Compounds and methods targeting human tau

Abstract

To provide antibodies for use in the treatment of neurodegenerative diseases, or pharmaceutical compositions thereof.SOLUTION: The present invention provides compounds and methods targeting human tau, particularly human tau phosphorylated at threonine (217) and isoforms of tau expressed only in the CNS, including therapeutic antibodies, pharmaceutical compositions and diagnostic applications useful in the field of neurodegenerative diseases such as AD, PSP and FTD.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention belongs to the field of pharmaceuticals. More specifically, the present invention relates to compounds, pharmaceutical compositions, diagnostic methods and methods comprising antibodies or fragments thereof against human tau. The compounds and methods of the present invention are expected to be useful in the field of neurodegenerative diseases, particularly tauopathy, including Alzheimer's disease (AD), progressive supranuclear palsy (PSP), and frontotemporal dementia (FTD), and include treatments and related diagnostic methods. [Background technology]

[0002] Tau is an axonal microtubule-associated protein expressed in both the central nervous system ("CNS") and the periphery, which promotes microtubule assembly and stability. Known isoforms of human tau expressed in the CNS are involved in the abnormal formation and aggregation of intraneuronal neurofibrillary tangles ("NFTs"). In neurodegenerative diseases such as Alzheimer's disease (AD), the density and neuroanatomical localization of CNS NFTs correlate with the severity of dementia, the extent of neuronal loss, and overall disease progression. In presbycusis, CNS NFT formation is also observed, and its density correlates with the severity of neuronal loss.

[0003] Alzheimer's disease (AD) is a neurodegenerative disorder characterized by dementia, causing problems with memory, thinking, and behavior. According to the Alzheimer's Association, there are an estimated 5.6 million Americans aged 65 and older with AD (i.e., roughly 1 in 10 people), and another 200,000 Americans under 65 with AD. The Alzheimer's Association also predicts that the number of Americans aged 65 and older with AD will increase by more than 26% by 2025. This represents significant healthcare expenditure, with direct AD-related healthcare costs in the United States estimated at $290 billion in 2019 alone, and this figure does not include unpaid caregiver costs. Despite the significant impact of AD on individuals and healthcare, to date there are no approved disease-modifying therapies for treating AD, leaving such treatments as an unmet healthcare need.

[0004] Furthermore, reliable and highly sensitive diagnostic methods for AD are needed to support the discovery and / or development of disease-modifying therapies. The approved diagnostic application for AD is Amyvid®. Flourtaucipir is a diagnostic application for AD currently under FDA review. Both Amyvid® and flortaucipir are radioisotope neurological imaging agents useful in the detection and staging of AD and other neurodegenerative diseases. In addition, a diagnostic assay targeting phosphorylated threonine ("hTau-pT181") at residue 181 (residue number based on SEQ ID NO: 1) of human tau in patient samples has recently been disclosed. However, the hTau-pT181 diagnostic application lacks the sensitivity required for diagnostic testing, such as identifying various AD stages in patients or patient prognosis in blood, plasma, and cerebrospinal fluid ("CSF") assays. Therefore, there is a need for a more inexpensive and less invasive diagnostic option that is applicable to testing using blood, plasma, and / or CSF, and that is also highly sensitive and reliable. Preferably, such diagnostic methods can identify and / or differentiate between AD patients (e.g., based on the stage or prognosis of AD). Such diagnostic methods can also preferably identify and / or differentiate between those with effective treatment responses. In some embodiments, such diagnostic methods can also preferably identify and / or differentiate between patients who require further diagnostic evaluation, e.g., patients for whom neurological imaging such as flouraucipir and / or amyvid is appropriate. [Overview of the project]

[0005] Accordingly, in one embodiment, the Disclosure provides an antibody against human tau phosphorylated with threonine at residue 217 (residue number based on SEQ ID NO: 1) ("hTau-pT217"), a pharmaceutical composition thereof, and methods and diagnostic applications using such an antibody and pharmaceutical composition. Furthermore, according to one embodiment of the Disclosure, an antibody against an isoform of human tau expressed in the CNS (e.g., one that recognizes an isoform expressed in the CNS but does not recognize an isoform of human tau expressed exclusively outside the CNS), and a pharmaceutical composition thereof. [Brief explanation of the drawing]

[0006] [Figure 1] The ROC curves for hTau-pT217 in the AD, CU-A+, and CU-A- control groups are shown. [Modes for carrying out the invention]

[0007] According to several embodiments, antibodies that specifically bind to hTau-pT217 are provided. In more specific embodiments, antibodies that bind to an epitope region of human tau comprising phosphorylated threonine at residue 217 of SEQ ID NO: 1 are provided, wherein if the threonine at residue 217 of SEQ ID NO: 1 is not phosphorylated, such antibodies do not bind to human tau. In even more specific embodiments of the present disclosure, such antibodies are provided comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity-determining regions (CDRs) LCDR1, LCDR2, and LCDR3, and the HCVR comprises the CDRs HCDR1, HCDR2, and HCDR3, wherein LCDR1 has the amino acid sequence of SEQ ID NO: 13, LCDR2 has the amino acid sequence of SEQ ID NO: 14, LCDR3 has the amino acid sequence of SEQ ID NO: 15, HCDR1 has the amino acid sequence of SEQ ID NO: 10, HCDR2 has the amino acid sequence of SEQ ID NO: 11, and HCDR3 has the amino acid sequence of SEQ ID NO: 12. In some embodiments, LCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 13, LCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 14, LCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 15, HCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 10, HCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 11, and HCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 12. According to some embodiments of the antibodies provided by this disclosure, LCVR has the amino acid sequence of SEQ ID NO: 5, and HCVR has the amino acid sequence of SEQ ID NO: 3. In some embodiments of the antibodies provided by this disclosure, LCVR has the amino acid sequence of SEQ ID NO: 8, and HCVR has the amino acid sequence of SEQ ID NO: 6.In some further embodiments, LCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 5, and HCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 3. In further embodiments, LCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 8, and HCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 6.

[0008] Embodiments of this disclosure provide an antibody that specifically binds to a CNS expression isoform of human tau (e.g., a known isoform of human tau expressed in the CNS), such an antibody that does not bind to a human tau isoform expressed exclusively in regions outside the CNS (including the peripheral nervous system). According to certain embodiments, such an antibody that specifically binds to a CNS expression isoform of human tau binds to the epitope region of human tau comprising 124 (glutamine) and 125 (alanine) of SEQ ID NO: 1. In certain embodiments, such an antibody is provided comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity-determining regions (CDRs) LCDR1, LCDR2, and LCDR3, and the HCVR comprises the CDRs HCDR1, HCDR2, and HCDR3, wherein LCDR1 has the amino acid sequence of SEQ ID NO: 23, LCDR2 has the amino acid sequence of SEQ ID NO: 24, LCDR3 has the amino acid sequence of SEQ ID NO: 25, HCDR1 has the amino acid sequence of SEQ ID NO: 20, HCDR2 has the amino acid sequence of SEQ ID NO: 21, and HCDR3 has the amino acid sequence of SEQ ID NO: 22. In some embodiments, LCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 23, LCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 24, LCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 25, HCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 20, HCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 21, and HCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 22. According to some embodiments of the antibodies provided by this disclosure, LCVR has the amino acid sequence of SEQ ID NO: 17, and HCVR has the amino acid sequence of SEQ ID NO: 19.In some further embodiments, LCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 17, and HCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 19.

[0009] According to some embodiments, the antibodies of the Disclosure may be humanized. In some embodiments, the antibodies of the Disclosure include an IgG quadruple chain. In some embodiments, the antibodies of the Disclosure include a kappa light chain. In further embodiments, the Disclosure provides a pharmaceutical composition comprising the antibody of the Disclosure and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0010] In further embodiments, the Disclosure provides a method for treating a neurodegenerative disease, comprising administering an effective amount of the antibody of the Disclosure, or a pharmaceutical composition thereof, to a patient in need thereof. In some such embodiments, the neurodegenerative disease is a tauopathy. In even more specific embodiments, the tauopathy is one of AD, PSP, and FTD.

[0011] According to some embodiments, the present disclosure provides antibodies or pharmaceutical compositions thereof for therapeutic use. Furthermore, antibodies or pharmaceutical compositions thereof are provided for use in the treatment of neurodegenerative diseases. In some such embodiments, the neurodegenerative disease is a tauopathy. In some more specific embodiments, the tauopathy is selected from the group consisting of AD, PSP, and FTD.

[0012] According to some embodiments of the present disclosure, antibodies or pharmaceutically active compositions thereof are provided for use in the manufacture of pharmaceuticals for the treatment of neurodegenerative diseases. In some such embodiments, the neurodegenerative disease is a tauopathy. In more specific embodiments, the tauopathy is selected from the group consisting of AD, PSP, and FTD.

