14 times, 13 times, 12 times, 11 times, 10.5 times, 10 times, 9.5 times, 9 times, 8.5 times, 8 times, 7.5 times, 7 times, 6.5 times, 6 times, 5.5 times, 5 times, 4.5 times, 4 times, 3.5 times, 3 times, 2.9 times, 2.8 times, 2.7 times, 2.6 times, 2.5 times, 2.4 times, 2.3 times, 2.2 times, 2 times, 1.9 times, 1.8 times, 1.7 times, 1.6 times, 1.5 times, 1.4 times, 1.3 times, 1.2, or 1.1 times higher than the predetermined threshold.

6. A biomarker for identifying a subject suitable for receiving an Alzheimer's disease drug, (a) Determine the amount of 2-aminoethyl dihydrogen phosphate in the sample obtained from the subject, and (b) If the amount of 2-aminoethyl dihydrogen phosphate in the sample exceeds the average or mean of 2-aminoethyl dihydrogen phosphate in a healthy control sample, it is determined that the sample is from a person with Alzheimer's disease or at risk of developing it. A biomarker used in a method that includes the following.

7. The aforementioned Alzheimer's disease drugs include L-serine, laritrin, phenytoin, lamotrigine, carbamazepine, lidocaine, tetrodotoxin, nitroindazole, sulforaphane or sulforaphane analogues, gabapentin, pregabalin, mirogabalin, gabapentin enacarbil, phenibut, imagabalin, atagabalin, 4-methylpregabalin, PD-217, 014, riluzole, edaravone, tetrabenazine, haloperidol, risperidone, quetiapine, amantadine, levetiracetam, clonazepam, citalopram, escitalopram, fluoxetine, sertraline, olanzapine, valoproate, carbamazepine, lamotrigine, vaccines, cholinesterase inhibitors, memantine, antidepressants, and N-methyl A biomarker according to claim 6, selected from D-aspartic acid (NMDA) antagonists, omega-3 fatty acids, curcumin or curcumin derivatives, vitamin E, hypnotics, anxiolytics, anticonvulsants, antipsychotics, carbidopa / levodopa, amantadine, dopamine agonists, monoamine oxidase B (MAO-B) inhibitors, catechol O-methyltransferase (COMT) inhibitors, and anticholinergics.

8. The biomarker according to claim 6, wherein the Alzheimer's disease drug is selected from L-serine, laritolin, phenytoin, lamotrigine, carbamazepine, lidocaine, tetrodotoxin, riluzole, edaravone, gabapentin, pregabalin, mirogabalin, gabapentin enacarbil, phenibut, imagabalin, atagabalin, 4-methylpregabalin, PD-217,014, trihexyphenidyl, amitriptyline, baclofen, diazepam, and CK-2127107.

9. The biomarker according to claim 6, wherein the Alzheimer's disease drug comprises L-serine.

10. The biomarker according to claim 1, wherein the sample comprises blood or a blood product.

11. The biomarker according to claim 10, wherein the blood product contains or consists of plasma.

12. The biomarker according to claim 1, wherein the subject is a mammal.

13. The biomarker according to claim 12, wherein the mammal is a non-human primate.

14. The biomarker according to claim 13, wherein the non-human primate is a savanna monkey.

15. The biomarker according to claim 1, wherein the subject is a human.

16. The biomarker according to claim 1, wherein the subject is asymptomatic for Alzheimer's disease.

17. A biomarker according to claim 1, further comprising monitoring the progression of Alzheimer's disease in the subject, wherein the method is performed two or more times.

18. The biomarker according to claim 1, wherein the subject has not been diagnosed with Alzheimer's disease prior to the determination in (b).

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