[0013] Further embodiments of the present disclosure provide a method for detecting hTau-pT217 in a patient sample. Such a method includes the steps of contacting the patient sample with an antibody of the present disclosure that specifically binds to hTau-pT217, and detecting a signal provided by the contact step.

[0014] According to several embodiments, a method is provided for detecting isoforms of human tau expressed exclusively in the CNS. Such a method includes the steps of contacting a patient sample with an antibody of the Disclosure that specifically binds to isoforms of human tau expressed in the CNS (i.e., does not bind to isoforms of human tau expressed exclusively outside the CNS), and detecting a signal provided by the contact step.

[0015] According to some embodiments, a method is provided for quantifying hTau-pT217 in a patient sample. Such a method includes the steps of contacting the patient sample with an antibody of the Disclosure that specifically binds to hTau-pT217, and detecting the signal provided by the contact step. In some embodiments, such a method further includes the steps of contacting a control standard with the antibody, and detecting the signal provided by the contact step with the control standard.

[0016] In some embodiments, methods for quantifying hTau-pT217 in patient samples are provided by the present disclosure. Such methods include contacting a patient sample with an antibody of the present disclosure that specifically binds to hTau-pT217, and contacting the patient sample with an antibody of the present disclosure that specifically binds to a CNS-expressed isoform of human tau, where the antibodies do not bind to overlapping epitope regions of the antibodies and one of the antibodies comprises a detectable label; detecting a signal provided by the detectable label upon formation of a complex comprising the antibody and hTau-pT217; contacting a control standard with the antibody; and detecting a signal provided by the detectable label upon formation of a complex comprising the antibody and the control standard.

[0017] According to some embodiments of the present disclosure, methods are provided for diagnosing a patient as one or more of (i) having a neurodegenerative disease, (ii) being at risk of having a neurodegenerative disease, (iii) having a need for treatment for a neurodegenerative disease, (iv) being in AD Braak stage I, II, III, IV, V or VI, or (v) having a need for neurological imaging. According to such embodiments, such methods include contacting a patient sample with an antibody of the present disclosure that specifically binds to hTau-pT217, and detecting binding between hTau-pT217 and the antibody in the patient sample. In some such embodiments, the method further includes diagnosing the patient as one of (i) having a neurodegenerative disease, (ii) being at risk of having a neurodegenerative disease, (iii) having a need for treatment for a neurodegenerative disease, (iv) being in AD Braak stage I, II, III, IV, V or VI, or (v) having a need for neurological imaging, if the level of hTau-pT217 detected in the patient sample exceeds a reference level.

[0018] In some embodiments of the present disclosure, methods for diagnosing and treating neurodegenerative diseases in patients are provided. According to such embodiments, the method comprises contacting a patient sample with an antibody of the present disclosure that specifically binds to hTau-pT217, detecting the binding between hTau-pT217 and the antibody in the patient sample, diagnosing a patient having a neurodegenerative disease, and administering a therapeutically effective amount of an anti-human tau antibody to the diagnosed patient. In some embodiments, the diagnosing step comprises diagnosing a patient as having a neurodegenerative disease if the presence of hTau-pT217 in the patient sample exceeds a reference level.

[0019] According to some embodiments of the method of the present disclosure, such method further comprises quantifying hTau-pT217 in a patient sample. In such embodiments, the step of quantifying hTau-pT217 comprises quantifying hTau-pT217 in the patient sample relative to a reference standard.

[0020] According to some embodiments of the method of the present disclosure, the patient sample is one of blood, plasma, serum or CSF.

[0021] According to some embodiments of the methods of the present disclosure, the method further includes the step of contacting a patient sample with an antibody that specifically binds to hTau-pT217 and a second antibody, wherein the second antibody specifically binds to a CNS expression isoform of human tau. In some such methods, one of the antibodies or the second antibody includes a detectable label, and the detection step includes detecting a signal provided by the detectable label upon the formation of a complex comprising the antibody, the second antibody and hTau-pT217. According to some such embodiments, one of the antibodies or the second antibody is immobilized on a substrate. In some embodiments of the methods of the present disclosure, the step of contacting the patient sample with the antibody and the step of contacting the patient sample with the second antibody are performed simultaneously. According to some more specific embodiments, the second antibody includes the antibody of the present disclosure that specifically binds to a CNS expression isoform of human tau disclosed herein.

[0022] As used herein, “antibody” is an immunoglobulin molecule containing two HCs and two LCs interconnected by disulfide bonds. The amino-terminal portion of each LC and HC contains a variable region of approximately 100-120 amino acids, primarily responsible for antigen recognition via the CDR contained within it. The CDR contains scattered, more conserved regions called framework regions ("FRs"). Each LCVR and HCVR consists of three CDRs and four FRs arranged from the amino-terminal to the carboxyl-terminal in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDRs of the LC are called “LCDR1, LCDR2, and LCDR3,” and the three CDRs of the HC are called “HCDR1, HCDR2, and HCDR3.” The CDRs contain the majority of the residues that form specific interactions with the antigen. The functional ability of an antibody to bind to a particular antigen is largely influenced by the six CDRs. The assignment of amino acids to the CDR domain within the LCVR and HCVR regions of the antibody of the present invention is based on the well-known Kabat numbering rules (Kabat, et al., Ann. NY Acad. Sci. 190:382-93 (1971), Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, USD Department of Health and Human Services, NIH Publication No. 91-3242 (1991)), and the North numbering rules (North et al., A New Clustering of Antibody CDR Loop Conformations, Journal of Molecular Biology, 406:228-256 (2011)).

[0023] According to some embodiments of the present disclosure, LCs are classified as kappa or lambda, each characterized by a specific constant region known in the art. According to some embodiments of the present disclosure, HCs are classified as gamma, mu, alpha, delta, or epsilon, each defining the antibody isotype as IgG, IgM, IgA, IgD, or IgE, respectively. According to some embodiments, the antibody comprises IgG HC, which can be further divided into subclasses, e.g., IgG1, IgG2, IgG3, IgG4. The carboxyl-terminal portion of each HC defines a constant region primarily responsible for effector function. In certain embodiments, the antibody of the present invention has one or more modifications within the constant region of each HC that reduce effector function.

[0024] The antibody of the present invention is a monoclonal antibody. A monoclonal antibody is an antibody derived from a single copy or clone (e.g., any eukaryote, prokaryote, or phage clone), not the method by which it is produced. Monoclonal antibodies can be produced, for example, by hybridoma technology, recombination technology, phage presentation technology, synthesis technology, e.g., CDR transplantation, or a combination of such technologies or other technologies known in the art.

[0025] Methods for producing and purifying antibodies are well known in the art and can be found, for example, in Harlow and Lane (1988), Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, chapters 5-8 and 15, ISBN 0-87969-314-2. For example, mice or rabbits may be immunized with hTau-pT217, the resulting antibodies may be recovered, purified, and their amino acid sequences determined using conventional methods well known in the art. Similarly, a phage library may be screened to screen thousands of Fab fragments for interaction with hTau-pT217, the resulting interactions may be recovered, purified, and their amino acid sequences determined using conventional methods well known in the art, thereby enabling the construction of an initial lead antibody. Embodiments of antibodies in this disclosure include antibodies engineered to include one or more human framework regions surrounding a CDR derived from a non-human antibody. The germline sequences of the human framework can be obtained, for example, from ImMunoGeneTics (INGT) through its website (http: / / imgt.cines.fr) or from The Immunoglobulin FactsBook by Marie-Paule Lefranc and Gerard Lefranc, Academic Press, 2001, ISBN 012441351.

[0026] In certain embodiments of the present invention, an antibody, or a nucleic acid encoding an antibody, is provided in an isolated form. As used herein, the term “isolated” means a protein, peptide, or nucleic acid that does not contain, or substantially contains, other macromolecular species found in the cellular environment.

[0027] The antibodies provided herein may be used in the treatment of patients. More specifically, embodiments of the antibodies herein may be useful in the treatment of neurodegenerative diseases or disorders, including tauopathies, including AD, PSP, and FTD. While the antibodies of the present invention may be useful in the treatment of AD, PSP, and FTD, such antibodies may also be useful in the treatment of other neurodegenerative diseases, particularly diseases involving tau pathologies such as NFT formation. Where used interchangeably herein, “treatment” and / or “to treat” and / or “to cure” are intended to refer to all processes in which there may be a slowing, interference, inhibition, control, cessation, or reversal of the progression of the disorders described herein, but not necessarily the complete elimination of the symptoms of all disorders. The treatment comprises administering an antibody of the present invention for the treatment of a disease or condition in humans, which benefits from a reduction in the propagation of at least one of tau aggregation formation, NFT formation, and neuronal defect, and includes (a) inhibiting further progression of the disease, i.e., inhibiting its development, and (b) alleviating the disease, i.e., causing regression of the disease or disorder, or alleviating its symptoms or complications.

[0028] Where used interchangeably herein, the terms “patient,” “subject,” and “individual” refer to a human being. In certain embodiments, a patient is further characterized as having a disease, disorder, or condition (e.g., a neurodegenerative disorder) that benefits from reduced propagation of at least one of tau aggregate formation, neurofibrillary tangle formation, and neuronal loss. In other embodiments, a patient is further characterized as being at risk of developing a neurodegenerative disorder, disease, or condition that benefits from reduced propagation of at least one of tau aggregate formation, NFT formation, and neuronal loss.

[0029] As used herein, the term “specifically binds to hTau-pT217” refers to the interaction of the antibody with the epitope region of human tau containing phosphorylated threonine at residue 217 of SEQ ID NO: 1. Such binding depends on the phosphorylation of threonine at residue 217 of SEQ ID NO: 1. It should be understood that there are known variations or isoforms of human tau, for example, arising from splice variants. It should also be understood that such known variants may result in changes in residue numbering for some amino acid residues of SEQ ID NO: 1, including phosphorylated threonine, as provided herein with reference to the human tau sequence described in SEQ ID NO: 1.

[0030] As used herein, the term "specifically binds to a human tau CNS expression isoform" refers to the interaction of the antibody of this disclosure with an epitope region common to or present on human tau isoforms expressed in the CNS, which is not present on human tau isoforms expressed exclusively outside the CNS. An antibody that specifically binds to a human tau CNS expression isoform does not bind to human tau isoforms expressed exclusively outside the CNS (e.g., isoforms expressed only in other parts of the body, such as the peripheral nervous system). According to some embodiments, an antibody that specifically binds to a human tau CNS expression isoform binds to or recognizes an epitope region of a human tau isoform expressed in the CNS, which includes glutamine at residue 124 (Q124) and alanine at residue 125 (A125), with the residue numbers referenced from Sequence ID No. 1. For example, it should be understood that there are known variations or isoforms of human tau arising from splice variants, and that such variants may result in changes to the residue numbering of several amino acid residues, including glutamine and alanine, as provided herein with reference to the human tau sequence described in Sequence ID No. 1.

[0031] As used herein, the term “epitope region” refers to a distinct three-dimensional site of an antigen that is recognized either entirely or partially by the antibody of the present invention. The amino acids of the epitope region may provide a chemically active surface group of human tau, forming a specific three-dimensional structure of human tau and providing specific charge properties. Conformational epitopes and non-conformational / linear epitopes can be distinguished in that binding to conformational epitope regions is lost in the presence of a denaturing solvent, whereas linear epitope regions are not.

[0032] The antibody of the present invention can be prepared by methods well known in the art and can be incorporated into a pharmaceutical composition comprising the antibody of the present invention and one or more pharmaceutically acceptable carriers and / or diluents (for example, Remington, The Science and Practice of Pharmacy, 22, which provides an overview of formulation techniques generally known to general practitioners). nd (Edition, Loyd V., Ed., Pharmaceutical Press, 2012). Suitable carriers for the pharmaceutical composition include any material that retains molecular activity when combined with the antibody of the present invention and is non-reactive with the patient's immune system. A pharmaceutical composition containing the antibody of the present invention can be administered by a parental route (e.g., subcutaneous, intravenous, intraperitoneal, intramuscular, or transdermal) to a patient who is at risk of or exhibits any of the diseases or disorders described herein. A pharmaceutical composition of the present invention contains an "effective" amount or a "therapeutic effective" amount (as used interchangeably herein) of the antibody of the present invention. The effective amount refers to the amount (in dose, duration, and means of administration) required to achieve the desired therapeutic outcome. The effective amount of antibody may vary depending on factors such as the individual's medical condition, age, sex, and weight, as well as the antibody's ability to induce a desired response in the individual. The effective amount is also the amount in which the therapeutically beneficial effect outweighs any toxic or adverse effect of the antibody of the present invention.

[0033] In the context of two or more amino acid sequences, the homology percentage as referred to herein refers to two or more sequences that have a specified percentage of the same amino acid residues when compared and aligned for maximum match, measured using a sequence comparison algorithm (e.g., BLASTP and BLASTN, or other algorithms available to those skilled in the art) or by visual inspection. Depending on the application, the homology percentage may exist across regions of the sequences being compared, for example, across functional domains, or across the entire length of the two sequences being compared. As an example, the homology percentage of a sequence can be compared to a reference sequence. For example, when using a sequence comparison algorithm, the test sequence and the reference sequence can be entered into a computer (subsequence coordinates may be further specified along with parameters of the sequence algorithm program, if necessary). The sequence comparison algorithm then calculates the sequence identity or homology percentage of the test sequence relative to the reference sequence based on the specified program parameters. Exemplary sequence alignment and / or homology algorithms are available through Smith & Waterman, Adv.Appl.Math.2:482(1981), Needleman & Wunsch, J.Mol.Biol.48:443(1970), Pearson & Lipman, Proc.Nat'l.Acad.Sci.USA 85:2444(1988), GAP, BESTFIT, FASTA, and TFASTA (the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see Ausubel et al., below). An example of a suitable algorithm for determining sequence identity and sequence similarity percentages is the BLAST algorithm, which is described in Altschul et al., J.Mol.Biol.215:403-410(1990).Software for performing BLAST analysis is available through the National Center for Biotechnology Information (www.ncbi.nlm.nih.gov / ).

[0034] As used herein, patient “sample” refers to a human sample. Non-limiting sources of sample for use in the present invention include blood, plasma, serum, and CSF. Furthermore, sample may also refer to lymph, biopsy aspirates, ascites, fluid extracts, solid tissue, external skin sections, respiratory tract, nasal passages, intestinal tract, and genitourinary tract, tears, saliva, lactation, tumors, organs, cell cultures, and / or cell culture components.

[0035] This disclosure also relates to methods of clinical diagnosis, prognosis, or theranosis of a subject performed by a healthcare professional using the methods disclosed herein. These methods may be performed, for example, by an individual, a healthcare professional, or a third party, such as a service provider interpreting information from the subject. As described herein, a healthcare professional may initiate or modify treatment after receiving information regarding the diagnostic methods of this disclosure. For example, a healthcare professional may recommend a treatment, a change in treatment, or additional diagnostic evaluation (e.g., neurological imaging).

[0036] Using the anti-tau antibody of this disclosure that specifically binds to hTau-pT217, hTau-pT217 can be isolated, detected, and / or quantified by techniques such as affinity chromatography, immunoprecipitation, immunohistochemistry, or ELISA-based assays. Using such assays, the expression of hTau-pT217 and / or its abundance and / or pattern can be detected and / or evaluated for diagnostic, prognostic, or theranostic purposes, by monitoring polypeptide levels (e.g., in serum, plasma, blood, or CSF) as part of a clinical trial procedure (e.g., to determine the efficacy of a given treatment regimen).

[0037] Using the anti-tau antibodies of this disclosure that specifically bind to CNS expression isoforms of human tau, isoforms of human tau expressed in the CNS (excluding isoforms of human tau expressed exclusively outside the CNS) can be isolated and / or detected by techniques such as affinity chromatography, immunoprecipitation, immunohistochemistry, or ELISA-based assays. Using such assays, the expression of CNS expression isoforms of human tau can be detected and / or evaluated for diagnostic, prognostic, or theranostic purposes, monitoring polypeptide levels (e.g., in serum, plasma, blood, or CSF) as part of a clinical trial procedure (e.g., to determine the efficacy of a given therapeutic regimen). As is understood in the art, the antibodies of the present invention may be conjugated to detectable substances or labels to facilitate their detection. Examples of detectable substances or labels include various enzymes, prosthetic groups, fluorescent substances, luminescent substances, bioluminescent substances, chemiluminescent substances, and radioactive substances. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent substances include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotnazinylamine, fluorescein, dansyl chloride, or phycoerythrin; examples of luminescent substances include luminol; examples of bioluminescent substances include luciferase, luciferin, ruthenium, and aequorin; and examples of suitable radioactive substances include 125 I, 131 I, 35 S, or 3The antibody of the present invention contains H. The antibody may also be useful in pharmacological genomic analysis. Such embodiments may be used to identify individuals who may benefit from specific or modified therapeutic approaches and / or to monitor the effectiveness of current therapeutic regimens.

[0038] The levels or measurements of hTau-pT217 provided by the assay of the present invention may be absolute values ​​(e.g., concentration in a biological sample) or relative values ​​(e.g., concentration compared to a reference). As used herein, if a method for detecting hTau-pT217 indicates that the level or concentration of hTau-pT217 in a patient sample is higher than a reference value, then hTau-pT217 is said to be "increased" in the patient sample. Conversely, if the level or concentration of hTau-pT217 in a patient sample is lower than a reference value, or, for example, an hTau-pT217 value measured in a previous patient sample, then hTau-pT217 is said to be "decreased" in the patient sample.

[0039] As used herein, “reference value” refers to a known or approximate concentration of hTau-pT217 associated with a particular condition. The concentration level at the reference value may be the absolute or relative amount of hTau-pT217, a range of amounts, or the minimum, average, and / or median amount. The reference value may also serve as a baseline for hTau-pT217 against which a patient sample is compared.

[0040] As used herein, “control standard” refers to a sample that may be used to compare results obtained from patient samples in the methods of the present invention. A control standard may be the concentration of a known protein spiked in cells, blood, plasma, CSF, tissue, or culture medium. The concentration level in the control standard may be the absolute or relative amount, range of amounts, or minimum, average, and / or median amount of hTau-pT217. The control standard may also act as a baseline for hTau-pT217 against which the patient sample is compared. The control standard may include concentration values ​​from the same patient or a known normal reference for hTau-pT217. Furthermore, in some embodiments, the control standard may represent the hTau-pT217 concentration in the form of a standard curve.

[0041] As used herein, the term “capture antibody” refers to an antibody that binds to hTau-pT217. In such embodiments, the capture antibody can, under appropriate conditions, bind to and capture hTau-pT217 in a patient sample, and can, for example, specifically bind to hTau-pT217 (e.g., it will not bind to human tau if the threonine at residue 217 of SEQ ID NO: 1 is not phosphorylated), and as a result, the capture antibody-hTau-pT217 complex can be separated from the rest of the sample. In some embodiments, the capture antibody may be an antibody that specifically binds to a CNS expression isoform of human tau (e.g., which may include hTau phosphorylated with threonine at residue 217), and the antibody that specifically binds to hTau-pT217 is used as a “second (or detection) antibody.” In some embodiments, the capture antibody is immobilized. In some embodiments, the detection antibody is labeled with a detectable label. In some embodiments, the capture antibody is immobilized in a “sandwich” immunoassay, and the capture or first antibody specifically binds to the epitope region of human tau containing phosphorylated threonine at residue 217 of SEQ ID NO: 1. In such a sandwich immunoassay, a “detection (or second) antibody” is also utilized. According to some embodiments, the detection or second antibody can specifically bind to the capture antibody and may be labeled with a detectable label. In some embodiments, the detection second antibody specifically binds to hTau-pT217, which is already bound or captured by the capture or first antibody. In such embodiments, the detection antibody binds to hTau-pT217 in a second epitope region that does not overlap with the first or capture antibody and may be labeled with a detectable label. In some such embodiments, the second antibody is the antibody of the present invention that specifically binds to the CNS expression isoform of human tau.

[0042] As used herein, “detectable label” is a portion, composition, or technique that can be used to detect the formation of a complex between the antibody of the present invention, which specifically binds to hTau-pT217, and hTau-pT217. According to some embodiments, the detectable label may be conjugated directly or indirectly to the antibody (which may, as applicable, either capture or detection). Exemplary embodiments of detectable labels include biotin, radioisotopes, fluorophores or other fluorescent moieties, and enzyme moieties.

[0043] Where used interchangeably herein, the terms “diagnose” or “to diagnose” refer to a method by which a person skilled in the art can estimate and / or determine the probability (“likelihood”) that a patient has a given disease or condition. In the present invention, “diagnosing” a patient includes using the results of the assay of the present invention to identify or diagnose a neurological disorder such as AD, PSP, or FTD, and to identify a patient (e.g., the presence or occurrence of a neurological disorder or condition or a need for treatment), or to identify the effectiveness of a treatment for a neurological disorder in a patient. According to the present invention, a diagnosis may also be based on a combination of other clinical signs understood by a healthcare professional to arrive at a diagnosis. According to some embodiments of the present invention, a diagnostic application of the present invention may be used to diagnose a patient as being in AD Braak stages I, II, III, IV, V, or VI. The AD Braak stages are known in the art and are described in Braak, et al., (2006) Acta Neuropathol 112(4):389-404. [Examples]

[0044] Anti-hTau-pT217 antibody The anti-hTau-pT217 antibody of this disclosure, or an antibody that specifically binds to hTau-pT217, is generated using a hybridoma method (for example, as first described by Kohler et al., Nature, 256:495 (1975)). Briefly, for example, rabbits are immunized with a peptide representing phosphorylated threonine and four or more amino acids at the N-terminus and C-terminus of such threonine, represented by SEQ ID NO: 1 (an example of a peptide that can be used in immunization is provided by SEQ ID NO: 26). Lymphocytes capable of producing antibodies that bind to hTau-pT217 are isolated and fused with myeloma cell lines using a fusion agent suitable for forming hybridoma cells (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)). The hybridomas are seeded in a suitable culture medium (preferably containing one or more substances that inhibit the survival of unfused myeloma cells) and grown. Next, the binding specificity of monoclonal antibodies produced by the hybridoma is determined by in vitro binding assays (e.g., immunoprecipitation, radioimmunoassay (RIA), or enzyme-linked immunosorbent assay (ELISA)) using both hTau-pT217 and recombinant tau not phosphorylated with threonine at residue 217 (numbered based on reference to SEQ ID NO: 1). Antibodies that specifically bind to hTau-pT217 (e.g., those that do not bind to human tau not phosphorylated at residue 217, numbered based on SEQ ID NO: 1) are identified. Preferred hybridomas can be subcloned using limiting dilution procedures and grown by standard methods, including in vivo as ascites tumors in animals (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)).Monoclonal antibodies secreted by hybridomas (and / or subclones) are purified using conventional procedures such as affinity chromatography (e.g., protein A or protein G-sepharose), ion exchange chromatography, hydroxyapatite chromatography, gel electrophoresis, or dialysis.

[0045] The cDNA encoding the antibody of the present invention is sequenced using conventional procedures. An exemplary rabbit anti-hTau-pT217 antibody ("mAb A") produced substantially according to the procedures described herein contains the heavy chain of SEQ ID NO: 2 and the light chain of SEQ ID NO: 4. The complementarity-determining region (CDR) or variable region of the sequenced antibody can be used in conversion to a chimeric or humanized antibody and / or to other mammalian IgG forms. For example, the clone can be converted to a mouse IgG chimeric antibody such as an exemplary rabbit variable region, mouse IgG constant region chimeric anti-hTau-pT217 antibody ("mAb B") having the heavy chain variable region of SEQ ID NO: 6 and the heavy chain of SEQ ID NO: 7 and the light chain variable region of SEQ ID NO: 8 and the light chain of SEQ ID NO: 9. The binding specificity can then be re-evaluated. The cDNA sequences encoding the heavy and light chains can be cloned and manipulated in a GS (glutamine synthetase) expression vector. The manipulated immunoglobulin expression vector can then be stably transfected into CHO cells. As those skilled in the art will understand, mammalian expression of antibodies will typically result in glycosylation at highly conserved N-glycosylation sites in the Fc region. Stable clones can be confirmed for the expression of antibodies that specifically bind to hTau-pT217. Positive clones can be expanded into serum-free culture medium for antibody production in a bioreactor. The culture medium from which the antibodies have secreted can be purified by conventional techniques. For example, the medium can be conveniently applied to a Protein A or G Sepharose FF column equilibrated with a suitable buffer such as phosphate-buffered saline. The column is washed to remove nonspecific binding components. The bound antibody is eluted, for example, by a pH gradient, and the antibody fraction is detected, for example, by SDS-PAGE and then pooled. The antibody can be concentrated and / or filtered sterile using common techniques. Soluble aggregates and macromers can be effectively removed by common techniques including size exclusion, hydrophobic interactions, ion exchange, or hydroxyapatite chromatography. The product can be immediately frozen, for example, at -70°C, or lyophilized.

[0046] Antibodies specific to the isoform of hTau expressed only in the CNS. Antibodies of this disclosure that specifically bind to human tau CNS expression isoforms can be generated using a hybridoma method (for example, as first described by Kohler et al., Nature, 256:495 (1975)). Briefly, exemplarily, non-human mammals (e.g., mice or rabbits) can be immunized with human tau protein (e.g., hTau, as given by SEQ ID NO: 1), which contains glutamine at residue 124 and alanine at residue 125, as numbered by SEQ ID NO: 1, or its peptides. Lymphocytes capable of producing antibodies that specifically bind to human tau CNS expression isoforms can be isolated and fused with myeloma cell lines using a suitable fusion agent for forming hybridoma cells (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)). The hybridomas can be seeded and grown in a suitable culture medium (preferably containing one or more substances that inhibit the survival of unfused myeloma cells). Next, the binding specificity of monoclonal antibodies produced by hybridomas is determined by in vitro binding assays (e.g., immunoprecipitation, radioimmunoassay (RIA), or enzyme-linked immunosorbent assay (ELISA)) against both the CNS expression isoform of human tau (e.g., a peptide having the sequence of SEQ ID NO: 1 and / or the peripheral expression isoform of human tau (e.g., a peptide having the sequence given by SEQ ID NO: 27, but without glutamine at residue 124 adjacent to alanine at residue 125 represented by SEQ ID NO: 1)). Antibodies that specifically bind to the CNS expression isoform of human tau (e.g., those that do not bind to peripherally expressed human tau) can be identified. Preferred hybridomas can be subcloned using limiting dilution procedures and grown by standard methods, including in vivo as ascites tumors in animals (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)).Monoclonal antibodies secreted by hybridomas (and / or subclones) can be purified using conventional procedures such as affinity chromatography (e.g., protein A or protein G-sepharose), ion exchange chromatography, hydroxyapatite chromatography, gel electrophoresis, and dialysis.

[0047] The cDNA encoding the antibody of the present invention can be sequenced using conventional procedures. An exemplary mouse antibody of this disclosure ("mAb C"), generated substantially according to the procedures described herein, specifically binds to a human tau CNS expression isoform and comprises the heavy chain of SEQ ID NO: 16 and the light chain of SEQ ID NO: 18. The complementarity-determining region (CDR) or variable region of the sequenced antibody can be used in conversion to a chimeric or humanized antibody, and / or other mammalian IgG forms, which can then be expressed in component host cells such as CHO cells.

[0048] Binding kinetics and affinity The binding of the antibody of the present invention, which specifically binds to hTau-pT217, to the recombinant hTau-pT217 protein having the amino acid sequence of SEQ ID NO: 1, is measured using a biolayer interferometry (BLI) assay (using HBS-EP + running buffer (GE Healthcare, 10 mM Hepes pH 7.4 + 150 mM NaCl + 3 mM EDTA + 0.05% surfactant P20) at 25°C) with an Octet Red96® instrument available from ForteBio.

[0049] All reagents and materials, except as noted, are from ForteBio (Freemont, CA). A Protein A biosensor is used to immobilize the antibody of interest for analysis. An exemplary antibody sample of the present invention (mAb A) is prepared to 5 μg / mL by dilution into running buffer. Recombinant hTau-pT217 protein is prepared to concentrations of 300, 100, 33.3, 11.1, 3.7, 1.24, 0.4115, and 0 (blank) nM by dilution into running buffer. Each analysis consists of: (1) capturing the antibody sample on the biosensor for 240 seconds; (2) establishing a baseline by incubating the antibody-loaded biosensor with running buffer for 60 seconds; (3) monitoring the association phase by incubating the antibody-loaded biosensor with serially diluted recombinant hTau-pT217 protein for 300 seconds; and (4) monitoring the dissociation phase by returning the biosensor to running buffer.

[0050] Binding data is processed using standard dual referencing and fit to a 1:1 binding model using Data Analysis v9.0 evaluation software to determine the association rate (k on , M -1 s -1 units), and the dissociation rate (k off , s -1 units). The equilibrium dissociation constant (K D ) is calculated from the relationship K off = k on / k D and is in molar units. The results are shown in Table 1.

Table 1

[0051] Binding Specificity to hTau-pT217 The specificity of the exemplary antibodies (mAb A and mAb B) of the present invention that specifically bind to hTau-pT217 is determined using a BLI assay (using HBS-EP + running buffer (GE Healthcare, 10 mM Hepes pH 7.4 + 150 mM NaCl + 3 mM EDTA + 0.05% surfactant P20) at 25°C) with synthetic peptides and an Octet Red96® instrument available from ForteBio. The N-terminal biotin-labeled peptide of SEQ ID NO: 26, with or without phosphorylated threonine at residue 7, is immobilized on a streptavidin biosensor (ForteBio). The peptides are incubated with IgG of mAb A and mAb B diluted to 5 μg / mL in running buffer for 300 seconds, followed by dissociation for 300 seconds. Binding data is determined using Data Analysis v9.0 evaluation software. The binding signals (nm) for each peptide at the end of dissociation are shown in Table 2. [Table 2]

[0052] hTau-pT217 immunoassay An immunoassay for measuring hTau-pT217 in plasma is designed to measure disease-related differences in AD patients. For example, the immunoassay of this disclosure is performed on a streptavidin small spot plate using the Meso Scale Discovery (MSD) platform. Either mAb A or mAb B is used as the capture antibody and biotinylated. A SULFO-TAG 2 antibody, such as mAb C (the antibody of this disclosure that specifically binds to the human tau CNS expression isoform), is used as the detection antibody. The antibody is conjugated with Sulfo-NHS-Biotin (Thermo Scientific, catalog no. 21329) or MSD GOLD SULFO-TAG NHS-Ester (MSD, catalog no. R91AO-1) according to the manufacturer's protocol. The assay is calibrated using recombinant tau (4R2N, NCBI tau v2) protein, phosphorylated in vitro using a reaction with glycogen synthase kinase 3 and characterized by mass spectrometry. Samples are thawed on moist ice, vortexed briefly, and plasma is refrozen in sample buffer (for mAb A: phosphate-buffered saline (PBS), 0.5% bovine serum albumin (BSA), 0.5% Tween20, 5 mM EDTA, 5 mM EGTA), mAb A. B For this, dilute 1:4 in 50mM HEPES, 300mM NaCl, 5mM EDTA, 5mM EGTA, 1% Triton X-100, 1% MSD Blocker A, and 2% PEG, and add Heterophilic Blocking Reagent 1 to a concentration of 200ug / mL (Scantibodies Inc, catalog number 3KC533). Prepare the calibrator diluent by mixing the sample buffer with Knock-Out Serum Replacement (Gibco, 10828-010) in a 50 / 50 ratio.

[0053] Plates coated with MSD small-spot streptavidin (MSD, L45SA) are blocked at room temperature for 1 hour with 200 μL of 3% BSA in PBS while shaking at 650 rpm on a plate shaker. The plates are washed three times with 200 μL of washing buffer (PBS + 0.05% Tween 20), and 0.464 ug / mL of biotinylated capture antibody (mAb A) is added (diluted in DPBS + 0.1% BSA + 0.05% Tween 20). B For this step, add 25 μL of (plus 2% PEG) to the hTau-pT217 plate and incubate at room temperature for 1 hour while shaking at 650 rpm on a plate shaker. Wash the plate again three times with 200 μL of wash buffer, add 50 μL of diluted calibrator or sample to the plate, and incubate at room temperature for 2 hours while shaking at 650 rpm on a plate shaker. Next, wash the plate three times with 200 μL of wash buffer, and add 25 μL of SULFO-tagged detection antibody (mAb C) at 0.25 ug / mL (diluted in MSD Diluent 35, mAb B For this, add 2% PEG)hTau-pT217 to a plate and incubate at room temperature for 1 hour while shaking at 650 rpm on a plate shaker. Finally, wash the plate with 200 μL of wash buffer, add 150 μL of 2X MSD Read Buffer T with Surfactant (MSD, R92TC) to each plate, and read the result on an MSD SQ120 within 10 minutes of adding the read buffer. The result is given by the formula Y = b1 + ((b2-b1) / (1+(x / b3)). b4 Using )), 4PL, the weights of the standard curve for interpolation are 1 / y 2 This is calculated using MSD software.

[0054] hTau-pT217 immunoassay as a prognostic assay for neurological imaging Levels of hTau-pT217 and hTau-pT181 will be assessed in the blood of subjects enrolled in the AD clinical trial. hTau-pT217 will be evaluated in the plasma of AD subjects from two studies (Study 1, N=42; Study 2, N=185) using the hTau-pT217 immunoassay described herein. hTau-pT181 will also be measured in the same patients using the hTau-pT181 immunoassay previously described in the art. All patients will have tau positron emission tomography (PET) measured by flortaucipir neurological imaging. Samples will be acquired between the two independent clinical trials (including baseline) and stored at -80°C for future biomarker studies. flortaucipir SUVR will be determined in the neocortical region of interest against a reference signal in white matter. The correlation between flortaucipir SUVR and plasma pTau (pT181 and pT217, respectively) will be evaluated using Spearman's rank correlation. Receiver operating characteristic (ROC) curve analysis will utilize a logistic regression model incorporating age and sex as covariates, along with a flortaucipir-positive cutoff of SUVR ≥ 1.1. Baseline pTau predicting future cognitive decline will be assessed using a mixed-effects model that evaluates pTau by quartiles.

[0055] The hTau-pT217 immunoassay showed a statistically significant higher correlation with Flortaucipir PET in both studies, as shown in Table 3 (p-value < 0.05). [Table 3]

[0056] hTau-pT217 immunoassay as a diagnostic indicator for AD and disease progression The levels of hTau-pT217 in CSF using the described immunoassays will be evaluated and compared to those of the hTau-pT181 immunoassay. In short, CSF samples from the Swedish BioFINDER study—including elderly individuals without disability (CU, n=65), patients with mild cognitive impairment due to AD (MCI-AD, n=29), AD dementia (n=43), and other neurodegenerative disorders (n=57)—will be evaluated using the hTau-pT217 and hTau-p181 immunoassays. 184 participants participated. 18 I underwent F-Flortaucipir positron emission tomography (PET). 18 We quantify F-Flortaucipir uptake in a priori defined regions and inferior temps associated with tau pathology in AD, including Taublag stages I-II, III-IV, and V-VI.

[0057] In CU patients, both the hTau-PT217 immunoassay and the hTau-pT181 immunoassay were black stage I-II. 18 F-Flortaucipir correlates, and in AD patients, both the hTau-pT217 immunoassay and the hTau-pT181 immunoassay correlate in the regions of Braak stages III-IV and V-VI, and in MCI patients, hTau-pT217 correlates in regions I-II of Braak stages III-IV and V-VI. 18 F-Flortaucipir is correlated with hTau-pT181, while hTau-pT181 is correlated only in the region of Braak stages I-II. Importantly, the region 18 The correlation between F-Flortaucipir and the hTau-pT217 immunoassay was observed across all three diagnostic groups (CU, MCI, and AD), and across all regions (Brack stages I-II, III-IV, and V-VI), and with respect to the hTau-pT181 immunoassay. 18 It shows a statistically significant improvement (p<0.001~0.016) that surpasses the correlation with F-Flortaucipir.

[0058] The correlation coefficient for the hTau-pT217 immunoassay was, across all regions, 18 For F-Flortaucipir, statistically significant (all p<0.001) values ​​were consistently higher compared to the hTau-pT181 immunoassay: hTau-pT217 (0.698~0.752) vs. hTau-pT181 (0.572~0.706). Furthermore, the hTau-pT217 immunoassay showed pathological results in all areas. 18 The hTau-pT217 immunoassay was found to be a statistically significant (p<0.001) more accurate predictor of F-Flortaucipir status (hTau-pT217, AUC 0.890~0.929; hTau-pT181 immunoassay, AUC 0.859~0.904). Furthermore, the hTau-pT217 immunoassay showed statistically significant (p=0.026) improved performance compared to the hTau-pT181 immunoassay in differentiating AD neurodegenerative diseases from non-AD neurodegenerative diseases (hTau-pT217, AUC 0.943; hTau-pT181, AUC 0.914). These results suggest that the hTau-pT217 immunoassay is superior in differentiating AD neurodegenerative diseases from non-AD neurodegenerative diseases. 18 F-Flortaucipir correlates with neurological imaging and can differentiate AD from other neurological disorders and stages, showing a significant improvement over the hTau-pT181 immunoassay.

[0059] The hTau-pT217 immunoassay is associated with tau-PET SUVr. Tau-PET SUVr has been shown in the literature to be associated with tau pathology. The hTau-pT217 immunoassay disclosed herein correlates with tau-PET SUVr in a study measuring hTau-pT217 in plasma, serum, and CSF from patients with mild AD. Briefly, 190 subjects underwent the hTau-pT217 immunoassay in plasma at baseline. Of these 190 subjects, 185 patients underwent the tau-PET SUVr assay. The data were analyzed using Spearman's test, and a significant correlation was observed with Spearman ρ = 0.49 and unadjusted p-value < 0.001. Furthermore, 187 subjects underwent the determination of hTau-pT217 in serum at baseline. Of these 187 subjects, 182 underwent the tau-PET SUVr assay. The data were analyzed using Spearman's test, and a significant correlation was observed with Spearman ρ = 0.41 and an unadjusted p-value < 0.001. Furthermore, 86 subjects underwent hTau-pT217 determination in CSF at baseline. Of these 86 subjects, 29 underwent tau PET SUVr assay. The data were analyzed using Spearman's test, and a significant correlation was observed with Spearman ρ = 0.70 and an unadjusted p-value < 0.001. The results support the use of pTau217 levels measured in CSF, plasma, or serum to identify tau pathology.

[0060] The hTau-pT217 immunoassay is associated with mild AD cognitive status. The mean values ​​of hTau-pT217 in plasma, serum, and CSF of patients with mild Alzheimer's disease were calculated according to the immunoassay described above. The results for each matrix are shown in Table 4. [Table 4]

[0061] The hTau-pT217 immunoassay disclosed herein is associated with cognitive status (based on the Mini-Mental State Examination, MMSE) and changes from baseline compared to placebo in MMSE scores in patients with mild Alzheimer's disease (AD). hTau-pT217 is evaluated by immunoassays in plasma, serum, and CSF from patients with mild AD. In short, for each matrix (plasma, serum, and CSF), subjects with hTau-pT217 measurements (as described herein), baseline MMSE assessments, and changes from baseline assessments are evaluated for significance using the Spearman test. The hTau-pT217 immunoassay shows a statistically significant association with both cognitive status and changes from baseline in plasma and serum (the CSF sample size is too low for statistical significance, but the data suggest that if the sample size is increased, as with the assessments for serum and plasma, CSF would likely show a statistically significant association with both MMSE scores and changes from baseline). The results are shown in Table 5. [Table 5]

[0062] These results demonstrate a cross-sectional association between hTau-pT217 and the MMSE, a cognitive measure, and highlight the usefulness of hTau-pT217 in determining the future risk of cognitive decline.

[0063] hTau-pT217 immunoassay is associated with amyloid status. The hTau-pT217 immunoassay disclosed herein is associated with amyloid status. In short, plasma samples from four different patient groups with known AD and amyloid status (based on PET neurological imaging) are evaluated for hTau-pT217 association: (i) unaffected elderly amyloid-positive (CU-A+), (ii) unaffected elderly amyloid-negative (CU-A-), (iii) elderly amyloid-positive AD (AD-A+), and (iv) clinically unaffected young adults (CUY-A-). Samples from each group are analyzed using the hTau-pT217 immunoassay described herein. The results are shown in Table 6. [Table 6]

[0064] The evaluation of the hTau-pT217 immunoassay for identifying amyloid-positive subjects is determined by evaluating the results for each group using the Student's t-test, as shown in Table 6. The results are shown in Table 7. [Table 7]

[0065] The data provided herein demonstrate that the hTau-pT217 assay of this disclosure can identify amyloid-positive subjects, act as a diagnostic method for AD, determine the cognitive status of subjects associated with AD, identify subjects at risk of AD and / or in the early stages of AD, and act as a diagnostic method for the progression of AD. The data also demonstrate that the hTau-pT217 assay of this disclosure correlates with neurological imaging, is functional in serum, plasma, and CSF matrices, and is superior to the known hTau-pT181 assay.

[0066] Exemplary Embodiments of the Present Disclosure 1. An antibody that specifically binds to human tau phosphorylated with threonine at residue 217 of Sequence ID No. 1 ("hTau-pT217").

[0067] 2. An antibody comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity-determining regions (CDRs) of LCDR1, LCDR2, and LCDR3, and the HCVR comprises CDRs of HCDR1, HCDR2, and HCDR3. The antibodies are as follows: LCDR1 has the amino acid sequence of SEQ ID NO: 13, LCDR2 has the amino acid sequence of SEQ ID NO: 14, LCDR3 has the amino acid sequence of SEQ ID NO: 15, HCDR1 has the amino acid sequence of SEQ ID NO: 10, HCDR2 has the amino acid sequence of SEQ ID NO: 11, and HCDR3 has the amino acid sequence of SEQ ID NO: 12.

[0068] 3. An antibody comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity-determining regions (CDRs) of LCDR1, LCDR2, and LCDR3, and the HCVR comprises CDRs of HCDR1, HCDR2, and HCDR3. Antibodies in which LCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 13, LCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 14, LCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 15, HCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 10, HCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 11, and HCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 12.

[0069] 4. a. LCVR having the amino acid sequence of SEQ ID NO: 5, and HCVR having the amino acid sequence of SEQ ID NO: 3, b. An antibody of Embodiment 2 or 3 comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR) selected from an LCVR having the amino acid sequence of SEQ ID NO: 8 and an HCVR having the amino acid sequence of SEQ ID NO: 6.

[0070] 5. a. LCVR having an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 5, and HCVR having an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 3, b. An antibody of Embodiment 2 or 3 comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR) selected from an LCVR having an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 8, and an HCVR having an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 6.

[0071] 6. An antibody according to any of Embodiments 1 to 5, wherein the antibody is humanized.

[0072] 7. An antibody according to any of Embodiments 1 to 6, wherein the antibody contains an IgG quadruplex.

[0073] 8. An antibody according to any of Embodiments 1 to 7, wherein the antibody contains a kappa light chain.

[0074] 9. A pharmaceutical composition comprising one antibody from any of Embodiments 1 to 8 and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0075] 10. An antibody that specifically binds to the CNS expression isoform of human tau.

[0076] 11. The antibody of Embodiment 10, wherein the antibody binds to the epitope region of human tau, which includes glutamine at residue 124 and alanine at residue 125 of SEQ ID NO: 1.

[0077] 12. An antibody comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity-determining regions (CDRs) of LCDR1, LCDR2, and LCDR3, and the HCVR comprises CDRs of HCDR1, HCDR2, and HCDR3. The antibodies are as follows: LCDR1 has the amino acid sequence of SEQ ID NO: 23, LCDR2 has the amino acid sequence of SEQ ID NO: 24, LCDR3 has the amino acid sequence of SEQ ID NO: 25, HCDR1 has the amino acid sequence of SEQ ID NO: 20, HCDR2 has the amino acid sequence of SEQ ID NO: 21, and HCDR3 has the amino acid sequence of SEQ ID NO: 22.

[0078] 13. An antibody comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity-determining regions (CDRs) of LCDR1, LCDR2, and LCDR3, and the HCVR comprises CDRs of HCDR1, HCDR2, and HCDR3. Antibodies in which LCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 23, LCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 24, LCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 25, HCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 20, HCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 21, and HCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 22.

[0079] 14. An antibody according to Embodiment 12 or 13, comprising a light chain variable region (LCVR) having the amino acid sequence of SEQ ID NO: 17 and a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO: 19.

[0080] 15. An antibody according to Embodiment 12 or 13, comprising a light chain variable region (LCVR) having an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 17, and a heavy chain variable region (HCVR) having an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 19.

[0081] 16. An antibody according to any of embodiments 10 to 15, wherein the antibody is humanized.

[0082] 17. An antibody according to any of Embodiments 10 to 16, wherein the antibody contains an IgG quadruplex.

[0083] 18. An antibody according to any of Embodiments 10 to 17, wherein the antibody contains a kappa light chain.

[0084] 19. A pharmaceutical composition comprising one antibody from any of embodiments 10 to 18 and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0085] 20. A method for treating a neurodegenerative disease, comprising administering an effective amount of one antibody or pharmaceutical composition thereof according to Embodiments 1 to 19 to a patient in need thereof.

[0086] 21. The method of Embodiment 20, wherein the neurodegenerative disease is a tauopathy.

[0087] 22. The method of Embodiment 21, wherein the tauopathy is one of AD, PSP, and FTD.

[0088] 23. Any one antibody according to Embodiments 1 to 19, or a pharmaceutical composition thereof, for use in therapeutic purposes.

[0089] 24. Any one antibody according to Embodiments 1 to 19, or a pharmaceutical composition thereof, for use in the treatment of neurodegenerative diseases.

[0090] 25. The antibody of Embodiment 24, or a pharmaceutical composition thereof, wherein the neurodegenerative disease is tauopathy.

[0091] 26. The antibody of Embodiment 25, or a pharmaceutical composition thereof, wherein the tauopathy is selected from the group consisting of AD, PSP, and FTD.

[0092] 27. Any one antibody according to Embodiments 1 to 19, or a pharmaceutical composition thereof, for use in the manufacture of a pharmaceutical for the treatment of neurodegenerative diseases.

[0093] 28. The antibody of Embodiment 27, or a pharmaceutical composition thereof, wherein the neurodegenerative disease is tauopathy.

[0094] 29. The antibody of Embodiment 28, or a pharmaceutical composition thereof, wherein the tauopathy is selected from the group consisting of AD, PSP, and FTD.

[0095] 30. A method for detecting hTau-pT217 in a patient sample, A method comprising the steps of contacting a patient sample with one antibody from any of embodiments 1 to 8, and detecting a signal provided by the contact step.

[0096] 31. A method for quantifying hTau-pT217 in patient samples, A method comprising the steps of contacting a patient sample with one antibody from any of embodiments 1 to 8, and detecting a signal provided by the contact step.

[0097] 32. The method of Embodiment 31, further comprising the steps of contacting a control standard with an antibody and detecting a signal provided by the step of contacting the control standard.

[0098] 33. A method for quantifying hTau-pT217 in a patient sample, comprising the steps of: contacting the patient sample with an antibody of Embodiments 1 to 8; contacting the patient sample with a second antibody, wherein the second antibody is an antibody of Embodiments 10 to 18, and one of the antibodies and the second antibody includes a detectable label; detecting a signal provided by the detectable label upon the formation of a complex comprising the antibody, the second antibody and hTau-pT217; contacting a control standard with the antibody; contacting a control standard with the second antibody, wherein the antibody and one of the second antibody includes a detectable label; and detecting a signal provided by the detectable label upon the formation of a complex comprising the antibody, the second antibody and the control standard.

[0099] 34. A method for diagnosing a patient as having one or more of the following conditions: (i) having a neurodegenerative disease, (ii) being at risk of having a neurodegenerative disease, (iii) needing treatment for a neurodegenerative disease, or (iv) needing neurological imaging, comprising the steps of: contacting a patient sample with one of the antibodies of Embodiments 1 to 8; and detecting the binding between hTau-pT217 and the antibody in the patient sample.

[0100] 35. The method of Embodiment 34, further comprising the step of diagnosing the patent as one of (i) having a neurodegenerative disease, (ii) being at risk of having a neurodegenerative disease, (iii) having a need for treatment for a neurodegenerative disease, or (iv) having a need for neurological imaging, if the level of hTau-pT217 detected in a patient sample exceeds a reference level.

[0101] 36. A method for diagnosing and treating a neurodegenerative disease in a patient, comprising the steps of: contacting a patient sample with one antibody from any one of Embodiments 1 to 18; detecting the binding between hTau-pT217 and the antibody in the patient sample; diagnosing a patient having a neurodegenerative disease; and administering a therapeutically effective amount of anti-human tau antibody to the diagnosed patient.

[0102] 37. The method of Embodiment 36, wherein the diagnostic step includes diagnosing a patient as having a neurodegenerative disease if the presence of hTau-pT217 in a patient sample exceeds a reference level.

[0103] 38. Any method of Embodiments 31 to 37, further comprising the step of quantifying hTau-pT217 in a patient sample.

[0104] 39. The method of Embodiment 38, wherein the step of quantifying hTau-pT217 includes quantifying hTau-pT217 in a patient sample against a reference standard.

[0105] 40. Any method of Embodiments 30 to 39, wherein the patient sample is one of blood, plasma, serum, or CSF.

[0106] 41. Any method of Embodiments 30-32 and 34-40, further comprising the step of contacting a patient sample with a second antibody, wherein the second antibody binds to an epitope region of hTau-pT217 that does not overlap with the antibody.

[0107] 42. The method of Embodiment 41, wherein one of the antibody or a second antibody includes a detectable label, and the detection step includes detecting a signal provided by the detectable label in the formation of a complex comprising the antibody, the second antibody and hTau-pT217.

[0108] 43. Any method according to Embodiments 41 to 42, wherein one of the antibodies and a second antibody is immobilized on a substrate.

[0109] 44. Any method according to Embodiments 30 to 43, wherein the steps of contacting the patient sample with an antibody and contacting the patient sample with a second antibody are performed simultaneously.

[0110] 45. The second antibody comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR), where the LCVR comprises complementarity-determining regions (CDRs) of LCDR1, LCDR2, and LCDR3, and the HCVR comprises CDRs of HCDR1, HCDR2, and HCDR3. The method according to any of Embodiments 32 and 41-44, wherein LCDR1 has the amino acid sequence of SEQ ID NO: 23, LCDR2 has the amino acid sequence of SEQ ID NO: 24, LCDR3 has the amino acid sequence of SEQ ID NO: 25, HCDR1 has the amino acid sequence of SEQ ID NO: 20, HCDR2 has the amino acid sequence of SEQ ID NO: 21, and HCDR3 has the amino acid sequence of SEQ ID NO: 22.

[0111] 46. ​​The second antibody comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR), where the LCVR comprises complementarity-determining regions (CDRs) of LCDR1, LCDR2, and LCDR3, and the HCVR comprises CDRs of HCDR1, HCDR2, and HCDR3. The method according to any one of Embodiments 33 and 41-44, wherein LCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 23, LCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 24, LCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 25, HCDR1 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 20, HCDR2 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 21, and HCDR3 has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 22.

[0112] 47. Any method according to Embodiments 46 to 47, wherein the second antibody comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR), the LCVR having the amino acid sequence of SEQ ID NO: 19 and the HCVR having the amino acid sequence of SEQ ID NO: 17.

[0113] 48. The second antibody contains a light chain variable region (LCVR) and a heavy chain variable region (HCVR), A method according to any of Embodiments 46 to 47, wherein LCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 19, and HCVR has an amino acid sequence having at least 95% homology to the amino acid sequence of SEQ ID NO: 17.

[0114] 49. A method for forming a complex between a first antibody, a second antibody, and human tau expressed in the CNS and phosphorylated with threonine residue 217 of SEQ ID NO: 1, comprising: contacting a patient sample with the first antibody, wherein the first antibody is one of the antibodies from Embodiments 1 to 8; and contacting a patient sample with the second antibody, wherein the second antibody is one of the antibodies from Embodiments 10 to 18.

[0115] 50. An assay for detecting human tau expressed in the CNS and phosphorylated with threonine at residue 217 of SEQ ID NO: 1, comprising one antibody from Embodiments 1 to 8 and one antibody from Embodiments 10 to 18.

[0116] 51. The assay of Embodiment 50, wherein one of the antibodies contains a detectable label.

[0117] Sequence List Sequence ID 1 (hTau-pT217) MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGD TPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPT REPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHH KPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL Sequence ID No. 2 (exemplary rabbit anti-hTau-pT217 antibody HC) QSVEESGGRLVTPGTPLTLTCTVSGLSPSWYGVHWVRQAPGKGLEWIGVLRAGSHTYYAGWAKGRFAISKTSTTVALKITSPTTEDTAIYFCGSVGRGIWGPGTLVTV SLGQPKAPSVFPLAPCCGDTPSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTTCSKPTCP PPELLGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISK ARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK Sequence ID 3 (exemplary rabbit anti-hTau-pT217 antibody against HCVR) QSVEESGGRLVTPGTPLTLTCTVSGLSPSWYGVHWVRQAPGKGLEWIGVLRAGSHTYYAGWAKGRFAISKTSTTVALKITSPTTEDTAIYFCGSVGRGIWGPGTLVTVSL Sequence ID No. 4 (LC of rabbit anti-hTau-pT217 antibody as an example) AQVLTQTASPVSATVGGTVTINCQASLAVYNNNYLAWYQQKPGQPPKRLIYLASSLSSGVSSHFKGSGSGTQFTLTISDVQADDAATYFCQGSYDCTIADCVAFGGGT EVVVKGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADNTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC Sequence ID No. 5 (LCVR of exemplary rabbit anti-hTau-pT217 antibody) AQVLTQTASPVSATVGGTVTINCQASLAVYNNNYLAWYQQKPGQPPKRLIYLASSLSSGVSSHFKGSGSGTQFTLTISDVQADDAATYFCQGSYDCTIADCVAFGGGTEVVVK Sequence ID 6 (exemplary chimeric anti-hTau-pT217 antibody for HCVR) QSVEESGGRLVTPGTPLTLTCTVSGLSPSWYGVHWVRQAPGKGLEWIGVLRAGSHTYYAGWAKGRFAISKTSTTVALKITSPTTEDTAIYFCGSVGRGIWGPGTLVTVSL Sequence ID 7 (HC of exemplary chimeric anti-hTau-pT217 antibody) QSVEESGGRLVTPGTPLTLTCTVSGLSPSWYGVHWVRQAPGKGLEWIGVLRAGSHTYYAGWAKGRFAISKTSTTVALKITSPTTEDTAIYFCGSVGRGIWGPGTLVTV SLAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPC ICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISK TKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK Sequence ID No. 8 (LCVR of exemplary chimeric anti-hTau-pT217 antibody) AQVLTQTASPVSATVGGTVTINCQASLAVYNNNYLAWYQQKPGQPPKRLIYLASSLSSGVSSHFKGSGSGTQFTLTISDVQADDAATYFCQGSYDCTIADCVAFGGGTEVVVK Sequence ID 9 (LC of exemplary chimeric anti-hTau-pT217 antibody) AQVLTQTASPVSATVGGTVTINCQASLAVYNNNYLAWYQQKPGQPPKRLIYLASSLSSGVSSHFKGSGSGTQFTLTISDVQADDAATYFCQGSYDCTIADCVAFGGGTEV VVKRADAAPTVSIFPPSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC Sequence ID 10 (exemplary HCDR1) GLSPSWYGVH Sequence ID 11 (exemplary HCDR2) VLRAGSHTYYAGWAKG Sequence ID 12 (exemplary HCDR3) VGRGI Sequence ID 13 (exemplary LCDR1) QASLAVYNNNYLA Sequence ID 14 (Example LCDR2) LASSLSS Sequence ID 15 (Example LCDR3) LASSLSS Sequence ID No. 16 (HC of a human tau-conjugated antibody expressed only in the exemplary CNS) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSPYYWSWIRQPPDKGLEWIGEINWSGDTNYNPSLKSRVTISLDTSKNQFSLNLSSVTAADTAVYYCARSFDRWGQGTLVTV SSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVEPRGPTIKPCP PCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAPIERTI SKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK Sequence ID No. 17 (HCVR of human tau-conjugated antibody expressed only in the exemplary CNS) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSPYYWSWIRQPPDKGLEWIGEINWSGDTNYNPSLKSRVTISLDTSKNQFSLNLSSVTAADTAVYYCARSFDRWGQGTLVTVSS Sequence ID No. 18 (LC of a human tau-conjugated antibody expressed only in the exemplary CNS) EIVLTQSPGTLSLSPGERATLSCRASQSVRSNYFAWYQQKPGQAPRLLIYGVSRRAFGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGASLITFGQGTRLEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID No. 19 (LCVR of a human tau-conjugated antibody expressed only in the exemplary CNS) EIVLTQSPGTLSLSPGERATLSCRASQSVRSNYFAWYQQKPGQAPRLLIYGVSRRAFGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGASLITFGQGTRLEIK Sequence ID No. 20 (HCDR1, a human tau-conjugated antibody expressed only in the exemplary CNS) AVYGGSFSPYYWS Sequence ID No. 21 (HCDR2, a human tau-conjugated antibody expressed only in the exemplary CNS) EINWSGDTN Sequence ID No. 22 (HCDR3 of a human tau-conjugated antibody expressed only in exemplary CNS) ARSFDR SEQ ID NO: 23 (LCDR1, a human tau-conjugated antibody expressed only in exemplary CNS) RASQSVRSNYFA Sequence ID No. 24 (LCDR2, a human tau-conjugated antibody expressed only in the exemplary CNS) YGVSRRAF SEQ ID NO: 25 (LCDR3, a human tau-conjugated antibody expressed only in exemplary CNS) QQYGASLIT Sequence ID No. 26 (Exemplary Peptide for Immunization) RTPSLPTPPTR Here, the T at residue 7 is phosphorylated. Sequence ID No. 27 (Exemplary Peptide for Immunization) AEPRQEFEVMEDHAGTYGLDGDRKDQGGYTMHQDQEGDTDAGLKESPLQT PTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPE GTTAEEAGIGDTPSLEDEAAGHVTQEPESGKVVQEGFLREPGPP GLSHQLMSGMPGAPLLPEGPREATRQP SGTGPEDTEGGRHAPELLKHQ LLGDLHQEGPPLKGAGGKE RPGSKEEVDEDRDVDESSPQ DSPPSKASPA QDGRPPQTAAREATSIPGFPAEGAIPLPVDFLSKVSTEIPASEPDGPSVG RAKGQDAPLEFTFHVEITPNVQKEQAHSEEHLGRAAFPGAPGEGPEARGP SLGEDTKEAD LPEPSEKQPAAPRGKPVSRVPQLKARMVS KSKDGTGSDD KKAKTSTRSSAKTLKNRPCLSPKHPTPGSSDPLIQPSSAAVCPEPPSSPKYVSSVT SRTG SSGAKEMKLKGADGKTKIAT PRGAAPPGQKGQANATRIPAKTPP APKTPPSSGEPPKSGDRSGYSSPGTPGSRSRTP SLPTPPTREP KKVAVVRTPKPSSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGG KVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKC GSLNIHHKPG GGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGG NKKIETHKLTFR ENAKAKTDHGAEIVYKSPVVSGDT SPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL

Claims

1. An antibody that specifically binds to a human tau CNS expression isoform comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity determining regions (CDRs) of LCDR1, LCDR2, and LCDR3, and the HCVR comprises the CDRs of HCDR1, HCDR2, and HCDR3. An antibody in which LCDR1 has the amino acid sequence of SEQ ID NO: 23, LCDR2 has the amino acid sequence of SEQ ID NO: 24, LCDR3 has the amino acid sequence of SEQ ID NO: 25, HCDR1 has the amino acid sequence of SEQ ID NO: 20, HCDR2 has the amino acid sequence of SEQ ID NO: 21, and HCDR3 has the amino acid sequence of SEQ ID NO:

22.

2. The antibody according to claim 1, comprising a light chain variable region (LCVR) having the amino acid sequence of SEQ ID NO: 17 and a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO:

19.

3. The antibody according to claim 1 or 2, wherein the antibody is humanized.

4. The antibody according to any one of claims 1 to 3, wherein the antibody comprises an IgG quadrilateral heavy chain.

5. The antibody according to any one of claims 1 to 4, wherein the antibody comprises a kappa light chain.

6. A pharmaceutical composition comprising an antibody according to any one of claims 1 to 5 and one or more pharmaceutically acceptable carriers, diluents, or excipients.

Images